TY  - CHAP
AU  - Stojiljković, Maja
AU  - Komazec, Jovana
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2248
AB  - Svaka bolest čija je učestalost manja od 1 u 2000 ljudi definiše se kao retka bolest. Iz tog razloga, broj retkih
bolesti je veliki. Do sada je opisano preko 6000 različitih retkih bolesti. Preko 80% retkih bolesti ima genetičku
osnovu i to je razlog zašto su znanja iz molekularne biologije od neprocenjivog značaja za
istraživanje molekularne osnove retkih bolesti, postavljanje tačne dijagnoze i razvoj inovativnih terapeutika.
Cilj ovog rada je da objasni važnost otkrivanja molekularno-genetičke osnove retkih bolesti i da prikaže
desetogodišnje iskustvo primene sekvenciranja nove generacije u Srbiji (2014.-2023.) u tu svrhu. U
prethodnom periodu za istraživanje retkih bolesti korišćeni su sekvenciranje kliničkog egzoma, sekvenciranje
kompletnog egzoma i sekvenciranje kompletnog genoma. Takođe, date su i perspektive za budućnost
gde će genomika biti kompletirana tehnologijom sekvenciranja dugačkih fragmenata i
komplementirana upotrebom transkriptomike, proteomike, metabolomike i drugih „omika“.
AB  - Any disease found in less than 1 person out of 2000 people is defined as a rare disease. For this reason, the
number of rare diseases is high. Over 6,000 different rare diseases have been described so far. More than
80% of rare diseases have a genetic basis, and this is the reason why knowledge of molecular biology is invaluable
for research into the molecular basis of rare diseases, establishing an accurate diagnosis and developing
innovative therapeutics. The aim of this paper is to explain the importance of discovering the
molecular genetic basis of rare diseases and to present the ten-year experience of applying new generation
sequencing in Serbia (2014-2023) for this purpose. During this period, clinical exome sequencing,
complete exome sequencing and complete genome sequencing were used for research of rare diseases.
In the future, it is expected that genomics, which until now was based mainly on the technology of shortread
fragments, will be broaden with the long-reads sequencing technology, and complemented by the
use of transcriptomics, proteomics, metabolomics and other omics.
PB  - Beograd : Institut za molekularnu genetiku i genetičko inženjerstvo
T2  - Trendovi u molekularnoj Biologiji
T1  - Sadašnjost i budućnost primene sekvenciranja nove generacije za retke bolesti
T1  - Present and future of next-generation sequencing application for rare diseases
EP  - 89
IS  - 3
SP  - 78
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2248
ER  - 

@inbook{
author = "Stojiljković, Maja and Komazec, Jovana",
year = "2023",
abstract = "Svaka bolest čija je učestalost manja od 1 u 2000 ljudi definiše se kao retka bolest. Iz tog razloga, broj retkih
bolesti je veliki. Do sada je opisano preko 6000 različitih retkih bolesti. Preko 80% retkih bolesti ima genetičku
osnovu i to je razlog zašto su znanja iz molekularne biologije od neprocenjivog značaja za
istraživanje molekularne osnove retkih bolesti, postavljanje tačne dijagnoze i razvoj inovativnih terapeutika.
Cilj ovog rada je da objasni važnost otkrivanja molekularno-genetičke osnove retkih bolesti i da prikaže
desetogodišnje iskustvo primene sekvenciranja nove generacije u Srbiji (2014.-2023.) u tu svrhu. U
prethodnom periodu za istraživanje retkih bolesti korišćeni su sekvenciranje kliničkog egzoma, sekvenciranje
kompletnog egzoma i sekvenciranje kompletnog genoma. Takođe, date su i perspektive za budućnost
gde će genomika biti kompletirana tehnologijom sekvenciranja dugačkih fragmenata i
komplementirana upotrebom transkriptomike, proteomike, metabolomike i drugih „omika“., Any disease found in less than 1 person out of 2000 people is defined as a rare disease. For this reason, the
number of rare diseases is high. Over 6,000 different rare diseases have been described so far. More than
80% of rare diseases have a genetic basis, and this is the reason why knowledge of molecular biology is invaluable
for research into the molecular basis of rare diseases, establishing an accurate diagnosis and developing
innovative therapeutics. The aim of this paper is to explain the importance of discovering the
molecular genetic basis of rare diseases and to present the ten-year experience of applying new generation
sequencing in Serbia (2014-2023) for this purpose. During this period, clinical exome sequencing,
complete exome sequencing and complete genome sequencing were used for research of rare diseases.
In the future, it is expected that genomics, which until now was based mainly on the technology of shortread
fragments, will be broaden with the long-reads sequencing technology, and complemented by the
use of transcriptomics, proteomics, metabolomics and other omics.",
publisher = "Beograd : Institut za molekularnu genetiku i genetičko inženjerstvo",
journal = "Trendovi u molekularnoj Biologiji",
booktitle = "Sadašnjost i budućnost primene sekvenciranja nove generacije za retke bolesti, Present and future of next-generation sequencing application for rare diseases",
pages = "89-78",
number = "3",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2248"
}

Stojiljković, M.,& Komazec, J.. (2023). Sadašnjost i budućnost primene sekvenciranja nove generacije za retke bolesti. in Trendovi u molekularnoj Biologiji
Beograd : Institut za molekularnu genetiku i genetičko inženjerstvo.(3), 78-89.
https://hdl.handle.net/21.15107/rcub_imagine_2248

Stojiljković M, Komazec J. Sadašnjost i budućnost primene sekvenciranja nove generacije za retke bolesti. in Trendovi u molekularnoj Biologiji. 2023;(3):78-89.
https://hdl.handle.net/21.15107/rcub_imagine_2248 .

Stojiljković, Maja, Komazec, Jovana, "Sadašnjost i budućnost primene sekvenciranja nove generacije za retke bolesti" in Trendovi u molekularnoj Biologiji, no. 3 (2023):78-89,
https://hdl.handle.net/21.15107/rcub_imagine_2248 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Tošić, Nataša
AU  - Pavlović, Sonja
AU  - Celic, Dejan
AU  - Vučenović, Jelica
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2277
AB  - Background/Objectives: Fabry disease (FD) is a rare X-linked
disorder caused by variants in the GLA gene leading to the deficiency
of lysosomal α-galactosidase-A and progressive accumulation
of globotriaosylceramide affecting the heart, nervous system,
and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, the precise molecular-genetic diagnosis
and the earliest possible treatment are essential to avoid significant
disease progression.
Methods: We analyzed 95 (34 female and 61 male) hemodialysis
patients with clinical suspicion of FD using Sanger sequencing
of all coding exons (7) and flanking intron regions of the GLA
gene, and measured the relative expression of the GLA gene in
available samples.
Results: The genetic analysis revealed 3 patients with a missense
variant (p.Asp313Tyr), and 10 patients with combinations of
non-coding variants, described as complex intronic haplotypes
(CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7
(7.4%) patients. Lyso-Gb3 biomarker levels were within the normal
range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of GLA gene in PBMC of 2 female
patients with CIH1 and one female patient carrying only c.-10C>T
variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that further
analyses are needed to confirm/exclude FD in these patients.
Conclusion: Because the effects of CIHs are not yet fully
understood, our work highlights the importance of analyzing
intronic regions of the GLA gene as genetic modifiers and the
need to include expression analysis in the diagnostic algorithm.
PB  - Springer Nature
C3  - European Journal of Human Genetic
T1  - Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology
EP  - 433
IS  - Supplement S1
SP  - 432
VL  - 31
DO  - 10.1038/s41431-023-01339-3
ER  - 

@conference{
author = "Parezanović, Marina and Stojiljković, Maja and Anđelković, Marina and Stevanović, Nina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Tošić, Nataša and Pavlović, Sonja and Celic, Dejan and Vučenović, Jelica and Skakić, Anita",
year = "2023",
abstract = "Background/Objectives: Fabry disease (FD) is a rare X-linked
disorder caused by variants in the GLA gene leading to the deficiency
of lysosomal α-galactosidase-A and progressive accumulation
of globotriaosylceramide affecting the heart, nervous system,
and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, the precise molecular-genetic diagnosis
and the earliest possible treatment are essential to avoid significant
disease progression.
Methods: We analyzed 95 (34 female and 61 male) hemodialysis
patients with clinical suspicion of FD using Sanger sequencing
of all coding exons (7) and flanking intron regions of the GLA
gene, and measured the relative expression of the GLA gene in
available samples.
Results: The genetic analysis revealed 3 patients with a missense
variant (p.Asp313Tyr), and 10 patients with combinations of
non-coding variants, described as complex intronic haplotypes
(CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7
(7.4%) patients. Lyso-Gb3 biomarker levels were within the normal
range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of GLA gene in PBMC of 2 female
patients with CIH1 and one female patient carrying only c.-10C>T
variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that further
analyses are needed to confirm/exclude FD in these patients.
Conclusion: Because the effects of CIHs are not yet fully
understood, our work highlights the importance of analyzing
intronic regions of the GLA gene as genetic modifiers and the
need to include expression analysis in the diagnostic algorithm.",
publisher = "Springer Nature",
journal = "European Journal of Human Genetic",
title = "Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology",
pages = "433-432",
number = "Supplement S1",
volume = "31",
doi = "10.1038/s41431-023-01339-3"
}

Parezanović, M., Stojiljković, M., Anđelković, M., Stevanović, N., Spasovski, V., Ugrin, M., Komazec, J., Tošić, N., Pavlović, S., Celic, D., Vučenović, J.,& Skakić, A.. (2023). Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology. in European Journal of Human Genetic
Springer Nature., 31(Supplement S1), 432-433.
https://doi.org/10.1038/s41431-023-01339-3

Parezanović M, Stojiljković M, Anđelković M, Stevanović N, Spasovski V, Ugrin M, Komazec J, Tošić N, Pavlović S, Celic D, Vučenović J, Skakić A. Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology. in European Journal of Human Genetic. 2023;31(Supplement S1):432-433.
doi:10.1038/s41431-023-01339-3 .

Parezanović, Marina, Stojiljković, Maja, Anđelković, Marina, Stevanović, Nina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Tošić, Nataša, Pavlović, Sonja, Celic, Dejan, Vučenović, Jelica, Skakić, Anita, "Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology" in European Journal of Human Genetic, 31, no. Supplement S1 (2023):432-433,
https://doi.org/10.1038/s41431-023-01339-3 . .

TY  - CONF
AU  - Stevanović, Nina
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Spasovski, Vesna
AU  - Stojiljković, Maja
AU  - Parezanović, Marina
AU  - Ugrin, Milena
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2275
AB  - Background/Objectives: Primary ciliary dyskinesia (PCD) is a
disease caused by impaired ciliary motility and mainly affects the
lungs and reproductive organs. Inheritance is autosomal recessive
and X-linked with more than 40 disease-causing genes, wherefore
PCD patients have diverse clinical manifestations, thus making
diagnosis difficult. The utility of next-generation sequencing (NGS)
technology for diagnostic purposes allows a better understanding
of the PCD genetic background. However, the identification of
specific disease-causing variants is challenging. The objective of
this study was to create a unique guideline that will enable the
standardization of the assessment of novel variants within PCD
associated genes.
Methods: The study included designing a pipeline for the classification
of the rare genetic variants detected using NGS. The pipeline
included in silico (translation, 3D-model, protein-protein interactions,
sequence conservation, posttranslational modifications) and functional
analysis (expressional analysis, Western Blot) of the variants.
Results: The designed pipeline consists of three steps: sequencing,
detection, and identification of genes/variants; classification of
variants according to their effect; and variant characterization using
in silico structural and functional analysis. The pipeline was validated
by the analysis of the variants detected in a disease-causing
gene (DNAI1) and the novel candidate gene (SPAG16).
Conclusion: The application of the pipeline resulted in the
identification of disease-causing variants, as well as pathogenicity
validation, through the analysis on transcriptional, translational,
and posttranslational levels.The application of created pipeline
leads to the confirmation of PCD diagnosis and enables a shift
from candidate to disease-causing gene.
PB  - Springer Nature
C3  - European Journal of Human Genetic
T1  - Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis
EP  - 383
IS  - Supplement S1
SP  - 383
VL  - 31
DO  - 10.1038/s41431-023-01338-4
ER  - 

@conference{
author = "Stevanović, Nina and Anđelković, Marina and Skakić, Anita and Spasovski, Vesna and Stojiljković, Maja and Parezanović, Marina and Ugrin, Milena and Pavlović, Sonja",
year = "2023",
abstract = "Background/Objectives: Primary ciliary dyskinesia (PCD) is a
disease caused by impaired ciliary motility and mainly affects the
lungs and reproductive organs. Inheritance is autosomal recessive
and X-linked with more than 40 disease-causing genes, wherefore
PCD patients have diverse clinical manifestations, thus making
diagnosis difficult. The utility of next-generation sequencing (NGS)
technology for diagnostic purposes allows a better understanding
of the PCD genetic background. However, the identification of
specific disease-causing variants is challenging. The objective of
this study was to create a unique guideline that will enable the
standardization of the assessment of novel variants within PCD
associated genes.
Methods: The study included designing a pipeline for the classification
of the rare genetic variants detected using NGS. The pipeline
included in silico (translation, 3D-model, protein-protein interactions,
sequence conservation, posttranslational modifications) and functional
analysis (expressional analysis, Western Blot) of the variants.
Results: The designed pipeline consists of three steps: sequencing,
detection, and identification of genes/variants; classification of
variants according to their effect; and variant characterization using
in silico structural and functional analysis. The pipeline was validated
by the analysis of the variants detected in a disease-causing
gene (DNAI1) and the novel candidate gene (SPAG16).
Conclusion: The application of the pipeline resulted in the
identification of disease-causing variants, as well as pathogenicity
validation, through the analysis on transcriptional, translational,
and posttranslational levels.The application of created pipeline
leads to the confirmation of PCD diagnosis and enables a shift
from candidate to disease-causing gene.",
publisher = "Springer Nature",
journal = "European Journal of Human Genetic",
title = "Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis",
pages = "383-383",
number = "Supplement S1",
volume = "31",
doi = "10.1038/s41431-023-01338-4"
}

Stevanović, N., Anđelković, M., Skakić, A., Spasovski, V., Stojiljković, M., Parezanović, M., Ugrin, M.,& Pavlović, S.. (2023). Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis. in European Journal of Human Genetic
Springer Nature., 31(Supplement S1), 383-383.
https://doi.org/10.1038/s41431-023-01338-4

Stevanović N, Anđelković M, Skakić A, Spasovski V, Stojiljković M, Parezanović M, Ugrin M, Pavlović S. Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis. in European Journal of Human Genetic. 2023;31(Supplement S1):383-383.
doi:10.1038/s41431-023-01338-4 .

Stevanović, Nina, Anđelković, Marina, Skakić, Anita, Spasovski, Vesna, Stojiljković, Maja, Parezanović, Marina, Ugrin, Milena, Pavlović, Sonja, "Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis" in European Journal of Human Genetic, 31, no. Supplement S1 (2023):383-383,
https://doi.org/10.1038/s41431-023-01338-4 . .

TY  - DATA
AU  - Spasovski, Vesna
AU  - Srzentić Dražilov, Sanja
AU  - Nikčević, Gordana
AU  - Baščarević, Zoran
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Spasovski, Duško
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2089
AB  - Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.
PB  - MDPI
T2  - Diagnostics
T1  - Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.
IS  - 3
SP  - 471
VL  - 13
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2089
ER  - 

@misc{
author = "Spasovski, Vesna and Srzentić Dražilov, Sanja and Nikčević, Gordana and Baščarević, Zoran and Stojiljković, Maja and Pavlović, Sonja and Spasovski, Duško",
year = "2023",
abstract = "Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.",
publisher = "MDPI",
journal = "Diagnostics",
title = "Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.",
number = "3",
pages = "471",
volume = "13",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2089"
}

Spasovski, V., Srzentić Dražilov, S., Nikčević, G., Baščarević, Z., Stojiljković, M., Pavlović, S.,& Spasovski, D.. (2023). Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.. in Diagnostics
MDPI., 13(3), 471.
https://hdl.handle.net/21.15107/rcub_imagine_2089

Spasovski V, Srzentić Dražilov S, Nikčević G, Baščarević Z, Stojiljković M, Pavlović S, Spasovski D. Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.. in Diagnostics. 2023;13(3):471.
https://hdl.handle.net/21.15107/rcub_imagine_2089 .

Spasovski, Vesna, Srzentić Dražilov, Sanja, Nikčević, Gordana, Baščarević, Zoran, Stojiljković, Maja, Pavlović, Sonja, Spasovski, Duško, "Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471." in Diagnostics, 13, no. 3 (2023):471,
https://hdl.handle.net/21.15107/rcub_imagine_2089 .

TY  - CONF
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Spasovski, Vesna
AU  - Stevanović, Nina
AU  - Parezanović, Marina
AU  - Stanković, Sara
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2139
AB  - Introduction: Thalassemia syndromes are heterogeneous group of hereditary anemias characterized
by defects in the synthesis of hemoglobin (Hb) polypeptide chains. These disorders comprise thalassemias and thalassemic hemoglobin variants which are predominantly caused by mutationsin a- and
b-globin genes (HBA and HBB genes). Clinical manifestations of thalassemia syndromes range from
asymptomatic thalassemia minor to severe anemia in thalassemia major cases. The aim of thisstudy was
to update our previous findings on frequency of thalassemia mutations which result from a 13-year-old
systematic survey in Serbia.
Methods: Two hundred and fourteen patients from 149 unrelated families presented with hematological parameters indicative of thalassemia syndromes were studied. Detection of α- and β-globin gene
mutations was performed using PCR and direct sequencing.
Results: Two Hb variants and twelve different β-thalassemia mutations, including two mutations previously not reported in Serbian population, were detected. Hb variant Lepore Boston-Washington wasthe
most common cause of thalassemia, with frequency of 24.3%, followed by HBB:c.316-106C>G mutation
detected in 18.1% of families. The third most frequent cause of β-thalassemia were HBB:c.118C>T and
HBB:c.93-21G>A mutations with 16.6% incidence each. Together, these four variants account for over
75% of all mutated β-globin alleles. In addition, five families affected with α-thalassemia were detected.
Conclusion: Despite the increase in cohort size by 50% between this and our previous studies, the frequency of mutations affecting HBB gene remained unchanged. Results presented in this study will update Serbian national mutation database and contribute to better understanding of geographic history
of South European and Balkan populations.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Molecular basis of thalassemia syndromes in Serbia: an update
EP  - 86
SP  - 86
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2139
ER  - 

@conference{
author = "Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Spasovski, Vesna and Stevanović, Nina and Parezanović, Marina and Stanković, Sara and Stojiljković, Maja and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: Thalassemia syndromes are heterogeneous group of hereditary anemias characterized
by defects in the synthesis of hemoglobin (Hb) polypeptide chains. These disorders comprise thalassemias and thalassemic hemoglobin variants which are predominantly caused by mutationsin a- and
b-globin genes (HBA and HBB genes). Clinical manifestations of thalassemia syndromes range from
asymptomatic thalassemia minor to severe anemia in thalassemia major cases. The aim of thisstudy was
to update our previous findings on frequency of thalassemia mutations which result from a 13-year-old
systematic survey in Serbia.
Methods: Two hundred and fourteen patients from 149 unrelated families presented with hematological parameters indicative of thalassemia syndromes were studied. Detection of α- and β-globin gene
mutations was performed using PCR and direct sequencing.
Results: Two Hb variants and twelve different β-thalassemia mutations, including two mutations previously not reported in Serbian population, were detected. Hb variant Lepore Boston-Washington wasthe
most common cause of thalassemia, with frequency of 24.3%, followed by HBB:c.316-106C>G mutation
detected in 18.1% of families. The third most frequent cause of β-thalassemia were HBB:c.118C>T and
HBB:c.93-21G>A mutations with 16.6% incidence each. Together, these four variants account for over
75% of all mutated β-globin alleles. In addition, five families affected with α-thalassemia were detected.
Conclusion: Despite the increase in cohort size by 50% between this and our previous studies, the frequency of mutations affecting HBB gene remained unchanged. Results presented in this study will update Serbian national mutation database and contribute to better understanding of geographic history
of South European and Balkan populations.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Molecular basis of thalassemia syndromes in Serbia: an update",
pages = "86-86",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2139"
}

Ugrin, M., Komazec, J., Klaassen, K., Skakić, A., Anđelković, M., Spasovski, V., Stevanović, N., Parezanović, M., Stanković, S., Stojiljković, M.,& Pavlović, S.. (2023). Molecular basis of thalassemia syndromes in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2139

Ugrin M, Komazec J, Klaassen K, Skakić A, Anđelković M, Spasovski V, Stevanović N, Parezanović M, Stanković S, Stojiljković M, Pavlović S. Molecular basis of thalassemia syndromes in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2139 .

Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Spasovski, Vesna, Stevanović, Nina, Parezanović, Marina, Stanković, Sara, Stojiljković, Maja, Pavlović, Sonja, "Molecular basis of thalassemia syndromes in Serbia: an update" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):86-86,
https://hdl.handle.net/21.15107/rcub_imagine_2139 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Stevanović, Nina
AU  - Anđelković, Marina
AU  - Ugrin, Milena
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2176
AB  - Background: The current therapy for glycogen
storage disease Ib (GSD Ib) fails to prevent the
development of renal dysfunction, and hepatocellular/
renal carcinoma in many patients. Therefore,
new therapies for the treatment of life-threatening
complications of GSD Ib are of great interest. Recent
studies revealed that chronic endoplasmic reticulum
(ER) stress and increased apoptosis are
involved in pathogenesis of GSD Ib, whereas small
molecule phenylbutyrate (4-PBA) showed the capability
of reducing ER stress-induced apoptosis.
Methods: To analyze the function of 4-PBA
as ER stress inhibitor, we created a G6PT-deficient
FlpInHEK293 cell line using the CRISPR/Cas9
knockout method and tested if 4-PBA could decrease
chronic metabolic stress and prevent cell
death. We analyzed molecular markers of unfolded
protein response (ATF4, DDIT3, HSPA5, XBP1s),
and apoptosis (BCL2/BAX, CASP3, CASP7) in
G6PT-deficient cells before and upon the treatment
using RT-qPCR method.Results: Treatment with the most effective
dose of 1 mM 4-PBA reduced the expression of
executioner caspases (CASP3, CASP7) and increased
the BCL2/BAX ratio, indicating a reduced
apoptosis level. Additionally, 4-PBA decreased
UPR marker expression in G6PT-deficient cells.
Our results proved the concept that 4-PBA could
alleviate markers of ER stress detected in the GSD
Ib in vitro model system and prevent cell death.
Conclusion: We demonstrated, for the first
time, the potential of 4-PBA to be repurposed for
patients with GSD Ib and open perspectives for
translational research that could contribute to a
knowledge of GSD Ib treatments and other genetic
diseases where chronic ER stress-induced apoptosis
contribute to the disease pathology.
PB  - International Journal of Medical Genetic
C3  - International Journal of Medical Genetics
T1  - PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM
EP  - 54
IS  - Supplement
SP  - 54
VL  - 26
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2176
ER  - 

@conference{
author = "Parezanović, Marina and Stevanović, Nina and Anđelković, Marina and Ugrin, Milena and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Background: The current therapy for glycogen
storage disease Ib (GSD Ib) fails to prevent the
development of renal dysfunction, and hepatocellular/
renal carcinoma in many patients. Therefore,
new therapies for the treatment of life-threatening
complications of GSD Ib are of great interest. Recent
studies revealed that chronic endoplasmic reticulum
(ER) stress and increased apoptosis are
involved in pathogenesis of GSD Ib, whereas small
molecule phenylbutyrate (4-PBA) showed the capability
of reducing ER stress-induced apoptosis.
Methods: To analyze the function of 4-PBA
as ER stress inhibitor, we created a G6PT-deficient
FlpInHEK293 cell line using the CRISPR/Cas9
knockout method and tested if 4-PBA could decrease
chronic metabolic stress and prevent cell
death. We analyzed molecular markers of unfolded
protein response (ATF4, DDIT3, HSPA5, XBP1s),
and apoptosis (BCL2/BAX, CASP3, CASP7) in
G6PT-deficient cells before and upon the treatment
using RT-qPCR method.Results: Treatment with the most effective
dose of 1 mM 4-PBA reduced the expression of
executioner caspases (CASP3, CASP7) and increased
the BCL2/BAX ratio, indicating a reduced
apoptosis level. Additionally, 4-PBA decreased
UPR marker expression in G6PT-deficient cells.
Our results proved the concept that 4-PBA could
alleviate markers of ER stress detected in the GSD
Ib in vitro model system and prevent cell death.
Conclusion: We demonstrated, for the first
time, the potential of 4-PBA to be repurposed for
patients with GSD Ib and open perspectives for
translational research that could contribute to a
knowledge of GSD Ib treatments and other genetic
diseases where chronic ER stress-induced apoptosis
contribute to the disease pathology.",
publisher = "International Journal of Medical Genetic",
journal = "International Journal of Medical Genetics",
title = "PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM",
pages = "54-54",
number = "Supplement",
volume = "26",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2176"
}

Parezanović, M., Stevanović, N., Anđelković, M., Ugrin, M., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM. in International Journal of Medical Genetics
International Journal of Medical Genetic., 26(Supplement), 54-54.
https://hdl.handle.net/21.15107/rcub_imagine_2176

Parezanović M, Stevanović N, Anđelković M, Ugrin M, Pavlović S, Stojiljković M, Skakić A. PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM. in International Journal of Medical Genetics. 2023;26(Supplement):54-54.
https://hdl.handle.net/21.15107/rcub_imagine_2176 .

Parezanović, Marina, Stevanović, Nina, Anđelković, Marina, Ugrin, Milena, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM" in International Journal of Medical Genetics, 26, no. Supplement (2023):54-54,
https://hdl.handle.net/21.15107/rcub_imagine_2176 .

TY  - JOUR
AU  - Spasovski, Vesna
AU  - Srzentić Dražilov, Sanja
AU  - Nikčević, Gordana
AU  - Baščarević, Zoran
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Spasovski, Duško
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2088
AB  - Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.
PB  - MDPI
T2  - Diagnostics
T1  - Molecular Biomarkers in Perthes Disease: A Review
IS  - 3
SP  - 471
VL  - 13
DO  - 10.3390/diagnostics13030471
ER  - 

@article{
author = "Spasovski, Vesna and Srzentić Dražilov, Sanja and Nikčević, Gordana and Baščarević, Zoran and Stojiljković, Maja and Pavlović, Sonja and Spasovski, Duško",
year = "2023",
abstract = "Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.",
publisher = "MDPI",
journal = "Diagnostics",
title = "Molecular Biomarkers in Perthes Disease: A Review",
number = "3",
pages = "471",
volume = "13",
doi = "10.3390/diagnostics13030471"
}

Spasovski, V., Srzentić Dražilov, S., Nikčević, G., Baščarević, Z., Stojiljković, M., Pavlović, S.,& Spasovski, D.. (2023). Molecular Biomarkers in Perthes Disease: A Review. in Diagnostics
MDPI., 13(3), 471.
https://doi.org/10.3390/diagnostics13030471

Spasovski V, Srzentić Dražilov S, Nikčević G, Baščarević Z, Stojiljković M, Pavlović S, Spasovski D. Molecular Biomarkers in Perthes Disease: A Review. in Diagnostics. 2023;13(3):471.
doi:10.3390/diagnostics13030471 .

Spasovski, Vesna, Srzentić Dražilov, Sanja, Nikčević, Gordana, Baščarević, Zoran, Stojiljković, Maja, Pavlović, Sonja, Spasovski, Duško, "Molecular Biomarkers in Perthes Disease: A Review" in Diagnostics, 13, no. 3 (2023):471,
https://doi.org/10.3390/diagnostics13030471 . .

TY  - CONF
AU  - Skakić, Anita
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Parezanović, Marina
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1903
AB  - Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and
reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than
40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and
adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation
sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and
Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene
were detected in one patient suspected of PCD. The results of in silico prediction showed that the
p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites
and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The
p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and
is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method
showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the
control group, while Western blot analysis showed the presence of two protein products (699 ak and
455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and
p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function
and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia
IS  - 2 (Special edition)
SP  - 110
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1903
ER  - 

@conference{
author = "Skakić, Anita and Stevanović, Nina and Spasovski, Vesna and Parezanović, Marina and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Anđelković, Marina",
year = "2023",
abstract = "Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and
reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than
40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and
adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation
sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and
Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene
were detected in one patient suspected of PCD. The results of in silico prediction showed that the
p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites
and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The
p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and
is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method
showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the
control group, while Western blot analysis showed the presence of two protein products (699 ak and
455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and
p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function
and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia",
number = "2 (Special edition)",
pages = "110",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1903"
}

Skakić, A., Stevanović, N., Spasovski, V., Parezanović, M., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Anđelković, M.. (2023). Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 110.
https://hdl.handle.net/21.15107/rcub_imagine_1903

Skakić A, Stevanović N, Spasovski V, Parezanović M, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Anđelković M. Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications. 2023;7(2 (Special edition)):110.
https://hdl.handle.net/21.15107/rcub_imagine_1903 .

Skakić, Anita, Stevanović, Nina, Spasovski, Vesna, Parezanović, Marina, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Anđelković, Marina, "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia" in Genetics & Applications, 7, no. 2 (Special edition) (2023):110,
https://hdl.handle.net/21.15107/rcub_imagine_1903 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Klaassen, Kristel
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Stanković, Sara
AU  - Jocić, Nikola
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2121
AB  - Introduction: Glycogen storage disease type Ib (GSD-Ib) is an autosomal recessive disorder characterized by fasting hypoglycemia and the accumulation of glycogen in the liver, kidneys and intestinal mucosa. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis
play a role in the progression of disease manifestations. Although dietary control is commonly utilized
to manage hypoglycemia, there is still a lack of effective pharmacological therapy. Therefore, the establishment of proper model system is essential for testing novel treatment approaches.
Methods: To create GSD-Ib in vitro model system, CRISPR/Cas9-knockout (KO) method was used to introduce a deletion in SLC37A4 gene in the FlpInHEK293 cells. Characterization of CRISPR/Cas9-KO model
system was performed using Sanger sequencing, RT-qPCR and Western Blot. Additionally, the expression analysis of ER stress and apoptotic markers was performed.
Results: Sanger sequencing confirmed the presence of c.14_100del in SLC37A4 gene. The expression
level of SLC37A4 was decreased to 26.8% in the SLC37A4-/- cell line compared to the SLC37A4 wild-type
along with Western blot analysis, which confirmed reduced target protein level in SLC37A4-/- clones. Furthermore, ER stress (ATF4, DDIT3, HSPA5, XBP1s) and apoptotic (BCL2, BAX, CASP3, CASP7) markers expression levels were significantly altered in SLC37A4-/- clones compared to wild-type, which proved that
we created a suitable GSD-Ib in vitro model system.
Conclusion: Utilizing CRISPR/Cas9 technology, we established cellular GSD-Ib modelsystem that mirrors
increased ER stress and apoptosis. This model system could be used to facilitate a comprehensive understanding of disease mechanisms and enable testing of potential treatment effectiveness.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB
EP  - 65
SP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2121
ER  - 

@conference{
author = "Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Klaassen, Kristel and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Stanković, Sara and Jocić, Nikola and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Introduction: Glycogen storage disease type Ib (GSD-Ib) is an autosomal recessive disorder characterized by fasting hypoglycemia and the accumulation of glycogen in the liver, kidneys and intestinal mucosa. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis
play a role in the progression of disease manifestations. Although dietary control is commonly utilized
to manage hypoglycemia, there is still a lack of effective pharmacological therapy. Therefore, the establishment of proper model system is essential for testing novel treatment approaches.
Methods: To create GSD-Ib in vitro model system, CRISPR/Cas9-knockout (KO) method was used to introduce a deletion in SLC37A4 gene in the FlpInHEK293 cells. Characterization of CRISPR/Cas9-KO model
system was performed using Sanger sequencing, RT-qPCR and Western Blot. Additionally, the expression analysis of ER stress and apoptotic markers was performed.
Results: Sanger sequencing confirmed the presence of c.14_100del in SLC37A4 gene. The expression
level of SLC37A4 was decreased to 26.8% in the SLC37A4-/- cell line compared to the SLC37A4 wild-type
along with Western blot analysis, which confirmed reduced target protein level in SLC37A4-/- clones. Furthermore, ER stress (ATF4, DDIT3, HSPA5, XBP1s) and apoptotic (BCL2, BAX, CASP3, CASP7) markers expression levels were significantly altered in SLC37A4-/- clones compared to wild-type, which proved that
we created a suitable GSD-Ib in vitro model system.
Conclusion: Utilizing CRISPR/Cas9 technology, we established cellular GSD-Ib modelsystem that mirrors
increased ER stress and apoptosis. This model system could be used to facilitate a comprehensive understanding of disease mechanisms and enable testing of potential treatment effectiveness.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB",
pages = "65-65",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2121"
}

Parezanović, M., Anđelković, M., Stevanović, N., Klaassen, K., Spasovski, V., Ugrin, M., Komazec, J., Stanković, S., Jocić, N., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 65-65.
https://hdl.handle.net/21.15107/rcub_imagine_2121

Parezanović M, Anđelković M, Stevanović N, Klaassen K, Spasovski V, Ugrin M, Komazec J, Stanković S, Jocić N, Pavlović S, Stojiljković M, Skakić A. Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:65-65.
https://hdl.handle.net/21.15107/rcub_imagine_2121 .

Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Klaassen, Kristel, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Stanković, Sara, Jocić, Nikola, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):65-65,
https://hdl.handle.net/21.15107/rcub_imagine_2121 .

TY  - CONF
AU  - Stojiljković, Maja
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Komazec, Jovana
AU  - Ugrin, Milena
AU  - Spasovski, Vesna
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2111
AB  - Introduction: All inborn metabolic diseases are rare, having a prevalence less than 1:2000. Vast majority of them are monogenic and finding pathogenic genetic variantsis needed to setthe correct diagnosis, enable adequate treatment and provide genetic counseling to members of affected family. Thisstudy is an overview of genomic studies of rare metabolic diseases in Serbia. Methods: Since 2005, more than 300 patients suspected to have a rare metabolic or neurometabolic disease have been analyzed using sanger sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed). Results: Disease-causing variants were found in more than 60 different genes associated with a metabolic or neurometabolic disease. The most frequent disease was phenylketonuria (109 patients), followed by glycogen storage disease Ib (30 patients), while majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified in a group of phenylketonuria patients with an unusual phenotype. Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis and allowsstudying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore, characterization of novel genetic targets boosts development of precision medicine
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Rare metabolic diseases in the genomics era
EP  - 36
SP  - 36
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2111
ER  - 

@conference{
author = "Stojiljković, Maja and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Komazec, Jovana and Ugrin, Milena and Spasovski, Vesna and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: All inborn metabolic diseases are rare, having a prevalence less than 1:2000. Vast majority of them are monogenic and finding pathogenic genetic variantsis needed to setthe correct diagnosis, enable adequate treatment and provide genetic counseling to members of affected family. Thisstudy is an overview of genomic studies of rare metabolic diseases in Serbia. Methods: Since 2005, more than 300 patients suspected to have a rare metabolic or neurometabolic disease have been analyzed using sanger sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed). Results: Disease-causing variants were found in more than 60 different genes associated with a metabolic or neurometabolic disease. The most frequent disease was phenylketonuria (109 patients), followed by glycogen storage disease Ib (30 patients), while majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified in a group of phenylketonuria patients with an unusual phenotype. Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis and allowsstudying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore, characterization of novel genetic targets boosts development of precision medicine",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Rare metabolic diseases in the genomics era",
pages = "36-36",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2111"
}

Stojiljković, M., Klaassen, K., Skakić, A., Anđelković, M., Komazec, J., Ugrin, M., Spasovski, V.,& Pavlović, S.. (2023). Rare metabolic diseases in the genomics era. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 36-36.
https://hdl.handle.net/21.15107/rcub_imagine_2111

Stojiljković M, Klaassen K, Skakić A, Anđelković M, Komazec J, Ugrin M, Spasovski V, Pavlović S. Rare metabolic diseases in the genomics era. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:36-36.
https://hdl.handle.net/21.15107/rcub_imagine_2111 .

Stojiljković, Maja, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Komazec, Jovana, Ugrin, Milena, Spasovski, Vesna, Pavlović, Sonja, "Rare metabolic diseases in the genomics era" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):36-36,
https://hdl.handle.net/21.15107/rcub_imagine_2111 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1901
AB  - Fabry disease (FD) is a rare X-linked disorder caused by variants in the GLA gene leading to the
deficiency of lysosomal α-galactosidase-A and progressive accumulation of globotriaosylceramide
affecting the heart, nervous system, and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, a precise molecular-genetic diagnosis and the earliest possible treatment are
essential to avoid significant disease progression. The study aimed to determine the strategy for
establishing routine molecular genetic diagnostics of FD in Serbia to provide an early application of
appropriate therapy and genetic advice to families with a high risk for the birth of a child with FD. We
analyzed 95 (34 female and 61 male) hemodialysis patients with clinical suspicion of FD using Sanger
sequencing of all coding exons (7) and flanking intron regions of the GLA gene and measured the
relative expression of the GLA gene in available samples. The genetic analysis revealed 3 patients with
a missense variant (p.Asp313Tyr), and 10 patients with combinations of non-coding variants, described
as complex intronic haplotypes (CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7 (7.4%) patients. Lyso-Gb3 biomarker
levels were within the normal range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of the GLA gene in PBMC of 2 female patients with CIH1 and one
female patient carrying only c.-10C>T variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that
further analyses are needed to confirm/exclude FD in these patients. Because the effects of CIHs are not
yet fully understood, our work highlights the importance of analyzing intronic regions of the GLA gene
as genetic modifiers and the need to include expression analysis in the diagnostic algorithm.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology
IS  - 2 (Special edition)
IS  - 107
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1901
ER  - 

@conference{
author = "Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Fabry disease (FD) is a rare X-linked disorder caused by variants in the GLA gene leading to the
deficiency of lysosomal α-galactosidase-A and progressive accumulation of globotriaosylceramide
affecting the heart, nervous system, and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, a precise molecular-genetic diagnosis and the earliest possible treatment are
essential to avoid significant disease progression. The study aimed to determine the strategy for
establishing routine molecular genetic diagnostics of FD in Serbia to provide an early application of
appropriate therapy and genetic advice to families with a high risk for the birth of a child with FD. We
analyzed 95 (34 female and 61 male) hemodialysis patients with clinical suspicion of FD using Sanger
sequencing of all coding exons (7) and flanking intron regions of the GLA gene and measured the
relative expression of the GLA gene in available samples. The genetic analysis revealed 3 patients with
a missense variant (p.Asp313Tyr), and 10 patients with combinations of non-coding variants, described
as complex intronic haplotypes (CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7 (7.4%) patients. Lyso-Gb3 biomarker
levels were within the normal range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of the GLA gene in PBMC of 2 female patients with CIH1 and one
female patient carrying only c.-10C>T variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that
further analyses are needed to confirm/exclude FD in these patients. Because the effects of CIHs are not
yet fully understood, our work highlights the importance of analyzing intronic regions of the GLA gene
as genetic modifiers and the need to include expression analysis in the diagnostic algorithm.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology",
number = "2 (Special edition), 107",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1901"
}

Parezanović, M., Anđelković, M., Stevanović, N., Spasovski, V., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)).
https://hdl.handle.net/21.15107/rcub_imagine_1901

Parezanović M, Anđelković M, Stevanović N, Spasovski V, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Skakić A. High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology. in Genetics & Applications. 2023;7(2 (Special edition)).
https://hdl.handle.net/21.15107/rcub_imagine_1901 .

Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology" in Genetics & Applications, 7, no. 2 (Special edition) (2023),
https://hdl.handle.net/21.15107/rcub_imagine_1901 .

TY  - CONF
AU  - Klaassen, Kristel
AU  - Šinžar, Ksenija
AU  - Stanković, Sara
AU  - Đorđević Milošević, Maja
AU  - Kecman, Božica
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Parezanović, Marina
AU  - Stevanović, Nina
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2142
AB  - Introduction: Phenylketonuria (PKU) is the most frequent inborn disorder of amino acid metabolism
caused by variants in human phenylalanine hydroxylase gene (PAH).
Methods: In thisstudy (an update for the time period of 10 years, with patientsfrom our previousstudies included) a total of 109 PKU patients from Serbia were analyzed. They were classified into three phenotypic categories in accordance with pre-treatment plasma phenylalanine level: classic PKU, mild PKU
and mild hyperphenylalaninemia. For genetic analyses, we combined Sanger sequencing, MLPA and
next generation sequencing to identify disease-causing variantsin PAH gene, which were further classified using ACMG classification. Additionally, we used in silico and/or eukaryotic expression studiesto assess the effect of novel genetic variants identified in our patients.
Results: Disease-causing variants were identified in 217 of 218 alleles, reaching detection rate of 99.5%.
We detected a total of 32 different variants, of which 29 previously described and three novel ones:
p.Gln226Lys, p.Pro244His and p.Pro416Leu. In silico and/or eukaryotic expression studies confirmed pathogenic effect of all novel genetic variants. The most frequent variant was p.Leu48Ser (31.2%), followed
by p.Arg408Trp (13.8%), p.Ile306Val (9.2%). p.Glu390Gly (5%), p.Pro281Leu (4.6%), and p.Arg261Gln
(3.2%). All detected disease-causing variants were classified as pathogenic using ACMG classification.
Conclusion: Our study brings the updated spectrum of molecular genetic data, variant classification
and detailed phenotypic characteristicsfor PKU patientsfrom Serbia. Therefore, ourstudy contributesto
better understanding of molecular landscape of PKU in Europe and to general knowledge on genotype–
phenotype correlation in PKU.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Molecular basis of phenylketonuria in Serbia: an update
EP  - 93
SP  - 93
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2142
ER  - 

@conference{
author = "Klaassen, Kristel and Šinžar, Ksenija and Stanković, Sara and Đorđević Milošević, Maja and Kecman, Božica and Anđelković, Marina and Skakić, Anita and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Parezanović, Marina and Stevanović, Nina and Pavlović, Sonja and Stojiljković, Maja",
year = "2023",
abstract = "Introduction: Phenylketonuria (PKU) is the most frequent inborn disorder of amino acid metabolism
caused by variants in human phenylalanine hydroxylase gene (PAH).
Methods: In thisstudy (an update for the time period of 10 years, with patientsfrom our previousstudies included) a total of 109 PKU patients from Serbia were analyzed. They were classified into three phenotypic categories in accordance with pre-treatment plasma phenylalanine level: classic PKU, mild PKU
and mild hyperphenylalaninemia. For genetic analyses, we combined Sanger sequencing, MLPA and
next generation sequencing to identify disease-causing variantsin PAH gene, which were further classified using ACMG classification. Additionally, we used in silico and/or eukaryotic expression studiesto assess the effect of novel genetic variants identified in our patients.
Results: Disease-causing variants were identified in 217 of 218 alleles, reaching detection rate of 99.5%.
We detected a total of 32 different variants, of which 29 previously described and three novel ones:
p.Gln226Lys, p.Pro244His and p.Pro416Leu. In silico and/or eukaryotic expression studies confirmed pathogenic effect of all novel genetic variants. The most frequent variant was p.Leu48Ser (31.2%), followed
by p.Arg408Trp (13.8%), p.Ile306Val (9.2%). p.Glu390Gly (5%), p.Pro281Leu (4.6%), and p.Arg261Gln
(3.2%). All detected disease-causing variants were classified as pathogenic using ACMG classification.
Conclusion: Our study brings the updated spectrum of molecular genetic data, variant classification
and detailed phenotypic characteristicsfor PKU patientsfrom Serbia. Therefore, ourstudy contributesto
better understanding of molecular landscape of PKU in Europe and to general knowledge on genotype–
phenotype correlation in PKU.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Molecular basis of phenylketonuria in Serbia: an update",
pages = "93-93",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2142"
}

Klaassen, K., Šinžar, K., Stanković, S., Đorđević Milošević, M., Kecman, B., Anđelković, M., Skakić, A., Spasovski, V., Ugrin, M., Komazec, J., Parezanović, M., Stevanović, N., Pavlović, S.,& Stojiljković, M.. (2023). Molecular basis of phenylketonuria in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 93-93.
https://hdl.handle.net/21.15107/rcub_imagine_2142

Klaassen K, Šinžar K, Stanković S, Đorđević Milošević M, Kecman B, Anđelković M, Skakić A, Spasovski V, Ugrin M, Komazec J, Parezanović M, Stevanović N, Pavlović S, Stojiljković M. Molecular basis of phenylketonuria in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:93-93.
https://hdl.handle.net/21.15107/rcub_imagine_2142 .

Klaassen, Kristel, Šinžar, Ksenija, Stanković, Sara, Đorđević Milošević, Maja, Kecman, Božica, Anđelković, Marina, Skakić, Anita, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Parezanović, Marina, Stevanović, Nina, Pavlović, Sonja, Stojiljković, Maja, "Molecular basis of phenylketonuria in Serbia: an update" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):93-93,
https://hdl.handle.net/21.15107/rcub_imagine_2142 .

TY  - CONF
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Stevanović, Nina
AU  - Parezanović, Marina
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Spasovski, Vesna
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1900
AB  - Rare lung diseases (RLDs) are a group of diseases that individually affect one in 2,000 people, with an
estimate that about 80% of RLDs have a genetic origin. Despite the variations among RLDs in clinical
characteristics and manifestations, most of these diseases similarly damage the lungs, making diagnosis
difficult. The utility of NGS technology in RLDs for diagnostic purposes allows a better understanding
of the genetic background, however, the identification and classification of disease-causing variants are
challenging. Further, numerous VUS (variants of uncertain significance) that cannot be precisely
defined and classified are produced. The main goal of this study was to create a unique guideline that
will enable the standardization of the assessment of novel genetic variants in RLDs causative genes.
The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of
genes/variants, (2) classification of variants, and (3) characterization of variants using in silico
structural and functional analysis. The pipeline validation was performed through the analysis of
variants detected in a disease-causing and candidate genes of one of the RLDSs, and detected VUS
variants have gained diagnostic significance. The application of this pipeline resulted in the
identification and classification of novel variants, through analysis at the transcriptional, translational,
and posttranslational levels, and led to accurate diagnosis.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data
IS  - 2 (Special edition)
SP  - 104
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1900
ER  - 

@conference{
author = "Anđelković, Marina and Skakić, Anita and Stevanović, Nina and Parezanović, Marina and Komazec, Jovana and Klaassen, Kristel and Spasovski, Vesna and Stojiljković, Maja and Pavlović, Sonja",
year = "2023",
abstract = "Rare lung diseases (RLDs) are a group of diseases that individually affect one in 2,000 people, with an
estimate that about 80% of RLDs have a genetic origin. Despite the variations among RLDs in clinical
characteristics and manifestations, most of these diseases similarly damage the lungs, making diagnosis
difficult. The utility of NGS technology in RLDs for diagnostic purposes allows a better understanding
of the genetic background, however, the identification and classification of disease-causing variants are
challenging. Further, numerous VUS (variants of uncertain significance) that cannot be precisely
defined and classified are produced. The main goal of this study was to create a unique guideline that
will enable the standardization of the assessment of novel genetic variants in RLDs causative genes.
The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of
genes/variants, (2) classification of variants, and (3) characterization of variants using in silico
structural and functional analysis. The pipeline validation was performed through the analysis of
variants detected in a disease-causing and candidate genes of one of the RLDSs, and detected VUS
variants have gained diagnostic significance. The application of this pipeline resulted in the
identification and classification of novel variants, through analysis at the transcriptional, translational,
and posttranslational levels, and led to accurate diagnosis.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data",
number = "2 (Special edition)",
pages = "104",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1900"
}

Anđelković, M., Skakić, A., Stevanović, N., Parezanović, M., Komazec, J., Klaassen, K., Spasovski, V., Stojiljković, M.,& Pavlović, S.. (2023). Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 104.
https://hdl.handle.net/21.15107/rcub_imagine_1900

Anđelković M, Skakić A, Stevanović N, Parezanović M, Komazec J, Klaassen K, Spasovski V, Stojiljković M, Pavlović S. Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data. in Genetics & Applications. 2023;7(2 (Special edition)):104.
https://hdl.handle.net/21.15107/rcub_imagine_1900 .

Anđelković, Marina, Skakić, Anita, Stevanović, Nina, Parezanović, Marina, Komazec, Jovana, Klaassen, Kristel, Spasovski, Vesna, Stojiljković, Maja, Pavlović, Sonja, "Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data" in Genetics & Applications, 7, no. 2 (Special edition) (2023):104,
https://hdl.handle.net/21.15107/rcub_imagine_1900 .

TY  - JOUR
AU  - Spasovski, Vesna
AU  - Anđelković, Marina
AU  - Parezanović, Marina
AU  - Komazec, Jovana
AU  - Ugrin, Milena
AU  - Klaassen, Kristel
AU  - Stojiljković, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2085
AB  - Systemic sclerosis (SSc) is a complex autoimmune inflammatory disorder with multiple organ involvement. Skin changes present the hallmark of SSc and coincide with poor prognosis. Interstitial lung diseases (ILD) are the most widely reported complications in SSc patients and the primary cause of death. It has been proposed that the processes of autophagy and apoptosis could play a significant role in the pathogenesis and clinical course of different autoimmune diseases, and accordingly in SSc. In this manuscript, we review the current knowledge of autophagy and apoptosis processes in the skin and lungs of patients with SSc. Profiling of markers involved in these processes in skin cells can be useful to recognize the stage of fibrosis and can be used in the clinical stratification of patients. Furthermore, the knowledge of the molecular mechanisms underlying these processes enables the repurposing of already known drugs and the development of new biological therapeutics that aim to reverse fibrosis by promoting apoptosis and regulate autophagy in personalized treatment approach. In SSc-ILD patients, the molecular signature of the lung tissues of each patient could be a distinctive criterion in order to establish the correct lung pattern, which directly impacts the course and prognosis of the disease. In this case, resolving the role of tissue-specific markers, which could be detected in the circulation using sensitive molecular methods, would be an important step toward development of non-invasive diagnostic procedures that enable early and precise diagnosis and preventing the high mortality of this rare disease.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis
IS  - 13
SP  - 11212
VL  - 24
DO  - 10.3390/ijms241311212
ER  - 

@article{
author = "Spasovski, Vesna and Anđelković, Marina and Parezanović, Marina and Komazec, Jovana and Ugrin, Milena and Klaassen, Kristel and Stojiljković, Maja",
year = "2023",
abstract = "Systemic sclerosis (SSc) is a complex autoimmune inflammatory disorder with multiple organ involvement. Skin changes present the hallmark of SSc and coincide with poor prognosis. Interstitial lung diseases (ILD) are the most widely reported complications in SSc patients and the primary cause of death. It has been proposed that the processes of autophagy and apoptosis could play a significant role in the pathogenesis and clinical course of different autoimmune diseases, and accordingly in SSc. In this manuscript, we review the current knowledge of autophagy and apoptosis processes in the skin and lungs of patients with SSc. Profiling of markers involved in these processes in skin cells can be useful to recognize the stage of fibrosis and can be used in the clinical stratification of patients. Furthermore, the knowledge of the molecular mechanisms underlying these processes enables the repurposing of already known drugs and the development of new biological therapeutics that aim to reverse fibrosis by promoting apoptosis and regulate autophagy in personalized treatment approach. In SSc-ILD patients, the molecular signature of the lung tissues of each patient could be a distinctive criterion in order to establish the correct lung pattern, which directly impacts the course and prognosis of the disease. In this case, resolving the role of tissue-specific markers, which could be detected in the circulation using sensitive molecular methods, would be an important step toward development of non-invasive diagnostic procedures that enable early and precise diagnosis and preventing the high mortality of this rare disease.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis",
number = "13",
pages = "11212",
volume = "24",
doi = "10.3390/ijms241311212"
}

Spasovski, V., Anđelković, M., Parezanović, M., Komazec, J., Ugrin, M., Klaassen, K.,& Stojiljković, M.. (2023). The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis. in International Journal of Molecular Sciences
MDPI., 24(13), 11212.
https://doi.org/10.3390/ijms241311212

Spasovski V, Anđelković M, Parezanović M, Komazec J, Ugrin M, Klaassen K, Stojiljković M. The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis. in International Journal of Molecular Sciences. 2023;24(13):11212.
doi:10.3390/ijms241311212 .

Spasovski, Vesna, Anđelković, Marina, Parezanović, Marina, Komazec, Jovana, Ugrin, Milena, Klaassen, Kristel, Stojiljković, Maja, "The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis" in International Journal of Molecular Sciences, 24, no. 13 (2023):11212,
https://doi.org/10.3390/ijms241311212 . .

TY  - CONF
AU  - Stevanović, Nina
AU  - Skakić, Anita
AU  - Parezanović, Marina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2140
AB  - Introduction: Primary ciliary dyskinesia is a rare and heterogeneous disorder primarily affecting the respiratory organs, with impaired mucociliary clearance being a common characteristic. Recently, the importance of the miR34/449 family in ciliogenesisin animal models has been described. Thisstudy aimed
to establish a modelsystem to study respiratory diseases and assessfor the first time the role of the miR34 family on the mucociliary process in humans.
Methods: We cultured the primary Normal Human Bronchial Epithelial (NHBE) cells in the air-liquid interface system, enabling the differentiation of multiciliated cells(MCCs) and goblet cells(GCs). During the
differentiation process, transient overexpression of miR-34a/b/c members was conducted. The model
system and treatments were validated through confocal microscopy (β-tubulin, MUC5B, MUC5AC antibodies) and qRT-PCR of miRNAs,specifically ciliogenesis markers(NOTCH1, MCIDAS, GEMC1, CCNO, RFX3),
and differentiated cell markers (FOXJ1 and TFF3).
Results: Expression levels of ciliogenesis and differentiated cells markers and detection of cilia and
mucins at confocal microscopy confirmed the successful establishment of cellular modelsystem. During
the initial differentiation stage, an overexpression of miR34a/b/c changed the expression profile of ciliogenesis and differentiated cell markers.
Conclusion: The established model system provides a valuable platform for exploring innovative treatment approaches for lung diseases. These findings suggest that overexpression of miR34a/b/c has impact on mucociliary process by reducing the duration required for the process of ciliogenesis.
Furthermore, the expression levels of differentiated cell markerssuggest increased number of MCCs and
decreased number of GCs, indicating the role of miR34a/b/c in enhancing mucociliary clearance.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system
EP  - 90
SP  - 90
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2140
ER  - 

@conference{
author = "Stevanović, Nina and Skakić, Anita and Parezanović, Marina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Anđelković, Marina",
year = "2023",
abstract = "Introduction: Primary ciliary dyskinesia is a rare and heterogeneous disorder primarily affecting the respiratory organs, with impaired mucociliary clearance being a common characteristic. Recently, the importance of the miR34/449 family in ciliogenesisin animal models has been described. Thisstudy aimed
to establish a modelsystem to study respiratory diseases and assessfor the first time the role of the miR34 family on the mucociliary process in humans.
Methods: We cultured the primary Normal Human Bronchial Epithelial (NHBE) cells in the air-liquid interface system, enabling the differentiation of multiciliated cells(MCCs) and goblet cells(GCs). During the
differentiation process, transient overexpression of miR-34a/b/c members was conducted. The model
system and treatments were validated through confocal microscopy (β-tubulin, MUC5B, MUC5AC antibodies) and qRT-PCR of miRNAs,specifically ciliogenesis markers(NOTCH1, MCIDAS, GEMC1, CCNO, RFX3),
and differentiated cell markers (FOXJ1 and TFF3).
Results: Expression levels of ciliogenesis and differentiated cells markers and detection of cilia and
mucins at confocal microscopy confirmed the successful establishment of cellular modelsystem. During
the initial differentiation stage, an overexpression of miR34a/b/c changed the expression profile of ciliogenesis and differentiated cell markers.
Conclusion: The established model system provides a valuable platform for exploring innovative treatment approaches for lung diseases. These findings suggest that overexpression of miR34a/b/c has impact on mucociliary process by reducing the duration required for the process of ciliogenesis.
Furthermore, the expression levels of differentiated cell markerssuggest increased number of MCCs and
decreased number of GCs, indicating the role of miR34a/b/c in enhancing mucociliary clearance.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system",
pages = "90-90",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2140"
}

Stevanović, N., Skakić, A., Parezanović, M., Spasovski, V., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Anđelković, M.. (2023). The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 90-90.
https://hdl.handle.net/21.15107/rcub_imagine_2140

Stevanović N, Skakić A, Parezanović M, Spasovski V, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Anđelković M. The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:90-90.
https://hdl.handle.net/21.15107/rcub_imagine_2140 .

Stevanović, Nina, Skakić, Anita, Parezanović, Marina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Anđelković, Marina, "The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):90-90,
https://hdl.handle.net/21.15107/rcub_imagine_2140 .

TY  - CONF
AU  - Jocić, Nikola
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Ugrin, Milena
AU  - Spasovski, Vesna
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Komazec, Jovana
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2141
AB  - Introduction: Glycogen storage disease type Ib (GSD-Ib) is characterized by a deficiency of glucose-6-
phosphate translocase (G6PT) encoded by the SLC37A4 gene, affecting glucose homeostasis and disrupting autophagy. Recent findings suggest that G6PT may also play a role in autophagy and
glycogen-selective autophagy (glycophagy) activation independent of its transport function. To investigate this hypothesis, two groups of GSD-Ib patients carrying variants with different effects on G6PT
transport activity and stability (p.Asn27Lys and p.Leu348Valfs*53), were compared to the control group
of subjects.
Methods: The relative expression levels of SLC37A4 gene, autophagy (mTOR, ULK1, PRKAG1), and glycophagy markers(GABARAPL1, GAA, STBD1) were assessed in mononuclear cells of GSD Ib patients(four
carrying p.Asn27Lys and four carrying p.Leu348Valfs*53 variant) compared to control group using RTqPCR. Statistical analysis was performed using one-way ANOVA followed by a post-hoc t-test.
Results: The p.Asn27Lys group exhibited 1.5-2.5 times higher expression of SLC37A4 and autophagy
markers, while the p.Leu348Valfs*53 group showed downregulation by approximately 50% compared to
the control group. Glycophagy markers were increased twofold in both patient groups, except for GAA,
which had similar expression levels as the control group.
Conclusion: Individuals carrying the p.Asn27Lys variant display the presence of SLC37A4 transcript in
their cells, which correlates with autophagy activation. Conversely, in patients with the p.Leu348Valfs*53
variant SLC37A4 is downregulated, indicating compromised autophagy activation. These findings support the role of G6PT in autophagy activation, independent of itstransport activity. Furthermore, the elevated expression of glycophagy markers observed in both patient groups can be attributed to the
accumulated glycogen.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients
EP  - 92
SP  - 92
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2141
ER  - 

@conference{
author = "Jocić, Nikola and Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Ugrin, Milena and Spasovski, Vesna and Klaassen, Kristel and Stanković, Sara and Komazec, Jovana and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Introduction: Glycogen storage disease type Ib (GSD-Ib) is characterized by a deficiency of glucose-6-
phosphate translocase (G6PT) encoded by the SLC37A4 gene, affecting glucose homeostasis and disrupting autophagy. Recent findings suggest that G6PT may also play a role in autophagy and
glycogen-selective autophagy (glycophagy) activation independent of its transport function. To investigate this hypothesis, two groups of GSD-Ib patients carrying variants with different effects on G6PT
transport activity and stability (p.Asn27Lys and p.Leu348Valfs*53), were compared to the control group
of subjects.
Methods: The relative expression levels of SLC37A4 gene, autophagy (mTOR, ULK1, PRKAG1), and glycophagy markers(GABARAPL1, GAA, STBD1) were assessed in mononuclear cells of GSD Ib patients(four
carrying p.Asn27Lys and four carrying p.Leu348Valfs*53 variant) compared to control group using RTqPCR. Statistical analysis was performed using one-way ANOVA followed by a post-hoc t-test.
Results: The p.Asn27Lys group exhibited 1.5-2.5 times higher expression of SLC37A4 and autophagy
markers, while the p.Leu348Valfs*53 group showed downregulation by approximately 50% compared to
the control group. Glycophagy markers were increased twofold in both patient groups, except for GAA,
which had similar expression levels as the control group.
Conclusion: Individuals carrying the p.Asn27Lys variant display the presence of SLC37A4 transcript in
their cells, which correlates with autophagy activation. Conversely, in patients with the p.Leu348Valfs*53
variant SLC37A4 is downregulated, indicating compromised autophagy activation. These findings support the role of G6PT in autophagy activation, independent of itstransport activity. Furthermore, the elevated expression of glycophagy markers observed in both patient groups can be attributed to the
accumulated glycogen.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients",
pages = "92-92",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2141"
}

Jocić, N., Parezanović, M., Anđelković, M., Stevanović, N., Ugrin, M., Spasovski, V., Klaassen, K., Stanković, S., Komazec, J., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 92-92.
https://hdl.handle.net/21.15107/rcub_imagine_2141

Jocić N, Parezanović M, Anđelković M, Stevanović N, Ugrin M, Spasovski V, Klaassen K, Stanković S, Komazec J, Pavlović S, Stojiljković M, Skakić A. Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:92-92.
https://hdl.handle.net/21.15107/rcub_imagine_2141 .

Jocić, Nikola, Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Ugrin, Milena, Spasovski, Vesna, Klaassen, Kristel, Stanković, Sara, Komazec, Jovana, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):92-92,
https://hdl.handle.net/21.15107/rcub_imagine_2141 .

TY  - CONF
AU  - Stojiljković, Maja
AU  - Ugrin, Milena
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Komazec, Jovana
AU  - Spasovski, Vesna
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2175
AB  - Introduction: Rare diseases are heterogeneous group of diseases, with one common characteristics, a
prevalence less than 1 in 2000 people. Vast majority of them are monogenic and finding pathogenic genetic
variants is needed to set the correct diagnosis, enable adequate treatment and provide genetic counselling to
members of affected family. This study is an overview of genomic studies of rare diseases in Serbia.
Methods: More than 1200 patients suspected to have a rare disease have been analyzed using sanger
sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to
find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling
or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed).
Results: Disease-causing variants were found in more than 150 different genes associated with a rare
disease. The most frequent were thalassemia syndromes (214 patients), followed by phenylketonuria (109
patients), congenital adrenal hyperplasia (>90 patients) and glycogen storage disease Ib (30 patients), while
majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively
characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified
in a group of phenylketonuria patients with an unusual phenotype.
Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis
and allows studying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore,
characterization of novel genetic targets boosts development of precision medicine.
PB  - Macedonian Academy of Sciences and Arts
C3  - International Journal of Medical Genetics
T1  - THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES
EP  - 38
IS  - Supplement
SP  - 38
VL  - 26
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2175
ER  - 

@conference{
author = "Stojiljković, Maja and Ugrin, Milena and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Komazec, Jovana and Spasovski, Vesna and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: Rare diseases are heterogeneous group of diseases, with one common characteristics, a
prevalence less than 1 in 2000 people. Vast majority of them are monogenic and finding pathogenic genetic
variants is needed to set the correct diagnosis, enable adequate treatment and provide genetic counselling to
members of affected family. This study is an overview of genomic studies of rare diseases in Serbia.
Methods: More than 1200 patients suspected to have a rare disease have been analyzed using sanger
sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to
find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling
or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed).
Results: Disease-causing variants were found in more than 150 different genes associated with a rare
disease. The most frequent were thalassemia syndromes (214 patients), followed by phenylketonuria (109
patients), congenital adrenal hyperplasia (>90 patients) and glycogen storage disease Ib (30 patients), while
majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively
characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified
in a group of phenylketonuria patients with an unusual phenotype.
Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis
and allows studying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore,
characterization of novel genetic targets boosts development of precision medicine.",
publisher = "Macedonian Academy of Sciences and Arts",
journal = "International Journal of Medical Genetics",
title = "THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES",
pages = "38-38",
number = "Supplement",
volume = "26",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2175"
}

Stojiljković, M., Ugrin, M., Klaassen, K., Skakić, A., Anđelković, M., Komazec, J., Spasovski, V.,& Pavlović, S.. (2023). THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES. in International Journal of Medical Genetics
Macedonian Academy of Sciences and Arts., 26(Supplement), 38-38.
https://hdl.handle.net/21.15107/rcub_imagine_2175

Stojiljković M, Ugrin M, Klaassen K, Skakić A, Anđelković M, Komazec J, Spasovski V, Pavlović S. THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES. in International Journal of Medical Genetics. 2023;26(Supplement):38-38.
https://hdl.handle.net/21.15107/rcub_imagine_2175 .

Stojiljković, Maja, Ugrin, Milena, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Komazec, Jovana, Spasovski, Vesna, Pavlović, Sonja, "THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES" in International Journal of Medical Genetics, 26, no. Supplement (2023):38-38,
https://hdl.handle.net/21.15107/rcub_imagine_2175 .

TY  - CONF
AU  - Stevanović, Nina
AU  - Skakić, Anita
AU  - Spasovski, Vesna
AU  - Stojiljković, Maja
AU  - Parezanović, Marina
AU  - Ugrin, Milena
AU  - Pavlović, Sonja
AU  - Anđelković, Marina
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1738
AB  - Primarna cilijarna diskinezija (PCD) je retka motorna ciliopatija, koja
predominantno utiče na pluća i reproduktivne organe. PCD ima heterogenu
genetičku osnovu, te je neophodno analizirati više od 40 gena uzročnika kako bi
se postavila precizna dijagnoza, što je od suštinskog značaja za optimalno
lečenje i adekvatno genetičko savetovanje. Analizirano je 5 pacijenata suspektnih
na PCD upotrebom metode sekvenciranja nove generacije (NGS). Patogenost
genetičkih varijanti ispitana je in silico, qRT-PCR i Western blot metodama. Kod
jednog pacijenta suspektnog na PCD detektovane su dve novootkrivene varijante
p.N450Lfs*6 i p.D562N u genu DNAI1. Rezultati in silico predikcije pokazali su da
varijanta p.N450Lfs*6 utiče na strukturu 3D modela proteina, da ukida mesta za
vezivanje liganada i posttranslacione modifikacije, čime dolazi do narušavanja
protein-protein interakcija (PPI). Varijanta p.D562N nema uticaja na 3D
strukturu proteina, ali utiče na mesto vezivanja liganda i nalazi se u WD-40
domenu čime najverovatnije narušava PPI. Rezultati metode qRT-PCR pokazali
su snižen nivo ekspresije iRNK DNAI1 za oko 50% kod pacijenta u odnosu na
kontrolnu grupu, dok je Western blot analizom pokazano prisustvo dva proteinska
produkta (699 ak i 455 ak). Analizom dobijenih rezultata, zaključeno je da
promene p.N450Lfs*6 i p.D562N utiču na dužinu i količinu proteina DNAI1
čime dovode do gubitka proteinske funkcije i odgovorne su za nastanak primarne
cilijarne diskinezije kod analiziranog pacijenta.
AB  - Примарна цилијарна дискинезија (ПЦД) је ретка моторна цилиопатија, која
предоминантно утиче на плућа и репродуктивне органе. ПЦД има хетерогену
генетичку основу, те је неопходно анализирати више од 40 гена узрочника како би
се поставила прецизна дијагноза, што је од суштинског значаја за оптимално
лечење и адекватно генетичко саветовање. Анализирано је 5 пацијената суспектних
на ПЦД употребом методе секвенцирања нове генерације (НГС). Патогеност
генетичких варијанти испитана је in silico, qRT-PCR и Western blot методама. Код
једног пацијента суспектног на ПЦД детектоване су две новооткривене варијанте
p.N450Lfs*6 и p.D562N у гену DNAI1. Резултати in silico предикције показали су да
варијанта p.N450Lfs*6 утиче на структуру 3Д модела протеина, да укида места за
везивање лиганада и посттранслационе модификације, чиме долази до нарушавања
протеин-протеин интеракција (ППИ). Варијанта p.D562N нема утицаја на 3Д
структуру протеина, али утиче на место везивања лиганда и налази се у WD-40
домену чиме највероватније нарушава ППИ. Резултати методе qRT-PCR показали
су снижен ниво експресије iRNK DNAI1 за око 50% код пацијента у односу на
контролну групу, док је Western blot анализом показано присуство два протеинска
продукта (699 ак и 455 ак). Анализом добијених резултата, закључено је да
промене p.N450Lfs*6 и p.D562N утичу на дужину и количину протеина DNAI1
чиме доводе до губитка протеинске функције и одговорне су за настанак примарне
цилијарне дискинезије код анализираног пацијента.
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Funkcionalna karatkerizacija novootkrivenih varijanti u genu DNAI1 kod pacijenta sa primarnom cilijarnom diskinezijom
T1  - Функционална караткеризација новооткривених варијанти у гену DNAI1 код пацијента са примарном цилијарном дискинезијом
SP  - 324
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1738
ER  - 

@conference{
author = "Stevanović, Nina and Skakić, Anita and Spasovski, Vesna and Stojiljković, Maja and Parezanović, Marina and Ugrin, Milena and Pavlović, Sonja and Anđelković, Marina",
year = "2022",
abstract = "Primarna cilijarna diskinezija (PCD) je retka motorna ciliopatija, koja
predominantno utiče na pluća i reproduktivne organe. PCD ima heterogenu
genetičku osnovu, te je neophodno analizirati više od 40 gena uzročnika kako bi
se postavila precizna dijagnoza, što je od suštinskog značaja za optimalno
lečenje i adekvatno genetičko savetovanje. Analizirano je 5 pacijenata suspektnih
na PCD upotrebom metode sekvenciranja nove generacije (NGS). Patogenost
genetičkih varijanti ispitana je in silico, qRT-PCR i Western blot metodama. Kod
jednog pacijenta suspektnog na PCD detektovane su dve novootkrivene varijante
p.N450Lfs*6 i p.D562N u genu DNAI1. Rezultati in silico predikcije pokazali su da
varijanta p.N450Lfs*6 utiče na strukturu 3D modela proteina, da ukida mesta za
vezivanje liganada i posttranslacione modifikacije, čime dolazi do narušavanja
protein-protein interakcija (PPI). Varijanta p.D562N nema uticaja na 3D
strukturu proteina, ali utiče na mesto vezivanja liganda i nalazi se u WD-40
domenu čime najverovatnije narušava PPI. Rezultati metode qRT-PCR pokazali
su snižen nivo ekspresije iRNK DNAI1 za oko 50% kod pacijenta u odnosu na
kontrolnu grupu, dok je Western blot analizom pokazano prisustvo dva proteinska
produkta (699 ak i 455 ak). Analizom dobijenih rezultata, zaključeno je da
promene p.N450Lfs*6 i p.D562N utiču na dužinu i količinu proteina DNAI1
čime dovode do gubitka proteinske funkcije i odgovorne su za nastanak primarne
cilijarne diskinezije kod analiziranog pacijenta., Примарна цилијарна дискинезија (ПЦД) је ретка моторна цилиопатија, која
предоминантно утиче на плућа и репродуктивне органе. ПЦД има хетерогену
генетичку основу, те је неопходно анализирати више од 40 гена узрочника како би
се поставила прецизна дијагноза, што је од суштинског значаја за оптимално
лечење и адекватно генетичко саветовање. Анализирано је 5 пацијената суспектних
на ПЦД употребом методе секвенцирања нове генерације (НГС). Патогеност
генетичких варијанти испитана је in silico, qRT-PCR и Western blot методама. Код
једног пацијента суспектног на ПЦД детектоване су две новооткривене варијанте
p.N450Lfs*6 и p.D562N у гену DNAI1. Резултати in silico предикције показали су да
варијанта p.N450Lfs*6 утиче на структуру 3Д модела протеина, да укида места за
везивање лиганада и посттранслационе модификације, чиме долази до нарушавања
протеин-протеин интеракција (ППИ). Варијанта p.D562N нема утицаја на 3Д
структуру протеина, али утиче на место везивања лиганда и налази се у WD-40
домену чиме највероватније нарушава ППИ. Резултати методе qRT-PCR показали
су снижен ниво експресије iRNK DNAI1 за око 50% код пацијента у односу на
контролну групу, док је Western blot анализом показано присуство два протеинска
продукта (699 ак и 455 ак). Анализом добијених резултата, закључено је да
промене p.N450Lfs*6 и p.D562N утичу на дужину и количину протеина DNAI1
чиме доводе до губитка протеинске функције и одговорне су за настанак примарне
цилијарне дискинезије код анализираног пацијента.",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Funkcionalna karatkerizacija novootkrivenih varijanti u genu DNAI1 kod pacijenta sa primarnom cilijarnom diskinezijom, Функционална караткеризација новооткривених варијанти у гену DNAI1 код пацијента са примарном цилијарном дискинезијом",
pages = "324",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1738"
}

Stevanović, N., Skakić, A., Spasovski, V., Stojiljković, M., Parezanović, M., Ugrin, M., Pavlović, S.,& Anđelković, M.. (2022). Funkcionalna karatkerizacija novootkrivenih varijanti u genu DNAI1 kod pacijenta sa primarnom cilijarnom diskinezijom. in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo., 324.
https://hdl.handle.net/21.15107/rcub_imagine_1738

Stevanović N, Skakić A, Spasovski V, Stojiljković M, Parezanović M, Ugrin M, Pavlović S, Anđelković M. Funkcionalna karatkerizacija novootkrivenih varijanti u genu DNAI1 kod pacijenta sa primarnom cilijarnom diskinezijom. in Treći kongres biologa Srbije. 2022;:324.
https://hdl.handle.net/21.15107/rcub_imagine_1738 .

Stevanović, Nina, Skakić, Anita, Spasovski, Vesna, Stojiljković, Maja, Parezanović, Marina, Ugrin, Milena, Pavlović, Sonja, Anđelković, Marina, "Funkcionalna karatkerizacija novootkrivenih varijanti u genu DNAI1 kod pacijenta sa primarnom cilijarnom diskinezijom" in Treći kongres biologa Srbije (2022):324,
https://hdl.handle.net/21.15107/rcub_imagine_1738 .

TY  - CONF
AU  - Stojiljković, Maja
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Parezanović, Marina
AU  - Stevanović, Nina
AU  - Pavlović, Sonja
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1747
AB  - Tokom poslednjih decenija, istraživanja na polju retkih bolesti napreduju
ogromnom brzinom usled sve veće upotrebe sekvenciranja nove generacije (engl.
Next-generation sequencing, NGS). Sekvenciranje kompletnog ljuskog genoma (engl.
Whole genome sequencing, WGS) osoba koje boluju od retkih bolesti je postalo
lako dostupno. Pored pronalaženja novih varijanti i novih gena koji leže u
osnovi retkih bolesti, genomika je omogućila i otkriće gena modifikatora koji
mogu da objasne uočene nedoslednosti u korelaciji genotipa i fenotipa.
Fenilketonurija je urođena metabolička retka bolest koja je uzrokovana
varijantama u genu za fenilalanin hidroksilazu (PAH). U ovoj studiji, sproveli
smo sekvenciranje kompletnog genoma 4 osobe iz nepovezanih nesrodnih porodica
koje su imale patogene varijante u PAH genu, ali nisu razvile fenilketonuriju
uprkos tome što nisu bile lečene. Otkrili smo dve nove varijante, p.Pro1591Ala
u SHANK1 i p.Asp18Asn u SHANK2 genima, kao i prethodno opisane
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro i SHANK3:p.Pro1716Thr
varijante. Računarske predikcije su pokazale da identifikovane varijante ne
ukidaju funkciju SHANK proteina. Međutim, promene u posttranslacionim
modifikacijama SHANK proteina mogu uticati na funkcionisanje
glutamatergičnih sinapsi, regulaciju citoskeleta i doprineti održavanju
optimalne sinaptičke gustine i broja dendritskih bodlji. Naši rezultati po
prvi put povezuju porodicu SHANK gena i osobine neuroloških promena kod
osoba sa fenilketonurijom.
AB  - Током последњих деценија, истраживања на пољу ретких болести напредују
огромном брзином услед све веће употребе секвенцирања нове генерације (енгл.
Next-generation sequencing, NGS). Секвенцирање комплетног љуског генома (енгл.
Whole genome sequencing, WGS) особа које болују од ретких болести је постало
лако доступно. Поред проналажења нових варијанти и нових гена који леже у
основи ретких болести, геномика је омогућила и откриће гена модификатора који
могу да објасне уочене недоследности у корелацији генотипа и фенотипа.
Фенилкетонурија је урођена метаболичка ретка болест која је узрокована
варијантама у гену за фенилаланин хидроксилазу (PAH). У овој студији, спровели
смо секвенцирање комплетног генома 4 особе из неповезаних несродних породица
које су имале патогене варијанте у PAH гену, али нису развиле фенилкетонурију
упркос томе што нису биле лечене. Открили смо две нове варијанте, p.Pro1591Ala
у SHANK1 и p.Asp18Asn у SHANK2 генима, као и претходно описане
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro и SHANK3:p.Pro1716Thr
варијанте. Рачунарске предикције су показале да идентификоване варијанте не
укидају функцију SHANK протеина. Међутим, промене у посттранслационим
модификацијама SHANK протеина могу утицати на функционисање
глутаматергичних синапси, регулацију цитоскелета и допринети одржавању
оптималне синаптичке густине и броја дендритских бодљи. Наши резултати по
први пут повезују породицу SHANK гена и особине неуролошких промена код
особа са фенилкетонуријом.
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Retke bolesti u eri genomike
T1  - Ретке болести у ери геномике
SP  - 287
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1747
ER  - 

@conference{
author = "Stojiljković, Maja and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Parezanović, Marina and Stevanović, Nina and Pavlović, Sonja",
year = "2022",
abstract = "Tokom poslednjih decenija, istraživanja na polju retkih bolesti napreduju
ogromnom brzinom usled sve veće upotrebe sekvenciranja nove generacije (engl.
Next-generation sequencing, NGS). Sekvenciranje kompletnog ljuskog genoma (engl.
Whole genome sequencing, WGS) osoba koje boluju od retkih bolesti je postalo
lako dostupno. Pored pronalaženja novih varijanti i novih gena koji leže u
osnovi retkih bolesti, genomika je omogućila i otkriće gena modifikatora koji
mogu da objasne uočene nedoslednosti u korelaciji genotipa i fenotipa.
Fenilketonurija je urođena metabolička retka bolest koja je uzrokovana
varijantama u genu za fenilalanin hidroksilazu (PAH). U ovoj studiji, sproveli
smo sekvenciranje kompletnog genoma 4 osobe iz nepovezanih nesrodnih porodica
koje su imale patogene varijante u PAH genu, ali nisu razvile fenilketonuriju
uprkos tome što nisu bile lečene. Otkrili smo dve nove varijante, p.Pro1591Ala
u SHANK1 i p.Asp18Asn u SHANK2 genima, kao i prethodno opisane
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro i SHANK3:p.Pro1716Thr
varijante. Računarske predikcije su pokazale da identifikovane varijante ne
ukidaju funkciju SHANK proteina. Međutim, promene u posttranslacionim
modifikacijama SHANK proteina mogu uticati na funkcionisanje
glutamatergičnih sinapsi, regulaciju citoskeleta i doprineti održavanju
optimalne sinaptičke gustine i broja dendritskih bodlji. Naši rezultati po
prvi put povezuju porodicu SHANK gena i osobine neuroloških promena kod
osoba sa fenilketonurijom., Током последњих деценија, истраживања на пољу ретких болести напредују
огромном брзином услед све веће употребе секвенцирања нове генерације (енгл.
Next-generation sequencing, NGS). Секвенцирање комплетног љуског генома (енгл.
Whole genome sequencing, WGS) особа које болују од ретких болести је постало
лако доступно. Поред проналажења нових варијанти и нових гена који леже у
основи ретких болести, геномика је омогућила и откриће гена модификатора који
могу да објасне уочене недоследности у корелацији генотипа и фенотипа.
Фенилкетонурија је урођена метаболичка ретка болест која је узрокована
варијантама у гену за фенилаланин хидроксилазу (PAH). У овој студији, спровели
смо секвенцирање комплетног генома 4 особе из неповезаних несродних породица
које су имале патогене варијанте у PAH гену, али нису развиле фенилкетонурију
упркос томе што нису биле лечене. Открили смо две нове варијанте, p.Pro1591Ala
у SHANK1 и p.Asp18Asn у SHANK2 генима, као и претходно описане
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro и SHANK3:p.Pro1716Thr
варијанте. Рачунарске предикције су показале да идентификоване варијанте не
укидају функцију SHANK протеина. Међутим, промене у посттранслационим
модификацијама SHANK протеина могу утицати на функционисање
глутаматергичних синапси, регулацију цитоскелета и допринети одржавању
оптималне синаптичке густине и броја дендритских бодљи. Наши резултати по
први пут повезују породицу SHANK гена и особине неуролошких промена код
особа са фенилкетонуријом.",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Retke bolesti u eri genomike, Ретке болести у ери геномике",
pages = "287",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1747"
}

Stojiljković, M., Klaassen, K., Skakić, A., Anđelković, M., Spasovski, V., Ugrin, M., Komazec, J., Parezanović, M., Stevanović, N.,& Pavlović, S.. (2022). Retke bolesti u eri genomike. in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo., 287.
https://hdl.handle.net/21.15107/rcub_imagine_1747

Stojiljković M, Klaassen K, Skakić A, Anđelković M, Spasovski V, Ugrin M, Komazec J, Parezanović M, Stevanović N, Pavlović S. Retke bolesti u eri genomike. in Treći kongres biologa Srbije. 2022;:287.
https://hdl.handle.net/21.15107/rcub_imagine_1747 .

Stojiljković, Maja, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Parezanović, Marina, Stevanović, Nina, Pavlović, Sonja, "Retke bolesti u eri genomike" in Treći kongres biologa Srbije (2022):287,
https://hdl.handle.net/21.15107/rcub_imagine_1747 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Tošić, Nataša
AU  - Pavlović, Sonja
AU  - Ćelić, Dejan
AU  - Vučenović, Jelica
AU  - Skakić, Anita
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1740
AB  - Fabrijeva bolest (FB) je retka, monogenska bolest, koja se odlikuje veoma
heterogenom kliničkom slikom izazvanom promenama u genu GLA, smeštenom na
dugom kraku hromozoma H. Usled narušene funkcije lizozomalnog enzima α-
galaktozidaze A dolazi do nagomilavanja Gb3 supstrata, što dovodi do
multisistemskih oštećenja. Precizna molekularno-genetička dijagnoza je
esencijalna kako bi se pacijentima omogućilo pravovremeno lečenje. U našoj
studiji, analizirano je 95 ispitanika suspektnih na FB Sangerovim
sekvenciranjem svih egzona i okolnih intronskih regiona gena GLA, nakon čega je
izmerena relativna ekspresija kod 6 pacijenata. Genetičkom analizom kod 3
pacijenta detektovana je kodirajuća varijanta (p.D313Y), dok je kod 10 pacijenata
detektovana kombinacija intronskih varijanti, opisanih kao kompleksni
intronski haplotip (KIH). KIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-
16A>G, c.1000-22C>T), najčešći haplotip, detektovan je kod 7 (7,4%) pacijenata.
Uprkos normalnim biohemijskim parametrima, rezultati RT-qPCR metode
pokazali su snižen nivo ekspresije gena GLA u perifernoj krvi za 9,1%, 7,4%,
46,3% kod dve pacijentkinje (KIH1) i jedne pacijentkinje kod koje je detektovana
promotorska varijanta c.-10C>T, respektivno, ukazujući na potrebu dodatnih
analiza za konačno uspostavljanje dijagnoze FB. Uzevši u obzir da efekat
intronskih haplotipova nije funkcionalno okarakterisan, naša studija ukazuje
na važnost analiziranja intronskih regiona gena GLA kao potencijalnih
genetičkih modifikatora FB.
AB  - Фабријева болест (ФБ) је ретка, моногенска болест, која се одликује веома
хетерогеном клиничком сликом изазваном променама у гену GLA, смештеном на
дугом краку хромозома Х. Услед нарушене функције лизозомалног ензима α-
галактозидазе А долази до нагомилавања Gb3 супстрата, што доводи до
мултисистемских оштећења. Прецизна молекуларно-генетичка дијагноза је
есенцијална како би се пацијентима омогућило правовремено лечење. У нашој
студији, анализирано је 95 испитаника суспектних на ФБ Сангеровим
секвенцирањем свих егзона и околних интронских региона гена GLA, након чега је
измерена релативна експресија код 6 пацијената. Генетичком анализом код 3
пацијента детектована је кодирајућа варијанта (p.D313Y), док је код 10 пацијената
детектована комбинација интронских варијанти, описаних као комплексни
интронски хаплотип (КИХ). КИХ1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-
16A>G, c.1000-22C>T), најчешћи хаплотип, детектован је код 7 (7,4%) пацијената.
Упркос нормалним биохемијским параметрима, резултати RT-qPCR методе
показали су снижен ниво експресије гена GLA у периферној крви за 9,1%, 7,4%,
46,3% код две пацијенткиње (КИХ1) и једне пацијенткиње код које је детектована
промоторска варијанта c.-10C>T, респективно, указујући на потребу додатних
анализа за коначно успостављање дијагнозе ФБ. Узевши у обзир да ефекат
интронских хаплотипова није функционално окарактерисан, наша студија указује
на важност анализирања интронских региона гена GLA као потенцијалних
генетичких модификатора ФБ.
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Molekularna dijagnostika Fabrijeve bolesti kod pacijenata sa hroničnom bubrežnom insuficijencijom nepoznate etiologije
T1  - Молекуларна дијагностика Фабријеве болести код пацијената са хроничном бубрежном инсуфицијенцијом непознате етиологије
SP  - 313
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1740
ER  - 

@conference{
author = "Parezanović, Marina and Stojiljković, Maja and Anđelković, Marina and Stevanović, Nina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Tošić, Nataša and Pavlović, Sonja and Ćelić, Dejan and Vučenović, Jelica and Skakić, Anita",
year = "2022",
abstract = "Fabrijeva bolest (FB) je retka, monogenska bolest, koja se odlikuje veoma
heterogenom kliničkom slikom izazvanom promenama u genu GLA, smeštenom na
dugom kraku hromozoma H. Usled narušene funkcije lizozomalnog enzima α-
galaktozidaze A dolazi do nagomilavanja Gb3 supstrata, što dovodi do
multisistemskih oštećenja. Precizna molekularno-genetička dijagnoza je
esencijalna kako bi se pacijentima omogućilo pravovremeno lečenje. U našoj
studiji, analizirano je 95 ispitanika suspektnih na FB Sangerovim
sekvenciranjem svih egzona i okolnih intronskih regiona gena GLA, nakon čega je
izmerena relativna ekspresija kod 6 pacijenata. Genetičkom analizom kod 3
pacijenta detektovana je kodirajuća varijanta (p.D313Y), dok je kod 10 pacijenata
detektovana kombinacija intronskih varijanti, opisanih kao kompleksni
intronski haplotip (KIH). KIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-
16A>G, c.1000-22C>T), najčešći haplotip, detektovan je kod 7 (7,4%) pacijenata.
Uprkos normalnim biohemijskim parametrima, rezultati RT-qPCR metode
pokazali su snižen nivo ekspresije gena GLA u perifernoj krvi za 9,1%, 7,4%,
46,3% kod dve pacijentkinje (KIH1) i jedne pacijentkinje kod koje je detektovana
promotorska varijanta c.-10C>T, respektivno, ukazujući na potrebu dodatnih
analiza za konačno uspostavljanje dijagnoze FB. Uzevši u obzir da efekat
intronskih haplotipova nije funkcionalno okarakterisan, naša studija ukazuje
na važnost analiziranja intronskih regiona gena GLA kao potencijalnih
genetičkih modifikatora FB., Фабријева болест (ФБ) је ретка, моногенска болест, која се одликује веома
хетерогеном клиничком сликом изазваном променама у гену GLA, смештеном на
дугом краку хромозома Х. Услед нарушене функције лизозомалног ензима α-
галактозидазе А долази до нагомилавања Gb3 супстрата, што доводи до
мултисистемских оштећења. Прецизна молекуларно-генетичка дијагноза је
есенцијална како би се пацијентима омогућило правовремено лечење. У нашој
студији, анализирано је 95 испитаника суспектних на ФБ Сангеровим
секвенцирањем свих егзона и околних интронских региона гена GLA, након чега је
измерена релативна експресија код 6 пацијената. Генетичком анализом код 3
пацијента детектована је кодирајућа варијанта (p.D313Y), док је код 10 пацијената
детектована комбинација интронских варијанти, описаних као комплексни
интронски хаплотип (КИХ). КИХ1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-
16A>G, c.1000-22C>T), најчешћи хаплотип, детектован је код 7 (7,4%) пацијената.
Упркос нормалним биохемијским параметрима, резултати RT-qPCR методе
показали су снижен ниво експресије гена GLA у периферној крви за 9,1%, 7,4%,
46,3% код две пацијенткиње (КИХ1) и једне пацијенткиње код које је детектована
промоторска варијанта c.-10C>T, респективно, указујући на потребу додатних
анализа за коначно успостављање дијагнозе ФБ. Узевши у обзир да ефекат
интронских хаплотипова није функционално окарактерисан, наша студија указује
на важност анализирања интронских региона гена GLA као потенцијалних
генетичких модификатора ФБ.",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Molekularna dijagnostika Fabrijeve bolesti kod pacijenata sa hroničnom bubrežnom insuficijencijom nepoznate etiologije, Молекуларна дијагностика Фабријеве болести код пацијената са хроничном бубрежном инсуфицијенцијом непознате етиологије",
pages = "313",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1740"
}

Parezanović, M., Stojiljković, M., Anđelković, M., Stevanović, N., Spasovski, V., Ugrin, M., Komazec, J., Tošić, N., Pavlović, S., Ćelić, D., Vučenović, J.,& Skakić, A.. (2022). Molekularna dijagnostika Fabrijeve bolesti kod pacijenata sa hroničnom bubrežnom insuficijencijom nepoznate etiologije. in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo., 313.
https://hdl.handle.net/21.15107/rcub_imagine_1740

Parezanović M, Stojiljković M, Anđelković M, Stevanović N, Spasovski V, Ugrin M, Komazec J, Tošić N, Pavlović S, Ćelić D, Vučenović J, Skakić A. Molekularna dijagnostika Fabrijeve bolesti kod pacijenata sa hroničnom bubrežnom insuficijencijom nepoznate etiologije. in Treći kongres biologa Srbije. 2022;:313.
https://hdl.handle.net/21.15107/rcub_imagine_1740 .

Parezanović, Marina, Stojiljković, Maja, Anđelković, Marina, Stevanović, Nina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Tošić, Nataša, Pavlović, Sonja, Ćelić, Dejan, Vučenović, Jelica, Skakić, Anita, "Molekularna dijagnostika Fabrijeve bolesti kod pacijenata sa hroničnom bubrežnom insuficijencijom nepoznate etiologije" in Treći kongres biologa Srbije (2022):313,
https://hdl.handle.net/21.15107/rcub_imagine_1740 .

TY  - CONF
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Stevanović, Nina
AU  - Parezanović, Marina
AU  - Stojiljković, Maja
AU  - Spasovski, Vesna
AU  - Pavlović, Sonja
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1744
AB  - Retke plućne bolesti (RPB) su grupa oboljenja koje pojedinačno pogađaju jednog od
2.000 ljudi, sa procenom da oko 80% RPB ima genetičku osnovu. Uprkos
varijacijama među RPB u kliničkim karakteristikama i manifestacijama, većina
ovih bolesti na sličan način oštećuje pluća, što otežava uspostavljanje
dijagnoze. Upotreba NGS tehnologije (engl. Next Generation Sequencing, NGS) u
dijagnostičke svrhe omogućava bolje razumevanje genetičke osnove bolesti,
međutim, identifikacija i klasifikacija varijanti koje izazivaju bolesti je
kompleksna zbog detekcije brojnih VUS varijanti (engl. Variants of uncertain
significance, VUS) koje se ne mogu precizno klasifikovati. Glavni cilj ove studije
bio je da se dizajnira jedinstveni vodič koji će omogućiti standardizaciju
analize novih genetičkih varijanti u genima uzročnicima RPB. Dizajnirane
smernice se sastoje od tri glavna koraka: (1) sekvenciranje, detekcija i
identifikacija gena/varijanti, (2) klasifikacija varijanti i (3) karakterizacija
varijanti korišćenjem in silico strukturne i funkcionalne analize (qRT-PCR i
Western blot analiza). Validacija smernica je izvršena analizom varijanti
otkrivenih u genu uzročniku i genu kandidatu za RPB, a detektovane VUS
varijante su dobile dijagnostički značaj. Primena ovih smernica je rezultirala
identifikacijom i klasifikacijom novootkrivenih genetičkih varijanti, kroz
analizu na transkripcionom, translacionom i posttranslacionom nivou, i
dovela do uspostavljanja precizne dijagnoze bolesti.
AB  - Ретке плућне болести (РПБ) су група обољења које појединачно погађају једног од
2.000 људи, са проценом да око 80% РПБ има генетичку основу. Упркос
варијацијама међу РПБ у клиничким карактеристикама и манифестацијама, већина
ових болести на сличан начин оштећује плућа, што отежава успостављање
дијагнозе. Употреба НГС технологије (енгл. Next Generation Sequencing, NGS) у
дијагностичке сврхе омогућава боље разумевање генетичке основе болести,
међутим, идентификација и класификација варијанти које изазивају болести је
комплексна због детекције бројних ВУС варијанти (енгл. Variants of uncertain
significance, VUS) које се не могу прецизно класификовати. Главни циљ ове студије
био је да се дизајнира јединствени водич који ће омогућити стандардизацију
анализе нових генетичких варијанти у генима узрочницима РПБ. Дизајниране
смернице се састоје од три главна корака: (1) секвенцирање, детекција и
идентификација гена/варијанти, (2) класификација варијанти и (3) карактеризација
варијанти коришћењем in silico структурне и функционалне анализе (qRT-PCR и
Western blot анализа). Валидација смерница је извршена анализом варијанти
откривених у гену узрочнику и гену кандидату за РПБ, а детектоване ВУС
варијанте су добиле дијагностички значај. Примена ових смерница је резултирала
идентификацијом и класификацијом новооткривених генетичких варијанти, кроз
анализу на транскрипционом, транслационом и посттранслационом нивоу, и
довела до успостављања прецизне дијагнозе болести.
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Dizajniranje jedinstvenih smernica za standardizaciju analize NGS podataka kod pacijenata sa retkim plućnim bolestima
T1  - Дизајнирање јединствених смерница за стандардизацију анализе НГС података код пацијената са ретким плућним болестима
SP  - 291
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1744
ER  - 

@conference{
author = "Anđelković, Marina and Skakić, Anita and Stevanović, Nina and Parezanović, Marina and Stojiljković, Maja and Spasovski, Vesna and Pavlović, Sonja",
year = "2022",
abstract = "Retke plućne bolesti (RPB) su grupa oboljenja koje pojedinačno pogađaju jednog od
2.000 ljudi, sa procenom da oko 80% RPB ima genetičku osnovu. Uprkos
varijacijama među RPB u kliničkim karakteristikama i manifestacijama, većina
ovih bolesti na sličan način oštećuje pluća, što otežava uspostavljanje
dijagnoze. Upotreba NGS tehnologije (engl. Next Generation Sequencing, NGS) u
dijagnostičke svrhe omogućava bolje razumevanje genetičke osnove bolesti,
međutim, identifikacija i klasifikacija varijanti koje izazivaju bolesti je
kompleksna zbog detekcije brojnih VUS varijanti (engl. Variants of uncertain
significance, VUS) koje se ne mogu precizno klasifikovati. Glavni cilj ove studije
bio je da se dizajnira jedinstveni vodič koji će omogućiti standardizaciju
analize novih genetičkih varijanti u genima uzročnicima RPB. Dizajnirane
smernice se sastoje od tri glavna koraka: (1) sekvenciranje, detekcija i
identifikacija gena/varijanti, (2) klasifikacija varijanti i (3) karakterizacija
varijanti korišćenjem in silico strukturne i funkcionalne analize (qRT-PCR i
Western blot analiza). Validacija smernica je izvršena analizom varijanti
otkrivenih u genu uzročniku i genu kandidatu za RPB, a detektovane VUS
varijante su dobile dijagnostički značaj. Primena ovih smernica je rezultirala
identifikacijom i klasifikacijom novootkrivenih genetičkih varijanti, kroz
analizu na transkripcionom, translacionom i posttranslacionom nivou, i
dovela do uspostavljanja precizne dijagnoze bolesti., Ретке плућне болести (РПБ) су група обољења које појединачно погађају једног од
2.000 људи, са проценом да око 80% РПБ има генетичку основу. Упркос
варијацијама међу РПБ у клиничким карактеристикама и манифестацијама, већина
ових болести на сличан начин оштећује плућа, што отежава успостављање
дијагнозе. Употреба НГС технологије (енгл. Next Generation Sequencing, NGS) у
дијагностичке сврхе омогућава боље разумевање генетичке основе болести,
међутим, идентификација и класификација варијанти које изазивају болести је
комплексна због детекције бројних ВУС варијанти (енгл. Variants of uncertain
significance, VUS) које се не могу прецизно класификовати. Главни циљ ове студије
био је да се дизајнира јединствени водич који ће омогућити стандардизацију
анализе нових генетичких варијанти у генима узрочницима РПБ. Дизајниране
смернице се састоје од три главна корака: (1) секвенцирање, детекција и
идентификација гена/варијанти, (2) класификација варијанти и (3) карактеризација
варијанти коришћењем in silico структурне и функционалне анализе (qRT-PCR и
Western blot анализа). Валидација смерница је извршена анализом варијанти
откривених у гену узрочнику и гену кандидату за РПБ, а детектоване ВУС
варијанте су добиле дијагностички значај. Примена ових смерница је резултирала
идентификацијом и класификацијом новооткривених генетичких варијанти, кроз
анализу на транскрипционом, транслационом и посттранслационом нивоу, и
довела до успостављања прецизне дијагнозе болести.",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Dizajniranje jedinstvenih smernica za standardizaciju analize NGS podataka kod pacijenata sa retkim plućnim bolestima, Дизајнирање јединствених смерница за стандардизацију анализе НГС података код пацијената са ретким плућним болестима",
pages = "291",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1744"
}

Anđelković, M., Skakić, A., Stevanović, N., Parezanović, M., Stojiljković, M., Spasovski, V.,& Pavlović, S.. (2022). Dizajniranje jedinstvenih smernica za standardizaciju analize NGS podataka kod pacijenata sa retkim plućnim bolestima. in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo., 291.
https://hdl.handle.net/21.15107/rcub_imagine_1744

Anđelković M, Skakić A, Stevanović N, Parezanović M, Stojiljković M, Spasovski V, Pavlović S. Dizajniranje jedinstvenih smernica za standardizaciju analize NGS podataka kod pacijenata sa retkim plućnim bolestima. in Treći kongres biologa Srbije. 2022;:291.
https://hdl.handle.net/21.15107/rcub_imagine_1744 .

Anđelković, Marina, Skakić, Anita, Stevanović, Nina, Parezanović, Marina, Stojiljković, Maja, Spasovski, Vesna, Pavlović, Sonja, "Dizajniranje jedinstvenih smernica za standardizaciju analize NGS podataka kod pacijenata sa retkim plućnim bolestima" in Treći kongres biologa Srbije (2022):291,
https://hdl.handle.net/21.15107/rcub_imagine_1744 .

TY  - JOUR
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Ugrin, Milena
AU  - Spasovski, Vesna
AU  - Klaassen, Kristel
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1524
AB  - Glycogen storage diseases (GSDs) are rare metabolic monogenic disorders characterized by an excessive accumulation of glycogen in the cell. However, monogenic disorders are not simple regarding genotype-phenotype correlation. Genes outside the major disease-causing locus could have modulatory effect on GSDs, and thus explain the genotype-phenotype inconsistencies observed in these patients. Nowadays, when the sequencing of all clinically relevant genes, whole human exomes, and even whole human genomes is fast, easily available and affordable, we have a scientific obligation to holistically analyze data and draw smarter connections between genotype and phenotype. Recently, the importance of glycogen-selective autophagy for the pathophysiology of disorders of glycogen metabolism have been described. Therefore, in this manuscript, we review the potential role of genes involved in glycogen-selective autophagy as modifiers of GSDs. Given the small number of genes associated with glycogen-selective autophagy, we also include genes, transcription factors, and non-coding RNAs involved in autophagy. A cross-link with apoptosis is addressed. All these genes could be analyzed in GSD patients with unusual discrepancies between genotype and phenotype in order to discover genetic variants potentially modifying their phenotype. The discovery of modifier genes related to glycogen-selective autophagy and autophagy will start a new chapter in understanding of GSDs and enable the usage of autophagy-inducing drugs for the treatment of this group of rare-disease patients.
PB  - MDPI, Basel
T2  - Life-Basel
T1  - Crosstalk between Glycogen-Selective Autophagy, Autophagy and Apoptosis as a Road towards Modifier Gene Discovery and New Therapeutic Strategies for Glycogen Storage Diseases
IS  - 9
VL  - 12
DO  - 10.3390/life12091396
ER  - 

@article{
author = "Anđelković, Marina and Skakić, Anita and Ugrin, Milena and Spasovski, Vesna and Klaassen, Kristel and Pavlović, Sonja and Stojiljković, Maja",
year = "2022",
abstract = "Glycogen storage diseases (GSDs) are rare metabolic monogenic disorders characterized by an excessive accumulation of glycogen in the cell. However, monogenic disorders are not simple regarding genotype-phenotype correlation. Genes outside the major disease-causing locus could have modulatory effect on GSDs, and thus explain the genotype-phenotype inconsistencies observed in these patients. Nowadays, when the sequencing of all clinically relevant genes, whole human exomes, and even whole human genomes is fast, easily available and affordable, we have a scientific obligation to holistically analyze data and draw smarter connections between genotype and phenotype. Recently, the importance of glycogen-selective autophagy for the pathophysiology of disorders of glycogen metabolism have been described. Therefore, in this manuscript, we review the potential role of genes involved in glycogen-selective autophagy as modifiers of GSDs. Given the small number of genes associated with glycogen-selective autophagy, we also include genes, transcription factors, and non-coding RNAs involved in autophagy. A cross-link with apoptosis is addressed. All these genes could be analyzed in GSD patients with unusual discrepancies between genotype and phenotype in order to discover genetic variants potentially modifying their phenotype. The discovery of modifier genes related to glycogen-selective autophagy and autophagy will start a new chapter in understanding of GSDs and enable the usage of autophagy-inducing drugs for the treatment of this group of rare-disease patients.",
publisher = "MDPI, Basel",
journal = "Life-Basel",
title = "Crosstalk between Glycogen-Selective Autophagy, Autophagy and Apoptosis as a Road towards Modifier Gene Discovery and New Therapeutic Strategies for Glycogen Storage Diseases",
number = "9",
volume = "12",
doi = "10.3390/life12091396"
}

Anđelković, M., Skakić, A., Ugrin, M., Spasovski, V., Klaassen, K., Pavlović, S.,& Stojiljković, M.. (2022). Crosstalk between Glycogen-Selective Autophagy, Autophagy and Apoptosis as a Road towards Modifier Gene Discovery and New Therapeutic Strategies for Glycogen Storage Diseases. in Life-Basel
MDPI, Basel., 12(9).
https://doi.org/10.3390/life12091396

Anđelković M, Skakić A, Ugrin M, Spasovski V, Klaassen K, Pavlović S, Stojiljković M. Crosstalk between Glycogen-Selective Autophagy, Autophagy and Apoptosis as a Road towards Modifier Gene Discovery and New Therapeutic Strategies for Glycogen Storage Diseases. in Life-Basel. 2022;12(9).
doi:10.3390/life12091396 .

Anđelković, Marina, Skakić, Anita, Ugrin, Milena, Spasovski, Vesna, Klaassen, Kristel, Pavlović, Sonja, Stojiljković, Maja, "Crosstalk between Glycogen-Selective Autophagy, Autophagy and Apoptosis as a Road towards Modifier Gene Discovery and New Therapeutic Strategies for Glycogen Storage Diseases" in Life-Basel, 12, no. 9 (2022),
https://doi.org/10.3390/life12091396 . .

TY  - CONF
AU  - Klaassen, Kristel
AU  - Stanković, Biljana
AU  - Zukić, Branka
AU  - Kotur, Nikola
AU  - Gašić, Vladimir
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1577
PB  - Springernature, London
C3  - European Journal of Human Genetics
T1  - Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection
EP  - 603
IS  - SUPPL 1
SP  - 603
VL  - 30
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1577
ER  - 

@conference{
author = "Klaassen, Kristel and Stanković, Biljana and Zukić, Branka and Kotur, Nikola and Gašić, Vladimir and Pavlović, Sonja and Stojiljković, Maja",
year = "2022",
publisher = "Springernature, London",
journal = "European Journal of Human Genetics",
title = "Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection",
pages = "603-603",
number = "SUPPL 1",
volume = "30",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1577"
}

Klaassen, K., Stanković, B., Zukić, B., Kotur, N., Gašić, V., Pavlović, S.,& Stojiljković, M.. (2022). Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection. in European Journal of Human Genetics
Springernature, London., 30(SUPPL 1), 603-603.
https://hdl.handle.net/21.15107/rcub_imagine_1577

Klaassen K, Stanković B, Zukić B, Kotur N, Gašić V, Pavlović S, Stojiljković M. Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection. in European Journal of Human Genetics. 2022;30(SUPPL 1):603-603.
https://hdl.handle.net/21.15107/rcub_imagine_1577 .

Klaassen, Kristel, Stanković, Biljana, Zukić, Branka, Kotur, Nikola, Gašić, Vladimir, Pavlović, Sonja, Stojiljković, Maja, "Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection" in European Journal of Human Genetics, 30, no. SUPPL 1 (2022):603-603,
https://hdl.handle.net/21.15107/rcub_imagine_1577 .

TY  - CHAP
AU  - Skakić, Anita
AU  - Stojiljković, Maja
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1728
AB  - CRISPR/Cas9 (eng. Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9) je prirodni
alat za editovanje genoma usvojen iz prokariotskog adaptivnog imunskog odbrambenog sistema.
Zbog svoje visoke efikasnosti i preciznosti, protein Cas9 koji pripada CRISPR sistemu klase II, pronašao je
primenu u mnogim poljima nauke. Editovanje genoma zasnovano na tehnologiji CRISPR/Cas9 predstavlja
jedan od najperspektivnijih alata za lečenje humanih bolesti sa genetičkom osnovom, uključujući kardiovaskularne
bolesti, neurodegenerativne poremećaje i različite vrste tumora. Genska terapija zasnovana na
CRISPR/Cas9 opsežno se proučava u pretkliničkim i kliničkim studijama. Editovanje genoma CRISPR/Cas9 je
takođe robustan alat za stvaranje in vitro ćelijskih i životinjskih model sistema za istraživanje i lečenje genetičkih
bolesti, posebno bolesti povezanih sa tačkastim promenama. U ovom radu, osvrnućemo se kratko na
istoriju i mehanizam sistema CRISPR/Cas9, različite metodološke pristupe, napraviti pregled primena u biomedicini
i opisati njegovu ulogu u razvoju novih molekularnih terapeutika za retke nasledne bolesti.
AB  - Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) is a
naturally occurring genome editing tool adopted from the prokaryotic adaptive immune defense system.
Due to its high efficiency and precision, the Cas9 protein derived from the type II CRISPR system, has found
applications in many fields of science. Currently, CRISPR/Cas9-based genome editing has become one of
the most promising tools for treating human genetic diseases, including cardiovascular diseases, neurodegenerative
disorders, and various types of tumors. CRISPR/Cas9-based gene therapy is extensively studied
in preclinic and clinic treatments. CRISPR/Cas9 genome editing is also a robust tool to create in vitro cellular
and animal model systems for investigating and treating human genetic disorders, particularly diseases
associated with point mutations. Therefore, in this review, we will present a brief history and mechanism of
the CRISPR/Cas9 system. We will also describe the different methodological approaches, review applications
in biomedicine and describe its role in the development of new molecular therapeutics for rare inherited
diseases.
PB  - Trendovi u molekularnoj Biologiji
T2  - Trendovi u molekularnoj Biologiji
T1  - Primena crISPr/cas9 tehnologije u otkrivanju novih molekularnih terapeutika
T1  - Application of crISPr/cas9 technology in the discovery of new molecular therapeutics
EP  - 52
SP  - 42
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1728
ER  - 

@inbook{
author = "Skakić, Anita and Stojiljković, Maja",
year = "2021",
abstract = "CRISPR/Cas9 (eng. Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9) je prirodni
alat za editovanje genoma usvojen iz prokariotskog adaptivnog imunskog odbrambenog sistema.
Zbog svoje visoke efikasnosti i preciznosti, protein Cas9 koji pripada CRISPR sistemu klase II, pronašao je
primenu u mnogim poljima nauke. Editovanje genoma zasnovano na tehnologiji CRISPR/Cas9 predstavlja
jedan od najperspektivnijih alata za lečenje humanih bolesti sa genetičkom osnovom, uključujući kardiovaskularne
bolesti, neurodegenerativne poremećaje i različite vrste tumora. Genska terapija zasnovana na
CRISPR/Cas9 opsežno se proučava u pretkliničkim i kliničkim studijama. Editovanje genoma CRISPR/Cas9 je
takođe robustan alat za stvaranje in vitro ćelijskih i životinjskih model sistema za istraživanje i lečenje genetičkih
bolesti, posebno bolesti povezanih sa tačkastim promenama. U ovom radu, osvrnućemo se kratko na
istoriju i mehanizam sistema CRISPR/Cas9, različite metodološke pristupe, napraviti pregled primena u biomedicini
i opisati njegovu ulogu u razvoju novih molekularnih terapeutika za retke nasledne bolesti., Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) is a
naturally occurring genome editing tool adopted from the prokaryotic adaptive immune defense system.
Due to its high efficiency and precision, the Cas9 protein derived from the type II CRISPR system, has found
applications in many fields of science. Currently, CRISPR/Cas9-based genome editing has become one of
the most promising tools for treating human genetic diseases, including cardiovascular diseases, neurodegenerative
disorders, and various types of tumors. CRISPR/Cas9-based gene therapy is extensively studied
in preclinic and clinic treatments. CRISPR/Cas9 genome editing is also a robust tool to create in vitro cellular
and animal model systems for investigating and treating human genetic disorders, particularly diseases
associated with point mutations. Therefore, in this review, we will present a brief history and mechanism of
the CRISPR/Cas9 system. We will also describe the different methodological approaches, review applications
in biomedicine and describe its role in the development of new molecular therapeutics for rare inherited
diseases.",
publisher = "Trendovi u molekularnoj Biologiji",
journal = "Trendovi u molekularnoj Biologiji",
booktitle = "Primena crISPr/cas9 tehnologije u otkrivanju novih molekularnih terapeutika, Application of crISPr/cas9 technology in the discovery of new molecular therapeutics",
pages = "52-42",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1728"
}

Skakić, A.,& Stojiljković, M.. (2021). Primena crISPr/cas9 tehnologije u otkrivanju novih molekularnih terapeutika. in Trendovi u molekularnoj Biologiji
Trendovi u molekularnoj Biologiji., 42-52.
https://hdl.handle.net/21.15107/rcub_imagine_1728

Skakić A, Stojiljković M. Primena crISPr/cas9 tehnologije u otkrivanju novih molekularnih terapeutika. in Trendovi u molekularnoj Biologiji. 2021;:42-52.
https://hdl.handle.net/21.15107/rcub_imagine_1728 .

Skakić, Anita, Stojiljković, Maja, "Primena crISPr/cas9 tehnologije u otkrivanju novih molekularnih terapeutika" in Trendovi u molekularnoj Biologiji (2021):42-52,
https://hdl.handle.net/21.15107/rcub_imagine_1728 .

TY  - JOUR
AU  - Stevanović, Nina
AU  - Skakić, Anita
AU  - Minić, Predrag
AU  - Sovtić, Aleksandar
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Anđelković, Marina
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1456
AB  - Primary ciliary dyskinesia (PCD) is a disease caused by impaired function of motile cilia. PCD mainly affects the lungs and reproductive organs. Inheritance is autosomal recessive and X-linked. PCD patients have diverse clinical manifestations, thus making the establishment of proper diagnosis challenging. The utility of next-generation sequencing (NGS) technology for diagnostic purposes allows for better understanding of the PCD genetic background. However, identification of specific disease-causing variants is difficult. The main aim of this study was to create a unique guideline that will enable the standardization of the assessment of novel genetic variants within PCD-associated genes. The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of genes/variants; (2) classification of variants according to their effect; and (3) variant characterization using in silico structural and functional analysis. The pipeline was validated through the analysis of the variants detected in a well-known PCD disease-causing gene (DNAI1) and the novel candidate gene (SPAG16). The application of this pipeline resulted in identification of potential disease-causing variants, as well as validation of the variants pathogenicity, through their analysis on transcriptional, translational, and posttranslational levels. The application of this pipeline leads to the confirmation of PCD diagnosis and enables a shift from candidate to PCD disease-causing gene.
PB  - MDPI, Basel
T2  - International Journal of Molecular Sciences
T1  - Identification and Classification of Novel Genetic Variants: En Route to the Diagnosis of Primary Ciliary Dyskinesia
IS  - 16
VL  - 22
DO  - 10.3390/ijms22168821
ER  - 

@article{
author = "Stevanović, Nina and Skakić, Anita and Minić, Predrag and Sovtić, Aleksandar and Stojiljković, Maja and Pavlović, Sonja and Anđelković, Marina",
year = "2021",
abstract = "Primary ciliary dyskinesia (PCD) is a disease caused by impaired function of motile cilia. PCD mainly affects the lungs and reproductive organs. Inheritance is autosomal recessive and X-linked. PCD patients have diverse clinical manifestations, thus making the establishment of proper diagnosis challenging. The utility of next-generation sequencing (NGS) technology for diagnostic purposes allows for better understanding of the PCD genetic background. However, identification of specific disease-causing variants is difficult. The main aim of this study was to create a unique guideline that will enable the standardization of the assessment of novel genetic variants within PCD-associated genes. The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of genes/variants; (2) classification of variants according to their effect; and (3) variant characterization using in silico structural and functional analysis. The pipeline was validated through the analysis of the variants detected in a well-known PCD disease-causing gene (DNAI1) and the novel candidate gene (SPAG16). The application of this pipeline resulted in identification of potential disease-causing variants, as well as validation of the variants pathogenicity, through their analysis on transcriptional, translational, and posttranslational levels. The application of this pipeline leads to the confirmation of PCD diagnosis and enables a shift from candidate to PCD disease-causing gene.",
publisher = "MDPI, Basel",
journal = "International Journal of Molecular Sciences",
title = "Identification and Classification of Novel Genetic Variants: En Route to the Diagnosis of Primary Ciliary Dyskinesia",
number = "16",
volume = "22",
doi = "10.3390/ijms22168821"
}

Stevanović, N., Skakić, A., Minić, P., Sovtić, A., Stojiljković, M., Pavlović, S.,& Anđelković, M.. (2021). Identification and Classification of Novel Genetic Variants: En Route to the Diagnosis of Primary Ciliary Dyskinesia. in International Journal of Molecular Sciences
MDPI, Basel., 22(16).
https://doi.org/10.3390/ijms22168821

Stevanović N, Skakić A, Minić P, Sovtić A, Stojiljković M, Pavlović S, Anđelković M. Identification and Classification of Novel Genetic Variants: En Route to the Diagnosis of Primary Ciliary Dyskinesia. in International Journal of Molecular Sciences. 2021;22(16).
doi:10.3390/ijms22168821 .

Stevanović, Nina, Skakić, Anita, Minić, Predrag, Sovtić, Aleksandar, Stojiljković, Maja, Pavlović, Sonja, Anđelković, Marina, "Identification and Classification of Novel Genetic Variants: En Route to the Diagnosis of Primary Ciliary Dyskinesia" in International Journal of Molecular Sciences, 22, no. 16 (2021),
https://doi.org/10.3390/ijms22168821 . .