Balkan Journal of Medical Genetics最新文献

Microcephaly, Epilepsy, and Diabetes Syndrome 1 (MEDS1) is a very rare autosomal recessive neurodevelopmental disorder (OMIM#614231) characterized by the triad microcephaly with simplified gyration, neonatal permanent diabetes and infantile epileptic encephalopathy. Its occurrence is due to biallelic mutations in the IER3IP1 gene, which encodes for the Immediate Early Response 3 Interacting Protein 1 (IER3IP1). To date, only eleven cases have been reported worldwide. Here, we describe the twelfth case from northern Morocco. This 4-month-old patient, born to a healthy non-consanguineous couple presented with microcephaly, epileptic seizures and insulin-requiring permanent neonatal diabetes. Brain MRI revealed simplified gyration with partial agenesis of the corpus callosum. Targeted next generation sequencing identified the patient as compound heterozygous for two IER3IP1 variants; a novel nonsense mutation, p. Leu79Ter, and a previously reported pathogenic variant, p. Val21Gly. This report extends the phenotypic and genotypic spectrum of MEDS1 syndrome and provides further evidence of the role of IER3IP1 in neural and pancreatic development, emphasizing the clinical importance of genetic screening in cases of early-onset diabetes and epilepsy especially when accompanied by significant head growth failure in a newborn infant. Further research into the pathophysiology of IER3IP1 may potentially lead to new therapeutic approaches.

Introduction: A group of primary bone tumors called chondrosarcomas are diverse and characterized by neoplastic tissue of hyaline cartilage. They are the second most typical primary osseous cancer. Recurrence is indicative of a poor prognosis because traditional chondrosarcomas are resistant to treatment. Our knowledge of the pathobiology of conventional chondrosarcomas has greatly expanded owing to recent findings in these tumors' biology, genetics, and epigenetics studies. These findings also provide information on possible treatment targets. CHEK2 encodes a checkpoint kinase involved in DNA damage response and cell cycle regulation, and it is an important cancer susceptibility gene.

Methods: The Gene2 drug and DSEA assisted with in silico screening. The antitumor activities of the candidate drugs were extracted from DepMap via the PRISM viability assay on eight chondrosarcoma cell lines.

Results and conclusion: Advances in studies show promise for discovering potential targeted treatments for cancer. Bisacodyl is a targeted G protein-coupled receptor 35. G protein-coupled receptors are widely known targets for cancer treatment. Here, we showed that bisacodyl can be a potential therapeutic agent in chondrosarcoma cell lines.

To characterize the prevalence of CYP2B6 and OPRM1 gene polymorphisms and evaluate their associations with serum methadone concentrations and dose requirements in Vietnamese patients undergoing methadone maintenance treatment (MMT). This cross-sectional study enrolled 200 patients with opioid dependence from multiple MMT clinics in Ninh Binh, Vietnam, between October 2023 and March 2024. Demographic, clinical, and behavioral data were collected using structured questionnaires. Blood samples were obtained for genotyping CYP2B6 (rs2279343 [785A>G], rs3745274 [516G>T], rs8192709 [64C>T]) and OPRM1 (A118G, rs1799971) using Sanger sequencing. Trough serum methadone concentrations were measured by validated liquid chromatography-tandem mass spectrometry (LC-MS/MS). Multivariable linear regression was performed to identify clinical and genetic factors associated with serum methadone concentrations, the concentration-to-dose ratio (CDR), and maintenance methadone dose. Among 200 participants (99% male, 96.9% Kinh ethnicity), the most common CYP2B6 diplotypes were *1/*4 (33.3%) and *4/*6 (23.7%), with the OPRM1 AG genotype present in 47.5%. Most patients (53.5%) had been on methadone treatment for over 3 years. In multivariable regression, the *1/*6 genotype was independently associated with lower maintenance dose (β = -26.33, 95% CI: -51.97, -0.69). The *2/*6 genotype was significantly associated with lower log serum methadone concentration (β = -2.485, 95% CI: -3.024, -1.947) and lower log CDR (β = -2.595, 95% CI: -3.145, -2.046). Most demographic, behavioral, and OPRM1 genotypes showed no significant associations. Specific CYP2B6 genotypes and SNPs significantly influence methadone pharmacokinetics in Vietnamese patients receiving MMT, although actual daily dose remains the most important determinant of serum levels. These findings highlight the potential role of genotype-guided methadone dosing in optimizing therapy for opioid dependence in Vietnamese populations.

A thorough functional evaluation of the immune system requires a method capable of measuring multiple cytokines to assess the adaptive immune response, as well as immunoglobulin isotypes and seroconversion to evaluate the humoral immune response following infections and vaccinations. Flow Cytometry Multiplex Bead Array (FCMBA) technology has been shown to be highly effective in detecting and quantifying numerous analytes simultaneously in a single reaction well. This technique is increasingly being used in immune profiling for cytokines in various disease contexts, as well as in adoptive cell transfer, immunotherapies, and more recently, in the characterization of humoral responses to newly developed COVID-19 vaccinations. FCMBA technology has shown advantages in assessing the depth of the humoral response mounted against COVID-19 vaccinations, compared to post-COVID infection status, and screening for the suitability of sera for use in COVID-19 convalescent plasma therapy. In this article, we present an overview of the FCMBA technology concept and its versatility, highlighting its ability to map many essential analytes and its superiority over conventional Enzyme Linked Immunosorbent Assay (ELISA) techniques. Furthermore, we discuss the current and potential clinical applications of this technology in deciphering COVID-19-triggered humoral immune responses, whether after contracting the infection or post-vaccination.

Pregnancy loss (PL), particularly early pregnancy loss (EPL), is a prevalent reproductive complication, with approximately 15% of confirmed pregnancies affected. Chromosomal abnormalities are implicated in more than half of EPLs, with trisomies being the most prevalent. Partial abnormalities, including segmental deletions, duplications, and unbalanced translocations, are detected in up to 10% of EPL cases. This study focuses on the precise characterization of partial chromosomal abnormalities, previously identified by Quantitative fluorescent polymerase chain reaction (QF-PCR) and multiplex ligation probe amplification (MLPA) analyses. By employing an array comparative genomic hybridization (aCGH), we analyzed 20 EPL samples, identifying 32 partial chromosomal abnormalities, including 18 deletions and 14 duplications, with an average size of 33.2 Mb. Notably, two abnormalities previously undetected by QF-PCR and MLPA were revealed (deletions in 7q36, and 1p36.32p36.31regions), emphasizing the necessity of high-resolution genomic tools. Chromosomes 1, 18, and 13 emerged as frequently involved, aligning with previous associations with recurrent pregnancy loss. Recurrent abnormalities were identified in six chromosomal regions, with chromosome 1p36.33-p36.32 exhibiting the highest frequency. Gene Ontology (GO) enrichment analysis of recurrent regions highlighted disruptions in critical biological processes, including molecular binding, enzymatic activity, and cellular development. Many genes in these regions are linked to multisystem syndromes, suggesting their involvement in early embryonic development and pregnancy viability. Our findings underscore the complexity of EPL's genetic landscape, demonstrating that large CNVs, may disrupt multiple genes critical for development. Although, subtelo-meric MLPA reliably detects telomeric partial chromosomal abnormalities in EPLs, aCGH is essential for detection and precise characterization of all CNVs, thus enhancing diagnostic and counseling strategies in affected couples.

The complex cytokine network plays an important role in disease susceptibility and development, therefore single-nucleotide polymorphisms (SNPs) in or near cytokine genes may be relevant to development of systemic sclerosis (SSc). We in this study investigated 22 SNPs in 13 cytokine genes of SSc patients, and their association with disease susceptibility. Twenty-three clinically diagnosed SSc patients were enrolled for this purpose along with 80 healthy volunteers for comparisons. Aseptically collected 2ml of peripheral venous blood from each subject was processed for DNA extraction. Cytokine genotyping was carried out using the extracted genomic DNA by PCR employing sequence-specific primers and data was analyzed for any association with SSc susceptibility. Variations in allele, genotype, or haplotype distribution between patients and healthy volunteers were observed for the following SNPs: IL-1β -511 C/T (rs16944) and +3962 T/C (rs1143634); IL-4Rα +1902 G/A (rs1801275); IL-12 -1188 C/A (rs3212227); TGF-β1 codon 25 G/C (rs1800471); TNF-α-308 G/A (rs1800629) and -238 G/A (rs361525); IL-4 -1098 T/G (rs2243248) and -590 T/C (rs2243250); IL-6 -174 G/C (rs1800795) and nt565 G/A (rs1800797); and IL-10 -1082 G/A (rs1800896), -819 C/T (rs1800871) and -592 C/A (rs1800872). However, only the SNPs in IL-1β -511 and +3962, and TNF-α -308 and -238 were found to be significantly associated with SSc susceptibility. Our findings suggest that IL-1β and TNF-α gene SNPs may play a role in development of SSc, although large observational and experimental studies are needed to substantiate these findings.

Vesicoureteral reflux (VUR) is an important factor in the etiology of recurrent urinary tract infections (UTIs). Permanent kidney damage may develop in children with high-grade VUR in the long term. This damage may progress with the development of scar tissue in some patients. The TLR4 gene is an important resistance mechanism, especially against UTIs. TLR4 gene polymorphism is associated with recurrent UTIs and kidney scar development in the long term. This study aimed to examine the relationship between scar development and TLR4 gene polymorphism in children with VUR. This cross-sectional study included 49 patients with recurrent UTIs and primary vesicoureteral reflux. Patients were divided into two groups (26 patients with the scar, and 23 patients without scar) according to the presence of scar tissue. TLR4 gene polymorphisms of the patients were evaluated by Next Generation Sequencing. The TLR4 gene polymorphism was significantly higher in the compound heterozygous group with scarring than in the group without scarring (p=0.03). Gene polymorphisms, c.958T>C, c.942A>G, c.776A>G, c.1076C>T, c.896AT, c.1078C>T were presented more commonly in the group with scarring. Moreover, gene polymorphisms c.942A>G and c.776A>G were defined for the first time in this study among patients with scar tissue. The higher incidence of some TLR4 gene polymorphisms in patients with scarring suggested that these variations might cause permanent kidney damage. In addition to genetic predisposition, environmental factors such as untreated UTIs might also contribute to scar formation.

Clopidogrel, a P2Y12 receptor antagonist, is widely used to prevent cardiovascular events, but significant variability in its efficacy persists among patients. AKR1D1, involved in bile acid synthesis and regulation of CYP enzymes, may contribute to this variability. This study aims to investigate whether clopidogrel and its inactive metabolite, 2-oxoclopidogrel, interact with AKR1D1 at the enzymatic or transcriptional level. Enzymatic activity assays demonstrated that neither clopidogrel nor 2-oxoclopidogrel acts as a substrate or inhibitor of AKR1D1. Expression studies in HepG2 cells further revealed no significant changes in AKR1D1 mRNA levels following treatment with these compounds. These findings indicate that clopidogrel does not directly influence AKR1D1's metabolic functions, including bile acid synthesis, steroid hormone clearance, or the production of 5β-reduced steroids, which regulate CYP enzyme expression. From a physiological perspective, the absence of interaction minimizes the risk of adverse effects on CYP-mediated drug metabolism, nutrient absorption, lipid digestion, and the absorption of lipophilic drugs. Future research should explore AKR1D1's broader substrate specificity, particularly focusing on non-steroidal compounds, and investigate the clinical implications of AKR1D1 polymorphisms in clopidogrel-treated patients to enhance personalized therapeutic strategies.

Although the identification of disease subtypes conveying prognostic significance along with minimal residual disease (MRD) assessment represent cornerstones for stratification in childhood acute lymphoblastic leukemia (ALL), approximately half of the relapses occur in patients from standard-risk groups. Identification of the drivers of treatment failure is crucial for detection of high-risk clones at diagnosis. We evaluated clinical variables and the most common genetic alterations in an unselected cohort of 55 patients with B-ALL treated according to the ALL-IC-BFM 2002 protocol, with a median follow-up of 46 months. Matched diagnosis-relapse samples underwent screening for additional alterations using whole-exome sequencing. Mutations in the CREBBP gene were found in 80% (4/5) of the patients with relapse, either present from the disease onset or acquired at relapse, while none of the examined patients in remission presented alterations in this gene. Deletions in TP53 and EBF1 (present in 2/5 and 1/5 of the patients with relapse, respectively) were infrequent or absent in the patients in remission, respectively. Screening for alterations in the CREBBP gene at diagnosis and/or at multiple time-points during chemotherapy could be incorporated into treatment protocols, as it may contribute to the identification of significant number of patients with predefined or acquired chemoresistant clones.

Optical genome mapping (OGM) is a novel method enabling the detection of structural genomic variants. The method is based on the laser image acquisition of single, labeled, high-molecular-weight DNA molecules and can detect structural genomic variants such as translocations, inversions, insertions, deletions, duplications, and complex structural rearrangements. We aim to present our experience with OGM at the Clinical Institute of Genomic Medicine, University Medical Centre Ljubljana, Slovenia. Since its introduction in 2021, we have used OGM for the testing of facioscapulohumeral muscular dystrophy 1, characterization and resolution of variants identified by other technologies such as microarrays, exome and genome next-generation sequencing, karyotyping, as well as testing of rare disease patients in whom no genetic cause could be identified using these methods. We present an example family case of two previously undiagnosed male siblings with an overlapping clinical presentation of thrombocytopenia, obesity, and presacral teratoma. After karyotyping, microarray analysis and next-generation sequencing, by using OGM, a maternally inherited cryptic translocation t(X;18)(q27.1;q12.2) was identified in both brothers. Despite an extended segregation analysis, based on strictly applied ACMG criteria and ClinGen guidelines, the identified translocation remains a variant of unknown significance. Despite the remaining limitations of OGM, which will hopefully be resolved by improvements in databases of known benign SV variation and the establishment of official guidelines on the clinical interpretation of OGM variants, our work highlights the complexity of the diagnostic journey, including this novel method, in rare disease cases.