To investigate the regulatory role and molecular mechanisms of TSPAN9-mediated mitocytosis in an interleukin-1β (IL-1β)-induced rat chondrocyte senescence model, and to identify novel therapeutic targets for osteoarthritis (OA). Primary knee articular chondrocytes were isolated from the cartilage of knee joints harvested from 1-week-old male Sprague-Dawley rats and maintained in culture. Cell viability was assessed using the cell counting kit 8 (CCK-8) assay, and cell-cycle distribution was analyzed by flow cytometry to determine the optimal concentration and exposure time of IL-1β for model induction, thereby establishing an in vitro chondrocyte senescence model [early-OA (E-OA), middle-OA (M-OA), late-OA (L-OA)] and grouped. Cellular senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining. Real-time quantitative PCR (qRT-PCR) was used to quantify the mRNA expression levels of senescence markers [cyclin-dependent kinase inhibitor 2A (Cdkn2a) and Cdkn1a], extracellular matrix (ECM) catabolic-anabolic genes [collagen type Ⅱ alpha 1 chain (Col2a1), Aggrecan (Acan), matrix metalloproteinase 13 (MMP-13), and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5)], and key mitocytosis-related genes [kinesin family member 5B (KIF5B), TSPAN9, and TSPAN4]. Mitochondrial ultrastructure and mitocytosis-related morphological features were examined by transmission electron microscopy, and changes in mitochondrial membrane potential were assessed using the JC-1 fluorescent probe. To activate TSPAN9 expression, chondrocytes were transduced with a TSPAN9-overexpressing lentivirus (LV-TSPAN9). The experiments included four groups: control group, M-OA group, lentivirus negative control group, and LV-TSPAN9 group. After confirming overexpression efficiency, differences in cellular senescence, mitochondrial homeostasis, and key ECM metabolic indices were compared among the groups. Using CCK-8 assays and cell-cycle analysis, the staged rat chondrocyte senescence model induced by IL-1β (5 ng/mL), and defined 12, 24, and 48 hours as the E-OA, M-OA, and L-OA senescence phenotypes were successfully established, respectively. Model validation demonstrated that, with prolonged induction, the proportion of SA-β-gal-positive cells increased markedly, mitochondrial membrane potential progressively declined ( P<0.05), and mitochondrial ultrastructural damage became increasingly severe. qRT-PCR showed progressive upregulation of the senescence markers Cdkn2a and Cdkn1a, as well as the ECM catabolic genes MMP-13 and ADAMTS5 ( P<0.05), together with sustained downregulation of the ECM anabolic genes Col2a1 and Acan ( P<0.05). Notably, the key mitocytosis-related genes KIF5B, TSPAN4, and TSPAN9 displayed a biphasic pattern, with early compensatory upregulation followed by decompensatory decline at the middle-to-late stages. In the M-OA model, TSPAN9 overexpression markedly reversed these pathological changes, restored mitochondrial membrane potential ( P<0.05), ameliorated the senescent phenotype and ECM metabolic imbalance, and significantly upregulated the expression of mitocytosis-related genes and proteins ( P<0.05), with transmission electron microscope revealing morphological structures suggestive of mitocytosis-related events. Impaired mitocytosis is an important pathological mechanism underlying IL-1β-induced chondrocyte senescence, and TSPAN9 overexpression may delay chondrocyte senescence by promoting mitocytosis in concert with KIF5B, thereby facilitating the clearance of damaged mitochondria and restoring intracellular homeostasis.

Osteogenesis imperfecta (OI) is a clinically heterogeneous genetic disorder whose primary clinical manifestations include susceptibility to recurrent pathological fractures and progressive skeletal deformities. In clinical practice, cases are observed where the proband’s parents show no overt signs of the disease despite having multiple affected children. This may suggest gonadal mosaicism – a condition in which the causative variant is present only in a subset of parent’s germ cells.Gonadal mosaicism remains an understudied inheritance mechanism in monogenic diseases, complicating genetic counseling and reproductive risk assessment. In OI, this phenomenon may account for sporadic cases or recurrent births of affected children to clinically healthy parents.This article presents a case report of a family in which the proband and his younger sister were diagnosed with COL1A1 -associated OI, while the parents and other children showed no disease manifestations. Based on clinical and genetic data, the likelihood of gonadal mosaicism in one parent is discussed, along with considerations for differential diagnosis, patient management strategies, and family genetic counseling.This case highlights the importance of molecular genetic testing not only for the probands but also for their parents to clarify the inheritance mechanism and predict the risks of disease recurrence in the family.

Cartilage defects are difficult to heal because articular cartilage is both avascular and aneural. Directing adipose-derived mesenchymal stem cells (ADSCs) toward a chondrogenic fate in vitro is regarded as a promising therapeutic strategy. However, the endogenous pathways that specify chondrogenic fate remain incompletely defined. Herein, we identify cholesterol-25-hydroxylase (CH25H) as a key inducer of early chondrogenic differentiation that upregulates the cartilage marker genes SOX9, ACAN, and COL2A1. In ADSCs, FPR1 knockdown and rescue experiments, together with pharmacological inhibition of FPR1 or AKT, supported a signalling model in which 25-HC engages FPR1 to promote AKT/GSK3β pathway activation and chondrogenic differentiation. Surface plasmon resonance assays show that 25-HC directly binds to FPR1 with a dissociation constant of 1.18 µM. Molecular docking and membrane MD/MM/GBSA analyses provided structural context, suggesting stable accommodation of 25-HC in an FPR1 pocket and nominating candidate contacts (including a F81/V105 hydrophobic clamp) for future mutagenesis. In a rat trochlear cartilage-defect model, intra-defect implantation of ADSC-laden hydrogel combined with weekly intra-articular 25-HC dosing was associated with improved repair features, while co-administration of Cyclosporin H/HCH6-1 reduced these benefits. Together, these findings delineate the CH25H/25-HC-FPR1 axis as an endogenous metabolite-GPCR signalling pathway that couples cholesterol metabolism to chondrogenic fate specification in ADSCs.

Objectives: The objective of the study was to investigate the mRNA expression of critical gene patterns, including IL-6, CCL2, IL-10, MAPK1, MAPK8, MMP9, COL1A1, COL3A1, and TGFB1, and their associations with adverse postoperative outcomes in reconstructive and plastic surgery patients, depending on age. Methods: A total of 95 women participated in this prospective longitudinal cohort study and underwent reconstructive/plastic surgery. The mean age was 35.48 ± 6.61 years (range: 19-57). mRNA expression of IL-6, CCL2, IL-10, MAPK1, MAPK8, MMP9, COL1A1, COL3A1, and TGFB1 genes was evaluated in peripheral blood leukocytes using a PCR-based method with reverse transcription of cDNA. Results: The risk of postoperative complications significantly increased with elevated expression levels of IL-6 and COL3A1 (7.5-fold, p = 0.007), CCL2 (6.2-fold, p = 0.012), and MAPK1 (25.5-fold, p < 0.001). Increased expression of MAPK8, IL-10, and MMP9 was associated with a 13.2-fold higher risk (p < 0.001). The strongest association was observed for COL1A1 overexpression, which increased complication risk by 58.33-fold (p < 0.001). Risk stratification using the Molecular Complication Risk Index (MCRI), incorporating weighted gene contributions, identified an unfavorable molecular profile predominantly among women aged ≥ 40 years. Receiver operating characteristic analysis confirmed the model's discriminative ability (AUC = 0.78; 95% CI 0.68-0.87), with an optimal cut-off value of MCRI ≥ 8.5 (sensitivity 76%, specificity 71%, p < 0.05). Conclusions: The transcriptional activity of IL-6, CCL2, IL-10, MAPK1, MAPK8, MMP9, COL1A1, COL3A1, and TGFB1 is associated with postoperative wound healing risk. Women aged over 40 years are at the highest risk of complications. Implementation of the MCRI model may enable early identification of high-risk patients, support targeted preventive strategies, and improve personalized surgical planning.

Alport syndrome is a hereditary nephropathy caused by pathogenic variants in the COL4A3, COL4A4, or COL4A5 genes, encoding type IV collagen chains that are essential for glomerular basement membrane integrity. The missense pathogenic variant p.Gly407Arg in COL4A3 has been reported in Portuguese families, but detailed phenotypic characterization remains limited. This study aimed to describe the renal and extra-renal manifestations and identify predictors of renal outcomes in a cohort of patients carrying the COL4A3 p.Gly407Arg variant. We conducted a retrospective observational study including 62 patients from 25 families followed at a tertiary nephrology center in northern Portugal between 2007 and 2025 with genetically confirmed Autosomal Dominant Alport syndrome due to monoallelic COL4A3 p.Gly407Arg pathogenic variant. Demographic, clinical, laboratory, audiologic, and ophthalmologic data were analyzed. Renal events were defined as incident kidney failure, estimated Glomerular Filtration Rate (eGFR) <15 mL/min/1.73 m2, or eGFR ≥30% decline from baseline sustained over time. Changes in eGFR over time were analyzed using a univariate linear mixed effects model. At diagnosis, 98% of patients presented with microscopic hematuria, and 53% had proteinuria (urinary protein-creatinine ratio (UPCR) 240mg/g, IQR 9-840). The mean eGFR was 94 ± 32 mL/min/1.73m2. Mean follow-up time in the study (from the initiation of clinical observation until the occurrence of the event or the end of follow-up) was 4.72 years (IQR 2.43-8.43). During a median follow-up of 4.72 years, 16 patients (26%) developed renal events, with 5 (8%) initiating KRT at a mean age of 58 (range 45-66) years. The mean renal survival was 67 years (63-72). Proteinuria ≥500mg/g was significantly associated with faster eGFR decline (p < 0.001) and a higher risk of renal events (imputed as a time-varying variable, HR 1.03, 95% CI 1.001-1.059, p= 0.04). Hearing loss occurred in 34% of patients and ocular changes in 18% of the patients evaluated. Patients carrying monoallelic COL4A3 p.Gly407Arg pathogenic variant exhibit variable phenotypic expression, with proteinuria representing the strongest predictor of renal function decline.

Umbilical cord-derived mesenchymal stem cells restore renal homeostasis in Alport Syndrome: Mechanistic insights and clinical translation.

Generation of an induced pluripotent stem cell and isogenic control line from a vascular Ehlers-Danlos Syndrome (vEDS) patient harboring a pathogenic c.755G>T in the COL3A1 gene.

Exosome-like nanovesicles (ELNs) are extracellular vesicles (50-200 nm) with a bilayer membrane derived from endosomes. They contain nucleic acids, proteins, lipids and secondary metabolites, and function as mediators of intercellular communication. ELNs isolated from plants and mushrooms have been reported to exhibit diverse biological activities. ELN-enriched vesicle fraction derived from Sparassis crispa was extracted using low-temperature pressure squeeze pre-treatment and tangential flow filtration system. The ethyl acetate portion was purified using multiple chromatography techniques to yield three compounds. Their structures were determined by spectroscopic analysis and comparison with reported data. The effects of the compounds on type I collagen gene (COL1A1) expression were evaluated in CCD-1064Sk human fibroblasts. Molecular docking was conducted to assess their interactions with collagenases (MMP-1, MMP-8 and MMP-13). Three compounds were isolated from ELN-enriched vesicle fraction derived from S. crispa and identified as sparalide C (1), 5-hydroxy-7-methoxyphthalide (2) and methyl orsellinate (3). All compounds increased COL1A1 gene expression in a dose-dependent manner. These compounds exhibited binding energies in the range of -5.8 to -7.0 kcal/mol against three collagenases. This study identified three bioactive compounds, sparalide C (1), 5-hydroxy-7-methoxyphthalide (2) and methyl orsellinate (3), from ELN-enriched vesicle fraction derived from S. crispa. All isolated compounds enhanced COL1A1 gene expression in CCD-1064Sk human fibroblasts, suggesting their potential to promote collagen biosynthesis. Molecular docking analysis further supported their relevance by indicating moderate binding affinities to collagenase enzymes (MMP-1, MMP-8 and MMP-13), which are known to degrade type I collagen. These findings suggest that ELN-enriched vesicle fraction derived from S. crispa may serve as a promising natural source for skin health applications, particularly in maintaining extracellular matrix homeostasis.

Alport syndrome (AS) is a prevalent inherited kidney disorder mainly caused by mutations in COL4A3, COL4A4, and COL4A5 genes. To elucidate the genetic variants of AS, we implemented a sequential sequencing strategy within a Chinese cohort of 555 patients, comprising whole-exome sequencing (WES) for all participants, followed by whole-genome sequencing (WGS), RNA sequencing (RNA-seq), and nanopore long-read sequencing (NLR-seq) for selected individuals. We identify 431 distinct variants in 509 (91.7%) patients, with 42.2% being novel. Beyond WES, additional sequencing approaches resolve 23 patients with noncoding, copy number, or structural variants. Notably, noncoding variants account for 16.2% of detected variants and exhibit ethnic-specific mutagenesis patterns. More interestingly, NLR-seq uncovers two novel types of structural variants, namely large insertions in intronic regions and complex duplication-inversion variants. This study provides deeper insights into the genetic architecture of AS and proposes a research paradigm for improving the genetic diagnosis of inherited diseases.

Type II, IX and XI collagenopathies encompass Stickler syndrome and a spectrum of related connective tissue disorders with diverse and overlapping phenotypes. This study evaluated outcomes of commercial gene panels to understand how genetic testing was used in these collagenopathies. A retrospective review was undertaken of genetic tests including genes COL2A1, COL11A1, COL11A2, COL9A1, COL9A2 and COL9A3 from a clinical diagnostic laboratory. Cases harbouring pathogenic/likely pathogenic (P/LP) variants were categorised by the ordering panel. Indications for testing were classified into ocular, orofacial, auditory, musculoskeletal, cardiovascular or other symptoms/signs. Between February 2020 and May 2024, 7798 diagnostic panels were ordered containing the six collagen genes of interest, with 214 unique cases reporting at least one P/LP variant. Overall, Stickler syndrome was the main indication for testing in 48% of cases. Family history and ocular signs were the most common indications in Stickler syndrome genetic testing, while musculoskeletal and neurological/developmental signs more frequently prompted testing with other panel types. The diagnostic yield of Stickler syndrome panels was 50%, with a higher rate among cases reporting a family history (OR: 2.6 (95% CI 1.3 to 5.4); p=0.005) or presenting with ocular signs (OR: 2.2 (95% CI 1.1 to 4.5), p=0.03). Stickler syndrome is the primary indication for half of the genetic tests ordered for type II, IX and XI collagenopathies, yet accurate diagnosis remains challenging due to overlapping collagenopathy phenotypes. Diagnosis and management could be enhanced through comprehensive documentation of multisystem signs, identification of syndrome-specific features and multidisciplinary care approaches.

"Alport spectrum disorder" describes a phenotypically and genotypically multifaceted disease entity encompassing classic autosomal recessive and X-linked Alport syndrome (AS) but also more heterogenous and typically milder, yet not benign, hematuric phenotypes like autosomal dominant AS, formerly also known as thin basement membrane nephropathy (TBMN). Alport spectrum disorder is associated with disease-causing variants in the type IV collagen genes COL4A3, COL4A4 and COL4A5. Variants and genotypes, reported by a genetic diagnostics lab between 2009 and 2014, of 91 index cases with the clinical tentative diagnosis of AS (66/91), TBMN (21/91), or AS/TBMN (not specified further; 4/91), were reassessed based on 2015 ACMG (American College of Medical Genetics and Genomics) criteria and current amendments. 80 different variants, all originally reported as "mutations", and their genotypes have been reassessed (COL4A3: 21/80, COL4A4: 15/80, COL4A5: 44/80). In 10/80 variants, classification changed from disease-causing to variant of uncertain significance (VUS). 69/91 (76%) index cases included in the analysis could be classified as solved. 22/91 (24%) index cases had an ambiguous result either on variant, genotype, or both variant and genotype level. VUS cases had a significantly more limited phenotype (e.g., isolated microscopic hematuria) compared to non-downgraded cases (e.g., additional extrarenal manifestations). Reassessment of variants/genotypes in this study showed a significant reduction in unequivocal genetic diagnoses highlighting variant and genotype interpretation as a dynamic process. Genetic reports of individuals with suspected Alport spectrum disorder, especially those obtained in the pre-ACMG criteria era, should therefore be critically evaluated.

Skeletal dysplasias comprise a heterogeneous group of genetic disorders characterized by abnormalities in bone and cartilage development, with wide clinical variability in terms of severity and prognosis. Advances in obstetric ultrasonography have enabled the prenatal suspicion of these conditions during pregnancy. Among these disorders, osteogenesis imperfecta stands out as a hereditary connective tissue disease frequently associated with mutations in the genes responsible for type I collagen synthesis, particularly COL1A1 and COL1A2. This study reports a case of prenatal suspicion of non-lethal skeletal dysplasia with postnatal molecular confirmation of osteogenesis imperfecta associated with the COL1A2 gene. The patient was a primigravida without significant comorbidities who received regular prenatal care. Obstetric ultrasonographic examinations performed from the second trimester revealed marked shortening of the long bones in a micromelic pattern, bowing of the limbs, suspected intrauterine fractures, and reduced mineralization of the cranial vault. Bilateral clubfoot and fetal growth restriction were also observed, while thoracic proportions remained preserved, suggesting the absence of ultrasonographic criteria for lethality. Delivery was performed by cesarean section due to breech presentation. The newborn required resuscitation at birth and was subsequently admitted to the Neonatal Intensive Care Unit. Physical examination revealed limb shortening and dysmorphic facial features, maintaining the clinical suspicion of skeletal dysplasia. Diagnostic investigation was complemented by a next-generation sequencing panel for skeletal dysplasias, which identified a pathogenic heterozygous variant in the COL1A2 gene, confirming the diagnosis of osteogenesis imperfecta. This case highlights the importance of detailed prenatal ultrasonographic evaluation for the early recognition of skeletal dysplasias and underscores the role of molecular testing in establishing the etiological diagnosis and guiding perinatal care planning.

Background: Sports injuries can be an important health problem that is difficult to compensate for athletes. Anterior cruciate ligament injury (ACLI) is a soft tissue injury frequently faced by both athletes and sedentary individuals. In early studies on ACLI, it was reported that these injuries were mostly hormonal, biomechanical and genetic. However, the inconsistencies in these studies are still not fully explained. It is also known that differences in the protein levels of collagen structures play a critical role in non-contact soft tissue injuries. In this sense, it is thought that gene variants may cause sensitivity on ACLI by affecting collagen tissues.ObjectiveThe aim of the study is to systematically examine the relationship between ACLI and genes. In addition, the current study aims to reveal that some genes that are frequently studied in the literature may be related to the structure of the ACL.MethodsStudies on the subject were examined in PubMed and PubMed-Central databases registered with The National Center for Biotechnology Information. Preferred Reporting Items for Systematic Reviews and Meta-Analyses were used in data selection and evaluation.Results11 articles that met the criteria were subjected to bias analysis. It was reported that 7 studies were at a critical level in terms of overall risk. Additionally, statistically significant differences were reported in 6 of the 11 studies.ConclusionThe genes COL1A1, COL5A1, MMP3, TGFBR3, TGFBI, VEGFA, β-fibrinogen and COL12A1 can protect individuals against ACLI by affecting the formation of soft tissues.

ObjectiveTo investigate the clinical application value of exome sequencing technology in fetuses with sonographically detected skeletal dysplasia.MethodsA total of 80 pregnant women whose fetuses were diagnosed with sonographically detected skeletal dysplasia at 11⁺0 to 38⁺0 weeks of gestation in our hospital from March 2021 to January 2022 were enrolled in this study. G-banded chromosomal karyotype analysis (KA), chromosomal microarray analysis (CMA) and whole exome sequencing (WES) were simultaneously performed for all cases. Clinical data including maternal age, gestational week, prenatal ultrasound findings, results of KA, CMA and WES, and pregnancy outcomes were collected and subjected to statistical analysis.ResultsLimb shortening was the most common phenotypic manifestation of fetal skeletal dysplasia. The diagnostic yield of KA combined with CMA for fetal skeletal dysplasia was only 7.5% (6/80), while the additional combination with WES identified 43 positive cases, with an overall diagnostic yield of 53.75% (43/80). There was a statistically significant difference in the diagnostic yield between the two detection protocols (χ2=50.19, P<0.001), with an absolute increase of 46.25% in the diagnostic yield after the integration of WES. The most common clinical disorders caused by pathogenic genes included osteogenesis imperfecta, achondroplasia and thanatophoric dysplasia. The majority of pathogenic variants were de novo mutations, mainly involving the COL1A1, FGFR2 and FGFR3 genes. Among the 43 WES-positive cases, 34 pregnancies opted for termination of pregnancy.ConclusionWhole-exome sequencing technology holds important application value in the prenatal diagnosis of fetal skeletal system diseases, and provides a more accurate evidence base for genetic counseling and clinical decision-making.

This study investigates the potential therapeutic effects of low-intensity extracorporeal shock wave therapy (Li- ESWT) in the treatment of chronic kidney disease (CKD) using a preclinical mice model, which was established by high adenine diet. Four experimental groups were formed: normal (NL), NL+Li-ESWT (NL+ESWT), CKD, and CKD+Li-ESWT (CKD+ESWT), and received appropriate interventions over a period of 3 weeks. Immunohistochemistry analysis revealed significant upregulation of HSP70 expression, a marker for cells affected by mechanical stimulus, in the NL+ESWT group, confirming successful delivery of Li-ESWT to kidney tissues. Body weight, kidney weight, blood urea nitrogen (BUN), and creatinine values showed no significant differences between the NL and NL+ESWT groups, indicating that Li-ESWT alone did not affect these parameters. However, in the CKD+ESWT group, although kidney weight did not significantly differ from the CKD group, both body weight and renal function parameters showed marked improvement compared to the CKD group, suggesting a positive impact of Li-ESWT on general condition and renal function in CKD. The following histological examination showed that the CKD+ESWT group exhibited significantly reduced tubulointerstitial injury and fibrosis compared to the CKD group, further supporting the therapeutic potential of Li-ESWT in CKD management. To reveal potential modulation of ESWT on the upstream of fibrosis, fibrosis-related gene expressions, i.e., Acta2, Col1a1, Col3a1, and Fn1, were examined. The results demonstrated a significant reduction in the expression of fibrosis-related genes in the CKD+ESWT group compared to the CKD group, highlighting the molecular effects of Li-ESWT in mitigating CKDassociated fibrosis.

Background and ObjectivesCongenital myopathies (CMYOs) and congenital muscular dystrophies (CMDs) are rare, clinically and genetically heterogeneous neuromuscular conditions characterized by muscle weakness, usually with onset at birth or in the first few months of life. Next-generation sequencing (NGS) has significantly enhanced diagnostic capabilities and transformed the diagnostic process for such rare conditions. The aim of this study was to describe the outcomes of NGS analysis and genotypic prevalence among patients with CMYO and CMD referred for diagnostic assessment to the National Highly Specialized Service (HSS) at the Dubowitz Neuromuscular Centre in London, United Kingdom, over a period of 10 years.MethodsDiagnostic outcomes of all referrals to the HSS for NGS analysis of CMYO and/or CMD gene panels from 2014 to 2023 were included and reviewed.ResultsA total of 1,927 patients were referred to the service, and a total of 2,352 genetic analyses were completed over 10 years. Overall, 553 of 1,927 unrelated individuals (28.7%) had a genetic diagnosis of CMYO or CMD, due to pathogenic variant/s in one of 59 genes. A total of 345 patients had a diagnosis of CMYO and 208 had CMD. The most common CMYOs were due to pathogenic variants in RYR1 (23.8%), TTN (10.7%), MTM1 (10.4%), NEB (8.7%), SELENON (7.5%), ACTA1 (6.7%), or DNM2 (4.6%) genes. Pathogenic changes in further 27 CMYO genes were also identified. The most common CMDs were due to pathogenic variants in COL6A1 (20.7%), LAMA2 (15.4%), COL6A2 (13.5%), COL6A3 (7.2%), GMPPB (6.7%), POMGnT1 (6.3%), FKRP (6.3%), or LMNA (4.8%) genes. Pathogenic changes in further 17 CMD genes were also identified. A total of 1,374 patients remained undiagnosed. Of these, 78 patients (5.7%) carried a heterozygous pathogenic change in a recessive gene and 419 patients (30.5%) carried a variant of unknown significance, with variants in RYR1 (17.1%), NEB (14.2%), and TTN (12.7%) genes being the most frequent.DiscussionThis large, real-world cohort provides a comprehensive overview of the genetic distribution of CMYO and CMD in routine practice. Our findings offer a robust framework to guide diagnostic strategies, inform variant interpretation, and support clinical decision making in the genomic era.

To explore the contribution of genetic risk factors to isolated superior mesenteric artery dissection (ISMAD), this study detected variations in known arterial dissection-related genes in a cohort of patients with symptomatic ISMAD. Thirty-two patients were recruited. Whole-exome sequencing was conducted on blood samples from all participants and 83 controls. Prevalence, clinical characteristics, treatments, and causative gene carriers were also studied. Eleven (34.4%) patients with ISMAD had pathogenic variants in COL3A1 (n = 3), COL12A1 (n = 3), TLN1 (n = 1), FN1 (n = 2), and TNXB (n = 2). Pathogenic variants in the COL3A1 and COL12A1 genes are strongly correlated with ISMAD. In 9.4% (3/32) of the patients with ISMAD, conservative treatment failed and further endovascular treatment was required. Among them, 2 (66.7%) harbored pathogenic mutations in COL3A1. Pathogenic variants in the COL3A1 gene are associated with a high predisposition to endovascular treatment (P = .02). COL3A1, COL12A1, and TLN1 mutations may be risk factors for ISMAD. A small proportion of patients with ISMAD and carrying pathogenic mutations in COL3A1 were more likely to experience more severe arterial tearing and require endovascular treatment.

Alport syndrome (AS) is a hereditary nephropathy caused by mutations in the COL4A3 , COL4A4 , and COL4A5 genes. Rare contiguous COL4A5–COL4A6 alterations cause AS with diffuse leiomyomatosis (AS-DL). Case report. A 16-year-old male had mild proteinuria, hematuria, hearing loss and myopia since childhood. Estimated glomerular filtration rate was 82.8 mL/min/1.73 m 2 . Kidney biopsy showed segmental mesangial sclerosis; immunofluorescence was negative. Electron microscopy demonstrated diffuse glomerular basement membrane thinning and podocyte foot-process effacement. Two deceased brothers had end stage kidney disease; the mother had hematuria, uterine myoma, and a benign bladder tumor. A diagnosis of X-linked AS was established according to the Flinter criteria and nephroprotective treatment was initiated. Since 2024, the patient had complained of epigastric symptoms. An endoscopy revealed a 2-cm gastric submucosal lesion. A whole-exome sequencing identified a hemizygous missense variant in COL4A5 and a COL4A6 exon 1–2 deletion, confirming AS-DL. Discussion. This case demonstrates the co-occurrence of AS-DL and shows that early pedigree assessment, combined with integrated clinicopathologic-genetic evaluation in a multidisciplinary framework, enables a timely diagnosis of atypical familial AS-DL and improves clinical management.

Globally, chronic kidney disease (CKD) affects over 800 million individuals and is characterized by significant genetic complexity. More than 600 genes are associated with hereditary kidney disease, which may manifest as isolated kidney issues or as part of a syndrome that also includes extrarenal manifestations. The aim of this study was to identify genetic variants in a group of ten patients who presented with clinical signs suggestive of genetic syndromes associated with CKD, or who were asymptomatic but had a positive family history of CKD. Extensive genetic testing (targeted gene panels and whole-exome sequencing-WES) identified a mutation in the PKD1 gene in 3 out of 10 cases. In one patient, a known mutation in the PKD2 gene was identified. Another four patients were diagnosed with Alport syndrome: three of these presented with de novo missense mutations in the COL4A5 gene, and one patient had a mutation in the COL4A3 gene. One patient was diagnosed with MODY5, caused by a known mutation in the HNF1B gene, and one patient was diagnosed with Bartter syndrome type 1, resulting from a known mutation in the SLC12A1 gene. We present genotype-phenotype correlations, highlighting the particularities of each patient within their family context. Our findings emphasize the importance of genotype-phenotype correlations in refining diagnosis, personalizing therapeutic management, and providing essential genetic counseling for at-risk relatives.

Collagen is the primary protein in the extracellular matrix of human cells and the body and is essential for cell structure and function. Here, for the first time, we report a method for producing recombinant triple-helical collagen type III (rhCOL3) in transgenic tobacco as a bioreactor. We constructed a pMDV-COL3A1 vector containing the human type III collagen gene COL3A1, as well as a pMDV-COL3A1:5E vector that coexpressed COL3A1 and the enzymes required for its posttranslational modification. These two vectors were used to transform tobacco genetically. The COL3A1 gene was successfully coexpressed in tobacco plants with four enzymes that promote its posttranslational modification. The transcriptional level of COL3A1 in the transgenic lines coexpressing posttranslational modification genes was greater than that in the transgenic lines expressing only COL3A1. The enzyme-modified recombinant collagen was subsequently purified from a COL3A1:5E transgenic line. Our experimental results demonstrated that the terminal propeptides of plant-derived rhCOL3 can be correctly cleaved through the enzymatic hydrolysis of procollagen by coexpressed procollagen C proteinase (PCP) and procollagen N proteinase (PNP). The plant-derived rhCOL3 was thermally stable because the purified peptide chains can form a triple helix structure. Experiments have shown that plant-derived rhCOL3 has biological activity. In this study, functional recombinant full-length mature type III collagen with a triple-helix structure was successfully expressed in tobacco, providing a foundational plant-made material for future applications of collagen in human skin and bone repair in regenerative medicine.