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  - Anđelković, Marina
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Marjanović, Irena
AU  - Kravljana, Ruzica
AU  - Đorđević, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2179
AB  - Background: Childhood epilepsies are
caused by heterogeneous underlying disorders where
approximately 40% can be attributed to genetic
factors. Application of next-generation sequencing
(NGS) has revolutionized diagnostics and therefore
has enabled the identification of disease-causing
genes and variants in childhood epilepsies.
Materials and Methods: Patients who presented
with epilepsy of unknown etiology in childhood,
with suspicion of a genetic cause were included
in this study. In total, 55 patients from unrelated
non-consanguineous families were included and analyzed
by NGS either using clinical-exome sequencing
(MiSeq, Illumina) or whole-exome sequencing
(DNBSEQ-G400, MGI). Variants were prioritized
using Variant Interpreter and VarSome and classified
according to the ACMG recommendations.
Results: Using CES we analyzed 38 patients,
and for 22 of them a diagnosis was established.
Using WES we analyzed 17 patients with childhood epilepsy, which led to the identification of
disease-causing genes in 11 patients. The diagnostic
success rate for CES was 55.3% (21/38) and the
diagnostic rate for WES was 64.7% (11/17), with
the overall diagnostic rate being 58.2% (32/55). For
these patients, we detected pathogenic, likely pathogenic
variants or VUS in 24 epilepsy genes that
correlate well to the observed phenotype. Sixteen
novel genetic variants were identified and characterized
using various in silico algorithms.
Conclusion: This is the first study reporting
the molecular-genetic basis of childhood epilepsy
in Serbia. The prompt establishment of a specific
diagnosis is essential in order to make available the
prognosis, optimize therapy, and enable counseling
on recurrence risk in future pregnancies.
PB  - Macedonian Academy of Sciences and Arts
C3  - International Journal of Medical Genetics
T1  - CHARACTERIZATION OF 16 NOVEL GENETIC VARIANTS IN GENES RELATED TO CHILDHOOD EPILEPSIES
EP  - 114
IS  - Supplement
SP  - 114
VL  - 26
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2179
ER  - 

@conference{
author = "Anđelković, Marina and Klaassen, Kristel and Skakić, Anita and Marjanović, Irena and Kravljana, Ruzica and Đorđević, Maja",
year = "2023",
abstract = "Background: Childhood epilepsies are
caused by heterogeneous underlying disorders where
approximately 40% can be attributed to genetic
factors. Application of next-generation sequencing
(NGS) has revolutionized diagnostics and therefore
has enabled the identification of disease-causing
genes and variants in childhood epilepsies.
Materials and Methods: Patients who presented
with epilepsy of unknown etiology in childhood,
with suspicion of a genetic cause were included
in this study. In total, 55 patients from unrelated
non-consanguineous families were included and analyzed
by NGS either using clinical-exome sequencing
(MiSeq, Illumina) or whole-exome sequencing
(DNBSEQ-G400, MGI). Variants were prioritized
using Variant Interpreter and VarSome and classified
according to the ACMG recommendations.
Results: Using CES we analyzed 38 patients,
and for 22 of them a diagnosis was established.
Using WES we analyzed 17 patients with childhood epilepsy, which led to the identification of
disease-causing genes in 11 patients. The diagnostic
success rate for CES was 55.3% (21/38) and the
diagnostic rate for WES was 64.7% (11/17), with
the overall diagnostic rate being 58.2% (32/55). For
these patients, we detected pathogenic, likely pathogenic
variants or VUS in 24 epilepsy genes that
correlate well to the observed phenotype. Sixteen
novel genetic variants were identified and characterized
using various in silico algorithms.
Conclusion: This is the first study reporting
the molecular-genetic basis of childhood epilepsy
in Serbia. The prompt establishment of a specific
diagnosis is essential in order to make available the
prognosis, optimize therapy, and enable counseling
on recurrence risk in future pregnancies.",
publisher = "Macedonian Academy of Sciences and Arts",
journal = "International Journal of Medical Genetics",
title = "CHARACTERIZATION OF 16 NOVEL GENETIC VARIANTS IN GENES RELATED TO CHILDHOOD EPILEPSIES",
pages = "114-114",
number = "Supplement",
volume = "26",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2179"
}

Anđelković, M., Klaassen, K., Skakić, A., Marjanović, I., Kravljana, R.,& Đorđević, M.. (2023). CHARACTERIZATION OF 16 NOVEL GENETIC VARIANTS IN GENES RELATED TO CHILDHOOD EPILEPSIES. in International Journal of Medical Genetics
Macedonian Academy of Sciences and Arts., 26(Supplement), 114-114.
https://hdl.handle.net/21.15107/rcub_imagine_2179

Anđelković M, Klaassen K, Skakić A, Marjanović I, Kravljana R, Đorđević M. CHARACTERIZATION OF 16 NOVEL GENETIC VARIANTS IN GENES RELATED TO CHILDHOOD EPILEPSIES. in International Journal of Medical Genetics. 2023;26(Supplement):114-114.
https://hdl.handle.net/21.15107/rcub_imagine_2179 .

Anđelković, Marina, Klaassen, Kristel, Skakić, Anita, Marjanović, Irena, Kravljana, Ruzica, Đorđević, Maja, "CHARACTERIZATION OF 16 NOVEL GENETIC VARIANTS IN GENES RELATED TO CHILDHOOD EPILEPSIES" in International Journal of Medical Genetics, 26, no. Supplement (2023):114-114,
https://hdl.handle.net/21.15107/rcub_imagine_2179 .

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  - Kotur, Nikola
AU  - Skakić, Anita
AU  - Klaassen, Kristel
AU  - Gašić, Vladimir
AU  - Jelovac, Marina
AU  - Ristivojević, Bojan
AU  - Zukić, Branka
AU  - Pavlović, Sonja
AU  - Stanković, Biljana
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1904
AB  - COVID-19 pandemic has proved to be an unrelenting health threat for more than a year now. The
emerging amount of data indicates that vitamin D could be important for clinical presentation of
COVID-19. Here, we investigated association of genetic variants related to the altered level and
bioavailability of vitamin D with clinical severity of COVID-19. We analyzed variants in genes
significant for the status of vitamin D (DHCR7/NADSYN1 rs12785878, GC rs2282679, and CYP2R1
rs10741657), and vitamin D effect (VDR rs2228570) in 120 Serbian adult and pediatric COVID-19
patients using allelic discrimination. Furthermore, we carried out comparative population genetic
analysis among European and other worldwide populations to investigate variation in allelic
frequencies of selected variants. The results showed that DHCR7/NADSYN rs12785878 and CYP2R1
rs10741657 variants were associated with severe COVID-19 in adults (p = 0.03, p = 0.017,
respectively); carriers of DHCR7/NADSYN TG+GG and CYP2R1 GG genotypes had 0.21 and 5.9 the
odds for developing severe disease, OR 0.21 (0.05–0.9) and OR 5.9 (1.4–25.2), respectively. There
were no associations between selected genetic variants and disease severity in pediatric patients.
Comparative population genetic analysis revealed that Serbian population had the lowest frequency of
CYP2R1 rs10741657 G allele compared to other non-Finish Europeans (0.58 compared to 0.69 and
0.66 in Spanish and Italian population, respectively), suggesting that other populations should also
investigate the relationship of CYP2R1 variant and the COVID-19 disease course. The results of the
study indicated that vitamin D related genetic variants were implicated in severe COVID-19 in adults.
This could direct prevention strategies based on population specific nutrigenetic profiles.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Covid-19 disease severity associated with vitamin d related genetic Variants
IS  - 2 (Special edition
SP  - 144
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1904
ER  - 

@conference{
author = "Kotur, Nikola and Skakić, Anita and Klaassen, Kristel and Gašić, Vladimir and Jelovac, Marina and Ristivojević, Bojan and Zukić, Branka and Pavlović, Sonja and Stanković, Biljana",
year = "2023",
abstract = "COVID-19 pandemic has proved to be an unrelenting health threat for more than a year now. The
emerging amount of data indicates that vitamin D could be important for clinical presentation of
COVID-19. Here, we investigated association of genetic variants related to the altered level and
bioavailability of vitamin D with clinical severity of COVID-19. We analyzed variants in genes
significant for the status of vitamin D (DHCR7/NADSYN1 rs12785878, GC rs2282679, and CYP2R1
rs10741657), and vitamin D effect (VDR rs2228570) in 120 Serbian adult and pediatric COVID-19
patients using allelic discrimination. Furthermore, we carried out comparative population genetic
analysis among European and other worldwide populations to investigate variation in allelic
frequencies of selected variants. The results showed that DHCR7/NADSYN rs12785878 and CYP2R1
rs10741657 variants were associated with severe COVID-19 in adults (p = 0.03, p = 0.017,
respectively); carriers of DHCR7/NADSYN TG+GG and CYP2R1 GG genotypes had 0.21 and 5.9 the
odds for developing severe disease, OR 0.21 (0.05–0.9) and OR 5.9 (1.4–25.2), respectively. There
were no associations between selected genetic variants and disease severity in pediatric patients.
Comparative population genetic analysis revealed that Serbian population had the lowest frequency of
CYP2R1 rs10741657 G allele compared to other non-Finish Europeans (0.58 compared to 0.69 and
0.66 in Spanish and Italian population, respectively), suggesting that other populations should also
investigate the relationship of CYP2R1 variant and the COVID-19 disease course. The results of the
study indicated that vitamin D related genetic variants were implicated in severe COVID-19 in adults.
This could direct prevention strategies based on population specific nutrigenetic profiles.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Covid-19 disease severity associated with vitamin d related genetic Variants",
number = "2 (Special edition",
pages = "144",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1904"
}

Kotur, N., Skakić, A., Klaassen, K., Gašić, V., Jelovac, M., Ristivojević, B., Zukić, B., Pavlović, S.,& Stanković, B.. (2023). Covid-19 disease severity associated with vitamin d related genetic Variants. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition), 144.
https://hdl.handle.net/21.15107/rcub_imagine_1904

Kotur N, Skakić A, Klaassen K, Gašić V, Jelovac M, Ristivojević B, Zukić B, Pavlović S, Stanković B. Covid-19 disease severity associated with vitamin d related genetic Variants. in Genetics & Applications. 2023;7(2 (Special edition):144.
https://hdl.handle.net/21.15107/rcub_imagine_1904 .

Kotur, Nikola, Skakić, Anita, Klaassen, Kristel, Gašić, Vladimir, Jelovac, Marina, Ristivojević, Bojan, Zukić, Branka, Pavlović, Sonja, Stanković, Biljana, "Covid-19 disease severity associated with vitamin d related genetic Variants" in Genetics & Applications, 7, no. 2 (Special edition (2023):144,
https://hdl.handle.net/21.15107/rcub_imagine_1904 .

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  - 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  - 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  - 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  - 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  - 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  - 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  - JOUR
AU  - Drljaca, Tamara
AU  - Zukić, Branka
AU  - Kovacević, Vladimir
AU  - Gemović, Branislava
AU  - Klaassen, Kristel
AU  - Perović, Vladimir
AU  - Lazarević, Mladen
AU  - Pavlović, Sonja
AU  - Veljković, Nevena
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1502
AB  - The complete understanding of the genomic contribution to complex traits, diseases, and response to treatments, as well as genomic medicine application to the well-being of all humans will be achieved through the global variome that encompasses fine-scale genetic diversity. Despite significant efforts in recent years, uneven representation still characterizes genomic resources and among the underrepresented European populations are the Western Balkans including the Serbian population. Our research addresses this gap and presents the first ever targeted sequencing dataset of variants in clinically relevant genes. By measuring population differentiation and applying the Principal Component and Admixture analysis we demonstrated that the Serbian population differs little from other European populations, yet we identified several novel and more frequent variants that appear as its unique genetic determinants. We explored thoroughly the functional impact of frequent variants and its correlation with the health burden of the population of Serbia based on a sample of 144 individuals. Our variants catalogue improves the understanding of genetics of modern Serbia, contributes to research on ancestry, and aids in improvements of well-being and health equity. In addition, this resource may also be applicable in neighboring regions and valuable in worldwide functional analyses of genetic variants in individuals of European descent.
PB  - Nature Portfolio, Berlin
T2  - Scientific Reports
T1  - The first insight into the genetic structure of the population of modern Serbia
IS  - 1
VL  - 11
DO  - 10.1038/s41598-021-93129-4
ER  - 

@article{
author = "Drljaca, Tamara and Zukić, Branka and Kovacević, Vladimir and Gemović, Branislava and Klaassen, Kristel and Perović, Vladimir and Lazarević, Mladen and Pavlović, Sonja and Veljković, Nevena",
year = "2021",
abstract = "The complete understanding of the genomic contribution to complex traits, diseases, and response to treatments, as well as genomic medicine application to the well-being of all humans will be achieved through the global variome that encompasses fine-scale genetic diversity. Despite significant efforts in recent years, uneven representation still characterizes genomic resources and among the underrepresented European populations are the Western Balkans including the Serbian population. Our research addresses this gap and presents the first ever targeted sequencing dataset of variants in clinically relevant genes. By measuring population differentiation and applying the Principal Component and Admixture analysis we demonstrated that the Serbian population differs little from other European populations, yet we identified several novel and more frequent variants that appear as its unique genetic determinants. We explored thoroughly the functional impact of frequent variants and its correlation with the health burden of the population of Serbia based on a sample of 144 individuals. Our variants catalogue improves the understanding of genetics of modern Serbia, contributes to research on ancestry, and aids in improvements of well-being and health equity. In addition, this resource may also be applicable in neighboring regions and valuable in worldwide functional analyses of genetic variants in individuals of European descent.",
publisher = "Nature Portfolio, Berlin",
journal = "Scientific Reports",
title = "The first insight into the genetic structure of the population of modern Serbia",
number = "1",
volume = "11",
doi = "10.1038/s41598-021-93129-4"
}

Drljaca, T., Zukić, B., Kovacević, V., Gemović, B., Klaassen, K., Perović, V., Lazarević, M., Pavlović, S.,& Veljković, N.. (2021). The first insight into the genetic structure of the population of modern Serbia. in Scientific Reports
Nature Portfolio, Berlin., 11(1).
https://doi.org/10.1038/s41598-021-93129-4

Drljaca T, Zukić B, Kovacević V, Gemović B, Klaassen K, Perović V, Lazarević M, Pavlović S, Veljković N. The first insight into the genetic structure of the population of modern Serbia. in Scientific Reports. 2021;11(1).
doi:10.1038/s41598-021-93129-4 .

Drljaca, Tamara, Zukić, Branka, Kovacević, Vladimir, Gemović, Branislava, Klaassen, Kristel, Perović, Vladimir, Lazarević, Mladen, Pavlović, Sonja, Veljković, Nevena, "The first insight into the genetic structure of the population of modern Serbia" in Scientific Reports, 11, no. 1 (2021),
https://doi.org/10.1038/s41598-021-93129-4 . .

TY  - JOUR
AU  - Kotur, Nikola
AU  - Skakić, Anita
AU  - Klaassen, Kristel
AU  - Gašić, Vladimir
AU  - Zukić, Branka
AU  - Skodrić-Trifunović, Vesna
AU  - Stjepanović, Mihailo
AU  - Zivković, Zorica
AU  - Ostojić, Olivera
AU  - Stevanović, Goran
AU  - Lavadinović, Lidija
AU  - Pavlović, Sonja
AU  - Stanković, Biljana
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1439
AB  - Background: COVID-19 pandemic has proved to be an unrelenting health threat for more than a year now. The emerging amount of data indicates that vitamin D, zinc and selenium could be important for clinical presentation of COVID-19. Here, we investigated association of genetic variants related to the altered level and bioavailability of vitamin D, zinc and selenium with clinical severity of COVID-19. Methods: We analyzed variants in genes significant for the status of vitamin D (DHCR7/NADSYN1 rs12785878, GC rs2282679, CYP2R1 rs10741657, and VDR rs2228570), zinc (PPCDC rs2120019) and selenium (DMGDH rs17823744) in 120 Serbian adult and pediatric COVID-19 patients using allelic discrimination. Furthermore, we carried out comparative population genetic analysis among European and other worldwide populations to investigate variation in allelic frequencies of selected variants. Results: Study showed that DHCR7/NADSYN rs12785878 and CYP2R1 rs10741657 variants were associated with severe COVID-19 in adults (p = 0.03, p = 0.017, respectively); carriers of DHCR7/NADSYN TG+GG and CYP2R1 GG genotypes had 0.21 and 5.9 the odds for developing severe disease, OR 0.21 (0.05-0.9) and OR 5.9 (1.4-25.2), respectively. There were no associations between selected genetic variants and disease severity in pediatric patients. Comparative population genetic analysis revealed that Serbian population had the lowest frequency of CYP2R1 rs10741657 G allele compared to other non-Finish Europeans (0.58 compared to 0.69 and 0.66 in Spanish and Italian population, respectively), suggesting that other populations should also investigate the relationship of CYP2R1 variant and the COVID-19 disease course. Conclusion: The results of the study indicated that vitamin D related genetic variants were implicated in severe COVID-19 in adults. This could direct prevention strategies based on population specific nutrigenetic profiles.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Nutrition
T1  - Association of Vitamin D, Zinc and Selenium Related Genetic Variants With COVID-19 Disease Severity
VL  - 8
DO  - 10.3389/fnut.2021.689419
ER  - 

@article{
author = "Kotur, Nikola and Skakić, Anita and Klaassen, Kristel and Gašić, Vladimir and Zukić, Branka and Skodrić-Trifunović, Vesna and Stjepanović, Mihailo and Zivković, Zorica and Ostojić, Olivera and Stevanović, Goran and Lavadinović, Lidija and Pavlović, Sonja and Stanković, Biljana",
year = "2021",
abstract = "Background: COVID-19 pandemic has proved to be an unrelenting health threat for more than a year now. The emerging amount of data indicates that vitamin D, zinc and selenium could be important for clinical presentation of COVID-19. Here, we investigated association of genetic variants related to the altered level and bioavailability of vitamin D, zinc and selenium with clinical severity of COVID-19. Methods: We analyzed variants in genes significant for the status of vitamin D (DHCR7/NADSYN1 rs12785878, GC rs2282679, CYP2R1 rs10741657, and VDR rs2228570), zinc (PPCDC rs2120019) and selenium (DMGDH rs17823744) in 120 Serbian adult and pediatric COVID-19 patients using allelic discrimination. Furthermore, we carried out comparative population genetic analysis among European and other worldwide populations to investigate variation in allelic frequencies of selected variants. Results: Study showed that DHCR7/NADSYN rs12785878 and CYP2R1 rs10741657 variants were associated with severe COVID-19 in adults (p = 0.03, p = 0.017, respectively); carriers of DHCR7/NADSYN TG+GG and CYP2R1 GG genotypes had 0.21 and 5.9 the odds for developing severe disease, OR 0.21 (0.05-0.9) and OR 5.9 (1.4-25.2), respectively. There were no associations between selected genetic variants and disease severity in pediatric patients. Comparative population genetic analysis revealed that Serbian population had the lowest frequency of CYP2R1 rs10741657 G allele compared to other non-Finish Europeans (0.58 compared to 0.69 and 0.66 in Spanish and Italian population, respectively), suggesting that other populations should also investigate the relationship of CYP2R1 variant and the COVID-19 disease course. Conclusion: The results of the study indicated that vitamin D related genetic variants were implicated in severe COVID-19 in adults. This could direct prevention strategies based on population specific nutrigenetic profiles.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Nutrition",
title = "Association of Vitamin D, Zinc and Selenium Related Genetic Variants With COVID-19 Disease Severity",
volume = "8",
doi = "10.3389/fnut.2021.689419"
}

Kotur, N., Skakić, A., Klaassen, K., Gašić, V., Zukić, B., Skodrić-Trifunović, V., Stjepanović, M., Zivković, Z., Ostojić, O., Stevanović, G., Lavadinović, L., Pavlović, S.,& Stanković, B.. (2021). Association of Vitamin D, Zinc and Selenium Related Genetic Variants With COVID-19 Disease Severity. in Frontiers in Nutrition
Frontiers Media Sa, Lausanne., 8.
https://doi.org/10.3389/fnut.2021.689419

Kotur N, Skakić A, Klaassen K, Gašić V, Zukić B, Skodrić-Trifunović V, Stjepanović M, Zivković Z, Ostojić O, Stevanović G, Lavadinović L, Pavlović S, Stanković B. Association of Vitamin D, Zinc and Selenium Related Genetic Variants With COVID-19 Disease Severity. in Frontiers in Nutrition. 2021;8.
doi:10.3389/fnut.2021.689419 .

Kotur, Nikola, Skakić, Anita, Klaassen, Kristel, Gašić, Vladimir, Zukić, Branka, Skodrić-Trifunović, Vesna, Stjepanović, Mihailo, Zivković, Zorica, Ostojić, Olivera, Stevanović, Goran, Lavadinović, Lidija, Pavlović, Sonja, Stanković, Biljana, "Association of Vitamin D, Zinc and Selenium Related Genetic Variants With COVID-19 Disease Severity" in Frontiers in Nutrition, 8 (2021),
https://doi.org/10.3389/fnut.2021.689419 . .

TY  - JOUR
AU  - Klaassen, Kristel
AU  - Đorđević, M.
AU  - Skakić, Anita
AU  - Kecman, B.
AU  - Drmanac, R.
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1466
AB  - Phenylketonuria (PKU) is an inborn error of metabolism caused by variants in the phenylalanine hydroxylase (PAH) gene and it is characterized by excessively high levels of phenylalanine in body fluids. PKU is a paradigm for a genetic disease that can be treated and majority of developed countries have a population-based newborn screening. Thus, the combination of early diagnosis and immediate initiation of treatment has resulted in normal intelligence for treated PKU patients. Although PKU is a monogenic disease, decades of research and clinical practice have shown that the correlation between the genotype and corresponding phenotype is not simple at all. Attempts have been made to discover modifier genes for PKU cognitive phenotype but without any success so far. We conducted whole genome sequencing of 4 subjects from unrelated non-consanguineous families who presented with pathogenic mutations in the PAH gene, high blood phenylalanine concentrations and near-normal cognitive development despite no treatment. We used cross sample analysis to select genes common for more than one patient. Thus, the SHANK gene family emerged as the only relevant gene family with variants detected in 3 of 4 analyzed patients. We detected two novel variants, p.Pro1591Ala in SHANK1 and p.Asp18Asn in SHANK2, as well as SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro and SHANK3:p.Pro1716Thr variants that were previously described. Computational analysis indicated that the identified variants do not abolish the function of SHANK proteins. However, changes in posttranslational modifications of SHANK proteins could influence functioning of the glutamatergic synapses, cytoskeleton regulation and contribute to maintaining optimal synaptic density and number of dendritic spines. Our findings are linking SHANK gene family and brain plasticity in PKU for the first time. We hypothesize that variant SHANK proteins maintain optimal synaptic density and number of dendritic spines under high concentrations of phenylalanine and could have protective modifying effect on cognitive development of PKU patients.
PB  - Elsevier, Amsterdam
T2  - Molecular Genetics and Metabolism Reports
T1  - Untreated PKU patients without intellectual disability: SHANK gene family as a candidate modifier
VL  - 29
DO  - 10.1016/j.ymgmr.2021.100822
ER  - 

@article{
author = "Klaassen, Kristel and Đorđević, M. and Skakić, Anita and Kecman, B. and Drmanac, R. and Pavlović, Sonja and Stojiljković, Maja",
year = "2021",
abstract = "Phenylketonuria (PKU) is an inborn error of metabolism caused by variants in the phenylalanine hydroxylase (PAH) gene and it is characterized by excessively high levels of phenylalanine in body fluids. PKU is a paradigm for a genetic disease that can be treated and majority of developed countries have a population-based newborn screening. Thus, the combination of early diagnosis and immediate initiation of treatment has resulted in normal intelligence for treated PKU patients. Although PKU is a monogenic disease, decades of research and clinical practice have shown that the correlation between the genotype and corresponding phenotype is not simple at all. Attempts have been made to discover modifier genes for PKU cognitive phenotype but without any success so far. We conducted whole genome sequencing of 4 subjects from unrelated non-consanguineous families who presented with pathogenic mutations in the PAH gene, high blood phenylalanine concentrations and near-normal cognitive development despite no treatment. We used cross sample analysis to select genes common for more than one patient. Thus, the SHANK gene family emerged as the only relevant gene family with variants detected in 3 of 4 analyzed patients. We detected two novel variants, p.Pro1591Ala in SHANK1 and p.Asp18Asn in SHANK2, as well as SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro and SHANK3:p.Pro1716Thr variants that were previously described. Computational analysis indicated that the identified variants do not abolish the function of SHANK proteins. However, changes in posttranslational modifications of SHANK proteins could influence functioning of the glutamatergic synapses, cytoskeleton regulation and contribute to maintaining optimal synaptic density and number of dendritic spines. Our findings are linking SHANK gene family and brain plasticity in PKU for the first time. We hypothesize that variant SHANK proteins maintain optimal synaptic density and number of dendritic spines under high concentrations of phenylalanine and could have protective modifying effect on cognitive development of PKU patients.",
publisher = "Elsevier, Amsterdam",
journal = "Molecular Genetics and Metabolism Reports",
title = "Untreated PKU patients without intellectual disability: SHANK gene family as a candidate modifier",
volume = "29",
doi = "10.1016/j.ymgmr.2021.100822"
}

Klaassen, K., Đorđević, M., Skakić, A., Kecman, B., Drmanac, R., Pavlović, S.,& Stojiljković, M.. (2021). Untreated PKU patients without intellectual disability: SHANK gene family as a candidate modifier. in Molecular Genetics and Metabolism Reports
Elsevier, Amsterdam., 29.
https://doi.org/10.1016/j.ymgmr.2021.100822

Klaassen K, Đorđević M, Skakić A, Kecman B, Drmanac R, Pavlović S, Stojiljković M. Untreated PKU patients without intellectual disability: SHANK gene family as a candidate modifier. in Molecular Genetics and Metabolism Reports. 2021;29.
doi:10.1016/j.ymgmr.2021.100822 .

Klaassen, Kristel, Đorđević, M., Skakić, Anita, Kecman, B., Drmanac, R., Pavlović, Sonja, Stojiljković, Maja, "Untreated PKU patients without intellectual disability: SHANK gene family as a candidate modifier" in Molecular Genetics and Metabolism Reports, 29 (2021),
https://doi.org/10.1016/j.ymgmr.2021.100822 . .

TY  - JOUR
AU  - Anastasovska, Violeta
AU  - Kocova, Mirjana
AU  - Zdraveska, Nikolina
AU  - Stojiljković, Maja
AU  - Skakić, Anita
AU  - Klaassen, Kristel
AU  - Pavlović, Sonja
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1411
AB  - Background Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder of adrenal steroidogenesis with a broad spectrum of clinical presentations, ranging from the severe classical salt-wasting (SW) and simple-virilizing (SV) form, to the mild nonclassical form. A large variety of CYP21A2 genotypes in correlation with phenotype have been described. Materials and methods DNA samples from a 14-day-old male newborn with clinical and laboratory signs of SW CAH and family members were subjected for molecular analysis of the nine most common point CYP21A2 mutations by ACRS/PCR method. Direct DNA sequencing of the whole CYP21A2 gene was performed to detect the second mutant allele in the patient. The in silico predicting analysis and the crystal structure analysis of the mutated CYP21A2 protein have been performed. Results Molecular analysis confirmed that the patient was compound heterozygote carrying p.Q318X mutation inherited from the mother and a novel c.1271_1279delGTGCCCGCG (p.G424_R426del) variant in exon 10 inherited from the father. The in silico predicting software tools classified the novel mutation as pathogenic. Crystal structure analysis showed that the three residues affected by the novel in-frame deletion form several hydrogen bonds that could lead to impaired stability and function of the CYP21A2 protein. These findings were concordant with the patient's phenotype. The need of several molecular methods to elucidate the genotype in this patient has also been discussed. Conclusions A novel 9 bp deletion in CYP21A2 gene with predicted pathogenic effect on the enzyme activity was detected in neonatal patient causing severe SW CAH.
PB  - Springer, New York
T2  - Endocrine
T1  - A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia
EP  - 202
IS  - 1
SP  - 196
VL  - 73
DO  - 10.1007/s12020-021-02680-7
ER  - 

@article{
author = "Anastasovska, Violeta and Kocova, Mirjana and Zdraveska, Nikolina and Stojiljković, Maja and Skakić, Anita and Klaassen, Kristel and Pavlović, Sonja",
year = "2021",
abstract = "Background Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder of adrenal steroidogenesis with a broad spectrum of clinical presentations, ranging from the severe classical salt-wasting (SW) and simple-virilizing (SV) form, to the mild nonclassical form. A large variety of CYP21A2 genotypes in correlation with phenotype have been described. Materials and methods DNA samples from a 14-day-old male newborn with clinical and laboratory signs of SW CAH and family members were subjected for molecular analysis of the nine most common point CYP21A2 mutations by ACRS/PCR method. Direct DNA sequencing of the whole CYP21A2 gene was performed to detect the second mutant allele in the patient. The in silico predicting analysis and the crystal structure analysis of the mutated CYP21A2 protein have been performed. Results Molecular analysis confirmed that the patient was compound heterozygote carrying p.Q318X mutation inherited from the mother and a novel c.1271_1279delGTGCCCGCG (p.G424_R426del) variant in exon 10 inherited from the father. The in silico predicting software tools classified the novel mutation as pathogenic. Crystal structure analysis showed that the three residues affected by the novel in-frame deletion form several hydrogen bonds that could lead to impaired stability and function of the CYP21A2 protein. These findings were concordant with the patient's phenotype. The need of several molecular methods to elucidate the genotype in this patient has also been discussed. Conclusions A novel 9 bp deletion in CYP21A2 gene with predicted pathogenic effect on the enzyme activity was detected in neonatal patient causing severe SW CAH.",
publisher = "Springer, New York",
journal = "Endocrine",
title = "A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia",
pages = "202-196",
number = "1",
volume = "73",
doi = "10.1007/s12020-021-02680-7"
}

Anastasovska, V., Kocova, M., Zdraveska, N., Stojiljković, M., Skakić, A., Klaassen, K.,& Pavlović, S.. (2021). A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia. in Endocrine
Springer, New York., 73(1), 196-202.
https://doi.org/10.1007/s12020-021-02680-7

Anastasovska V, Kocova M, Zdraveska N, Stojiljković M, Skakić A, Klaassen K, Pavlović S. A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia. in Endocrine. 2021;73(1):196-202.
doi:10.1007/s12020-021-02680-7 .

Anastasovska, Violeta, Kocova, Mirjana, Zdraveska, Nikolina, Stojiljković, Maja, Skakić, Anita, Klaassen, Kristel, Pavlović, Sonja, "A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia" in Endocrine, 73, no. 1 (2021):196-202,
https://doi.org/10.1007/s12020-021-02680-7 . .

TY  - JOUR
AU  - Klaassen, Kristel
AU  - Stanković, Biljana
AU  - Zukić, Branka
AU  - Kotur, Nikola
AU  - Gašić, Vladimir
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1350
AB  - New coronavirus SARS-CoV-2 is capable to infect humans and cause a novel disease COVID-19. Aiming to understand a host genetic component of COVID-19, we focused on variants in genes encoding proteases and genes involved in innate immunity that could be important for susceptibility and resistance to SARS-CoV-2 infection. Analysis of sequence data of coding regions of FURIN, PLG, PRSS1, TMPRSS11a, MBL2 and OAS1 genes in 143 unrelated individuals from Serbian population identified 22 variants with potential functional effect. In silico analyses (PolyPhen-2, SIFT, MutPred2 and Swiss-Pdb Viewer) predicted that 10 variants could impact the structure and/or function of proteins. These protein-altering variants (p.Gly146Ser in FURIN; p.Arg261His and p.Ala494Val in PLG; p.Asn54Lys in PRSS1; p.Arg52Cys, p.Gly54Asp and p.Gly57Glu in MBL2; p.Arg47Gln, p.Ile99Val and p.Arg130His in OAS1) may have predictive value for inter-individual differences in the response to the SARS-CoV-2 infection. Next, we performed comparative population analysis for the same variants using extracted data from the 1000 Genomes project. Population genetic variability was assessed using delta MAF and Fst statistics. Our study pointed to 7 variants in PLG, TMPRSS11a, MBL2 and OAS1 genes with noticeable divergence in allelic frequencies between populations worldwide. Three of them, all in MBL2 gene, were predicted to be damaging, making them the most promising population-specific markers related to SARS-CoV-2 infection. Comparing allelic frequencies between Serbian and other populations, we found that the highest level of genetic divergence related to selected loci was observed with African, followed by East Asian, Central and South American and South Asian populations. When compared with European populations, the highest divergence was observed with Italian population. In conclusion, we identified 4 variants in genes encoding proteases (FURIN, PLG and PRSS1) and 6 in genes involved in the innate immunity (MBL2 and OAS1) that might be relevant for the host response to SARS-CoV-2 infection.
PB  - Elsevier, Amsterdam
T2  - Infection Genetics and Evolution
T1  - Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection
VL  - 84
DO  - 10.1016/j.meegid.2020.104498
ER  - 

@article{
author = "Klaassen, Kristel and Stanković, Biljana and Zukić, Branka and Kotur, Nikola and Gašić, Vladimir and Pavlović, Sonja and Stojiljković, Maja",
year = "2020",
abstract = "New coronavirus SARS-CoV-2 is capable to infect humans and cause a novel disease COVID-19. Aiming to understand a host genetic component of COVID-19, we focused on variants in genes encoding proteases and genes involved in innate immunity that could be important for susceptibility and resistance to SARS-CoV-2 infection. Analysis of sequence data of coding regions of FURIN, PLG, PRSS1, TMPRSS11a, MBL2 and OAS1 genes in 143 unrelated individuals from Serbian population identified 22 variants with potential functional effect. In silico analyses (PolyPhen-2, SIFT, MutPred2 and Swiss-Pdb Viewer) predicted that 10 variants could impact the structure and/or function of proteins. These protein-altering variants (p.Gly146Ser in FURIN; p.Arg261His and p.Ala494Val in PLG; p.Asn54Lys in PRSS1; p.Arg52Cys, p.Gly54Asp and p.Gly57Glu in MBL2; p.Arg47Gln, p.Ile99Val and p.Arg130His in OAS1) may have predictive value for inter-individual differences in the response to the SARS-CoV-2 infection. Next, we performed comparative population analysis for the same variants using extracted data from the 1000 Genomes project. Population genetic variability was assessed using delta MAF and Fst statistics. Our study pointed to 7 variants in PLG, TMPRSS11a, MBL2 and OAS1 genes with noticeable divergence in allelic frequencies between populations worldwide. Three of them, all in MBL2 gene, were predicted to be damaging, making them the most promising population-specific markers related to SARS-CoV-2 infection. Comparing allelic frequencies between Serbian and other populations, we found that the highest level of genetic divergence related to selected loci was observed with African, followed by East Asian, Central and South American and South Asian populations. When compared with European populations, the highest divergence was observed with Italian population. In conclusion, we identified 4 variants in genes encoding proteases (FURIN, PLG and PRSS1) and 6 in genes involved in the innate immunity (MBL2 and OAS1) that might be relevant for the host response to SARS-CoV-2 infection.",
publisher = "Elsevier, Amsterdam",
journal = "Infection Genetics and Evolution",
title = "Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection",
volume = "84",
doi = "10.1016/j.meegid.2020.104498"
}

Klaassen, K., Stanković, B., Zukić, B., Kotur, N., Gašić, V., Pavlović, S.,& Stojiljković, M.. (2020). Functional prediction and comparative population analysis of variants in genes for proteases and innate immunity related to SARS-CoV-2 infection. in Infection Genetics and Evolution
Elsevier, Amsterdam., 84.
https://doi.org/10.1016/j.meegid.2020.104498

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 Infection Genetics and Evolution. 2020;84.
doi:10.1016/j.meegid.2020.104498 .

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 Infection Genetics and Evolution, 84 (2020),
https://doi.org/10.1016/j.meegid.2020.104498 . .

TY  - JOUR
AU  - Stanković, Biljana
AU  - Kotur, Nikola
AU  - Gašić, Vladimir
AU  - Klaassen, Kristel
AU  - Ristivojević, Bojan
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Zukić, Branka
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1362
AB  - Uvod: Kako ne postoje odobreni terapeutici za lečenje pacijenata sa COVID-19, mogućnost upotrebe postojećih lekova je postala važna. U nedostatku vremena za testiranje farmakogenomskih markera kod pojedinaca, populaciona farmakogenomika bi mogla biti od koristi u predviđanju povećanog rizika za pojavu neželjenih reakcija i neuspeha lečenja kod pacijenata sa COVID-19. Cilj naše studije bio je identifikovanje farmakogena i farmakogenomskih markera povezanih sa lekovima koji se preporučuju za lečenje COVID-19, hlorokin/hidroksihlorokin, azitromicin, lopinavir i ritonavir, u populaciji Srbije i drugim svetskim populacijama. Metode: Podaci o genotipu 143 osobe srpskog porekla dobijeni su iz baze podataka prethodno formirane analizama genoma korišćenjem TruSight One Gene Panel (Illumina). Podaci o genotipu pojedinaca iz različitih svetskih populacija dobijeni su iz Projekta 1000 genoma. Fišerov egzaktni test korišćen je za poređenje učestalosti alela. Rezultati: Identifikovali smo 11 potencijalnih farmakogenomskih markera u 7 farmakogena značajnih za lečenje COVID-19. Na osnovu visoke alterativne učestalosti alela u populaciji Srbije i funkcionalnog efekta varijanti, ABCB1 rs1045642 i rs2032582 mogu biti značajne za smanjeni klirens lekova azitromicina, lopinavira i ritonavira, a varijanta UGT1A7 rs17868323 za hiperbilirubinemiju kod bolesnika sa COVID-19 koji se leče ritonavirom. SLCO1B1 rs4149056 je potencijalni marker odgovora na lopinavir, posebno u populaciji Italije. Naši rezultati potvrdili su da se farmakogenomski profil afričke populacije razlikuje od ostatka sveta. Zaključak: Uzimajući u obzir farmakogenomski profil specifičan za populaciju, preventivno testiranje farmakogena značajnih za lekove koji se koriste u lečenju COVID-19 moglo bi doprineti boljem razumevanju interindividualnih razlika u odgovorima na terapiju i poboljšanju ishoda lečenja pacijenata sa COVID-19.
AB  - Background: Since there are no certified therapeutics to treat COVID-19 patients, drug repurposing became important. With lack of time to test individual pharmacogenomics markers, population pharmacogenomics could be helpful in predicting a higher risk of developing adverse reactions and treatment failure in COVID-19 patients. Aim of our study was to identify pharmacogenes and pharmacogenomics markers associated with drugs recommended for COVID-19 treatment, chloroquine/hydroxychloroquine, azithromycin, lopinavir and ritonavir, in population of Serbia and other world populations. Methods: Genotype information of 143 individuals of Serbian origin was extracted from database previously obtained using TruSight One Gene Panel (Illumina). Genotype data of individuals from different world populations were extracted from the 1000 Genome Project. Fisher's exact test was used for comparison of allele frequencies. Results: We have identified 11 potential pharmacogenomics markers in 7 pharmacogenes relevant for COVID-19 treatment. Based on high alternative allele frequencies in population and the functional effect of the variants, ABCB1 rs1045642 and rs2032582 could be relevant for reduced clearance of azithromycin, lopinavir and ritonavir drugs and UGT1A7 rs17868323 for hyperbilirubinemia in ritonavir treated COVID-19 patients in Serbian population. SLCO1B1 rs4149056 is a potential marker of lopinavir response, especially in Italian population. Our results confirmed that pharmacogenomics profile of African population is different from the rest of the world. Conclusions: Considering population specific pharmacogenomics landscape, preemptive testing for pharmacogenes relevant for drugs used in COVID-19 treatment could contribute to better understanding of the inconsistency in therapy response and could be applied to improve the outcome of the COVID-19 patients.
PB  - Društvo medicinskih biohemičara Srbije, Beograd i Versita
T2  - Journal of Medical Biochemistry
T1  - Farmakogenomski profil odgovora na terapiju za COVID-19 u populaciji Srbije i poređenje sa populacijama širom sveta
T1  - Pharmacogenomics landscape of COVID-19 therapy response in Serbian population and comparison with worldwide populations
EP  - 499
IS  - 4
SP  - 488
VL  - 39
DO  - 10.5937/jomb0-26725
ER  - 

@article{
author = "Stanković, Biljana and Kotur, Nikola and Gašić, Vladimir and Klaassen, Kristel and Ristivojević, Bojan and Stojiljković, Maja and Pavlović, Sonja and Zukić, Branka",
year = "2020",
abstract = "Uvod: Kako ne postoje odobreni terapeutici za lečenje pacijenata sa COVID-19, mogućnost upotrebe postojećih lekova je postala važna. U nedostatku vremena za testiranje farmakogenomskih markera kod pojedinaca, populaciona farmakogenomika bi mogla biti od koristi u predviđanju povećanog rizika za pojavu neželjenih reakcija i neuspeha lečenja kod pacijenata sa COVID-19. Cilj naše studije bio je identifikovanje farmakogena i farmakogenomskih markera povezanih sa lekovima koji se preporučuju za lečenje COVID-19, hlorokin/hidroksihlorokin, azitromicin, lopinavir i ritonavir, u populaciji Srbije i drugim svetskim populacijama. Metode: Podaci o genotipu 143 osobe srpskog porekla dobijeni su iz baze podataka prethodno formirane analizama genoma korišćenjem TruSight One Gene Panel (Illumina). Podaci o genotipu pojedinaca iz različitih svetskih populacija dobijeni su iz Projekta 1000 genoma. Fišerov egzaktni test korišćen je za poređenje učestalosti alela. Rezultati: Identifikovali smo 11 potencijalnih farmakogenomskih markera u 7 farmakogena značajnih za lečenje COVID-19. Na osnovu visoke alterativne učestalosti alela u populaciji Srbije i funkcionalnog efekta varijanti, ABCB1 rs1045642 i rs2032582 mogu biti značajne za smanjeni klirens lekova azitromicina, lopinavira i ritonavira, a varijanta UGT1A7 rs17868323 za hiperbilirubinemiju kod bolesnika sa COVID-19 koji se leče ritonavirom. SLCO1B1 rs4149056 je potencijalni marker odgovora na lopinavir, posebno u populaciji Italije. Naši rezultati potvrdili su da se farmakogenomski profil afričke populacije razlikuje od ostatka sveta. Zaključak: Uzimajući u obzir farmakogenomski profil specifičan za populaciju, preventivno testiranje farmakogena značajnih za lekove koji se koriste u lečenju COVID-19 moglo bi doprineti boljem razumevanju interindividualnih razlika u odgovorima na terapiju i poboljšanju ishoda lečenja pacijenata sa COVID-19., Background: Since there are no certified therapeutics to treat COVID-19 patients, drug repurposing became important. With lack of time to test individual pharmacogenomics markers, population pharmacogenomics could be helpful in predicting a higher risk of developing adverse reactions and treatment failure in COVID-19 patients. Aim of our study was to identify pharmacogenes and pharmacogenomics markers associated with drugs recommended for COVID-19 treatment, chloroquine/hydroxychloroquine, azithromycin, lopinavir and ritonavir, in population of Serbia and other world populations. Methods: Genotype information of 143 individuals of Serbian origin was extracted from database previously obtained using TruSight One Gene Panel (Illumina). Genotype data of individuals from different world populations were extracted from the 1000 Genome Project. Fisher's exact test was used for comparison of allele frequencies. Results: We have identified 11 potential pharmacogenomics markers in 7 pharmacogenes relevant for COVID-19 treatment. Based on high alternative allele frequencies in population and the functional effect of the variants, ABCB1 rs1045642 and rs2032582 could be relevant for reduced clearance of azithromycin, lopinavir and ritonavir drugs and UGT1A7 rs17868323 for hyperbilirubinemia in ritonavir treated COVID-19 patients in Serbian population. SLCO1B1 rs4149056 is a potential marker of lopinavir response, especially in Italian population. Our results confirmed that pharmacogenomics profile of African population is different from the rest of the world. Conclusions: Considering population specific pharmacogenomics landscape, preemptive testing for pharmacogenes relevant for drugs used in COVID-19 treatment could contribute to better understanding of the inconsistency in therapy response and could be applied to improve the outcome of the COVID-19 patients.",
publisher = "Društvo medicinskih biohemičara Srbije, Beograd i Versita",
journal = "Journal of Medical Biochemistry",
title = "Farmakogenomski profil odgovora na terapiju za COVID-19 u populaciji Srbije i poređenje sa populacijama širom sveta, Pharmacogenomics landscape of COVID-19 therapy response in Serbian population and comparison with worldwide populations",
pages = "499-488",
number = "4",
volume = "39",
doi = "10.5937/jomb0-26725"
}

Stanković, B., Kotur, N., Gašić, V., Klaassen, K., Ristivojević, B., Stojiljković, M., Pavlović, S.,& Zukić, B.. (2020). Farmakogenomski profil odgovora na terapiju za COVID-19 u populaciji Srbije i poređenje sa populacijama širom sveta. in Journal of Medical Biochemistry
Društvo medicinskih biohemičara Srbije, Beograd i Versita., 39(4), 488-499.
https://doi.org/10.5937/jomb0-26725

Stanković B, Kotur N, Gašić V, Klaassen K, Ristivojević B, Stojiljković M, Pavlović S, Zukić B. Farmakogenomski profil odgovora na terapiju za COVID-19 u populaciji Srbije i poređenje sa populacijama širom sveta. in Journal of Medical Biochemistry. 2020;39(4):488-499.
doi:10.5937/jomb0-26725 .

Stanković, Biljana, Kotur, Nikola, Gašić, Vladimir, Klaassen, Kristel, Ristivojević, Bojan, Stojiljković, Maja, Pavlović, Sonja, Zukić, Branka, "Farmakogenomski profil odgovora na terapiju za COVID-19 u populaciji Srbije i poređenje sa populacijama širom sveta" in Journal of Medical Biochemistry, 39, no. 4 (2020):488-499,
https://doi.org/10.5937/jomb0-26725 . .

TY  - JOUR
AU  - Dragasević, Sanja
AU  - Stanković, Biljana
AU  - Kotur, Nikola
AU  - Sokić-Milutinović, Aleksandra
AU  - Milovanović, Tamara
AU  - Lukić, Snežana
AU  - Milosavljević, Tomica
AU  - Srzentić Dražilov, Sanja
AU  - Klaassen, Kristel
AU  - Pavlović, Sonja
AU  - Popović, Dragan
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1398
AB  - Background: This study analyzed poorly understood relationship of two overlapping conditions: metabolic syndrome (MeS) and inflammatory bowel disease (IBD), both associated with inflammation in the visceral adipose tissue. Methods: Newly diagnosed 104 IBD patients, of which 50 Crohn's disease (CD) and 54 ulcerative colitis (UC), and 45 non-IBD controls were examined for MeS-related obesity and lipid markers. Th-17 immune genes IL17A, IL17F, IL23A, and TLR9 mRNAs were measured in intestinal mucosa by qRT-PCR. Subjects were genotyped for obesity-associated FTO variant rs9939609 by polymerase chain reaction-amplification refractory mutation system. Results: CD was associated with MeS (P = 0.01), while both CD and UC were associated with central obesity (P = 10(-5), P = 0.002, respectively) and low levels of high-density lipoprotein (HDL) cholesterol (P = 5 x 10(-6), P = 6 x 10(-6), respectively). IBD lipid profile was characterized by decreased total and HDL cholesterol, while low-density lipoprotein cholesterol was reduced only in CD. Negative correlations were found between total cholesterol and CD activity index (P = 0.005), waist circumference and IL17A as well as IL17F mRNA levels in inflamed CD colon (P = 0.003, P = 0.001, respectively). Carriers of FTO rs9939609 AA genotype showed increased risk of CD (OR 2.6, P = 0.01). Conclusions: MeS, central obesity, and dyslipidemia could be important for IBD pathogenesis. This could influence therapeutic approaches and prevention strategies in high-risk groups.
PB  - Mary Ann Liebert, Inc, New Rochelle
T2  - Metabolic Syndrome and Related Disorders
T1  - Metabolic Syndrome in Inflammatory Bowel Disease: Association with Genetic Markers of Obesity and Inflammation
EP  - 38
IS  - 1
SP  - 31
VL  - 18
DO  - 10.1089/met.2019.0090
ER  - 

@article{
author = "Dragasević, Sanja and Stanković, Biljana and Kotur, Nikola and Sokić-Milutinović, Aleksandra and Milovanović, Tamara and Lukić, Snežana and Milosavljević, Tomica and Srzentić Dražilov, Sanja and Klaassen, Kristel and Pavlović, Sonja and Popović, Dragan",
year = "2020",
abstract = "Background: This study analyzed poorly understood relationship of two overlapping conditions: metabolic syndrome (MeS) and inflammatory bowel disease (IBD), both associated with inflammation in the visceral adipose tissue. Methods: Newly diagnosed 104 IBD patients, of which 50 Crohn's disease (CD) and 54 ulcerative colitis (UC), and 45 non-IBD controls were examined for MeS-related obesity and lipid markers. Th-17 immune genes IL17A, IL17F, IL23A, and TLR9 mRNAs were measured in intestinal mucosa by qRT-PCR. Subjects were genotyped for obesity-associated FTO variant rs9939609 by polymerase chain reaction-amplification refractory mutation system. Results: CD was associated with MeS (P = 0.01), while both CD and UC were associated with central obesity (P = 10(-5), P = 0.002, respectively) and low levels of high-density lipoprotein (HDL) cholesterol (P = 5 x 10(-6), P = 6 x 10(-6), respectively). IBD lipid profile was characterized by decreased total and HDL cholesterol, while low-density lipoprotein cholesterol was reduced only in CD. Negative correlations were found between total cholesterol and CD activity index (P = 0.005), waist circumference and IL17A as well as IL17F mRNA levels in inflamed CD colon (P = 0.003, P = 0.001, respectively). Carriers of FTO rs9939609 AA genotype showed increased risk of CD (OR 2.6, P = 0.01). Conclusions: MeS, central obesity, and dyslipidemia could be important for IBD pathogenesis. This could influence therapeutic approaches and prevention strategies in high-risk groups.",
publisher = "Mary Ann Liebert, Inc, New Rochelle",
journal = "Metabolic Syndrome and Related Disorders",
title = "Metabolic Syndrome in Inflammatory Bowel Disease: Association with Genetic Markers of Obesity and Inflammation",
pages = "38-31",
number = "1",
volume = "18",
doi = "10.1089/met.2019.0090"
}

Dragasević, S., Stanković, B., Kotur, N., Sokić-Milutinović, A., Milovanović, T., Lukić, S., Milosavljević, T., Srzentić Dražilov, S., Klaassen, K., Pavlović, S.,& Popović, D.. (2020). Metabolic Syndrome in Inflammatory Bowel Disease: Association with Genetic Markers of Obesity and Inflammation. in Metabolic Syndrome and Related Disorders
Mary Ann Liebert, Inc, New Rochelle., 18(1), 31-38.
https://doi.org/10.1089/met.2019.0090

Dragasević S, Stanković B, Kotur N, Sokić-Milutinović A, Milovanović T, Lukić S, Milosavljević T, Srzentić Dražilov S, Klaassen K, Pavlović S, Popović D. Metabolic Syndrome in Inflammatory Bowel Disease: Association with Genetic Markers of Obesity and Inflammation. in Metabolic Syndrome and Related Disorders. 2020;18(1):31-38.
doi:10.1089/met.2019.0090 .

Dragasević, Sanja, Stanković, Biljana, Kotur, Nikola, Sokić-Milutinović, Aleksandra, Milovanović, Tamara, Lukić, Snežana, Milosavljević, Tomica, Srzentić Dražilov, Sanja, Klaassen, Kristel, Pavlović, Sonja, Popović, Dragan, "Metabolic Syndrome in Inflammatory Bowel Disease: Association with Genetic Markers of Obesity and Inflammation" in Metabolic Syndrome and Related Disorders, 18, no. 1 (2020):31-38,
https://doi.org/10.1089/met.2019.0090 . .

TY  - JOUR
AU  - Stanković, Biljana
AU  - Dragasević, Sanja
AU  - Klaassen, Kristel
AU  - Kotur, Nikola
AU  - Srzentić Dražilov, Sanja
AU  - Zukić, Branka
AU  - Sokic Milutinović, Aleksandra
AU  - Milovanović, Tamara
AU  - Lukić, Snežana
AU  - Popović, Dragan
AU  - Pavlović, Sonja
AU  - Nikčević, Gordana
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1329
AB  - Background: Crohn's disease (CD) is chronic inflammatory bowel disease with different phenotypic characteristics influencing disease prognosis and therapeutic strategies. The aim of this pilot study was to analyze selected inflammatory and apoptotic markers in non-inflamed and inflamed samples of ileal mucosa of non-stricturing/non-penetrating (NS/NP) and stricturing (S) CD mucosal phenotypes in order to characterize their distinct profiles. Methods: From twenty CD patients (9 NS/NP, 11 S) paired non-inflamed and inflamed ileal biopsies were collected and used for analysis of cytokine (TNF and IL6) and apoptotic (Bcl2, Box, Fas and FasL) genes' expression levels by real-time PCR, while NF kappa B transcriptional potency was assessed by electromobility gel shift assay. Results: Our results demonstrated significant upregulation of TNF and IL6 in inflamed area of both NS/NP (p = 0.03, p = 0.01) and S phenotypes (p = 0.04, p = 0.04), respectively. However, TNF increase was more prominent in NS/NP compared to S inflamed mucosa (p = 0.02). Also, level of proapoptotic Box was significantly higher in NS/NP compared to S inflamed mucosa (p = 0.01). Opposing transcription potency of NF kappa B has been detected between two phenotypes: being decreased in NS/NP (p = 0.07) and increased in S (p = 0.1) inflamed compared to non-inflamed mucosa, demonstrating trend towards statistical significance. Conclusions: We found that two distinct CD phenotypes have specific molecular signatures. Obtained results could direct improvement of current and development of new therapeutic strategies based on more specific molecular stratification of CD patients.
PB  - Elsevier Gmbh, Munich
T2  - Pathology Research and Practice
T1  - Exploring inflammatory and apoptotic signatures in distinct Crohn's disease phenotypes: Way towards molecular stratification of patients and targeted therapy
IS  - 6
VL  - 216
DO  - 10.1016/j.prp.2020.152945
ER  - 

@article{
author = "Stanković, Biljana and Dragasević, Sanja and Klaassen, Kristel and Kotur, Nikola and Srzentić Dražilov, Sanja and Zukić, Branka and Sokic Milutinović, Aleksandra and Milovanović, Tamara and Lukić, Snežana and Popović, Dragan and Pavlović, Sonja and Nikčević, Gordana",
year = "2020",
abstract = "Background: Crohn's disease (CD) is chronic inflammatory bowel disease with different phenotypic characteristics influencing disease prognosis and therapeutic strategies. The aim of this pilot study was to analyze selected inflammatory and apoptotic markers in non-inflamed and inflamed samples of ileal mucosa of non-stricturing/non-penetrating (NS/NP) and stricturing (S) CD mucosal phenotypes in order to characterize their distinct profiles. Methods: From twenty CD patients (9 NS/NP, 11 S) paired non-inflamed and inflamed ileal biopsies were collected and used for analysis of cytokine (TNF and IL6) and apoptotic (Bcl2, Box, Fas and FasL) genes' expression levels by real-time PCR, while NF kappa B transcriptional potency was assessed by electromobility gel shift assay. Results: Our results demonstrated significant upregulation of TNF and IL6 in inflamed area of both NS/NP (p = 0.03, p = 0.01) and S phenotypes (p = 0.04, p = 0.04), respectively. However, TNF increase was more prominent in NS/NP compared to S inflamed mucosa (p = 0.02). Also, level of proapoptotic Box was significantly higher in NS/NP compared to S inflamed mucosa (p = 0.01). Opposing transcription potency of NF kappa B has been detected between two phenotypes: being decreased in NS/NP (p = 0.07) and increased in S (p = 0.1) inflamed compared to non-inflamed mucosa, demonstrating trend towards statistical significance. Conclusions: We found that two distinct CD phenotypes have specific molecular signatures. Obtained results could direct improvement of current and development of new therapeutic strategies based on more specific molecular stratification of CD patients.",
publisher = "Elsevier Gmbh, Munich",
journal = "Pathology Research and Practice",
title = "Exploring inflammatory and apoptotic signatures in distinct Crohn's disease phenotypes: Way towards molecular stratification of patients and targeted therapy",
number = "6",
volume = "216",
doi = "10.1016/j.prp.2020.152945"
}

Stanković, B., Dragasević, S., Klaassen, K., Kotur, N., Srzentić Dražilov, S., Zukić, B., Sokic Milutinović, A., Milovanović, T., Lukić, S., Popović, D., Pavlović, S.,& Nikčević, G.. (2020). Exploring inflammatory and apoptotic signatures in distinct Crohn's disease phenotypes: Way towards molecular stratification of patients and targeted therapy. in Pathology Research and Practice
Elsevier Gmbh, Munich., 216(6).
https://doi.org/10.1016/j.prp.2020.152945

Stanković B, Dragasević S, Klaassen K, Kotur N, Srzentić Dražilov S, Zukić B, Sokic Milutinović A, Milovanović T, Lukić S, Popović D, Pavlović S, Nikčević G. Exploring inflammatory and apoptotic signatures in distinct Crohn's disease phenotypes: Way towards molecular stratification of patients and targeted therapy. in Pathology Research and Practice. 2020;216(6).
doi:10.1016/j.prp.2020.152945 .

Stanković, Biljana, Dragasević, Sanja, Klaassen, Kristel, Kotur, Nikola, Srzentić Dražilov, Sanja, Zukić, Branka, Sokic Milutinović, Aleksandra, Milovanović, Tamara, Lukić, Snežana, Popović, Dragan, Pavlović, Sonja, Nikčević, Gordana, "Exploring inflammatory and apoptotic signatures in distinct Crohn's disease phenotypes: Way towards molecular stratification of patients and targeted therapy" in Pathology Research and Practice, 216, no. 6 (2020),
https://doi.org/10.1016/j.prp.2020.152945 . .

TY  - JOUR
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Tošić, Nataša
AU  - Klaassen, Kristel
AU  - Đorđević, Maja
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1254
AB  - Glycogen storage disease type Ib (GSD Ib) is an autosomal recessive disorder, caused by a deficiency of ubiquitously expressed SLC37A4 protein. Deficiency of SLC37A4 leads to abnormal storage of glycogen in the liver and kidneys, resulting in long-term complications of renal disease and hepatocellular adenomas, whose mechanisms are poorly understood. Molecular markers of the adaptive responses to the metabolic stress caused by a deficiency of SLC37A4, such as markers related to the endoplasmic reticulum (ER) stress and unfolded protein response (UPR), have not been extensively studied. The aim of this study was to investigate the expression of molecular markers of the UPR response and apoptosis related to a deficiency of SLC37A4 in kidney cells. For that purpose, we intended to establish a human kidney cell model system for GSD Ib. The novel variant c.248G  gt  A, found in GSD Ib patients, was introduced into the Flp-In (TM) T-REx (TM)-293 cell line using CRISPR/Cas9-mediated precise gene editing method, resulting in significant decrease of SLC37A4 gene expression. In this model system we used RT-qPCR analysis to investigate the expression of molecular markers of the UPR response (ATF4, DDIT3, HSPA5, and XBP1s) and apoptosis (BCL2, BAX). We demonstrated that under chronic metabolic stress conditions caused by SLC37A4 deficiency, the ER stress-induced UPR was triggered, resulting in suppression of the UPR molecular markers and cell survival promotion (decreased expression levels of ATF4, DDIT3, HSPA5, with the exception of XHE1s). However, persistent metabolic stress overrides an adaptation and induces apoptosis through increased expression of pro-apoptotic markers (decreased ratio of BCL2/BAX genes). We established a cellular model system characterized by a deficiency of SLC37A4, which presents pathological manifestations of GSD Ib in the kidney. Expression analysis in a novel model system supports the hypothesis that renal dysfunction in the GSD Ib is partly due to the ER stress and increased apoptosis.
PB  - Elsevier Science Bv, Amsterdam
T2  - Gene
T1  - CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib
EP  - 25
SP  - 17
VL  - 703
DO  - 10.1016/j.gene.2019.04.002
ER  - 

@article{
author = "Skakić, Anita and Anđelković, Marina and Tošić, Nataša and Klaassen, Kristel and Đorđević, Maja and Pavlović, Sonja and Stojiljković, Maja",
year = "2019",
abstract = "Glycogen storage disease type Ib (GSD Ib) is an autosomal recessive disorder, caused by a deficiency of ubiquitously expressed SLC37A4 protein. Deficiency of SLC37A4 leads to abnormal storage of glycogen in the liver and kidneys, resulting in long-term complications of renal disease and hepatocellular adenomas, whose mechanisms are poorly understood. Molecular markers of the adaptive responses to the metabolic stress caused by a deficiency of SLC37A4, such as markers related to the endoplasmic reticulum (ER) stress and unfolded protein response (UPR), have not been extensively studied. The aim of this study was to investigate the expression of molecular markers of the UPR response and apoptosis related to a deficiency of SLC37A4 in kidney cells. For that purpose, we intended to establish a human kidney cell model system for GSD Ib. The novel variant c.248G  gt  A, found in GSD Ib patients, was introduced into the Flp-In (TM) T-REx (TM)-293 cell line using CRISPR/Cas9-mediated precise gene editing method, resulting in significant decrease of SLC37A4 gene expression. In this model system we used RT-qPCR analysis to investigate the expression of molecular markers of the UPR response (ATF4, DDIT3, HSPA5, and XBP1s) and apoptosis (BCL2, BAX). We demonstrated that under chronic metabolic stress conditions caused by SLC37A4 deficiency, the ER stress-induced UPR was triggered, resulting in suppression of the UPR molecular markers and cell survival promotion (decreased expression levels of ATF4, DDIT3, HSPA5, with the exception of XHE1s). However, persistent metabolic stress overrides an adaptation and induces apoptosis through increased expression of pro-apoptotic markers (decreased ratio of BCL2/BAX genes). We established a cellular model system characterized by a deficiency of SLC37A4, which presents pathological manifestations of GSD Ib in the kidney. Expression analysis in a novel model system supports the hypothesis that renal dysfunction in the GSD Ib is partly due to the ER stress and increased apoptosis.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Gene",
title = "CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib",
pages = "25-17",
volume = "703",
doi = "10.1016/j.gene.2019.04.002"
}

Skakić, A., Anđelković, M., Tošić, N., Klaassen, K., Đorđević, M., Pavlović, S.,& Stojiljković, M.. (2019). CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib. in Gene
Elsevier Science Bv, Amsterdam., 703, 17-25.
https://doi.org/10.1016/j.gene.2019.04.002

Skakić A, Anđelković M, Tošić N, Klaassen K, Đorđević M, Pavlović S, Stojiljković M. CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib. in Gene. 2019;703:17-25.
doi:10.1016/j.gene.2019.04.002 .

Skakić, Anita, Anđelković, Marina, Tošić, Nataša, Klaassen, Kristel, Đorđević, Maja, Pavlović, Sonja, Stojiljković, Maja, "CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib" in Gene, 703 (2019):17-25,
https://doi.org/10.1016/j.gene.2019.04.002 . .