This report presents a case of pontine autosomal dominant microangiopathy with leukoencephalopathy (PADMAL) in a 35 year-old male patient. The patient exhibited a consistent history of recurrent ischemic strokes, concentrated primarily in the pons region, accompanied by concurrent manifestations of leukoencephalopathy and microbleeds. Genetic evaluation revealed a heterozygous missense mutation consistent with c.3431C>G, p. Thr1144Arg substitution within exon 40 of the COL4A1 gene. This mutation was also identified in the patient\'s mother, affirming an autosomal dominant inheritance model. Our findings serve as testament to the potential role of mutation in the exon 40 of COL4A1 in the pathogenesis and progression of PADMAL, contributing to ongoing efforts aimed at better understanding the genetic basis of this debilitating disorder.

UNASSIGNED: Inflammatory bowel disease (IBD) and periodontitis (PD) are correlated, although the pathogenic mechanism behind their correlation has not been clarified. This study aims to explore the common signature genes and potential therapeutic targets of IBD and PD using transcriptomic analysis.
UNASSIGNED: The GEO database was used to download datasets of IBD and PD, and differential expression analysis was used to identify DEGs. We then conducted GO and KEGG enrichment analyses of the shared genes. Next, we applied 4 machine learning (ML) algorithms (GLM, RF, GBM, and SVM) to select the best prediction model for diagnosing the disease and obtained the hub genes of IBD and PD. The diagnostic value of the signature genes was verified by a validation set and qRT‒PCR experiments. Subsequently, immune cell infiltration in IBD samples and PD samples was analyzed by ssGSEA. Finally, we investigated and validated the response of hub genes to infliximab therapy.
UNASSIGNED: We identified 43 upregulated genes as shared genes by intersecting the DEGs of IBD and PD. Functional enrichment analysis suggested that the shared genes were closely associated with immunity and inflammation. The ML algorithm and qRT‒PCR results indicated that IGKC and COL4A1 were the hub genes with the most diagnostic value for IBD and PD. Subsequently, through immune infiltration analysis, CD4 T cells, NK cells and neutrophils were identified to play crucial roles in the pathogenesis of IBD and PD. Finally, through in vivo and in vitro experiments, we found that IGKC and COL4A1 were significantly downregulated during the treatment of patients with IBD using infliximab.
UNASSIGNED: We investigated the potential association between IBD and PD using transcriptomic analysis. The IGKC and COL4A1 genes were identified as characteristic genes and novel intervention targets for these two diseases. Infliximab may be used to treat or prevent IBD and PD.

OBJECTIVE: To evaluate the regulation of the aberrant expression of collagen type IV alpha 1 chain (COL4A1) in the development of age-related cataract (ARC).
METHODS: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot analysis were employed to evaluate the expression of COL4A1 in ARC patients and healthy controls. The proliferation, apoptosis, cell cycle and epithelial-mesenchymal transition (EMT) of human lens epithelial cell (HLE-B3) were further analyzed under the condition of COL4A1 gene silence. Alteration of gene expression at mRNA level after knockdown COL4A1 were also evaluated by qRT-PCR on HLE-B3 cells.
RESULTS: The aberrant expression of COL4A1 was identified a clinically associated with the ARC. Silencing of COL4A1 promoted the apoptosis and inhibited the proliferation of HLE-B3 by blocking the cell cycle. Moreover, COL4A1 gene silence didn\'t affect the cytoskeleton of HLE-B3 but down-regulated the Collagen type IV Alpha 2 Chain (COL4A2), paired box 6 (PAX6), procollagen-lysine 2-oxoglutarate 5-dioxygenases 1 (PLOD1) and procollagen-lysine 2-oxoglutarate 5-dioxygenases 2 (PLOD2) expression levels in HLE-B3 cells. Silencing the COL4A1 gene induced EMT of the HLE-B3 cells by promoting the transforming growth factor beta (TGF-β) expression.
CONCLUSIONS: Silencing of COL4A1 induces S-phase arrest, also inhibits the proliferation and enhance HLE-B3 apoptosis and EMT, and down-regulates the expression of COL4A2, PAX6, PLOD1 and PLOD2. Thus, the expression alteration of COL4A1 may play a critical role in the pathogenesis of ARC.

Background: The abnormal expression of genes in serum may be associated with early diagnosis of patients with malignant tumors. This study was designed to screen for significantly differentially expressed genes (DEGs) that may be associated with gastric cancer using bioinformatic methods. Methods: RNA-seq data from gastric cancers were downloaded from the TCGA and GEO databases, and 1903 secretory genes were downloaded from the HPA database. The diagnostic secretory RNAs of gastric cancer were screened using least absolute shrinkage and selection operator regression analysis. Univariate Cox regression analysis was used to evaluate the prognostic significance of the results. Biological functions were performed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Then, 640 cases of gastric cancer and paracancerous tissues were collected, and immunohistochemistry (IHC) was used to detect the expression of COL4A1. Results: In total, 25 upregulated differentially expressed genes (DEGs) were identified, which were secreted mainly in the blood and cell matrices. Six secretory genes (OLFM4, CEMIP, APOC1, CST1, COL4A1, and CD55) with diagnostic significance were identified, and the enrichment scores of these six genes were significantly associated with tumor stage. In addition, we found that increased COL4A1 expression might be associated with a poor prognosis in patients with gastric cancer. Based on GO and KEGG analyses, we found COL4A1-related DEGs were mainly enriched in connective tissue development, collagen fibrous tissue-related processes, extracellular structure, extracellular matrix (ECM) tissue, and related to the ECM receptor-related pathway, focal adhesion, and PI3K-Akt signaling pathway. Moreover, the results of immunohistochemical analyses showed that the COL4A1 protein level in gastric cancers was also higher than in the matched paracancerous tissues. Conclusions: In this study, we found six upregulated secretory genes, including OLFM4, CEMIP, APOC1, CST1, COL4A1, and CD55 which we hypothesized to be significant DEGs for the diagnosis of gastric cancer. Our data also suggest that COL4A1 may play an important role in the diagnosis and prognosis of gastric cancer.

Here, we screened the COL4A1 variants in Chinese intracerebral hemorrhage (ICH) patients to summarize the relationship between the variants and clinical characteristics. Targeted sequencing of a 65-gene panel including COL4A1 was performed to detect all the coding regions and ±10-bp splicing sites. In total, 568 patients were included. Regarding rare nonsynonymous variants with a minor allele frequency (MAF) <0.5%, 6 missense variants and five suspicious splice site variants, absent in 573 healthy controls, were found in 11 patients. The subgroup carrying rare variants did not show specific phenotype compared with non-variant carriers. For the single nucleotide polymorphism (SNP) loci with an MAF> 5%, we did not find a significant association between the allele or genotype distribution of the SNP loci and the risk of ICH. Rs3742207 was nominally associated with death at 1-year follow-up (p = 0.02027, OR 1.857, 95% CI 1.101-3.133) after adjusted by age, hypertension history, hematoma volume and recurrent ICH history. Nevertheless, after the Bonferroni correction, the association was no longer significant. In conclusion, rare nonsynonymous variants in COL4A1 were identified in 1.94% (11/568) of Chinese ICH patients, while rs3742207 maybe indicate a worse prognosis of ICH.

Gastric cancer (GC), which features high prevalence and mortality rate, remains the third most lethal cancer worldwide. The paper was designed to explore the impacts of collagen type IV alpha 1 (COL4A1) on GC, along with its potential mechanism. The mRNA and protein expressions of COL4A1 in GC cells were assessed using RT-qPCR and Western blot. After depleting COL4A1, RT-qPCR and Western blot were conducted again to check the transfection efficacy. With the application of CCK-8, wound healing and transwell, the capabilities of cells to proliferate, migrate and invade were appraised, respectively. Moreover, Western blot tested the protein levels of factors involved in migration, proliferation, epithelial-mesenchymal transition (EMT) and Hedgehog signaling. As a result, COL4A1 displayed elevated expression in GC tissues and cells, while its knockdown inhibited the cell viability, migration, invasion and EMT in GC. According to Gene Set Enrichment Analysis (GSEA), COL4A1 was involved in the regulation of Hedgehog signaling pathway, which was then further verified by the detection of Hedgehog-related proteins. To figure out the relationship between COL4A1 and Hedgehog signaling pathway, we used purmorphamine, an agonist of Hedgehog, to treat GC cells, finding that COL4A1 blocked Hedgehog signaling to inhibit the aggressive phenotypes of GC cells. In short, COL4A1 silence was testified to exhibit suppressive effects on the malignant process of GC, suggesting that COL4A1 might be a potent hallmark of GC therapy.

Forkhead box L2 (FOXL2), one member in the superfamily of forkhead transcription factors, is a core transcription factor specifically expressed in ovarian granulosa cells and is essential for the development of follicles. FOXL2 has been shown to regulate the transcription of genes encoding enzymes that synthesize steroid hormones and estrogen receptors and regulate the expression of collagen genes in granulosa cells. This study explored the effect of FOXL2 on collagen gene expression in granulosa cells by overexpressing Foxl2 in pregranulosa cells, prehierarchical follicles and preovulation follicle granulosa cells. The results showed that FOXL2 regulated the expression of several genes encoding collagens in chicken granulosa cells and that overexpression of Foxl2 significantly reduced the messenger RNA and protein levels of Col4a1 in different granulosa cells. Moreover, luciferase reporter and chromatin immunoprecipitation assays were performed to study how FOXL2 regulates the expression of collagen genes, and the results showed that FOXL2 directly regulated the expression of Col4a1 by binding to the motif of CAGCAGCACCAGCAG between -640 and -625 bp upstream of the coding region. The results indicated that FOXL2 could regulate the components of the extracellular matrix; however, the biological significance of this regulation needs further clarification.

Abdominal aortic aneurysm (AAA) is a potentially fatal vascular disease, and the dysregulated circular RNAs (circRNAs) play key roles in AAA progression. Circ_0092291 was downregulated in AAA patients, but its function in AAA remains unclear. This research was performed for the functional analysis of circ_0092291 and its mechanism exploration with mircoRNA-626 (miR-626) and collagen type IV alpha1 chain (COL4A1) in AAA. Human aortic vascular smooth muscle cells (T/G HA-VSMC) were treated with angiotensin II (Ang II). Levels of circ_0092291, miR-626, and COL4A1 were determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Inflammatory cytokines were examined by enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was measured using caspase3 activity assay and flow cytometry. Angiopoiesis was assessed via tube formation assay. The protein analysis was conducted by western blot. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and RNA pull-down assays were used to validate the molecular binding. Circ_0092291 downregulation was found in AAA samples and Ang II-treated cells. Inflammatory response and cell apoptosis were reduced while angiopoiesis and ECM level were facilitated after overexpression of circ_0092291 in Ang II-treated T/G HA-VSMC cells. MiR-626 was a miRNA target for circ_0092291, and miR-626 inhibition protected T/G HA-VSMC from Ang II-induced cell injury. Moreover, the regulation of circ_0092291 was achieved by serving as a miR-626 sponge in Ang II-treated cells. COL4A1 was affirmed as a target for miR-626 and circ_0092291 resulted in the level change of COL4A1 by sponging miR-626. Additionally, miR-626 downregulation inhibited the cell damages caused by Ang II through increasing the level of COL4A1 and the function of circ_0092291 was attributed to the upregulation of COL4A1. The evidence indicated that circ_0092291 could suppress the Ang II-induced cell dysfunction by targeting the miR-626/COL4A1 signaling axis. Circ_0092291 might improve the diagnosis and treatment of AAA.Key Points.Biological mechanism, Apoptosis, Molecular target.

Circular RNA hsa_circ_0088364 (circ_0088364) is a contributory factor in the malignancy of hepatocellular carcinoma (HCC). We aimed to elaborate its role and competing endogenous RNA (ceRNA) mechanism in HCC cell growth and motility. Expression of circ_0088364, microRNA (miR)-1270 and Collagen type IV alpha 1 chain (COL4A1) was measured by real-time quantitative PCR and Western blotting, and their relationships were determined by dual-luciferase reporter assay, RNA immunoprecipitation, biotinylated RNA pull-down, and Spearman\'s rank correlation analysis. Cellular programs were measured by cell counting kit-8 assay, flow cytometry and transwell assays, Western blotting, and xenograft experiment. Expression of circ_0088364 and COL4A1 was upregulated, and miR-1270 was downregulated in HCC patients\' tumors; moreover, there were linear correlations among circ_0088364, miR-1270, and COL4A1 expression. Essentially, circ_0088364 and COL4A1 were ceRNAs for miR-1270 via target binding. In function, silencing circ_0088364 or upregulating miR-1270 could suppress cell proliferation, cell cycle entrance, transwell migration and invasion in Huh7 and HCCLM3 cells, as well as promote apoptosis rate. Moreover, above-mentioned effects were accompanied with reduced B-cell lymphoma (Bcl)-2, N-cadherin and Vimentin levels, and elevated Bcl-2-associated X protein (Bax) and E-cadherin levels. Contrarily, exhausting miR-1270 and restoring COL4A1 could severally abrogate the tumor-suppressive roles of circ_0088364 knockdown and miR-1270 overexpression in HCC cells in vitro. In vivo, silencing circ_0088364 retarded xenograft tumor growth in nude mice induced by Huh7 cells by upregulating miR-1270 and downregulating COL4A1. Blocking circ_0088364 suppressed HCC by inhibiting cell growth and motility via targeting miR-1270-COL4A1 axis.

Pontine autosomal dominant microangiopathy and leukoencephalopathy (PADMAL) is a rare hereditary cerebral small vessel disease. We report a novel collagen type IV alpha 1 (COL4A1) gene mutation in a Chinese family with PADMAL. The index case was followed up for 6 years. Neuroimaging, whole-exome sequencing, skin biopsy, and pedigree analysis were performed. She initially presented with minor head injury at age 38. MRI brain showed chronic lacunar infarcts in the pons, left thalamus, and right centrum semiovale. Extensive workup was unremarkable except for a patent foramen ovale (PFO). Despite anticoagulation, PFO closure, and antiplatelet therapy, the patient had recurrent lacunar infarcts in the pons and deep white matter, as well as subcortical microhemorrhages. Whole-exome sequencing demonstrated a novel c.*34G > T mutation in the 3\' untranslated region of COL4A1 gene. Skin biopsy subsequently demonstrated thickening of vascular basement membrane, proliferation of endothelial cells, and stenosis of vascular lumen. Three additional family members had gene testing and 2 of them were found to have the same heterozygous mutation. Of the 18 individuals in the pedigree of 3 generations, 12 had clinical and MRI evidence of PADMAL. The mechanisms of both ischemic and hemorrhagic stroke are likely the overexpression of COLT4A1 in the basement membrane and frugality of the vessel walls. Our findings suggest that the novel c.*34G > T mutation appears to have the same functional consequences as the previously reported COL4A1 gene mutations in patients with PADMAL and multi-infarct dementia of Swedish type.