ITGB3 Research Articles

GDNF Promotes Astrocyte Abnormal Proliferation and Migration Through the GFRα1/RET/MAPK/pCREB/LOXL2 Signaling Axis.

Glial cell-line derived neurotrophic factor (GDNF) is a powerful astroglioma (AG) proliferation and migration factor that is highly expressed in AG cells derived from astrocytes. However, it is still unclear whether high levels of GDNF promote AG occurrence or if they are secondary to AG formation. We previously reported that high concentrations of GDNF (200 and 500ng/mL) can inhibit DNA damage-induced rat primary astrocytes (RA) apoptosis, suggesting that high concentrations of GDNF may be involved in the malignant transformation of astrocytes to AG cells. Here we show that 200ng/mL GDNF significantly increased the proliferation and migration ability of RA cells and human primary astrocytes (HA). This treatment also induced RA cells to highly express Pgf, Itgb2, Ibsp, Loxl2, Lif, Cxcl10, Serpine1, and other genes that enhance AG proliferation and migration. LOXL2 is an important AG occurrence and development promotion factor and was highly expressed in AG tissues and cells. High concentrations of GDNF promote LOXL2 expression and secretion in RA cells through GDNF family receptor alpha-1(GFRα1)/rearranged during transfection proto-oncogene (RET)/mitogen-activated protein kinase (MAPK)/phosphorylated cyclic AMP response element binding protein (pCREB) signaling. GDNF-induced LOXL2 significantly promotes RA and HA cell proliferation and migration, and increases the expression of Ccl2, Gbp5, MMP11, TNN, and other genes that regulate the extracellular microenvironment in RA cells. Our results demonstrate that high concentrations of GDNF activate LOXL2 expression and secretion via the GFRα1/RET/MAPK/pCREB signal axis, which leads to remodeling of the astrocyte extracellular microenvironment through molecules such as Ccl2, Gbp5, MMP11, TNN. This ultimately results in abnormal astrocyte proliferation and migration. Collectively, these findings suggest that high GDNF concentrations may promote the malignant transformation of astrocytes to AG cells.

Preclinical safety and efficacy of lentiviral-mediated gene therapy for leukocyte adhesion deficiency type I

Leukocyte adhesion deficiency type I (LAD-I) is a primary immunodeficiency caused by mutations in the ITGB2 gene, which encodes for the CD18 subunit of β2-integrins. Deficient expression of β2-integrins results in impaired neutrophil migration in response to bacterial and fungal infections. Using a lentiviral vector (LV) that mediates a preferential myeloid expression of human CD18 (Chim.hCD18-LV), we first demonstrated that gene therapy efficiently corrected the phenotype of mice with severe LAD-I. Next, we investigated if the ectopic hCD18 expression modified the phenotypic characteristics of human healthy donor hematopoietic stem cells and their progeny. Significantly, transduction of healthy CD34+ cells with the Chim.hCD18-LV did not modify the membrane expression of CD18 nor the adhesion of physiological ligands to transduced cells. Additionally, we observed that the repopulating properties of healthy CD34+ cells were preserved following transduction with the Chim.hCD18-LV, and that a safe polyclonal repopulation pattern was observed in transplanted immunodeficient NOD scid gamma (NSG) mice. In a final set of experiments, we demonstrated that transduction of CD34+ cells from a severe LAD-I patient with the Chim.hCD18-LV restores the expression of β2-integrins in these cells. These results offer additional preclinical safety and efficacy evidence supporting the gene therapy of patients with severe LAD-I.

The effects of Juglone-Selenium combination on invasion and metastasis in pancreatic cancer cell lines.

Pancreatic cancer does not show any symptoms in the early period and metastatic process is already passed when the diagnosis is done. Therefore, in the battle with pancreatic cancer, novel treatment strategies, particularly antiinvasive and antimetastatic strategies, are needed. The cytotoxic and anticancer effects of juglone and sodium selenite (NaSe) have been showed in various cancer cells. In this study, it is aimed to investigate the synergistic effects of juglone and selenium on PANC-1 and BxPC-3 pancreatic cancer cells. Antimetastatic effects of juglone-NaSe were carried out by adhesion and invasion assays and the genes and protein expressions. Expression analysis of the CDH1, ITGB3 and COL4A3 genes and their proteins E-cadherin, β3 integrin and tumstatin which play role in metastasis and angiogenesis processes, were done by qPCR and immunohistochemical analysis, respectively. Study findings have provided evidences that the juglone-selenium has a cytotoxic and dose dependent suppressive effect on invasion and metastasis in PANC-1 and BxPC-3 cells. The juglone-NaSe has the potential to be a promising agent especially to inhibit invasion and metastasis in pancreatic cancer treatment. However, more in depth studies are needed to more clearly demonstrate the effects of juglone-selenium.

CRISPR-mediated activation of autism gene Itgb3 restores cortical network excitability via mGluR5 signaling

Many mutations in autism spectrum disorder (ASD) affect a single allele, indicating a key role for gene dosage in ASD susceptibility. Recently, haplo-insufficiency of ITGB3, the gene encoding the extracellular matrix receptor β3 integrin, was associated with ASD. Accordingly, Itgb3 knockout (KO) mice exhibit autism-like phenotypes. The pathophysiological mechanisms of Itgb3 remain, however, unknown, and the potential of targeting this gene for developing ASD therapies uninvestigated. By combining molecular, biochemical, imaging, and pharmacological analyses, we establish that Itgb3 haplo-insufficiency impairs cortical network excitability by promoting extra-synaptic over synaptic signaling of the metabotropic glutamate receptor mGluR5, which is similarly dysregulated in fragile X syndrome, the most frequent monogenic form of ASD. To assess the therapeutic potential of regulating Itgb3 gene dosage, we implemented CRISPR activation and compared its efficacy with that of a pharmacological rescue strategy for fragile X syndrome. Correction of neuronal Itgb3 haplo-insufficiency by CRISPR activation rebalanced network excitability as effectively as blockade of mGluR5 with the selective antagonist MPEP. Our findings reveal an unexpected functional interaction between two ASD genes, thereby validating the pathogenicity of ITGB3 haplo-insufficiency. Further, they pave the way for exploiting CRISPR activation as gene therapy for normalizing gene dosage and network excitability in ASD.

Study the Association of Nucleotide Polymorphisms in Platelet Receptor and Cytochrome P450 Genes with the Development of Resistance to Antiplatelet Drugs in Patients with Coronary Artery Disease

Aim. To study the association of nucleotide polymorphisms in platelet receptor and cytochrome P450 genes with the development of resistance to antiplatelet drugs in CHD patients.Material and Methods. The study included 243 patients diagnosed with CHD after coronary artery bypass surgery (CABG), including 140 patients in the acetylsalicylic acid (ASA) treatment group and 103 patients in the dual antiplatelet therapy (DAT) group. All patients were tested for platelet aggregation using an optical aggregometer with inducers: 5 mM ADP and 1 mM arachidonic acid (AA). DNA samples were analyzed by allele-specific PCR for the presence of polymorphisms rs2046934, rs1126643, rs5918, rs6065, rs4244285 in the platelet receptor and cytochrome P450 genes.Results. No statistically significant differences were found during comparison of the prevalence of the studied polymorphisms in the platelet receptor and cytochrome P450 genes between the groups of aspirin-sensitive and aspirin-resistant patients, as well as between the groups of clopidogrelsensitive and clopidogrel-resistant patients. No association between carriage of the minor and major alleles of the polymorphisms studied and the development of antiplatelet drug resistance was found. In the group of patients on ASA therapy, carriers of the C allele of the T1565C (rs5918) ITGB3 polymorphism had a higher rate of AA-induced platelet aggregation compared to carriers of the T allele (18,49±25,92 vs 10,43±17,34, р=0,004).Conclusion. Polymorphisms of P2RY12 (rs2046934), ITGA2 (rs1126643), ITGB3 (rs5918), GP1BA (rs6065), CYP2C19*2 (rs4244285) genes are not associated with antiplatelet drug resistance in both patients on ASC therapy and on DAT. The presence of minor alleles of the rs2046934, rs1126643, rs6065, rs4244285 polymorphisms are not associated with increased platelet aggregation activity before CABG.However, in the group of patients on ASA therapy C-allele carriers of the rs5918 polymorphism of the ITGB3 gene had a higher rate of AA-induced platelet aggregation compared to T-allele carriers.

Leukocyte adhesion defect: Clinical correlation with integrin expression and genetics

Objectives: Leucocyte adhesion defect 1 (LAD1) is an autosomal recessive defect in integrin expression. In this study we present a case series of 19 children with LAD1. We analysed their clinical and laboratory data and correlated it with integrin expression and underlying genetic defect. Material and Methods: Retrospective case record study of all children diagnosed with LAD1 between 2012 and 2021. Results: Ten females and 9 males were included. Eight cases were born of consanguineous marriage. 78% (15/19) had disease onset within one year of life and 8 of them were neonates. Neonates with LAD1 commonly presented with omphalitis, late-onset sepsis and respiratory infections (RTI). Most common organism isolated from blood was pseudomonas (4/8). CD11/CD18-expression ranged from severely reduced (6/8) to moderately reduced (2/8) but disease was uniformly fatal in the 7/8 patients who could not undergo hematopoietic stem cell transplant (HSCT). Seven children presented between 1month and 1year of age with RTI, bacterial sepsis and soft tissue abscess. Four expired, 1 survived after HSCT and 2 were alive at last follow up. CD11/CD18-expression ranged between 0-1.6% except in 2 cases. One child with 11%-expression is alive without transplant. Another child had 0%-CD11 but 17%-CD18-expression with very low MFI (mean florescence intensity). She succumbed at 20 months of age with perianal abscess, sepsis and RTI but prior to this fatal episode she developed autoimmune cutaneous lesions with lymphocytic infiltration. Four children presented beyond infancy. Median age at onset of symptom was 2 years with skin abscess, oral ulcers, severe periodontitis with tooth loss and pyogenic arthritis. Three of them survived without HSCT, 1 was alive at last follow up. CD11/CD18-expression ranged from 20-32% in these children. Details of cord separation was available in 17 cases and it was delayed (beyond 2 weeks of life) in 9 children. Of the 17 cases with genetic analysis of ITGB2 gene, 16 had a homozygous and 1 had compound heterozygous variant. (nonsense: 2; splice site variants:5; 8: missense variants; 2: deletion). Conclusion: Disease onset less than 1 year age was fatal, irrespective of CD11/CD18 expression. Integrin expression levels beyond 20% and splice site mutation were associated with 100% survival without HSCT but were not event free. Pseudomonas sepsis is the commonest infection in LAD1 neonates. Between 1month to 1year, RTI and gram-negative sepsis dominated and beyond 1st-year skin/perianal abscess was common. Delayed separation of cord was present only in 52% of cases.

Identification of the causative mutation in the ITGB2 gene in a LAD1 patient by whole exome sequencing

Leukocyte adhesion deficiency type 1 (LAD1) is a rare congenital immunodeficiency disease. The cause of disease is determined to be the mutations in the ITGB2 gene that codes for CD18, the beta chain of beta-2 integrins, leads to decreased expression or functioning of CD18. This deficiency leads to severe impairment of leukocyte adhesion to the vascular wall and leukocyte migration to sites of infection and inflammation. LAD1 has also been associated with inhibition of interleukin-23 and interleukin-17 resulting in a hyperinflammatory and chronic inflammation. Patients with LAD1 typically present in early infancy with recurrent, life threatening infections that are frequently fatal before 2 years of age without hematopoietic stem cell transplant (HSCT). However, LAD1 is difficult to diagnose and many LAD1 patients die at a young age despite intensive antibiotic therapy. Accurate diagnosis requires detailed clinical information (delayed umbilical cord loss, severe periodontitis, delayed wound healing and sores, skin abscesses, and recurrent infection), and confirmation the absence of integrins by flow cytometric analysis. A better understanding of the molecular characteristics of this disease is needed to raise awareness and definitive diagnosis infants with LAD1. To definitive diagnosis, whole exome sequencing and Sanger sequencing were performed in an eighteen-month-old boy with severe leukocytosis, recurrent infections, delayed wound healing, and hepatosplenomegaly associated with an acquired cytomegalovirus infection. Two variants: One previously reported mutation (c.533C>T, p.Pro178Leu) and one novel variant (c.59-1G>A), in the ITGB2 gene were detected. These results can be used for definitive genetic diagnosis, genetic counseling, as well as a prenatal diagnosis in LAD1 patients.

Identification of differentially expressed long noncoding RNAs in the ovarian tissue of Shal and Sangsari ewes using RNA-seq.

BackgroundThe ovary has an important role in reproductive function. Animal reproduction is dominated by numerous coding genes and noncoding elements. Although long noncoding RNAs (LncRNAs) are important in biological activity, little is known about their role in the ovary and fertility.MethodsThree adult Shal ewes and three adult Sangsari ewes were used in this investigation. LncRNAs in ovarian tissue from two breeds were identified using bioinformatics analyses, and then target genes of LncRNAs were discovered. Target genes were annotated using the DAVID database, and their interactions were examined using the STRING database and Cytoscape software. The expression levels of seven LncRNAs with their target genes were assessed by real‐time PCR to confirm the RNA‐seq.ResultsAmong all the identified LncRNAs, 124 LncRNAs were detected with different expression levels between the two breeds (FDR < 0.05). According to the DAVID database, target genes were discovered to be engaged in one biological process, one cellular component, and 21 KEGG pathways (FDR < 0.05). The PES1, RPS9, EF‐1, Plectin, SURF6, CYC1, PRKACA MAPK1, ITGB2 and BRD2 genes were some of the most crucial target genes (hub genes) in the ovary.ConclusionThese results could pave the way for future efforts to address sheep prolificacy barriers.

CRESTONE: Initial efficacy and safety of seribantumab in solid tumors harboring NRG1 fusions.

3006 Background: NRG1 fusions are rare oncogenic drivers found in ̃0.2% of all solid tumors. These fusions elicit ERBB3/HER3 overactivation to drive tumor growth and cancer cell survival. Currently there are no approved targeted therapies for NRG1 fusion-positive tumors. Furthermore, patients (pts) with tumors harboring NRG1 fusions have poor outcomes with standard therapies. Seribantumab is a fully human anti-HER3 IgG2 monoclonal antibody that suppressed tumor growth in NRG1 fusion-driven preclinical models. Here, we present initial clinical data from the CRESTONE study (NCT04383210). Methods: CRESTONE is a Phase 2, global, multicenter, open-label study of seribantumab in adult pts with locally advanced or metastatic solid tumors harboring NRG1 fusions. A dose ranging phase established the RP2D as a 3g once weekly (QW) intravenous dose administered until treatment discontinuation criteria are met. In the expansion phase, cohort 1 will enroll at least 55 pts who had received at least one prior therapy and are naïve to ERBB-targeted therapy. Exploratory cohorts 2 or 3 will enroll pts previously treated with ERBB-targeted therapies and/or tumors harboring additional molecular alterations. The primary endpoint is objective response rate (ORR) by independent central review per RECIST v1.1. Initial data from cohort 1 pts who received seribantumab 3g QW with investigator (INV)-assessed response per RECIST v1.1 are reported. Results: By JAN-13-2022, 12 pts have received seribantumab 3g QW in cohort 1. Median age was 65 years (range 44–76), 67% were female, and median number of prior therapies was 1 (range 1–5). 92% (11/12) of pts had non-small cell lung cancer (NSCLC); 5 different NRG1 fusion partners ( ATP1B1, CD74, ITG B1, SDC4, SLC3 A2) were reported by local next-generation sequencing tests. Among 10 pts evaluable for INV-assessed response, the confirmed ORR was 30%, and the disease control rate was 90% (1 complete response, 2 partial responses, 6 stable disease, 1 progressive disease). 58% (7/12) of pts remain on study treatment, including 2 pts with NSCLC who achieved objective responses with an ongoing duration of response of 6 and 8.5 months. Seribantumab 3g QW was well tolerated with no drug discontinuations or dose reductions. Across all cohorts (n = 29), the most frequently (≥20%) reported treatment-related adverse events (TRAEs) were diarrhea (38%), fatigue (34%), and rash (24%), all were grade 1 or 2. One grade 3 TRAE of vomiting occurred; there were no Grade 4 or 5 TRAEs. Efficacy analysis is ongoing and updated efficacy data from evaluable pts in cohort 1 will be presented. Conclusions: Initial data indicate seribantumab induced durable responses in advanced solid tumors harboring NRG1 fusions and has a favorable safety profile. These data support the continued evaluation of seribantumab in NRG1 fusion-positive solid tumors in the ongoing CRESTONE study. Clinical trial information: NCT04383210.

Potential Role of LYN, CCL2, ITGB3 and IL6 Genes in the Immune Response of Porcine Buccal Mucosa Cells

Abstract The genes considered in this study, namely, LYN, CCL2, ITGB3 and IL6 can be related to immune response in porcine buccal mucosa cells primary cultured in vitro. These genes are also responsible for, inter alia, cell migration, differentiation, proliferation and apoptosis, intracellular signal transduction, extracellular matrix binding and wound healing. A total of 20 pubertal crossbred Landrace gilts bred on commercial farms were used to obtain buccal mucosa cell cultures, which were harvested on the 7th, 15th and 30th day after initiation of the culture. Expression levels of LYN, CCL2, ITGB3 and IL6 were evaluated employing Real-Time Quantitative Polymerase Chain Reaction. All studied genes showed expression. The expression of CCL2 on day 15 was the highest of all factors measured. The greatest difference between the measurements occurred in gene IL6, between 7th and 15th day, while the least difference between the measurements occurred in gene LYN, between 7th and 15th day. Moreover, on the 7th day, LYN presented the lowest expression among all studied genes. Although in vitro conditions are much more controlled than in vivo conditions, all the factors that may positively or negatively affect cultured cells still cannot be fully predicted. Nevertheless, LYN, CCL2, ITGB3 and IL6 are a valuable starting point for studying further immunological processes in oral mucosal epithelial cells. Given their high regenerative potential, research into them is a highly valuable source of information for future wound healing therapies, where immunological processes should be carefully considered.

Mechanism of ketotifen fumarate inhibiting renal calcium oxalate stone formation in SD rats

ObjectivesTo investigate the inhibitory effect of ketotifen fumarate (KFA), a mast cell membrane stabilizer, on renal calcium oxalate stone (CaOx) formation and its possible molecular mechanism. MethodsWe used the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database for functional and pathway enrichment analyses of osteopontin (OPN), CD44 and fibronectin (FN). Blood biochemistry, reactive oxygen species ratio (ROS), mast cells, proteins (CD44, OPN and FN) and OPN receptor integrin family genes were detected by ELISA, flow cytometry, immunohistochemistry and RT-QPCR, respectively. ResultsThe crystal area of CaOx in the KFA and Control group was significantly smaller than that in the Model group. The number of activated mast cells, the expression levels of OPN and CD44 in the Control and KFA groups were significantly lower than those in the Model group, and the percentage of ROS in the KFA group was also significantly lower than that in the Model group. The mRNA expression levels of ITGB1, ITGA9, ITGAV and ITGA4 genes in the prominent OPN receptor integrin family increased significantly in the Model group. ConclusionsKetotifen can effectively inhibit the crystal formation of CaOx and reduce the inflammatory response of tissue in SD rats. The mechanism may be to reduce the infiltration and activation of mast cells in renal tissue and down-regulate the expression of OPN, CD44 and FN in renal tubules and renal interstitium. And affect the synthesis of integrins (ITGA9, ITGA4, ITGAV, ITGB1, ITGB3 and ITGB5) and ROS.

Molecular-genetic substantiation of combined therapy in the early postoperative period in diffuse peritonitis

The OBJECTIVE was to give a molecular genetic foundation of combination therapy in the early postoperative period for peritonitis.METHODS AND MATERIALS. 70 patients with diffuse peritonitis were studied: the first (n=35) – patients underwent basic treatment, the second (n=35) – combined treatment with the inclusion of Remaxol and laser irradiation. The severity of endotoxemia and oxidative stress, the state of the hemostasis system and the functional status of the liver were determined. Molecular genotyping of the following genes was performed: beta subunits of platelet fibrinogen receptor (T1565C, ITGB3), fibrinogen (G(-455)A, FGB), catalase (-262C/T, CAT), superoxide dismutase (C47T, SOD2).RESULTS. It was found that the early postoperative period in patients with peritonitis is accompanied by endotoxemia, oxidative stress, impaired functional status of the liver, hypercoagulation and hypofibrinolysis. In patients with conditionally «mutant» genotypes of the studied genes, peritonitis is more severe, the severity of changes in the studied indicators of homeostasis is greater. At the same time, the effectiveness of therapy decreased, the number of complications increased, and the stay of patients in the hospital increased. The early inclusion of Remaxol and laser radiation in basic therapy increases the overall effectiveness of treatment: the severity of endotoxemia and oxidative stress decreases, the functional state of the liver and hemostasis system is restored relatively quickly, and as a result, an improvement in clinical results has been registered (a 37.5 % decrease in the frequency of complications (χ2=3.360, p=0.047) and a reduction in the length of stay of patients in the hospital by 16.6 % (p&lt;0.05)).CONCLUSION. The inclusion of Remaxol and laser therapy in the early postoperative period in the treatment regimen of patients with peritonitis makes it possible to influence pathogenetically important links of the disease relatively quickly, which improves treatment results. This type of therapy is particularly effective in patients with polymorphic genotypes T1565C and C1565C of the ITGB3 gene.

Molecular dissection of two Pakistani families segregating Leukocyte Adhesion Deficiency Type-I

BACKGROUND &amp; OBJECTIVE: Leukocyte adhesion deficiency (LAD1) is an autosomal recessive type of inherited disorder caused by total or partial deficiency of CD18 expression. LAD1 is characterized by recurrent bacterial and fungal infections, in some cases delayed umbilical cord separation, delayed wound healing due to blockade in leukocyte migration to site of inflammation and infection. The present study involves genetic analysis of two unrelated families suffering from LAD1.&#x0D; METHODOLOGY: In the present study, two separated and unrelated Pakistani families are included suffering from Leukocyte Adhesion Deficiency type -1 (PAD1). After detailed clinical evaluation, whole blood samples were collected from patients, parents and available healthy siblings. Genomic DNA was extracted from all the blood samples, and using a specific primer all the coding ITGB2 gene exons were PCR amplified.&#x0D; RESULTS: The amplified products were sanger sequenced. DNA sequencing analysis revealed a nonsense mutation c.186C &gt; A, p.(Cys62*) in exon four and a missense mutation c.382G&gt;T, p.(Asp128Tyr) in exon five of the gene. The mutation is segregating in autosomal recessive pattern in the family.&#x0D; CONCLUSION: Recurrent mutations on a specific locus (gene) changes the allele frequency from a healthy allele to a disease allele and hence play role in new genotype. This research study demonstrates the allelic heterogeneity of the ITGB2 gene in Pakistani patients diagnosed with LAD1. The research findings in the present study suggest that every population should develop national registry of patients suffering from primary immune deficiencies and a mutation database for rare genetic disorders. This will facilitate early diagnosis and genetic counseling to the patient family.

Extracellular Vesicle-Derived circITGB1 Regulates Dendritic Cell Maturation and Cardiac Inflammation via miR-342-3p/NFAM1.

Acute myocardial infarction (AMI) is a complication of atherosclerosis-related cardiovascular illness that is caused by prolonged ischemia. Circular RNAs (circRNAs) are concentrated in extracellular vesicles (EVs) and have been linked to cardiovascular disease. However, additional research is needed into the expression and function of circRNAs in AMI. In this study, circITGB1 (has_circRNA_0018146), derived from exon 1 of the ITGB1 gene localized on chromosome 10, was shown to be considerably increased in plasma from patients with AMI compared to healthy controls, as demonstrated by the comparison of EV-circRNA expression patterns. Using a luciferase screening assay and a biotin-labeled circITGB1 probe to identify microRNA(s) complementary to circITGB1 sequences, we discovered that circITGB1 competitively binds to miR-342-3p and inhibits its expression, which in turn increase the expression of NFAT activating molecule 1 (NFAM1). Based on western blotting and immunological studies, circITGB1 controls dendritic cell maturation by targeting miR-342-3p and NFAM1. circITGB1 also exacerbated cardiac damage and regulated miR-342-3p and NFAM1 expression in a mouse AMI model. This implies that EV-circITGB1 is involved in dendritic cell maturation and cardiac damage via miR-342-3p/NFAM1, and that is linked to AMI-associated pathogenic processes.

Association between the 5-methyl tetrahydrofolate reductase (5-MTHFR) and integrin Β3 subunit (ITGB3) genes polymorphism and major depressive disorder

Background and objectivesMajor depression disorder (MDD) is a common condition and syndromic sickness with devastating psychological, social, and economic effects. In general, it is a widespread problem, and the prevalence of MDD varies by country. The genetic contribution to MDD is estimated to be around 35%. This study was done to explore the association between 5-methyl tetrahydrofolate reductase (5-MTHFR) and integrin β3 subunit (ITGB3) genes polymorphism for development of major depressive disorder. Materials and methodsThe current study comprised thirty MDD patients, ranging in age from 18 to 52 years. All of the individuals were diagnosed using the Diagnostic and Statistical Manual of Mental Axis I Disorder in the outpatient departments of Taif Psychiatry Health Hospital (DSM-IV). The three polymorphic markers, C667T and A1298C of the 5-MTHFR gene and T1565C of the ITGB3 gene, were genotyped using the PCR-RFLP method. ResultsThe C667T single nucleotide polymorphism was found to be in heterozygous state in ten patients, while the A1298C single nucleotide polymorphism was not found in any of them. Twenty-one patients had the T1565C single nucleotide polymorphism in the ITGB3 gene, five of whom were homozygous state and 16 of whom were heterozygous state. Seven patients were discovered to have compound mutations in both genes. Interpretation & conclusionThe current findings could point to a link between MDD and the T allele of the C667T polymorphism in the 5-MTHFR gene and the C allele of the T1565C polymorphism in the ITGB3 gene.

Identification and functional characterization of imbalanced osteoarthritis-associated fibronectin splice variants.

To identify FN1 transcripts associated with OA pathophysiology and investigate the downstream effects of modulating FN1 expression and relative transcript ratio. FN1 transcriptomic data was obtained from our previously assessed RNA-seq dataset of lesioned and preserved OA cartilage samples from the Research osteoArthritis Articular Cartilage (RAAK) study. Differential transcript expression analysis was performed on all 27 FN1 transcripts annotated in the Ensembl database. Human primary chondrocytes were transduced with lentiviral particles containing short hairpin RNA (shRNA) targeting full-length FN1 transcripts or non-targeting shRNA. Subsequently, matrix deposition was induced in our 3D in vitro neo-cartilage model. Effects of changes in the FN1 transcript ratio on sulphated glycosaminoglycan (sGAG) deposition were investigated by Alcian blue staining and dimethylmethylene blue assay. Moreover, gene expression levels of 17 cartilage-relevant markers were determined by reverse transcription quantitative polymerase chain reaction. We identified 16 FN1 transcripts differentially expressed between lesioned and preserved cartilage. FN1-208, encoding migration-stimulating factor, was the most significantly differentially expressed protein coding transcript. Downregulation of full-length FN1 and a concomitant increased FN1-208 ratio resulted in decreased sGAG deposition as well as decreased ACAN and COL2A1 and increased ADAMTS-5, ITGB1 and ITGB5 gene expression levels. We show that full-length FN1 downregulation and concomitant relative FN1-208 upregulation was unbeneficial for deposition of cartilage matrix, likely due to decreased availability of the classical RGD (Arg-Gly-Asp) integrin-binding site of fibronectin.

Association of Genetic Polymorphisms with Complications of Implanted LVAD Devices in Patients with Congestive Heart Failure: A Kazakhstani Study.

The left ventricular assist device (LVAD) is one of the alternative treatments for heart failure (HF) patients. However, LVAD support is followed by thrombosis, and bleeding complications which are caused by high non-physiologic shear stress and antithrombotic/anticoagulant therapy. A high risk of complications occurs in the presence of the genotype polymorphisms which are involved in the coagulation system, hemostasis function and in the metabolism of the therapy. The aim of the study was to investigate the influence of single-nucleotide polymorphisms (SNP) in HF patients with LVAD complications. We analyzed 21 SNPs in HF patients (n = 98) with/without complications, and healthy controls (n = 95). SNPs rs9934438; rs9923231 in VKORC1, rs5918 in ITGB3 and rs2070959 in UGT1A6 demonstrated significant association with HF patients’ complications (OR (95% CI): 3.96 (1.42–11.02), p = 0.0057), (OR (95% CI): 3.55 (1.28–9.86), p = 0.011), (OR (95% CI): 5.37 (1.79–16.16), p = 0.0056) and OR (95% CI): 4.40 (1.06–18.20), p = 0.044]. Genotype polymorphisms could help to predict complications at pre- and post-LVAD implantation period, which will reduce mortality rate. Our research showed that patients can receive treatment with warfarin and aspirin with a personalized dosage and LVAD complications can be predicted by reference to their genotype polymorphisms in VKORC1, ITGB3 and UGT1A6 genes.

Identification of Biomarkers of Autophagy-Related Genes Between Early and Advanced Carotid Atherosclerosis.

BackgroundAccumulating evidence demonstrates that autophagy is important in inhibiting inflammation and cholesterol efflux. It suggested the autophagy may be a treatment of atherosclerosis. Thus, we screened autophagy-related mRNA to explore their mechanism of scientific basis for early diagnosis and therapy of atherosclerosis.MethodsThe GSE28829 datasets were assessed to analyze differentially expressed genes by GEO2R. And autophagy-related hub genes were identified by HADb. The biological function of autophagy-related DEmRNAs was examined by Metascape. The construction of a protein–protein network was explored by String. Cytohubba was utilized to screen hub genes. Analysis of DEmiRNA-mRNA pairs was executed by DIANA microT-CDS database. Finally, correlation analysis was carried out to identify the relationship between DEARGs and clinical and prognostic factors.ResultsA number of 1087 DEGs and 19 autophagy-related DEmRNAs were identified in advanced carotid atherosclerotic plaque compared with the early. The biological function containing development and growth was enriched. Moreover, we screened the top hub nodes with the highest degrees. MicroRNAs (miRNAs) are confirmed to participate in genesis and progression of atherosclerosis, so we further analyzed the miRNA–mRNA regulatory network genes with four hub genes to explore their potential mechanism in atherosclerosis. Then, we revealed co-expression of four key genes CTSB, ITGB1, CXCR4, TNFSF10 and autophagy-related genes. As for the clinical factors, hypertension factor showed higher expression of ITGB1. The probability of coronary heart disease factor was significantly increased with high expression of CTSB and CXCR4, as well as low expression of ITGB1 and TNFSF10. Diabetes factor tended to express distinguished levels of CTSB and ITGB1. TNFSF10 was highly expressed in both hyperlipidemia and ischemic stroke factor.ConclusionCTSB, ITGB1, CXCR4 and TNFSF10 may be critical in atherosclerosis development and were thought to be potential diagnostic biomarkers for atherosclerosis.

MP46-13 TEX11 SUPPRESS THE MIGRATION AND INVASION IN TESTICULAR GERM CELL TUMOR BY DOWN-REGULATED ITGB2

You have accessJournal of UrologyCME1 May 2022MP46-13 TEX11 SUPPRESS THE MIGRATION AND INVASION IN TESTICULAR GERM CELL TUMOR BY DOWN-REGULATED ITGB2 Wenjun Zhou Wenjun ZhouWenjun Zhou More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002612.13AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Testis expressed gene 11 (TEX11) is a X-linked gene and mainly involved in meiotic arrest and male infertility, its role in the testicular germ cell tumor (TGCT) remains unknown. Thus, we aim to investigate the role of TEX11 in the pathogenesis of TGCT. METHODS: In this study, the overexpression plasmid of TEX11 (pcDNA-3.1+/TEX11) was constructed and transfected into NCCIT cell and Tcam-2 cell respectively. After transfection, the change of cell function was detected, including cell cycle, proliferation and apoptosis, migration and invasion. Furthermore, RNA sequence was used to investigated the gene expression profile alternation after TEX11 over-expression in Tcam-2 cell, genes with P value ≤ 0.05 and |log2FC| ≥ 1 were considered as differentially expressed genes (DEGs). GO and KEGG enrichment analyses were used to revealed the function of TEX11. RESULTS: The results showed that after overexpressed TEX11 in NCCIT and Tcam-2, its mRNA expression level was significantly increased. The Transwell indicated that TEX11 inhibited the migration and invasion ability of both NCCIT and Tcam-2 cells. Furthermore, the bioinformatics analysis showed that a total of 131 DEGs were detected in the sequence data, including 75 up-regulated DEGs and 56 down-regulated DEGs. The GO analysis showed that the down-regulated DEGs were significantly enriched in cellular process, biological regulation, protein binding, cell and cell part. KEGG pathway analysis revealed that these genes were mainly enriched in cell adhesion molecules (CAMs), ovarian steroidogenesis, NF-kappa B pathway and leukocyte transendothelial migration. Based on the phenotype change of migration and invasion ability, we chosen the differentially expressed gene ITGB2 from CAMs pathway, as a target gene. Using GEPIA database, we found that ITGB2 was significantly up-regulated in the TGCT tissue, and negatively related with the expression level of TEX11. CONCLUSIONS: In conclusion, we believed that TEX11 is a novel gene in the pathogenesis of TGCT. TEX11 may down-regulate ITGB2 gene through cell adhesion molecules pathway, thereby inhibit the invasion and migration ability of testicular germ cell tumor. Source of Funding: The work was supported by Graduate Independent Innovation Project Fund of Central South University (2020zzts229) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e778 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Wenjun Zhou More articles by this author Expand All Advertisement PDF DownloadLoading ...

ITGB1 Suppresses Autophagy Through Inhibiting The mTORC2/AKT Signaling Pathway In H9C2 Cells.

Cardiomyocyte autophagy is closely related to myocardial infarction and hypertrophy. To study the molecular mechanism of autophagy is helpful for the prevention and treatment of these diseases. As a cell surface receptor, the function of ITGB1 gene in cardiomyocyte autophagy is not clear. The purpose of this research was to investigate the function and molecular mechanism of ITGB1 on autophagy. The autophagy-related marker proteins and signaling molecules were detected using western blot with knockdown and overexpression of ITGB1 in H9C2 cells. The results suggested that ITGB1 could inhibit autophagy and the mTORC2/Akt pathway molecules. To further investigate whether the effect of ITGB1 on autophagy might affect myocardial hypertrophy, we constructed AngII induced H9C2 cells and TAC induced rats models. The results showed that ITGB1 inhibited myocardial hypertrophy in both H9C2 cells and heart tissues of disease model. These data highlight the regulation mechanism on autophagy by ITGB1 and the potential usefulness of the gene as a potential target for preventing heart disease.