Extracellular Matrix Metalloproteinase Inducer Research Articles

The Roles of GDF-9, BMP-15, BMP-4 and EMMPRIN in Folliculogenesis and In Vitro Fertilization.

Growth differentiation factor 9 (GDF-9) contributes to early ovarian development and oocyte survival. Higher concentrations of GDF-9 in follicular fluid (FF) are associated with oocyte nuclear maturation and optimal embryo development. In in vitro fertilization (IVF), GDF-9 affects the ability of the oocyte to fertilize and subsequent embryonic development. Bone morphogenetic protein 15 (BMP-15) is involved in the regulation of ovarian function and affects oocyte development. During IVF, BMP-15 contributes to the formation of competent blastocysts. BMP-15 may play a role in embryo implantation by affecting endometrial receptivity. Bone morphogenetic protein 4 (BMP-4) is involved in the regulation of follicle growth and development and affects granulosa cell (GC) differentiation. In relation to IVF, BMP-4 is important for embryonic development, influences cell fate and differentiation, and plays a role in facilitating embryo-endometrial interactions during the implantation process. Extracellular matrix metalloproteinase inducer (EMMPRIN) is associated with ovulation and follicle rupture, promotes the release of mature eggs, and affects the modification of the extracellular matrix of the follicular environment. In IVF, EMMPRIN is involved in embryo implantation by modulating the adhesive properties of endometrial cells and promotes trophoblastic invasion, which is essential for pregnancy to occur. The purpose of the current article is to review the studies and recent findings of GDF-9, BMP-15, BMP-4 and EMMPRIN as fundamental factors in normal follicular development and in vitro fertilization.

BSG Isoform 2 (ENST00000353555) Is a Better Component Than Total BSG Expression in Generating Prognostic Signature for Overall Survival of Liver Cancer.

The basigin (BSG) gene, also known as CD147, has been implicated in the progression and prognosis of various cancers, including liver cancer. This study aimed to comprehensively evaluate the prognostic value of total BSG expression and its specific transcript variants, ENST00000353555 and ENST00000545507, in a large cohort of patients with primary liver cancer. The prognostic values of total BSG, ENST00000353555, and ENST00000545507 expression in overall survival (OS) and progression-free interval (PFI) of patients with primary liver cancer were assessed using The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset. Survival analysis, receiver operating characteristic (ROC) analysis, and validation of an extracellular matrix (ECM)-related prognostic signature were performed. In univariate and multivariate analyses, total BSG, ENST00000353555, and ENST00000545507 expression were associated with poor OS in liver cancer patients. ENST00000353555 showed the highest hazard ratio among the three prognostic indicators. ROC analysis revealed that ENST00000353555 had better prognostic performance than total BSG expression. Replacing total BSG with ENST00000353555 in an existing ECM-related prognostic signature marginally increased the area under the curve values for one year from 0.79 to 0.80, and five-year OS from 0.72 to 0.73. ENST00000353555 showed isoform-specific positive correlations with EDNRB, IL10, C10orf54, and VEGFA. ENST00000353555 serves as a better prognostic biomarker than total BSG expression in liver cancer, either as an individual marker or as a component of an ECM-related gene signature. Additionally, ENST00000353555 exhibited isoform-specific positive correlations with several immunosuppressive genes, suggesting a potential role in regulating the tumor microenvironment.

Abstract 4465: Galectin-1 regulates CD147 and downstream pathways critical for glioma progression

Abstract Galectin-1 (Gal-1) is an important protein for glioma progression and glioma stem cell maintenance. Gal-1 may act through CD147, a potential Gal-1 receptor. CD147, also known as Basigin or extracellular matrix metalloproteinase inducer (EMMPRIN), is a transmembrane glycoprotein highly expressed in various cancer cells. An analysis utilizing the Gliovis database and encompassing 667 Glioblastoma Multiforme (GBM) patients segregated based on median CD147 expression levels delineated cohorts with above-median (n=339) and below-median (n=328) CD147 expression. Kaplan-Meier survival curves unveiled a significant association between heightened CD147 expression and reduced overall survival (OS) in this patient population. Experimental approaches employing immunoblotting and immunohistochemistry techniques on shGal-1 treated cell lysates and mouse brain samples exhibited decreased CD147 expression compared to respective controls. Additionally, immunocytochemical studies corroborated an augmented colocalization of Gal-1 and CD147, with a subsequent reduction in this association following shGal-1 treatment. Next, we used AC-73, a selective CD147 inhibitor, to determine the levels of CD147 and Gal-1. Treatment with 20μM AC-73 and siRNA for CD147 showed reduced CD147 levels in GSC33 and GSC20 cells while maintaining unaltered Gal-1 expression. Inhibiting CD147 did not affect Gal-1 levels, further suggesting Gal-1 acts upstream. CD147 activates matrix metalloproteinase-9 (MMP9), and CD147 knockdown reduced MMP9 levels, elucidating a Gal-1-CD147-MMP9 pathway. Overall, Gal-1 appears to act upstream of CD147 to regulate MMP9, integrins, and other factors important for glioma progression. Further elucidation of these pathways will provide insights into glioma biology and potential therapeutic targets. Citation Format: Maheedhara R. Guda, Andrew J. Tsung, Swapna Asuthkar, Kiran Velpula. Galectin-1 regulates CD147 and downstream pathways critical for glioma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4465.

A comprehensive investigation on the receptor BSG expression reveals the potential risk of healthy individuals and cancer patients to 2019-nCoV infection.

Coronavirus disease-2019 (COVID-19) pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a newly emerging coronavirus. BSG (basigin) is involved in the tumorigenesis of multiple tumors and recently emerged as a novel viral entry receptor for SARS-CoV-2. However, its expression profile in normal individuals and cancer patients are still unclear. We performed a comprehensive analysis of the expression and distribution of BSG in normal tissues, tumor tissues, and cell lines via bioinformatics analysis and experimental verification. In addition, we investigated the expression of BSG and its isoforms in multiple malignancies and adjacent normal tissues, and explored the prognostic values across pan-cancers. Finally, we conducted function analysis for co-expressed genes with BSG. We found BSG was highly conserved in different species, and was ubiquitously expressed in almost all normal tissues and significantly increased in some types of cancer tissues. Moreover, BSG at mRNA expression level was higher than ACE2 in normal lung tissues, and lung cancer tissues. High expression of BSG indicated shorter overall survival (OS) in multiple tumors. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that BSG is mostly enriched in genes for mitochondria electron transport, oxidoreduction-driven active transmembrane transporter activity, mitochondrial inner membrane, oxidative phosphorylation, and genes involving COVID-19. Our present work emphasized the value of targeting BSG in the treatment of COVID-19 and cancer, and also provided several novel insights for understanding the SARS-CoV-2 pandemic.

Paradoxical Changes: EMMPRIN Tissue and Plasma Levels in Marfan Syndrome-Related Thoracic Aortic Aneurysms.

Background: Thoracic aortic aneurysms (TAAs) associated with Marfan syndrome (MFS) are unique in that extracellular matrix metalloproteinase inducer (EMMPRIN) levels do not behave the way they do in other cardiovascular pathologies. EMMPRIN is shed into the circulation through the secretion of extracellular vesicles. This has been demonstrated to be dependent upon the Membrane Type-1 MMP (MT1-MMP). We investigated this relationship in MFS TAA tissue and plasma to discern why unique profiles may exist. Methods: Protein targets were measured in aortic tissue and plasma from MFS patients with TAAs and were compared to healthy controls. The abundance and location of MT1-MMP was modified in aortic fibroblasts and secreted EMMPRIN was measured in conditioned culture media. Results: EMMPRIN levels were elevated in MFS TAA tissue but reduced in plasma, compared to the controls. Tissue EMMPRIN elevation did not induce MMP-3, MMP-8, or TIMP-1 expression, while MT1-MMP and TIMP-2 were elevated. MMP-2 and MMP-9 were reduced in TAA tissue but increased in plasma. In aortic fibroblasts, EMMPRIN secretion required the internalization of MT1-MMP. Conclusions: In MFS, impaired EMMPRIN secretion likely contributes to higher tissue levels, influenced by MT1-MMP cellular localization. Low EMMPRIN levels, in conjunction with other MMP analytes, distinguished MFS TAAs from controls, suggesting diagnostic potential.

NOS3 prevents MMP-9, and MMP-13 induced extracellular matrix proteolytic degradation through specific microRNA-targeted expression of extracellular matrix metalloproteinase inducer in hypertension-related atherosclerosis.

Endothelial nitric oxide synthase (NOS3) elicits atheroprotection by preventing extracellular matrix (ECM) proteolytic degradation through inhibition of extracellular matrix metalloproteinase inducer (EMMPRIN) and collagenase MMP-13 by still unknown mechanisms. C57BL/6 mice lacking ApoE , NOS3, and/or MMP13 were fed with a high-fat diet for 6 weeks. Entire aortas were extracted and frozen to analyze protein and nucleic acid expression. Atherosclerotic plaques were detected by ultrasound imaging, Oil Red O (ORO) staining, and Western Blot. RNA-seq and RT-qPCR were performed to evaluate EMMPRIN, MMP-9, and EMMPRIN-targeting miRNAs. Mouse aortic endothelial cells (MAEC) were incubated to assess the role of active MMP-13 over MMP-9. One-way ANOVA or Kruskal-Wallis tests were performed to determine statistical differences. Lack of NOS3 in ApoE null mice fed with a high-fat diet increased severe plaque accumulation, vessel wall widening, and high mortality, along with EMMPRIN-induced expression by upregulation of miRNAs 46a-5p and 486-5p. However, knocking out MMP-13 in ApoE/NOS3 -deficient mice was sufficient to prevent mortality (66.6 vs. 26.6%), plaque progression (23.1 vs. 8.8%), and MMP-9 expression, as confirmed in murine aortic endothelial cell (MAEC) cultures, in which MMP-9 was upregulated by incubation with active recombinant MMP-13, suggesting MMP-9 as a new target of MMP-13 in atherosclerosis. We describe a novel mechanism by which the absence of NOS3 may worsen atherosclerosis through EMMPRIN-induced ECM proteolytic degradation by targeting the expression of miRNAs 146a-5p and 485-5p. Focusing on NOS3 regulation of ECM degradation could be a promising approach in the management of atherosclerosis.

Expression profiles of MMP-9 and EMMPRIN in chronic rhinosinusitis with nasal polyps.

Metalloproteinases (MMPs) are implicated in tissue remodeling in chronic rhinosinusitis with nasal polyps (CRSwNP). This study aimed to evaluate the expression profiles of MMP-9 and the extracellular matrix metalloproteinase inducer (EMMPRIN) in nasal polyps compared to healthy mucosa. Tissue samples from 37 CRSwNP patients undergoing functional endoscopic sinus surgery and mucosal specimens from 12 healthy controls were obtained intra-operatively. MMP-9 and EMMPRIN mRNA levels were assessed by reverse transcription-polymerase chain reaction and their protein expression by Western blot analysis. MMP-9 mRNA expression levels were significantly elevated in CRSwNP compared to controls (p < 0.05). MMP-9 protein levels presented an increasing trend but with no statistical significance (p > 0.05). No statistically significant difference in EMMPRIN mRNA and protein levels was identified. Upregulation of MMP-9 in nasal polyps is evident and highlights its role in the pathogenesis of CRSwNP. The lack of concordance between MMP-9 mRNA and protein levels may be attributed to post-translational gene expression regulation. Although EMMPRIN expression was not significantly different between the two groups, its role remains to be elucidated. MMP-9 may be a valuable biomarker and treatment target in CRSwNP.

Abstract A069: Generation of llama-derived phage display library specific to hepatocellular carcinoma tumor targets

Abstract Background: Primary liver cancer is the third leading cause of cancer-related deaths worldwide, with its incidence and mortality continuing to increase. Hepatocellular carcinoma (HCC) accounts for approximately 90% of these cases, and outcomes for most patients are poor. Unlike many cancer types, liver cancer cannot reliably be detected by 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) due to its limited tumor uptake. Curiously, despite the advent of precision oncology, no HCC-selective agents are clinically approved for the diagnosis or treatment of HCC in the United States. There are multiple tumor targets, including Glypican-3(GPC3) and Basigin (BSG) which are overexpressed on HCC tumors. GPC3 is a heparin sulfate proteoglycan expressed in 75% of HCC tumors, and rarely presented on other tissue types, making it a promising HCC-specific target. Similarly, BSG is a transmembrane glycoprotein that is expressed in 60% of HCC tumors, and is closely associated with tumor invasion, metastasis, and angiogenesis. Accordingly, radiopharmaceutical agents specific to GPC3 or BSG could allow for HCC-selective imaging and therapy. Methods:Using standard methods, a young, naïve llama was immunized four times with human and murine GPC3, along with BSG . Nanobody cDNA sequences were isolated from the B cells of the llama, amplified by PCR, cloned into phagemids, and transformed into E. Coli cells to create a nanobody library with over 10⁶ clones. Phage, displaying the nanobody protein, are then produced and used for immunopanning to enrich for specific nanobodies with affinity against each antigen. Using antigen-coated high binding tubes, phage particles displaying a specific nanobody will bind to the antigen and be eluted following stringent wash conditions, while non-specific phage particles are discarded. The eluted phage is then amplified, and a repeat round of phage display is performed. Enriched clones will then be screened by ELISA to identify high affinity binders. Lead candidate nanobodies are then sequenced and unique clones expressed to determine yield, stability, and purity. Once specific nanobodies are identified, additional biophysical characterization will be performed. Results: An antibody titer was performed by ELISA, demonstrating a successful immune response to all antigens. A phage display library was successfully created that is specific to human GPC3, murine GPC3 and BSG, with over 10⁶ clones. It is anticipated that after a few rounds of immunopanning, specific nanobodies will be identified. Conclusion: Thus far, a phage display library was created that is specific to mGPC3, hGPC3 and BSG. Immunopanning is currently underway to select out clones that are specific to the targeted antigens. Once enriched clones are isolated, orthogonal in vitro binding affinity assays, such as ELISA, flow cytometry and biolayer interferometry, will be used to measure specificity and strength of antigen binding. Subsequently, assessment of in vivo target binding will be performed using biodistribution studies and PET imaging. Citation Format: Divya Nambiar, Thomas J Esparza, Peter L Choyke, Freddy E Escorcia. Generation of llama-derived phage display library specific to hepatocellular carcinoma tumor targets [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A069.

Lifestyle Modification Exerts Protection Against Obesity‐Induced Neurodegeneration Via CD147‐Related Mechanism

AbstractBackgroundCD147, also known as extracellular matrix metalloproteinase inducer, is a key mediator of inflammatory and immune responses in various kinds of neurodegenerative disease (1). However, the role of CD147 in obesity‐induced neurodegeneration has not been previously investigated. Therefore, this study aims to investigate the alteration of CD147 expression in the hippocampus of obese rats, both with and without lifestyle modification.MethodSeven‐week‐old male Wistar rats were divided into 4 groups (n = 5 per group), including 1) rats fed with a normal diet for 28 weeks [ND], 2) rats fed with a high‐fat diet (HFD) for 28 weeks [HFD], 3) rats fed with a HFD for 12 weeks followed by 40% caloric restriction for an additional 16 weeks [HFD‐CR], or 4) rats fed with a HFD for 12 weeks followed by exercise plus HFD for an additional 16 weeks [HFD‐EX]. At the end of week 28, cognitive function was determined by a novel object location test. The rats were then euthanized on the following day for molecular studies in the hippocampus.ResultWe found that hippocampal CD147 level exerted a strongly negative correlation with the level of cognitive function (Figure A). In addition, CD147 expression and its downstream consequences – p‐NFκB, MMP‐9, and TNF‐α – were upregulated in HFD‐fed rats (Figure B‐E). The HFD‐fed rats also exhibited an increase in activated microglia (CD68), along with decreased blood‐brain barrier protein (claudin‐5), dendritic spine density, and cognitive performance (Figure F‐I). Strikingly, both caloric restriction and exercise effectively reduced the expression of CD147 and its downstream consequences, as well as restored CD68, claudin‐5, dendritic spine density, and cognitive function in HFD‐fed rats (Figure B‐I).ConclusionOur findings suggested that CD147 is one of the detrimental mediators in obesity‐induced neurodegeneration (Figure J). Interestingly, an attenuation of obesity by lifestyle modification exerts protection against cognitive impairment via CD147‐related mechanisms (Figure J). Therefore, an inhibition of CD147 is considered as a therapeutic target to prevent cognitive impairment in obese individuals. Reference 1. Kaushik DK, Hahn JN, Yong VW. EMMPRIN, an upstream regulator of MMPs, in CNS biology. Matrix Biol. 2015;44‐46:138‐46.

The possible effects of COVID-19 on the human reproductive system

Spike surface glycoprotein and small envelope matrix-nucleocapsid proteins, is from the Coronaviridae family and binds to host receptors via spike surface proteins. Although it shows its symptoms especially on the respiratory tract, various studies have been carried out considering that it also affects other systems in the body. For the virus to enter the host cell, it must bind to ACE2 (angiotensin converting enzyme 2). ACE2 is a key protein involved in balancing Ang I and Ang II levels. With receptors such as TMPRSS2 (transmembrane serine protease 2), the effects of the virus on the human reproductive system are much better understood. Since human germ cells and early embryos express ACE2, there is a potential risk of the Coronavirus associated with germ cells. Studies show that the coronavirus changes the amount and density of hormones in the human reproductive system. The fact that most of the partners of 35 female patients who had SARS-CoV-2 in the studies were infected individuals suggests that sexual transmission may be possible. It was determined that TMPRSS4, Cathepsin B and L, FURIN, MX1 and BSG gene expressions were high in the menstrual cycle, while ACE 2 and TMPRSS2 were moderately expressed. It has been shown that the ACE2 enzyme is most intensely expressed is the testes. Studies have shown that sperm DNA (deoxyribonucleic acid) fragmentation, changes in hormone levels and the formation of anti-sperm antibodies are an important cause of male infertility. Infected men have been found to have an impaired spermatogenesis. This rewiew; it aims to draw attention to the possible effects of the corona virus on the human reproductive system and to reveal new mechanisms for new research to be done.

The renoprotective effects of taurine against diabetic nephropathy via the p38 MAPK and TGF-β/Smad2/3 signaling pathways.

Diabetic nephropathy (DN), a severe diabetes complication, causes kidney morphological and structural changes due to extracellular matrix accumulation. This accumulation is caused mainly by oxidative stress. Semi-essential amino acid derivative taurine has powerful antioxidant and antifibrotic effects. The aim of this study was to investigate the renoprotective effects of taurine through its possible roles in oxidative stress, extracellular matrix proteins, and the signaling pathways associated with the accumulation of extracellular matrix proteins in DN rats. 29 Wistar albino rats were randomly separated into control, taurine, diabetes, and diabetes + taurine groups. Diabetes animals were injected 45mg/kg streptozosine. Taurine is given by adding to drinking water as 1% (w/v). Urine, serum, and kidney tissue were collected from rats for biochemical and histological analysis after 12weeks. According to the studies, taurine significantly reduces the levels of malondialdehyde (MDA), total oxidant status (TOS), and protein expression of NADPH oxidase 4 (NOX4) that increase in diabetic kidney tissue. Also, decreased superoxide dismutase (SOD) activity levels significantly increased with taurine in diabetic rats. Moreover, increased mRNA and protein levels of fibronectin decreased with taurine. The matrix metalloproteinase (MMP)-2 and MMP-9 activities and their mRNA levels increased significantly, and this increase was significantly summed with taurine. There was a decrease in mRNA expression of Extracellular matrix metalloproteinase inducer (EMMPRIN). Taurine significantly increased this decrease. Diabetes increased mRNA expressions of transforming growth factor (TGF)-β and Smad2/3. Taurine significantly reduced this induction. TGF-β protein expression, p38, and Smad2/3 activations were also inhibited, but taurine was suppressed significantly. All these findings indicate that taurine may be an effective practical strategy to prevent renal diabetic injury.

CD98 heavy chain as a prognostic biomarker and target for cancer treatment.

The SLC3A2 gene encodes for a cell-surface transmembrane protein CD98hc (4F2). CD98hc serves as a chaperone for LAT1 (SLC7A5), LAT2 (SLC7A8), y+LAT1 (SLC7A7), y+LAT2 (SLC7A6), xCT (SLC7A11) and Asc1 (SLC7A10) providing their recruitment to the plasma membrane. Together with the light subunits, it constitutes heterodimeric transmembrane amino acid transporters. CD98hc interacts with other surface molecules, such as extracellular matrix metalloproteinase inducer CD147 (EMMPRIN) and adhesion receptors integrins, and regulates glucose uptake. In this way, CD98hc connects the signaling pathways sustaining cell proliferation and migration, biosynthesis and antioxidant defense, energy production, and stem cell properties. This multifaceted role makes CD98hc one of the critical regulators of tumor growth, therapy resistance, and metastases. Indeed, the high expression levels of CD98hc were confirmed in various tumor tissues, including head and neck squamous cell carcinoma, glioblastoma, colon adenocarcinoma, pancreatic ductal adenocarcinoma, and others. A high expression of CD98hc has been linked to clinical prognosis and response to chemo- and radiotherapy in several types of cancer. In this mini-review, we discuss the physiological functions of CD98hc, its role in regulating tumor stemness, metastases, and therapy resistance, and the clinical significance of CD98hc as a tumor marker and therapeutic target.

124I-labeled anti-CD147 antibody for noninvasive detection of CD147-positive pan-cancers: construction and preclinical studies.

Extracellular matrix metalloproteinase inducer CD147 is a glycoprotein on the cell surface. There is minimal expression of CD147 in normal epithelial and fetal tissues, but it is highly expressed in a number of aggressive tumors. CD147 has been implicated in pan-cancer immunity and progression. With the development of CD147-targeting therapeutic strategy, accurate detection of CD147 expression in tumors and its changes during the therapy is necessary. In this study we constructed a novel radiotracer by labeling the anti-CD147 mAb with radionuclide 124/125I (124/125I-anti-CD147) for noninvasive detection of CD147 expression in pan-cancers, and characterized its physicochemical properties, affinity, metabolic characteristics, biodistribution and immunoPET imaging with 124I-IgG and 18F-FDG as controls. By examining the expression of CD147 in cancer cell lines, we found high CD147 expression in colon cancer cells LS174T, FADU human pharyngeal squamous cancer cells and 22RV1 human prostate cancer cells, and low expression of CD147 in human pancreatic cancer cells ASPC1 and human gastric cancer cells BGC823. 124/125I-anti-CD147 was prepared using N-bromine succinimide (NBS) as oxidant and purified by PD-10 column. Its radiochemical purity (RCP) was over 99% and maintained over 85% in saline or 5% human serum albumin (HSA) for more than 7 d; the RCP of 125I-anti-CD147 in blood was over 90% at 3 h post injection (p.i.) in healthy mice. The Kd value of 125I-anti-CD147 to CD147 protein was 6.344 nM, while that of 125I-IgG was over 100 nM. 125I-anti-CD147 showed much greater uptake in CD147 high-expression cancer cells compared to CD147 low-expression cancer cells. After intravenous injection in healthy mice, 125I-anti-CD147 showed high initial uptake in blood pool and liver, the uptake was decreased with time. The biological half-life of distribution and clearance phases in healthy mice were 0.63 h and 19.60 h, respectively. The effective dose of 124I-anti-CD147 was estimated as 0.104 mSv/MBq. We conducted immunoPET imaging in tumor-bearing mice, and demonstrated a significantly higher tumor-to-muscle ratio of 124I-anti-CD147 compared to that of 124I-IgG and 18F-FDG in CD147 (+) tumors. The expression levels of CD147 in cells and tumors were positively correlated with the maximum standardized uptake value (SUVmax) (P < 0.01). In conclusion, 124/125I-anti-CD147 displays high affinity to CD147, and represents potential for the imaging of CD147-positive tumors. The development of 124I-anti-CD147 may provide new insights into the regulation of tumor microenvironment and formulation of precision diagnosis and treatment programs for tumors.

S-nitrosylation of EMMPRIN influences the migration of HSCs and MMP activity in liver fibrosis.

The mechanism of extracellular matrix metalloproteinase inducer (EMMPRIN) in the regulation of liver fibrosis has not been clarified. This study aims to investigate the role of EMMPRIN S-nitrosylation (SNO) in the regulation of hepatic stellate cell (HSC) migration and matrix metalloproteinase (MMP) activities in liver fibrosis. The results from the tissue microarrays and rat/mouse liver tissues suggest that EMMPRIN mRNA and protein levels in the fibrotic livers are lower than those in the corresponding normal control livers, but higher SNO level of EMMPRIN in fibrotic liver area was shown by immunohistochemistry, immunofluorescence staining, and biotin-switch assay conversely in vivo. Primary EMMPRIN comes from hepatocytes and liver sinus epithelial cells (LSECs) rather than quiescent HSCs. To mimic the uptake of extrinsic EMMPRIN, supernatants from mouse primary hepatocytes/293 cells transfected with EMMPRIN wild-type plasmids (WT) and EMMPRIN SNO site (cysteine 87) mutation plasmids (MUT) were collected and added to JS-1/LX2 cell medium. The MUT EMMPRIN diminishes SNO successfully, enhances the activities of MMP2 and MMP9, and subsequently increases HSC migration. In conclusion, SNO of EMMPRIN influences HSC migration and MMP activities in liver fibrosis. This finding may shed light on the possible regulatory mechanism of MMPs in ECM accumulation in liver fibrosis.

Extracellular matrix metalloproteinases inducer gene polymorphism and reduced serum matrix metalloprotease-2 activity in preeclampsia patients.

Preeclampsia increases the risk of pregnancy-related complications, nevertheless a successful spiral vessel remodeling, and trophoblast invasion reduces disorders of pregnancy. Matrix metalloproteinase-2 (MMP-2) clears the path for trophoblast invasion, and activation of MMP-2 largely depends on extracellular matrix metalloproteinases inducer (EMMPRIN) protein. This study aimed to investigate EMMPRIN gene polymorphism and MMP-2 activity in preeclampsia patients. Archival whole blood and serum samples of 74 preeclampsia and 66 normotensive pregnant women age-matched were used in this case-control study. Genomic DNA was extracted from the whole blood samples and EMMPRIN gene amplified with specific primers following fragments sequence mutation analysis. Serum MMP-2 activity was determined using enzyme-linked immunosorbent assay (ELISA) and socio-demographic data of participants retrieved from the database. Age of preeclampsia patients (32.78 ± 6.39) years and body mass index (BMI) (33.09 ± 7.27) kg/m2 compared with the normotensive counterparts (32.33 ± 5.56) years and (32.33 ± 5.56) kg/m2,respectively, were not statistically significant (P > 0.05). Serum matrix metalloprotease-2 (MMP-2) activity was significantly reduced in preeclampsia group (16.34 ± 7.07) compared with the normotensives (25.63 ± 4.56) (P < 0.001), and rs424243T/G variant (55.6%) was overrepresented among the cases compared with the normotensives (16.7%). The single-nucleotide polymorphism T/G was found to be associated with preeclampsia (odds ratio [OR] = 7.63; 95% confidence interval [CI] = 3.95-14.75; P < 0.0001). Decreased activity of MMP-2 and rs424243T/G SNP of EMMPRIN gene was reported in preeclampsia. These preliminary data warrant a further investigation into the relationship between EMMPRIN gene polymorphism and MMP-2 activity in preeclampsia.

Abstract P1047: The Extracellular Matrix Metalloproteinase Inducer And The Membrane Type 1 Matrix Metalloproteinase In Marfan Syndrome Related Thoracic Aortic Aneurysm

Objectives: Marfan Syndrome (MFS) is a connective tissue disorder commonly associated with Thoracic Aortic Aneurysms (TAAs). TAAs occur because of abnormal remodeling of the aortic extracellular matrix (ECM). The balance between matrix metalloproteinases (MMPs) and the Tissue Inhibitors of MMPs (TIMPs), regulates these processes. The extracellular MMP inducer (EMMPRIN), is a widely expressed glycoprotein capable of stimulating the production of various MMPs, suggesting a direct role in driving pathology. The Membrane Type-1 MMP (MT1-MMP) can cause proteolytic shedding of EMMPRIN. Therefore, we investigated this relationship in MFS TAA tissue and plasma and tested the hypothesis that a unique proteolytic profile may exist. Methods: Levels of EMMPRIN, MT1-MMP, MMP-2, MMP-9, MMP-3, MMP-8, TIMP-1, and TIMP-2 were measured in aortic tissue and plasma from MFS patients with TAA and compared to healthy controls. Following modification of the abundance and location of MT1-MMP in isolated primary aortic fibroblasts, secreted EMMPRIN was measured in conditioned culture media. Results: Compared to controls, EMMPRIN is elevated in MFS TAA tissue and reduced in the plasma. Elevation of EMMPRIN abundance in tissue does not induce MMP-3, MMP-8, or TIMP-1 expression. MT1-MMP and TIMP-2 are elevated in MFS TAA tissue. MMP-2 and MMP-9 are reduced in MFS TAA tissue while increased in the plasma. In aortic fibroblasts we found that MT1-MMP must be internalized from the plasma membrane for EMMPRIN secretion. Conclusions: Our findings indicate that the pathological consequences of MFS cause the shedding of EMMPRIN and its secretion from cells to be blocked, which leads to higher EMPRINN abundance in MFS TAA tissue. This process depends upon the cellular location of MT1-MMP. Taken together, these observations suggest that internalization of MT1-MMP from the plasma membrane is required for the cellular secretion of EMMPRIN from aortic fibroblasts.

Clathrin light chain A-enriched small extracellular vesicles remodel microvascular niche to induce hepatocellular carcinoma metastasis.

Small extracellular vesicles (sEVs) play a key role in exchanging cargoes between cells in tumour microenvironment. This study aimed to elucidate the functions and mechanisms of hepatocellular carcinoma (HCC) derived sEV-clathrin light chain A (CLTA) in remodelling microvascular niche. CLTA level in the circulating sEVs of HCC patients was analysed by enzyme-linked immunosorbent assay (ELISA). The functions of sEV-CLTA in affecting HCC cancerous properties were examined by multiple functional assays. Mass spectrometry was used to identify downstream effectors of sEV-CLTA in human umbilical vein endothelial cells (HUVECs). Tube formation, sprouting, trans-endothelial invasion and vascular leakiness assays were performed to determine the functions of sEV-CLTA and its effector, basigin (BSG) in HUVECs. BSG inhibitor, SP-8356, was tested in a mouse model of patient-derived xenografts (PDXs). Circulating sEVs of HCC patients had markedly enhanced CLTA levels than control individuals and were reduced in patients after surgery. HCC derived sEV-CLTA enhanced HCC cancerous properties, disrupted endothelial integrity and induced angiogenesis. Mechanistically, CLTA remodels microvascular niche by stabilizing and upregulating BSG. Last, SP-8356 alone or in combination with sorafenib attenuated PDXs growth. The study reveals the role of HCC derived sEV-CLTA in microvascular niche formation. Inhibition of CLTA and its mediated pathway may illuminate a new therapeutic strategy for HCC patients.

CD147 rs8259T>A Variant Confers Susceptibility to COVID-19 Infection within the Mexican Population.

Coronavirus disease 2019 (COVID-19) is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Clinical manifestations of COVID-19 range from mild flu-like symptoms to severe respiratory failure. Nowadays, extracellular matrix metalloproteinase inducer (EMMPRIN), also known as cluster of differentiation 147 (CD147) or BASIGIN, has been studied as enabling viral entry and replication within host cells. However, the impact of the CD147 rs8259T>A single nucleotide variant (SNV) on SARS-CoV-2 susceptibility remains poorly investigated. To investigate the impact of rs8259T>A on the CD147 gene in individuals from Mexico with COVID-19 disease. We genotyped the CD147 rs8359T>A SNV in 195 patients with COVID-19 and 185 healthy controls from Mexico. In addition, we also measured the expression levels of CD147 and TNF mRNA and miR-492 from whole blood of patients with COVID-19 through RT-q-PCR. We observed a significant association between the CD147 rs8259T>A SNV and susceptibility to COVID-19: T vs. A; OR 1.36, 95% CI 1.02-1.81; p = 0.037; and TT vs. AA; OR 1.77, 95% CI 1.01-3.09; p = 0.046. On the other hand, we did not find differences in CD147, TNF or miR-492 expression levels when considering the genotypes of the CD147 rs8259T>A SNV. Our results suggest that the CD147 rs8259T>A variant is a risk factor for COVID-19.

MMP-2 Regulation of Emmprin on Tumour Cells and CD73 on Fibroblasts During Tumour-Stromal Interaction.

We previously found that binding between CD73 and extracellular matrix metalloproteinase (MMP) inducer (emmprin) and suppression of CD73 in both tumour cells and fibroblasts suppressed MMP-2 production when co-cultured. However, the importance of CD73 expression in either fibroblasts or cancer cells for cancer invasion remains unknown. In this study, we used siRNA to separately down-regulate CD73 in individual cells, and then performed a 3D co-culture to investigate tumour invasion. siRNA was used for suppression of CD73 in either fibroblasts (ST353i, HDF) or tumour cells (FU-EPS-1, A431, CRL-2095). Immunoblotting was performed for detecting MMP-2 production after CD73 suppression. 3D-co-cultures were performed for examining tumour invasion. CD73 suppression revealed that CD73 expression on fibroblasts and emmprin on tumour cells were important in regulating MMP-2 production, suggesting that emmprin on tumour cells does not bind CD73 at the cis-manner, but rather at the trans-manner to CD73 present on fibroblasts. CD73 suppression also reduced MMP-2 production at the transcription level and reduced tumour invasion. CD73 on fibroblasts acts as a receptor for emmprin, which forms a complex that increases MMP-2 production, possibly resulting in increased invasiveness.

Intervention with extracellular matrix metalloproteinase inducer in osteoclasts attenuates periodontitis-induced bone resorption.

Extracellular matrix metalloproteinase inducer (EMMPRIN) plays critical roles in the regulation of inflammation and bone metabolism. The roles of EMMPRIN signaling in osteoclasts are worthy of deep study. The present study aimed to investigate bone resorption in periodontitis through the intervention of EMMPRIN signaling. The distribution of EMMPRIN in human periodontitis was observed. RANKL-induced osteoclast differentiation of mouse bone marrow-derived macrophages (BMMs) were treated with EMMPRIN inhibitor in vitro. Rats with ligation-induced periodontitis were treated with EMMPRIN inhibitor and harvested for microcomputed tomography scanning, histologic observation, immunohistochemistry, and double immunofluorescence analysis. Positive expressions of EMMPRIN could be found in the CD68+-infiltrating cells. Downregulated EMMPRIN restrained osteoclast differentiation of BMMs in vitro, which also inhibited MMP-9 expression (*P < 0.05). In vivo, EMMPRIN inhibitor restrained ligation-induced bone resorption by decreasing tartrate-resistant acid phosphatase-positive osteoclasts. Both EMMPRIN-positive and MMP-9-positive osteoclasts were less common in the EMMPRIN inhibitor groups than in the control groups. Intervention of EMMPRIN signaling in osteoclasts could probably provide a potential therapeutic target for attenuating ligation-induced bone resorption.