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Recent research shows that non-coding RNA transcripts of protein-coding genes could be an emerging novel class of diagnostic biomarkers. This study aimed to identify the most promising biomarkers for colorectal cancer (CRC) screening among upregulated non-coding transcripts of protein-coding genes in malignant CRC cells in comparison to non-malignant cells grown in 3D. Malignant CRC cell lines HCT116, DLD-1 and SW620, and a non-malignant human colon epithelial cell line HCEC-1CT, were cultivated in 3D as spheroids in 24-well plates with low attachment surface for 7 days. RNA sequencing was performed on ribosomal-depleted total RNA using Illumina’s NovaSeq6000 platform that generated paired-end 150bp reads. Highly upregulated transcripts (>10 FPKM) present in all malignant cell lines and absent in non-malignant cell line were filtered and analyzed by a set of in silico tools to filter the best candidates for further validation studies. The publicly available GSE164541 set consisting of triplicate tissue samples of normal, adenoma and primary CRC tissues collected from five patients with CRC was used for validation. As a result, 5 transcripts with retained introns ANXA3-204, LLGL2- 207, KRT19-204, KRT18-206 and KRT8-213 were analyzed by in silico tools. Only ANXA3-204 was classified as non-coding according to both CPC2 and LGC online coding potential prediction tools. Nucleus was predicted as subcellular localization for ANXA3- 204 by AnnoLnc2. Additionally, ANXA3-204 expression was upregulated in adenoma (p=0.04) and CRC tissue samples, although statistical significance was not reached for CRC, in comparison to normal tissue samples in the validation set. Transcriptomic analysis revealed that non-coding ANXA3-204 transcript was highly upregulated in malignant CRC cell lines and adenomas compared to control samples, making ANXA3-204 a potential candidate for early CRC screening. Further studies are needed to confirm diagnostic potential and regulatory role of ANXA3-20

While the incidence of late-onset (≥50 years) colon cancer (LOCC) has been decreased worldwide, the number of early-onset (<50 years) colon cancer (EOCC) is increasing. Cluster of differentiation (CD) markers are widely and successfully used for identification of leukocyte population using flow cytometry in order to differentiate phenotypes among blood cancers and to address appropriate treatments. However, the alternations in their expression levels have been observed in non-blood cancers as well, including colon cancer, which enhance their diagnostic, prognostic and therapeutic biomarker potential. The aim of this research was to identify potential CD biomarkers for EOCC and LOCC using open access to expression profiling by high throughput sequencing in order to conduct further experimental exploration. Transcriptomic data of dataset GSE240623, obtained from formalin-fixed paraffinembedded tumor tissue samples from 13 EOCC and 13 LOCC patients, and their pairedadjacent normal colon tissues, from Gene Expression Omnibus (GEO) database was used. Differentially expressed genes for EOCC and LOCC paired groups were obtained using DESeq2 package in R software with log2 fold change threshold set to 1. Up and downregulated genes were subsequently filtered only by CD markers for both comparisons. In EOCC patients, CD79B, CD22, CD19, CD79A, CD37 and CD48 were downregulated, while CD276 was upregulated in tumor tissue compared to control tissue. On the other hand, only CD33 was downregulated in LOCC tumor tissue compared to normal colon tissue. Finally, CD27 was downregulated in tumor tissues of both groups, EOCC and LOCC, in comparison to their counterparts. In this study, we have identified nine CD markers that have potential diagnostic, prognostic and/or therapeutic significance for colon cancer and will be further examined in in silico and in vitro study.

IntroductionBiomarkers capable of reflecting disease onset and short- and long-term therapeutic effects in individuals with spinal muscular atrophy (SMA) are still an unmet need and phosphorylated neurofilament heavy chain (pNF-H) holds significant promise.MethodsWe conducted a longitudinal prospective study to evaluate pNF-H levels in the cerebrospinal fluid (CSF) and plasma of 29 individuals with childhood-onset SMA treated with Nuinersen (SMA type 1: n = 6, 2: n = 17, 3: n = 6). pNF-H levels before and during treatment were compared with the levels of controls (n = 22), patients with Duchenne muscular dystrophy (n = 17), myotonic dystrophy type 1 (n = 11), untreated SMA individuals with chronic type 3 disease (n = 8), and children with presymptomatic SMA (n = 3).ResultsSMA type 1 showed the highest mean CSF pNF-H levels before treatment initiation. All Nusinersen-treated individuals (types 1, 2, and 3) showed significantly elevated mean baseline CSF pNF-H compared to controls, which inversely correlated with age at disease onset, age at first dose, disease duration and the initial CHOP INTEND result (SMA type 1 and 2). During 22 months of treatment, CSF pNF-H levels declined during loading doses, stabilizing at reduced levels from the initial maintenance dose in all individuals. Baseline plasma pNF-H levels in type 1 and 2 SMA were significantly increased compared to other cohorts and decreased notably in type 1 after 2 months of treatment and type 2 after 14 months. Conversely, SMA type 3, characterized by lower baseline pNF-H levels, did not show significant fluctuations in plasma pNF-H levels after 14 months of treatment.ConclusionOur findings suggest that CSF pNF-H levels in untreated SMA individuals are significantly higher than in controls and that monitoring of CSF pNF-H levels may serve as an indicator of rapid short-term treatment response in childhood-onset SMA individuals, irrespective of the subtype of the disease, while also suggesting its potential

Cysteine cathepsins F and W are members of the papain-like cysteine protease family, which have distinct structural features and functional roles in various physiological and pathological processes. This review provides a comprehensive overview of the current understanding of the structure, biological functions, and pathological implications of cathepsins F and W. Beginning with an introduction to these proteases, we delve into their structural characteristics and elucidate their unique features that dictate their enzymatic activities and substrate specificity. We also explore the intricate involvement of cathepsins F and W in malignancies, highlighting their role as potential biomarkers and therapeutic targets in cancer progression. Furthermore, we discuss the emerging roles of these enzymes in immune response modulation and neurological disorders, shedding light on their implications in autoimmune and neurodegenerative diseases. Finally, we review the landscape of inhibitors targeting these proteases, highlighting their therapeutic potential and challenges in clinical translation. This review brings together the diverse facets of cysteine cathepsins F and W, providing insights into their roles in health and disease and guiding future investigations for therapeutic advances.

Genome-wide association studies (GWAS) have provided strong evidence that early- and late-onset MG have different genetic backgrounds. Recent in silico analysis based on GWAS results revealed rs231735 and rs231770 variants within CTLA-4 locus as possible MG causative genetic factors. We aimed to explore the association of rs231735 and rs231770 with MG in a representative cohort of Serbian patients. We conducted an age-, sex-, and ethnicity-matched case–control study. Using TaqMan allele discrimination assays, the frequency of rs231735 and rs231770 genetic variants was examined in 447 AChR-MG patients and 447 matched controls. There was no significant association of rs231735 and rs231770 with the entire MG cohort (P > 0.05). Nevertheless, when stratifying patients into early-onset (n = 183) and late-onset MG (n = 264), we found early-onset patients had a significantly lower frequency of the rs231735 allele T compared to controls (OR = 0.734, 95% CI = 0.575–0.938, p10e6 permutation < 0.05), and rs231735 genotype TT and rs231770 genotype TT had a protective effect on early-onset MG (OR = 0.548, 95% CI = 0.339–0.888, and OR = 0.563, 95% CI = 0.314–1.011, p10e6 permutation < 0.05). Consequently, we found that individuals with the rs231735-rs231770 haplotype GC had a higher risk for developing early-onset MG (OR = 1.360, P = 0.027, p10e6 permutation < 0.05). Our results suggest that CTLA-4 rs231735 and rs231770 may be risk factors only for patients with early-onset MG in Serbian population.

Spinal muscular atrophy (SMA) is the prevalent genetic cause of childhood mortality. Pioneering treatments yield utmost advantages only within the presymptomatic phase, underlining the medical and ethical significance of newborn screening. In 2022, the Centre for Human Molecular Genetics initiated a pilot study of the newborn screening for SMA, working closely alongside the University Children’s Hospital Tirsova and Association SMA Serbia. The aim was to lay the foundation for the implementation of statewide newborn screening for SMA in Serbia by conducting screening for ~8000 infants from the Obstetrics and Gynaecology Clinic Narodni Front over the course of a year. In the subsequent year, we expanded the initiative to include another maternity hospital located outside Belgrade, introducing sample shipment via postal services and extending screening accessibility to a greater number of infants. In the initial year, 6 950 newborns underwent testing, revealing SMA in two unrelated infants. Subsequently, an older sibling of the first newborn, although asymptomatic at the time, was also tested at the age of 16 months, and SMA diagnosis was confirmed in this child as well. All three children received therapeutic interventions in <1 month from birth. To date, they have exhibited no signs of SMA, and there have been no false negative outcomes among the newborns who tested negative during the screening. In the second year, an additional 5 000 newborns underwent testing. As frontrunners in this field in Serbia, we orchestrated harmonized efforts across various tiers of healthcare, established screening and diagnostic algorithms and follow-up protocols. Our extensive efforts were primarily aimed at elevating awareness among all stakeholders about the critical importance of early disease detection. In this transformative journey, we transitioned from being isolated individuals and visionaries who championed a singular idea to an entire community and nation that now acknowledg

Dobrijević Z., J. Karanović, D. Savić-Pavićević, G. Brajušković (2023). The effect of epistatic interactions between genetic variants located in microRNA and silencing complex genes on prostate cancer progression risk.- Genetika, Vol 55, No.1, 263-275. Previous studies conducted in Asian and European populations have provided evidence of the association between microRNA-related genetic variants and prostate cancer (PCa) risk and/or progression. Nevertheless, the results obtained in these studies are inconsistent, which could be explained by the limitations of single-locus main effect evaluations to detect joint effects of multiple genetic variants, reflected in statistical epistases. Therefore, we conducted the analysis of potential epistatic interactions between variants located in microRNA genes and in genes encoding the components of RNA-induced silencing complex (RISC) in relation with PCa risk/aggressiveness. Raw data on genotyping results from our previous studies involving four microRNA polymorphisms and five variants in RISC genes were subjected to the exclusion of samples based on missing data criterion, followed by the re-evaluation of Hardy-Weinberg equilibrium. Afterwards, these genotyping results were included in the Multifactor dimensionality reduction (MDR) analysis. Permutation testing was conducted in order to assess statistical significance of the best models from MDR tests. MDR tests on the risk of developing PCa yielded statistically insignificant results. Nevertheless, the MDR results for comparison of PCa patients with high and low cancer progression risk were statistically significant for the analysis that included rs11614913, with the 3-locus best model comprising this genetic variant, rs7813 and rs784567. We conclude that statistical epistasis between rs11614913 in hsa-miR-196a2, rs7813 in GEMIN4 and rs784567 in TARBP2 shows association with the invasiveness of PCa., Ranije studije u evropskim i azijskim populacijama ukazale su na značajnu asocijaciju genetičkih varijanti sa efektom na funkciju mikroRNK sa rizikom za razvoj i/ili progresiju karcinoma prostate (KP). Međutim, rezultati navedenih studija su nepodudarni, za šta je jedan od mogućih razloga nemogućnost pristupa baziranih na proceni pojedinačnih efekata genetičkih varijanti da detektuju značajne zajedničke efekte više varijanti, a koji se reflektuju u statističke epistaze. Iz tog razloga, cilj naše studije bila je analiza potencijalnih epistatičkih interakcija između varijanti lociranih u genima za mikroRNK molekule i u genima za komponente utišavajućeg kompleksa indukovanog sa RNK (RISC) kao faktora rizika za razvoj i/ili progresiju KP. Rezultati genotipizacije dobijeni tokom sprovođenja naših ranijih studija, a koji uključuju podatke za četiri varijante u genima za mikroRNK i pet u genima za komponente RICS, podvrgnuti su inicijalnoj obradi podataka u smislu isključivanja uzoraka sa nedostajućim rezultatima, nakon čega je procenjeno odstupanje od Hardi-Vajnbergove ravnoteže. Rezultati su zatim analizirani metodom redukcije dimenzionalnosti višestrukih faktora (Multifactor dimensionality reduction - MDR analysis). Permutacioni testovi su sprovedeni sa ciljem procene statističke značajnosti najboljih modela iz MDR analize. Dobijeni rezultati MDR testova koji su se odnosili na rizik za razvoj KP nisu bili statistički značajni. S druge strane, MDR rezultati koji se odnose na rizik za progresiju KP bili su značajni za model koji uključuje tri lokusa: rs11614913, rs7813 and rs784567. Stoga, zaključci naše studije ukazuju na značaj epistatičkih interakcija između varijanti rs11614913 u hsa-miR-196a2, rs7813 u GEMIN4 i rs784567 u TARBP2 kao faktora koji ispoljavaju efekat na invazivnost KP.

PRMT6, PEX10 and SOX5 genetic variants were identified as male infertility-associated loci in a genome-wide association study and further validated in various populations. Still, the results of previous case-control studies varied, which could be due to differences in participants’ ethnic backgrounds. The main purpose of the present study was to evaluate the supposed association of these variants with idiopathic male infertility in North Macedonian population. Furthermore, we aimed to conduct the systematic quantitative data synthesis which includes the results of previous studies on the same issue in other European and non-European populations. A total of 137 men from North Macedonia diagnosed with idiopathic infertility and 130 age-matched fertile controls were included in the present case-control study. PCR-RFLP method was used for genotyping. Meta-analysis was performed by OpenMeta-analyst statistical software. Variants rs10842262 in SOX5, rs2477686 in PEX10 and rs12097821 in PRMT6 showed the lack of statistically significant differences in genotype distributions between men diagnosed with idiopathic infertility and the control group. Still, rs10842262 allele G frequency was significantly increased in men with poor sperm concentration (P= 0.024, OR = 2.10, 95%CI 1.08-4.06). Meta-analysis further showed the association of rs10842262 and rs12097821 with the risk of idiopathic male infertility. Our results obtained in North Macedonian population supported the previous reports on the involvement of rs10842262 in the genetic basis of male infertility. The meta-analysis confirmed the association of rs10842262 and rs12097821 with male infertility occurrence. Still, additional studies are needed to support the present findings., Asocijacija varijanti u genima PRMT6, PEX10 i SOX5 sa muškim sterilitetom identifikovana je u studiji genetičke asocijacije na čitavom genomu i kasnije analizirana u studijama slučajeva i kontrola u različitim populacijama. Rezultati prethodnih studija su pokazali značajnu varijabilnost, što može biti posledica razlika u etničkom poreklu studijskih grupa. Osnovni cilj ovog istraživanja je analiza asocijacije navedenih genetičkih varijanti sa rizikom za pojavu idiopatskog muškog steriliteta u populaciji Severne Makedonije. Takođe, naš cilj je bio i sprovođenje sistematske kvantitativne sinteze podataka iz studija sa istom ili sličnom temom istraživanja sprovedenim u drugim evropskim i neevropskim populacijama. Ukupno 137 muškaraca sa idiopatskim sterilitetom iz Severne Makedonije i 130 fertilnih kontrola slične starosti uključeno je u studiju slučajeva i kontrola. Genotipizacaija je vršena PCR-RFLP metodom, dok je za meta-analizu korišćen statistički softver OpenMeta-analyst. Za varijante rs10842262 u SOX5, rs2477686 u PEX10 i rs12097821 u PRMT6 nije utvrđena statistički značajna razlika u distribucijama genotipova između grupe ispitanika sa idiopatskim sterilitetom i kontrolne grupe. Međutim, učestalost alela G varijante rs10842262 bila je značajno povećana kod muškaraca sa niskom koncentracijom spermatozoida (P= 0.024, OR = 2.10, 95%CI 1.08– 4.06). Meta-analizom pokazana je asocijacija rs10842262, ali i rs12097821, sa rizikom za razvoj idiopatskog muškog steriliteta. Naši rezultati ustanovljeni u populaciji Severne Makedonije idu u prilog prethodnim navodima o učešću rs10842262 u genetičkoj osnovi muškog steriliteta. Ipak, dodatne studije su neophodne kako bi potvrdile značaj rezultata ovog istraživanja.

Myasthenia gravis (MG) is a rare autoimmune disease mediated by antibodies against components of the neuromuscular junction, particularly the acetylcholine receptor (AChR). The prevalence of MG in Belgrade has been estimated at 189 cases per 1,000,000 inhabitants, which is among the highest prevalence reported to date. Genetic studies have mainly pointed to specific HLA alleles associated with MG. However, CTLA-4 and TNFRSF11A, playing a role in the immune response, have recently been associated with MG in genome-wide association studies. Since CTLA-4 and TNFRSF11A promote other autoimmune diseases, the main objective of this casecontrol study was to determine the association between these candidate genes and the risk for developing MG in Serbian population. Genotyping of rs231735 and rs231770 within the CTLA-4 gene and rs4263037 within TNFRSF11A in 447 AChR-MG patients and 447 individually sex- and age-matched controls revealed no association with MG (p=0.344, p=0.923 and p=0.557, respectively). However, when stratifying patients into those with early-onset (n=183) and late-onset MG (n=264), we found an association of minor rs231735 allele T with early-onset MG under the recessive genetic model (OR=0.548, 95% CI=0.339-0.888, p=0.014, p10e6 permutation=0.014). Haplotype analysis revealed that individuals with the GC haplotype rs231735-rs231770 had a higher risk for developing earlyonset MG (OR =1.360, p=0.027, p10e6 permutation =0.027). Considering the sufficient statistical power of the study (>90%) and the selection criteria for controls, our results suggest that the CTLA-4 may be associated with early-onset MG in Serbian population. Analysis of additional variants is needed to understand the association of CTLA-4 with MG

Spinal muscular atrophy (SMA) is the most common genetic cause of death in childhood. Innovative therapies show the greatest benefit only when administered in the presymptomatic period, making newborn screening an ethical and medical priority in many countries. In 2022 Centre for Human Molecular Genetics initiated a feasibility study of the newborn screening for SMA in close collaboration with the University Children's Hospital Tirsova, Association SMA Serbia and with financial support from Novartis Gene Therapies, Roche and Biogen/Medis Pharma aiming to screen up to 8000 babies from the Obstetrics and Gynaecology Clinic Narodni Front during one year. A total of 6950 newborns were tested and SMA was confirmed in two unrelated newborns from families with no history of SMA. A 16-month old sibling of the first baby was also tested, even though he was completely asymptomatic, and SMA was also confirmed. Average time between birth and the first screen-positive result was 5 days, and 8 days between birth and final confirmation of diagnosis. All three children received modifying therapies in less than 10 days from final diagnosis. So far, no false-negatives have been reported among the newborns who tested negative in the screening. As pioneers and leaders in this field, we created synchronised work at different levels of healthcare system, established screening and diagnostic algorithms and follow-up protocols. We are currently involved in scaling up screening to include an additional maternity hospital and preparing the ground for the implementation of the newborn screening for SMA as the official national screening program.

Introduction:Myotonic dystrophy type 1 (DM1) is a rare, incurable multisystemic disease, with the main symptoms being skeletal muscle weakness, atrophy, and myotonia. It is caused by CTG expansion in the 3' UTR of the DMPK gene whose RNA acquires toxic functions and sequesters MBNL proteins, resulting in globally altered RNA metabolism. To better understand the DM1 transcriptome, we systematically analyzed gene expression in the skeletal muscles of various mouse DM1 models. Methods:We retrieved 13 publicly available RNA-seq datasets from mouse models expressing expanded CTG repeats (HSALR, CTG480KI, TREDT960I) and Mbnl knockout models (SKO, DKO, TKO). Our bioinformatic pipeline with unified parameters consisted of preprocessing, differential expression (DESeq2), gene network analysis (WGCNA), comparison of gene network interactions with the STRING database, and network node enrichment analysis (Cytoscape). Results: In models expressing CTG repeats, the average number of up-regulated genes was 787, compared to 676 in Mbnl knockout models, while there was 642 and 380 down-regulated genes, respectively (log2FC>1, padj>0.05). Both model groups had network modules whose nodes were enriched for muscle and secretory functions (FDR<0.05). There were modules related to immune response, lipid transfer, and insulin in models expressing repeats and modules related to immunoglobulins and extracellular matrix in knockout models. Conclusion: Gene expression patterns separated Mbnl knockouts from models expressing CTG repeats that had a greater number of smaller functionally distinct network modules. Our results revealed novel pathway changes in DM1 skeletal muscles, among which immunological and secretory are particularly interesting as molecular targets for further investigation.

Serotonin neurotransmission is sensitive to environmental stimuli. Serotonin receptor 2C (HTR2C) undergoes dynamic A-to-I editing that fine-tunes cell response to serotonin and is altered in depressed suicide victims and by pharmacological treatments. We aimed to explore a mediating role of Htr2c mRNA editing in response to enriched environment and factors involved in this response. Three-week-old BALB/c and C57BL/6 male mice were housed in enriched and standard conditions for four weeks. Htr2c mRNA editing pattern and expression, serotonin level and expression of Adar and Adarb1 mRNAs (coding enzymes catalyzing A-to-I editing) and Snord115 RNA (regulating Htr2c mRNA alternative splicing and editing) were measured in prefrontal cortex (PFC) and hippocampus (HC), brain regions implicated in suicidal behavior. BALB/c mice, a “stress-sensitive” strain due to genetically determined lower serotonin level, responded to enriched conditions by adapting the Htr2c editing pattern to a slight serotonin decrease in PFC and a significant increase in HC. C57BL/6 mice, a “stress-resilient” strain, responded to enriched environment by increasing the serotonin level and changing Adar and Adarb1 mRNAs expression in HC, and without changes in PFC. Our findings suggest that the enriched environment effect on the serotonin level and a mediating role of Htr2c mRNA editing in PFC depend on the genetic background and its interactions with the environment. On the other hand, changes in HC are primarily driven by enriched environment. Our results imply usefulness of enriched environment paradigm for understanding interactions of genetic and environmental factors underlying suicidal behavior, which might improve psychological treatments.

Considering the substantial impact of genetic therapies for spinal muscular atrophy (SMA), longitudinal follow-up of patients undergoing treatment is crucial to effectively monitor treatment response. While functional rating scales are commonly used as primary outcome measures, they may not fully capture all the therapeutic benefits. To address this limitation, the phosphorylated neurofilament heavy chain (pNFH) protein has emerged as a promising biomarker for evaluating treatment response. pNF-H is a neuron- specific filament that exhibits increased levels in the cerebrospinal fluid (CSF) and plasma in the presence of neuronal degeneration. Our study includes individuals treated with Nusinersen (CSF and plasma samples) and Risdiplam (plasma), as well as age- and sex-matched control subjects (CSF and plasma). By examining the dynamics of pNF-H levels in these groups, we sought to identify significant differences indicative of treatment response. Before treatment, SMA individuals typically exhibit higher levels of pNF-H compared to non-SMA individuals. Elevated levels of pNF-H are associated with more severe clinical manifestations of the disease. During Nusinersen treatment, a notable decline in pNF-H levels during the first 2 months can be observed. Current findings suggest that genetic therapies have a notable impact on reducing pNF-H levels over time. By examining the changes in pNF-H levels, our study offers valuable insights into the underlying biochemical alterations associated with these therapies. Furthermore, it supports the use of pNF-H as a complementary measure to functional rating scales and as a potential biomarker for evaluating treatment effectiveness and monitoring disease progression in SMA.

Introduction: Genome-wide association studies (GWAS) identify genomic loci that contain genetic determinants of complex diseases. Subsequent functional genomic approaches, such as bioinformatic finemapping and transcriptome-wide association studies (TWAS), can reveal potentially causal single nucleotide variants (SNVs) that can be tested on patient samples. We applied this approach to study causal SNVs for acetylcholine receptor (AChR) seropositive myasthenia gravis (MG). We focused on CHRNA1 and CHRNB1 loci, coding AChR subunits, and CTLA-4 locus, coding protein transmitting an inhibitory signal to T cells. Methods: CHRNA1 was fine-mapped by PAINTOR using data from GWAS summary statistics, 1000 genome and RegulomeDB. Alongside, rs4151121 identified by TWAS in CHRNB1, and rs231735 and rs231770 identified by fine-mapping in CTLA-4 were studied. SNVs were genotyped using allele discrimination assays in 447 Serbian AChR-MG patients (183 early-onset and 264 late-onset) and 447 sex- and age-matched controls. Results: CHRNA1 rs35274388 was fine-mapped as a potentially causal variant (PIP2=92%) exhibiting transcription factor binding and chromatin accessibility peaks. CHRNA1 rs35274388 minor allele A and CHRNAB1 rs4151121 minor allele G increased the risk for late-onset MG (OR=1.669, 95% CI=1.05-2.638, p=0.027, p10e6 permutation=0.031 and OR=1.322, 95% CI=1.063-1.644, p10e6 permutation=0.014, respectively). On the other hand, CTLA-4 rs231735 recessive genotype TT decreased, while rs231735- rs231770 haplotype GC increased the susceptibility to early-onset MG (OR=0.548, 95% CI=0.339-0.888, p=0.014, p10e6 permutation=0.014 and OR=1.360, p=0.027, p10e6 permutation=0.027, respectively). Conclusion: CHRNA1 rs35274388 and CHRNAB1 rs4151121 loci could be causal genetic factors for lateonset MG while CTLA-4 rs231735 and rs231770 could be causal genetic factors for early-onset MG in Serbian population.

Background: Spinal muscular atrophy (SMA) is the prevalent genetic cause of childhood mortality. Pioneering treatments yield utmost advantages only within the presymptomatic phase, underlining the significance of newborn screening. Materials and methods: In 2022, the Centre for Human Molecular Genetics initiated a pilot study of the newborn screening for SMA, working closely alongside the University Children’s Hospital Tirsova and Association SMA Serbia. The aim was to lay the foundation for the implementation of statewide newborn screening for SMA in Serbia by conducting screening for ~8000 infants from the Obstetrics and Gynaecology Clinic Narodni Front over the course of a year. Results: In the initial year, 6950 newborns underwent testing, revealing SMA in two unrelated infants and in an asymptomatic 16-month old sibling of the first newborn. All three children received therapeutic interventions in <1 month from birth. To date, they have exhibited no signs of SMA, and there have been no false-negative outcomes among the newborns who tested negative during the screening. As frontrunners in this field in Serbia, we orchestrated harmonized efforts across various tiers of healthcare, established screening and diagnostic algorithms and follow-up protocols. In the second year, we included a maternity hospital beyond Belgrade, introducing sample shipping via mail and extending screening accessibility to a greater number of infants. This resulted in 9800 infants undergoing testing within 16 months. Currently, we are actively preparing for the official incorporation of newborn screening for SMA into the national screening program. Conclusions: Timely detection and treatment can transform SMA into a manageable condition.

Samoubilačko ponašanje predstavlja globalni zdravstveni problem s obzirom da oko milion ljudi godišnje izvrši samoubistvo, a efikasne mere prevencije ili lečenja ne postoje. Kao i drugi složeni fenotipovi, etiologija samoubilačkog ponašanja vezuje se za brojne genetičke i sredinske faktore rizika i njihove međusobne interakcije. Pokušaj samoubistva je često, ali ne i nužno, povezan sa psihijatrijskim oboljenjima i stresnim događajima tokom života, kao što su stresni događaji iz detinjstva i akutni stresni događaji. Neurobiološka istraživanja ukazuju na hipofunkciju serotoninskog sistema kod osoba sa samoubilačkim ponašanjem. Editovanje RNK deaminacijom adenozina u inozin, katalizovan enzimima ADAR i ADARB1, doprinosi finoj i dinamičnoj regulaciji i održavanju optimalne serotoninske transmisije kroz editovanje transkripata za triptofanhidroksilazu 2 (TPH2), enzima koji katalizuje sintezu serotonina u mozgu, i transkripata za postsinaptički serotoninski receptor 2C (HTR2C). Cilj ove doktorske teze je identifikacija varijanti u genima TPH2, HTR2C, ADAR i ADARB1, stresnih životnih događaja i njihovih međusobnih interakcija koje mogu napraviti razliku između psihijatrijskih bolesnika koji su pokušali samoubistvo i onih koji to nikada nisu uradili. Studija je uključila 353 psihijatrijska bolesnika sa dijagnozom depresivnog poremećaja, bipolarnog afektivnog poremećaja i shizofrenije, od kojih je 165 pokušalo samoubistvo, dok njih 188 nije. Stresni događaji iz detinjstva i akutni stresni događaji procenjeni su korišćenjem upitnika Early Trauma Inventory–Self Report i List of Threatening Experiences Questionnaire, redom. Primenom Pirsonovog 2–testa i logističke regresije testirane su potencijalne pojedinačne asocijacije stresnih događaja tokom detinjstva, akutnih stresnih događaja, 23 tag varijante gena ADAR i ADARB1, varijanta rs6318 gena HTR2C i varijante rs7305115 i rs4290270 gena TPH2 sa pokušajem samoubistva. Generalizovani linearni modeli i selekcija unazad primenjeni, Suicidal behavior is a global health problem since about one million people per year commit suicide and there is no effective prevention or treatment. Like other complex phenotypes, etiology of suicidal behavior is associated with numerous genetic and environmental risk factors and their interactions. A suicide attempt is often, although not necessarily, associated with psychiatric disorders and stressful life events, such as childhood abuse and acute stressful life events. Neurobiological studies have indicated a hypofunction of the serotonin system in patients with suicidal behavior. RNA editing by deamination of adenosine to inosine, catalysed by ADAR and ADARB1 enzymes, contributes to fine and dynamic tuning and maintenance of an optimal serotonin transmission through the editing of transcripts encoded by the tryptophan-hydroxylase 2 (TPH2) and serotonin receptor 2C (HTR2C) genes. TPH2 is a rate limiting enzyme for serotonin synthesis in the brain, while HTR2C is a postsynaptic receptor. The aim of this doctoral thesis was to identify variants in the TPH2, HTR2C, ADAR and ADARB1 genes, individual stressful life events, and possible interaction among these factors that might help to differentiate between psychiatric patients who attempted suicide and those who did not. The study included 353 patients diagnosed with major depressive disorder, bipolar affective disorder and schizophrenia, of which 165 attempted suicide, while 188 did not. Childhood abuse and acute stressful life events were evaluated using the Early Trauma Inventory Self-Report questionnaire and the List of Threatening Experiences Questionnaire, respectively. Pearson’s 2–test and logistic regression were performed to test potential individual association of various domains of child abuse, acute stressful events, 23 ADAR and ADARB1 tag variants, HTR2C variant rs6318 and TPH2 variants rs7305115 and rs4290270 with attempted suicide. Generalized linear models and backward selection were applied to tes