Primary immunodeficiency diseases (PIDs) are monogenic inborn immune disorders. Of currently listed 555 genetic inborn errors of immunity (IEI) in 10 IUIS categories, PIDs range from life-threatening severe combined immunodeficiency (SCID) to milder antibody deficiencies. PIDs increase mortality risk and demand genetic/immunological testing. In Iran, high consanguinity rates may warrant influence of genetic factors. AQ1 Diagnostic challenges due to heterogeneity necessitate application of advanced tools like whole exome sequencing (WES). This study was carried out on Eastern Iran’s PIDs and similar approaches should be employed worldwide in populations that are highly consanguineous. This study focused on 99 patients from Eastern Iran clinically diagnosed with PIDs who were referred for genetic evaluations between 2016 and 2025. Genetic analyses involved DNA extraction from blood samples, WES withusing Illumina platforms, and in-house bioinformatic pipelines for variant identification and classification. Sanger sequencing and co-segregation studies further validated findings. Medical records, family histories, and pedigrees were thoroughly analyzed. Also we have delineated syndromic and non-syndromic PID disorders. The significant findings were as follows: Finding of 47 novel diseasecausing variants; High consanguinity rates (76%) correlated with an 82.8% diagnostic yield and a mortality rate of 8% amongst patients. SCID-associated mutations, as the most common discovered PIDs, found in 16 cases; other common disorders were AR LRBA LOF, AR ATM LOF, and AR EPG5 (possibly hypomorphic LOF). Additionally, certain mutations may link to pregnancy loss which may need further functional studies. Around 70% of unresolved cases were syndromic. The most frequently suspected pathogenic variants (n = 5), all novel and in homozygosity with matching phenotype, were noted in EPG5 and linked to Vici syndrome. In highly consanguineous populations, WES (which does miss some 5% of cases only verifiable by WGS, especially in primer sets are not the newest ones in WES) may reach a definite or highly probable genetic diagnosis in over 80% of cases, and that some IEIs may be associated with pregnancy loss. Methods: nrCFs were treated with angiotensin II, ascorbic acid, and dextran sulfate. Key features of cardiac fibrosis were evaluated using Alizarin Red, Picrosirius Red, Masson\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\'s trichrome staining, quantitative reverse transcription polymerase chain reaction, and immunocytochemistry and scratch assay. Results: Although dextran sulfate increased the expression of alpha-smooth muscle actin (α-SMA), it did not increase collagen deposition and cell migration. Therefore, it seems that the combination of 500 nM angiotensin II with 100 μM ascorbic acid would be effective in induction of cardiac fibrosis in vitro, which increased (1) the expression of collagen, α-SMA, and vimentin at protein level, (2) the expression of collagen type I alpha 1 chain, collagen type III alpha 1 chain, matrix metalloproteinase 2, and transforming growth factor-beta 1 at ribonucleic acid level, and (3) cell proliferation and migration. Conclusion: In this study, 72 hour-treatment of nrCFs with 500 nM angiotensin II and 100 μM ascorbic acid was effective in the creation of an in vitro model of cardiac fibrosis. It is hoped that this model will be useful for screening antifibrotic treatments.

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