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  - 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  - 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  - 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 .