Systemic sclerosis (SSc) is a rare connective tissue disorder with highest morbidity and mortality among rheumatologic diseases. Disease progression is highly heterogeneous between patients, implying a strong need for individualization of therapy. Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133 and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS) model. Association was found between MTHFR rs1801133 and higher risk for elevated systolic pressure in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS rank and elevated systolic pressure. Our results open a door wide for more extensive research on pharmacogenomics markers in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with SSc and help in prevention of adverse drug reactions.

Objectives: This study aims to show the distribution of angiotensin-converting enzyme (ACE) rs1799752 (I>D) gene insertion/deletion (I/D) polymorphism and angiotensin II receptor type 1 (AGTR1) rs5186 (A>C) gene polymorphism in adolescents with hypertension (HT) and type 1 diabetes (T1D), as well as its association with hypertension and the diurnal variation of mean blood pressure (dipping phenomenon). Methods: A cross-sectional study was conducted involving 118 adolescents diagnosed with T1D who underwent clinical and laboratory investigations, genetic analyses, and 24 h ambulatory blood pressure monitoring. The genotype frequencies were compared between adolescents with HT and those with normal blood pressure. Additionally, the genotype frequencies were compared between dippers and non-dippers. Results: Patients with HT were more likely to be female and exhibited significantly poorer glycemic control and higher triglycerides, along with increased body mass index and daily insulin dosage. The prevalence of ACE rs1799752 genotypes in the hypertensive group was 20% II, 66.7% ID, and 13.3% DD, which did not significantly differ from the normal blood pressure group with 29.1% II, 53.4% ID, and 17.5% DD (p = 0.625). The prevalence of AGTR1 rs5186 genotypes in the hypertensive group was 53.3% AC, 40% AA, and 6.7% CC, which also did not significantly differ from the normal blood pressure group with 39.8% AC, 52.4% AA, and 7.8% CC (p = 0.608). A total of 46% of the patients exhibited non-dipping phenomena. The prevalence of non-dippers among the ACE genotypes was 13% DD, 33.3% II, and 53.7% ID (p = 0.369), while for the AGTR1 genotypes, it was 50% AA, 42.6% AC, and 7.4% CC (p = 0.976). Conclusions: Our results indicate that in our adolescents with T1D, clinical and metabolic factors such as higher body mass index, triglycerides, suboptimal glycemic control, and female gender are more indicative of the development of hypertension than ACE and AGTR1 gene polymorphisms. A potential reason for this finding could be the young age of the patients or the relatively small size of the study group. Future research involving larger sample sizes is needed to further investigate the genetic predisposition for the development of hypertension.

A minority of individuals who consume excessive alcohol develop cirrhosis. Variants in the patatin-like phospholipase domain-containing protein 3 gene (PNPLA3) and the transmembrane 6 superfamily member 2 gene (TM6SF2) have been previously identified as associated with alcohol-related cirrhosis (ALC). This study aimed to examine the variants of PNPLA3 and TM6SF2 and to develop and assess a polygenic risk score (PRS) for ALC. We enrolled 118 patients diagnosed with ALC and 131 control subjects, who were either abstainers or low-level alcohol consumers without evidence of liver disease. Genotyping of risk variants was performed using PCR-RFLP methodology. PRS, based on independent allelic effect size estimates from genotyped genetic loci, were computed and compared across groups. The development of ALC was significantly associated with CG and GG genotypes of PNPLA3 (CG: OR: 1.82; 95% CI: 1.05-3.17; p=0.033; GG: OR: 7.64; 95% CI: 3.06-19.07; p<0.001) and the CT genotype of TM6SF2 (OR: 2.43; 95% CI: 1.27-4.63; p=0.007), controlling for age and sex. Patients with cirrhosis exhibited a significantly higher mean PRS compared to controls (0.32 vs. 0.167, p = 1.8e-07). The odds ratios (ORs) and 95% confidence intervals for the group with the highest PRS score compared to the reference group were 6.707; 95% CI: 3.313-13.581, p<0.001. In our ALC patient cohort, the PNPLA3 rs738409 and TM6SF2 rs58542926 variants were associated with an increased risk of ALC development. Moreover, the PRS derived from these two variants effectively identified the genetic components linked to cirrhosis within the study population.

Background: Pharmacogenomic variation contributes to interindividual differences in drug response, yet population-specific pharmacogenomic profiles remain insufficiently characterized in the Western Balkans (WB). This study aimed to comprehensively characterize pharmacogenomic variability in the WB populations using whole-genome sequencing (WGS), assess population differences in pharmacogenomic profiles, evaluate concordance among star allele calling tools, and identify rare and previously unreported variants in pharmacogenes. Methods: WGS data from 250 healthy individuals from five WB populations were analyzed across 88 pharmacogenes. Star alleles were assigned using Aldy4, PharmCAT, PyPGx, and Stargazer. Principal component analysis (PCA) was used to assess population structure, and pharmacogenomic allele and phenotype frequencies were compared within five European subpopulations. Unreported pharmacovariants were annotated using Ensembl VEP, gnomAD, ClinVar, and ClinPGx, with functional impact assessed using multiple in silico prediction tools. Results: A total of 357 star alleles were identified, with high tool concordance except for complex genes such as CYP2D6. WB populations clustered within the European genetic space, with limited subpopulation differentiation. Significant differences were observed within WB and compared with European populations, including clinically relevant genes CYP2B6, CYP2C19, RYR1, and VKORC1. Among 33 clinically relevant pharmacogenes, 22,638 SNVs were identified, including six previously unreported variants. Conclusions: The WB populations show substantial pharmacogenomic similarity to other European populations, while also exhibiting clinically relevant population-specific differences. Comprehensive WGS-based profiling revealed both established and previously unreported pharmacogenomic variation, highlighting the importance of including underrepresented populations in precision medicine research.

Interindividual variability in the efficacy and toxicity of 6-mercaptopurine (6-MP) and methotrexate (MTX) drugs remains a major challenge in the treatment of pediatric acute lymphoblastic leukemia (ALL). Germline variation in pharmacogenes involved in drug metabolism, transport, and folate pathways may contribute to this variability. In this retrospective cohort study, 43 pediatric patients with ALL treated at a tertiary referral center in Croatia according to ALL IC-BFM 2002 or 2009 protocols were genotyped for eleven variants in TPMT, ITPA, NUDT15, PACSIN2, MTHFR, TYMS, SLC19A1, and SLCO1B1 genes. Associations with average 6-MP dose during maintenance therapy, MTX pharmacokinetics during consolidation and MTX-related toxicity were analyzed. A polygenic risk score (PRS) was constructed to assess cumulative genetic risk of developing toxicity when administering these two drugs. Carriers of the ITPA rs1127354 variant required significantly lower average 6-MP doses compared with wild-type carriers. The NUDT15 rs61973267 variant was associated with higher tolerated 6-MP doses after exclusion of outliers. No significant associations were observed between individual variants and MTX clearance. A trend toward reduced risk of oral mucositis was observed in carriers of the SLCO1B1 rs4149056 variant. PRS analyses that included carriers of the TPMT rs1142345 and ITPA rs1127354 variants showed a borderline association with the requirement for 6-MP dose reduction. Our findings support a role for ITPA and NUDT15 variants in modulating 6-MP dose requirements, while single-variant associations with MTX pharmacokinetics and toxicity were limited and not supported. These results underscore the complexity of antimetabolite pharmacogenetics and suggest that integrative polygenic approaches may better capture clinically relevant pharmacogenetic risk.

Introduction/Objective. Alcoholic liver cirrhosis (ALC) is the primary cause of alcohol abuse-related mortality, resulting from alcohol-induced oxidative stress. Glutathione S-transferases (GSTM1, GSTT1) are enzymes essential for detoxification, and their deficiency may contribute to the onset of chronic inflammation and disease progression. This study sought to investigate the relationship between GSTM1 and GSTT1 deletions and ALC, as well as alcohol consumption patterns in cirrhosis development. Methods. The analysis included 114 ALC patients and 262 controls, with GSTM1 and GSTT1 deletions assessed via multiplex PCR. Results. Findings indicated that individuals with the GSTM1 null genotype had a three-fold increased risk of developing ALC (95% CI, 1.87–4.81; p < 0.0001), whereas GSTT1 null genotypes showed no significant impact. Individuals with both GSTM1 null and GSTT1 null genotypes exhibited an 11-fold heightened risk of ALC (OR = 11.21, 95% CI = 3.30–38.14, p < 0.001). Furthermore, patients who commenced alcohol consumption at 22.5 years or older developed cirrhosis more rapidly than their younger counterparts (p < 0.001). Conclusion. GSTM1 null and combined GSTM1/GSTT1 null genotypes constitute significant risk factors for ALC, with patients who started drinking at an older age experiencing accelerated disease progression irrespective of alcohol intake levels.