Matrix Metalloproteinase-9 Gene Research Articles

MP03-04 SECRETION OF NEUROTROPHINS AND THEIR PRECURSORS IS UNDER CONTROL OF NATRIURETIC PEPTIDE A IN BLADDER CELLS IN CULTURE

You have accessJournal of UrologyCME1 Apr 2023MP03-04 SECRETION OF NEUROTROPHINS AND THEIR PRECURSORS IS UNDER CONTROL OF NATRIURETIC PEPTIDE A IN BLADDER CELLS IN CULTURE Claudia Covarrubias, Philippe Cammisotto, and Lysanne Campeau Claudia CovarrubiasClaudia Covarrubias More articles by this author , Philippe CammisottoPhilippe Cammisotto More articles by this author , and Lysanne CampeauLysanne Campeau More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003214.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Urine storage and bladder voiding are under control of the peripheral and central nervous systems. Neurotrophins, such as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), controls neuroregeneration while their respective precursor proNGF and proBDNF trigger inflammatory processes and apoptosis. The ratio between neurotrophins to proneurotrophins must therefore be tighly controlled and constitutes a viable biomarker for overactive bladder syndrome (OAB). On the other hand, natriuretic peptide A (ANP) that is increased during aging, has been linked to OAB. We here examine how ANP is involved in the secretion of neurotrophins by cells of the bladder. METHODS: Smooth muscle (SMC) and urothelial cells (URO) were grown from rat bladder after collagenase digestion. After confluency, they were incubated for 24 hours with ANP (100 nM). NGF, proNGF, BDNF, proBDNF and matrix metalloproteinase-9 (MMP-9) were measured using specific kits. RESULTS: We found that both cell types are major source of NGF and proNGF while BDNF and proBDNF were mainly released by SMC. ANP decreases NGF and proBDNF secretion by SMC, while proNGF and BDNF levels were not affected, mimicking the observations obtained in urine samples of a female cohort of patients diagnosed with OAB. Interestingly, ANP also increased MMP-9 activity in the same cell culture medium. In UROs, no changes in the secretion of neurotrophins or proneurotrophins could be observed, however, the secretion of active MMP-9 was found potently enhanced by ANP with final levels around 20 times higher than the ones found in SMC medium. Finally, using CrisprCas9 to knockdown MMP-9 gene, we confirmed the essential role of this enzyme in the proteolysis of NGF into peptides and in the conversion of proBDNF to BDNF. CONCLUSIONS: Bladder cells exhibit unique pattern of neurotrophin secretions. Both are affected by ANP, SMC mostly at the level of neurotrophin secretions and urothelial cells in the amount of MMP-9 released in the extracellular medium. We suggest that the imbalance in neutrophin ratios observed in the urine of patients with OAB could result, at least in part, from a direct action of ANP on bladder cells. Source of Funding: Canadian Urologist Association and Quebec Network for Research on Aging (RQRV) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e22 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Claudia Covarrubias More articles by this author Philippe Cammisotto More articles by this author Lysanne Campeau More articles by this author Expand All Advertisement PDF downloadLoading ...

Risk factor of elevated matrix metalloproteinase-3 gene expression in synovial fluid in knee osteoarthritis women.

Metalloproteinases-3 (MMP3) are the main enzymes involved in cartilage degradation. Several genetic and non-genetic factors can increase the expression of MMP3 in patients with osteoarthritis (OA). This study aims to analyze the risk factors associated with the expression of the MMP3 gene rs679620 fluid synovial knee OA patients. A cross-sectional study was conducted at the orthopedic polyclinic Arifin Achmad Riau Province and Ibn Sina Hospital in Pekanbaru City. Ninety women who experienced knee OA were taken as samples by consecutive sampling and then signed the informed consent. Data were obtained through interviews using a questionnaire about characteristics, followed by weight and height measurements. Interleukin-1 β (IL-1β) and Tumor Necrosis Factor (TNF-α) were examined from the synovial fluid using the enzyme-linked immunosorbent assay (ELISA) method. The Metalloproteinases-3 (MMP3) gene polymorphism rs679620 was obtained from the DNA analysis of joint fluid results in the Biomedical Laboratory of the Faculty of Medicine, Andalas University. The data was processed computerized and then analyzed using the correlation Spearman-Rank, and chi-square tests. The results of statistical analysis are considered significant if the p-value is 0.05. The MMP3 rs679620 gene polymorphism of the mutant type was 88.9%, with the same proportion of AG and GG alleles (44.4%). Subjects aged ≥ 60 years were 53.3%, 85.6% did not work and 84.4% had menopause. The highest degree of OA was grade 2 (53.3%), most of whom had a risky nutritional status (84.4%). The median expression of the MMP3 rs679620 gene was 5.28 copies number. There is a significant relationship between MMP3 gene polymorphism rs679620, age, IL-1β, and TNF-α with MMP3 gene expression rs679620. There is no significant relationship between BMI, work status, and menopausal status with MMP3 gene expression rs679620. Conclusion. MMP3 gene polymorphism rs679620, age, levels of IL-1β and TNF-α are risk factors for increased MMP3 gene rs679620 expression in female knee OA.

Topical Application of Peptide Nucleic Acid Antisense Oligonucleotide for MMP-1 and Its Potential Anti-Aging Properties

Matrix metalloproteinase-1 (MMP-1) is a zinc-containing endopeptidase that degrades dermal collagen and other extracellular matrix molecules. It is recognized as one of the most important indicators of cellular senescence and age-related skin changes. Here, we introduced a novel MMP-1 peptide nucleic acid (PNA) derivative-PNA-20 carboxyethyl fluorene (CEF)-which can interact with and consequently silence the MMP-1 gene sequence. The investigation on the efficacy of PNA-20 CEF in MMP-1 silencing in human dermal fibroblasts revealed significantly decreased expression of MMP-1 at both gene and protein levels. Treatment with PNA-20 CEF showed significantly increased expression of collagen I protein, indicating its potential role in preventing the degradation of collagen I and consequently combating the skin aging process. Its topical application on 3D human skin tissue showed successful absorption into the epidermis and the upper dermis. Furthermore, the additional 4-week single-arm prospective study on 21 Asian women revealed improvements in facial wrinkles, skin moisture, elasticity, and density after the use of the topical PNA-20 CEF cosmeceutical formulation. Additional in-vitro and ex-vivo studies are needed for a comprehensive understanding of the skin anti-aging effects of MMP-1 PNA.

Earlier proteoglycan turnover promotes higher efficiency matrix remodeling in MRL/MpJ tendons.

While most mammalian tissue regeneration is limited, the Murphy Roths Large (MRL/MpJ) mouse has been identified to regenerate several tissues, including tendon. Recent studies have indicated that this regenerative response is innate to the tendon tissue and not reliant on a systemic inflammatory response. Therefore, we hypothesized that MRL/MpJ mice may also exhibit a more robust homeostatic regulation of tendon structure in response to mechanical loading. To assess this, MRL/MpJ and C57BL/6J flexor digitorum longus tendon explants were subjected to stress-deprived conditions in vitro for up to 14 days. Explant tendon health (metabolism, biosynthesis, and composition), matrix metalloproteinase (MMP) activity, gene expression, and tendon biomechanics were assessed periodically. We found a more robust response to the loss of mechanical stimulus in the MRL/MpJ tendon explants, exhibiting an increase in collagen production and MMP activity consistent with previous in vivo studies. This greater collagen turnover was preceded by an early expression of small leucine-rich proteoglycans and proteoglycan-degrading MMP-3, promoting efficient regulation and organization of newly synthesized collagen and allowing for more efficient overall turnover in MRL/MpJ tendons. Therefore, mechanisms of MRL/MpJ matrix homeostasis may be fundamentally different from that of B6 tendons and may indicate better recovery from mechanical microdamage in MRL/MpJ tendons. We demonstrate here the utility of the MRL/MpJ model in elucidating mechanisms of efficient matrix turnover and its potential to shed light on new targets for more effective treatments for degenerative matrix changes brought about by injury, disease, or aging.

Relationship between interferon-induced transmembrane protein 3 and matrix metalloproteinase-9 gene polymorphisms in patients with hepatocellular carcinoma

BackgroundHepatocellular carcinoma originates from hepatocytes as a result of the effects of numerous genetic variations. Interferon-Induced Transmembrane protein 3 (IFITM3) is involved in the processes of cellular differentiation, apoptosis, cell adhesion, and immune cell regulation. Matrix Metalloproteinase-9 (MMP-9) are zinc dependent endopeptidases that cleave extracellular matrix contents and play an important role in the progression of cancer. ObjectiveThe study aimed to outline the key molecular biology progression in hepatocellular carcinoma and the relationship between hepatocellular cancer and genetic polymorphisms of IFITM3 and MMP-9. MethodsIn total 200 patients with hepatocellular carcinoma patients (n=100) and a control group with Hepatitis C virus (n=100) which collected randomly from the EL-Mansoura oncology center during the interval between June 2020 and October 2021. The expression of MMP-9 and the IFITM3 SNP was investigated. MMP-9 gene polymorphisms were estimated by using PCR-RFLP and IFITM3 gene was detected using DNA sequencing, ELISA was used to measure protein levels of MMP-9 and IFITM3. ResultsThe T allele of MMP-9 was more frequent among patients (n=121) than control subjects (n=71). The C allele of IFITM3 was more frequent among patients (n=112) than control subjects (n=83), polymorphisms of the genes linked to a high risk of disease development, patients of MMP-9 (TT genotype), odd ratio (OR) = 2.63, IFITM3 (CC genotype), OR= 2.43. ConclusionsWe found that the genetic polymorphisms of MMP-9 and IFITM3 are related to the occurrence and development of hepatocellular carcinoma. This study might be utilized in clinical diagnosis and therapy and to provide a baseline for prevention.

Single-nucleotide polymorphisms of matrix metalloproteinase genes are associated with graft fibrosis after kidney transplantation.

Further research needs to be conducted on the role of genetic variables in kidney transplantation fibrosis. In this study, we used next-generation sequencing (NGS) to examine the relationship between matrix metalloproteinase (MMP) genes and single-nucleotide polymorphisms (SNPs) in renal allograft fibrosis. This study comprised 200 patients, whose complete DNA samples were taken. The SNPs in MMP genes were identified using targeted NGS. Hardy-Weinberg equilibrium (HWE) and minor allele frequency (MAF) tests were conducted, followed by a linkage disequilibrium (LD) analysis. Finally, the SNPs and severity of kidney allograft fibrosis were evaluated using different inheritance models. In total, 41 MMP gene-related SNPs were identified using targeted sequencing, and 20 tagger SNPs were retained for further study. The general linear models (GLMs) revealed that sirolimus treatment had a substantial effect on kidney graft fibrosis. The multiple inheritance model analyses revealed that SNP rs9059 of the MMP9 gene was strongly associated with kidney graft fibrosis. The in-vitro experiments showed the MMP9 rs9509 mutation promotes the process of epithelial-mesenchymal transition (EMT) in the human kidney 2 (HK2) cells. The SNP rs9059 is associated with significant kidney allograft pathological changes by promoting EMT progression. Our findings provide insights into the etiology of renal allograft interstitial fibrosis and the MMP9 could be used as a potential treatment target in the future.

EF-24, a Curcumin Analog, Inhibits Cancer Cell Invasion in Human Nasopharyngeal Carcinoma through Transcriptional Suppression of Matrix Metalloproteinase-9 Gene Expression.

Cancer metastasis is a main cause of failure in treating subjects with nasopharyngeal carcinoma (NPC) and is frequently linked to high death rates. EF-24, an analog of curcumin, has exhibited many anti-cancer properties and enhanced bioavailability over curcumin. Nevertheless, the effects of EF-24 on the invasiveness of NPC are poorly understood. In this study, we demonstrated that EF-24 effectively inhibited TPA-induced motility and invasion responses of human NPC cells but elicited very limited cytotoxicity. In addition, the TPA-induced activity and expression of matrix metalloproteinase-9 (MMP-9), a crucial mediator of cancer dissemination, were found to be reduced in EF-24-treated cells. Our reporter assays revealed that such a reduction in MMP-9 expression by EF-24 was transcriptionally mediated by NF-κB via impeding its nuclear translocation. Further chromatin immunoprecipitation assays displayed that the EF-24 treatment decreased the TPA-induced interaction of NF-κB with the MMP-9 promoter in NPC cells. Moreover, EF-24 inhibited the activation of JNK in TPA-treated NPC cells, and the treatment of EF-24 together with a JNK inhibitor showed a synergistic effect on suppressing TPA-induced invasion responses and MMP-9 activities in NPC cells. Taken together, our data demonstrated that EF-24 restrained the invasiveness of NPC cells through the transcriptional suppression of MMP-9 gene expression, implicating the usefulness of curcumin or its analogs in controlling the spread of NPC.

Effect of combination treatment of protocatechuic acid with 5-fluorouracil and oxaliplatin on colon cancer Caco-2 cell line

Among the most common antitumor drugs used in the treatment of colon cancer are 5-fluorouracil and oxaliplatin (5-FU and OXA). However, both these drugs have many side effects, and hence there is a need for new treatment\approach to reduce the side effects aas well as drug concentration. In this context, here, we investigated the effect of addition of protocatechuic acid (PCA) onto either monotherapies or combination therapies of 5-FU and OXA on the human colon cancer (Caco-2) cell line. In addition, we did evaluate the synergistic effect of PCA with 5-FU and OXA. Further, we determined the suppressive effects of different doses of PCA alone or in combination with 5-FU/OXA on cell proliferation after 24 and 48 hours. We identified a suppressive effect of PCA on cell viability at 48 h starting from the dose of 50 µM Matrix metalloproteinase-2 (MMP-2) and MMP-9 gene expression levels and apoptotic effects showed significant increases and decreases depending on the dose and time applied in the experimental groups. The highest synergistic activity was seen at 2:1 concentration of 5-FU+ PCA. Our findings indicate the presence of the cytotoxic and apoptotic effects of PCA in Caco-2 cells at 48 h, increasing with a dose- and time-dependent manner.

Identification of biomarkers and prediction of upstream miRNAs in diabetic nephropathy.

To explore biomarkers of diabetic nephropathy (DN) and predict upstream miRNAs. The data sets GSE142025 and GSE96804 were obtained from Gene Expression Omnibus database. Subsequently, common differentially expressed genes (DEGs) of renal tissue in DN and control group were identified and protein-protein interaction network (PPI) was constructed. Hub genes were screened from in DEGs and made an investigation on functional enrichment and pathway research. Finally, the target gene was selected for further study. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of target gene and predicted its upstream miRNAs. 130 common DEGs were obtained through analysis, and 10 Hub genes were further identified. The function of Hub genes was mainly related to extracellular matrix (ECM), collagen fibrous tissue, transforming growth factor (TGF) -β, advanced glycosylation end product (AGE) -receptor (RAGE) and so on. Research showed that the expression level of Hub genes in DN group was significantly higher than that in control group. (all P<0.05). The target gene matrix metalloproteinase 2 (MMP2) was selected for further study, and it was found to be related to the fibrosis process and the genes regulating fibrosis. Meanwhile, ROC curve analysis showed that MMP2 had a good predictive value for DN. miRNA prediction suggested that miR-106b-5p and miR-93-5p could regulate the expression of MMP2. MMP2 can be used as a biomarker for DN to participate in the pathogenesis of fibrosis, and miR-106b-5p and miR-93-5p may regulate the expression of MMP2 as upstream signals.

Integrative analyses of RNA-seq and ChIP-seq Reveal MITF as a Target Gene of TFPI-2 in MDA231 Cells.

Breast cancer is the most prevalent cancer in female patients worldwide. Tissue factor pathway inhibitor 2 (TFPI-2) is identified as an important tumor suppressor in various cancers. Recent studies have shown that TFPI-2 translocates into the nucleus, where it modulates the transcription of the matrix metalloproteinase-2 (MMP-2) gene. However, its biological role and molecular mechanisms in the progression of breast cancer remain unclear. In this study, we identified 5125 differentially expressed genes (DEGs) from RNA sequencing (RNA-seq) in TFPI-2-overexpressing MDA231 cells compared with control cells. Gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) analysis shown that cell cycle, cell differentiation, proteoglycans in cancer, and pathways associated with cancer were highly enriched in downregulated DEGs. Integration of the RNA-seq and ChIP-sequencing (ChIP-seq) data identified 73 genes directly controlled by TFPI-2 in MDA231 cells. Among them, melanocyte inducing transcription factor (MITF) gene expression was repressed by TFPI-2, which was further verified by a luciferase reporter assay and ChIP-quantitative PCR. Our study provides evidence of a novel role of TFPI-2 in human breast cancer involving targeting of the MITF.

Associations of Polymorphic Loci of Matrix Metalloproteinase Genes with the Development of Breast Cancer in Women Central Chernozem Region of Russia

The association of polymorphic variants of matrix metalloproteinase (MMP) genes with breast cancer (BC) in women of the Central Chernozem region of Russia, taking into account the presence/absence of a burdened family history, was studied. The study included 358 patients with breast cancer (68 patients had a burdened family history) and 746 women of the control group. Genotyping of 10 polymorphic loci of MMP genes (rs1799750 MMP1; rs243865 MMP2; rs679620 MMP3; rs1940475 MMP8; rs17576, rs17577, rs3918242, rs2250889, rs3787268, rs3918249 MMP9) was performed. To study the associations of polymorphic variants of MMP genes with breast cancer in two subgroups, taking into account the presence/absence of a burdened family history (the control group for these two subgroups was the same), the method of logistic regression analysis was used. It was revealed that a low risk of developing the disease among women with a burdened family history is associated with polymorphisms rs243865 MMP2 (OR = 0.53–0.54, pperm ≤ 0.03) and rs2250889 MMP9 (OR = 0.36–0.37, pperm ≤ 0.04). In women without burdened heredity, an increased risk of developing breast cancer is associated with rs3787268 MMP9 (OR = 2.16, pperm = 0.03) and haplotypes of polymorphic loci of the MMP9 gene (pperm ≤ 0.05): CA rs3918249–rs17576 (OR = 2.15), CCA rs3918242–rs3918249–rs17576 (OR = 1.69), CCAG rs3918242–rs3918249–rs17576–rs3787268 (OR = 1.69), CAGCG rs3918249–rs17576–rs3787268–rs2250889–rs17577 (OR = 3.06). Three haplotypes are associated with a low risk of breast cancer in women without burdened heredity: GG rs17576–rs3787268 (OR = 0.60), GGC rs17576–rs3787268–rs2250889 (OR = 0.63), and CGG rs3918249–rs17576–rs3787268 (OR = 0.62).

The promoter methylations of the autoimmune regulator (AIRE) gene and matrix metalloproteinase-3 (MMP-3) gene may have a role in gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) is characterized by new-onset glucose intolerance and is most common in the second and third trimesters of pregnancy. Epigenetic modifications regulate glucose and its cellular interactions with metabolic pathways. Emerging evidence suggests that epigenetic changes contribute to the pathophysiology of GDM. Since these patients have high glucose levels, the metabolic profiles of the fetus and the mother can affect these epigenetic changes. Therefore, we aimed to examine the potential alterations in the methylation profiles of three gene promoters: the autoimmune regulator (AIRE) gene, matrix metalloproteinase-3 (MMP-3), and calcium voltage-gated channel subunit alpha1 G (CACNA1G). A total of 44 patients diagnosed with GDM and 20 controls were involved in the study. DNA isolation and bisulfite modification were performed from peripheral blood samples of all patients. Then, the promoter methylation status of the AIRE, MMP-3, and CACNA1G genes was determined by methylation-specific polymerase chain reaction (PCR) methylation-specific (MSP). Our results demonstrated that the methylation status of AIRE and MMP-3 changed to unmethylated in the GDM patients compared to healthy pregnant women (p<0.001). However, CACNA1G promoter methylation status failed to show a significant change between experimental groups (p>0.05). Our results indicated that AIRE and MMP-3 are the genes affected by epigenetic modification, which could be one of the causes of the long-term metabolic effects in maternal and fetal health and can be a target for prevention, diagnosis, or treatment for GDM in future studies.

Associations of Polymorphic Loci of Matrix Metalloproteinase Genes with Breast Cancer in Women of the Central Black Earth Region of Russia

Associations of Polymorphic Loci of Matrix Metalloproteinase Genes with Breast Cancer in Women of the Central Black Earth Region of Russia

Polymorphisms of the Matrix Metalloproteinase Genes are Associated with Acute Ischemic Stroke in Chinese Han Population.

Studies have shown that matrix metalloproteinase (MMP-2,3,9) plays an important role in the pathologic process of ischemic stroke (IS). The aim of this study was to investigate the relationship between C1306T, 1612-5A/6A, C-1562T polymorphisms of MMP-2,3,9 genes and IS in Chinese Han population. The polymorphisms of MMP-2(C1306T), -3(1612-5A/6A), -9(C-1562T) gene were detected by PCR-RFLP and SNaPshot sequencing. Then, stratified analysis was used to study the relationship between IS subtypes and MMP-2,3,9 polymorphisms. For the MMP-2 gene C1306T polymorphism, TT genotype and T allele were significantly associated with a reduced risk of IS (P = 0.015, P = 0.003, respectively). T allele was significantly associated with a reduced risk of small artery occlusion (SAO) subtype compared with the control group (P = 0.012, OR = 0.550, 95% CI = 0.065-1.291). For the MMP-3 gene-1612 (5A/6A) polymorphism, 5A/5A genotype was significantly increased in the IS group (P = 0.011, OR = 0.370, 95% CI = 0.168-0.814), especially in the large-artery atherosclerosis (LAA) subtype (P = 0.001, OR = 2.345) as compared to the control group. Our study suggested that the T allele of MMP-2 may be a protective factor of IS, especially in SAO subtype, while the 5A/5A gene of MMP-3 may increase the risk of IS, especially in LAA subtype in Chinese Han population.

Abstract B038: Remodeling the extracellular matrix environment and establishing distinct immune cell profiles enables the formation of brain metastasis in non-small cell lung cancer adenocarcinoma

Abstract Dissemination of tumor cells from the primary site of the lung to distant sites in non-small cell lung cancer (NSCLC) is poorly understood. With distinct mutation signatures in patients, altering pathways is critical to preventing tumor cell dissemination to distant sites. Towards this, we characterized 21 "trios" patient-matched brain metastases (BrM), primary tumors (LT), and normal samples (LN), revealing a pattern of extracellular matrix remodeling which suppressed immune cell infiltration. Our objectives were to (i) determine if clinically sampled primary lung tumors encode gene expression patterns distinct from normal tissue, suggestive of increased tumor invasiveness including brain metastasis, and (ii) determine the biologic underpinning of immune-suppressive information carryover from primary lung tumor to brain metastasis. Transformation of the normal lung cells to primary tumor cells suggested a breakdown in ECM axes primarily governed by upregulation of collagen (COL1A1, COL1A2, COL3A1, COL10A1), fibronectin (FN1), and matrix metalloproteinases (MMP7, MMP9, MMP11, MMP12, MMP13, MMP14) genes. Collectively, the primary tumor cells used MMPs to break down the main components of lung ECM and reorganized collagen and fibronectin in the tumor microenvironment to facilitate migration and invasion. Migration and subsequent cell adhesion in the brain microenvironment revealed upregulation of neural cell adhesion molecule (NCAM1) and phosphoprotein 1 (SPP1). These changes in ECM profiles between LT and BrM signify primary tumor cells' adaptation in the brain microenvironment. Further computational deconvolution revealed reduced HLA class II gene expression (HLA-DMA, HLA-DOB, HLA-DPA1, HLA-DPB1, HLA-DQA2, HLA-DQB2, HLA-DRA) correlated with lower CIBERSORTx enrichment scores of immune cell populations in BrM. Overall, using the matched sequencing information, we will present the underlying signaling gradients that dictate dissemination of primary lung tumor cells to the brain environment in the background of altered ECM environment using cellular models harboring EGFR, KRAS, and ALK driver mutations. Citation Format: Albert R. Wang, Jared Brown, Andrew Baschnagel, Darya Buehler, Christina Kendziorski, Gopal Iyer. Remodeling the extracellular matrix environment and establishing distinct immune cell profiles enables the formation of brain metastasis in non-small cell lung cancer adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr B038.

Downregulation of COL12A1 and COL13A1 by a selective EP2 receptor agonist, omidenepag, in human trabecular meshwork cells.

Omidenepag isopropyl (OMDI) is an intraocular pressure (IOP)-lowering drug used to treat glaucoma. The active form of OMDI, omidenepag (OMD), lowers elevated IOP, the main risk factor for glaucoma, by increasing the aqueous humor outflow; however, a detailed understanding of this mechanism is lacking. To clarify the IOP-lowering mechanism of OMDI, the effects of OMD on the mRNA expression of the extracellular matrix, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs) were evaluated in human trabecular meshwork cells. Under 2D culture conditions, the mRNA expression of FN1, COL1A1, COL1A2, COL12A1, and COL13A1 decreased in a concentration-dependent manner after 6 or 24 h treatment with 10 nM, 100 nM, and 1 μM OMD, while that of COL18A1 decreased after 6 h treatment with 1 μM OMD. Significant changes in expression were observed for many MMP and TIMP genes. Under 3D culture conditions, the extracellular matrix-related genes COL12A1 and COL13A1 were downregulated by OMD treatment at all three concentrations. Under both 2D and 3D culture conditions, COL12A1 and COL13A1 were downregulated following OMD treatment. Reduction in the extracellular matrix contributes to the decrease in outflow resistance, suggesting that the downregulation of the two related genes may be one of the factors influencing the IOP-lowering effect of OMDI. Our findings provide insights for the use of OMDI in clinical practice.

Association of Matrix Metalloproteinase-3 (MMP-3) Gene Methylation Status with the Risk of Developing Achilles Tendonitis: A Preliminary Study

Association of Matrix Metalloproteinase-3 (MMP-3) Gene Methylation Status with the Risk of Developing Achilles Tendonitis: A Preliminary Study

The effect of ascorbic acid on histopathological, biochemical, pharmacological, and immunological toxicity of chronic lead acetate exposure on the spleen in a rat model

Objective To evaluate the effect of vitamin C on histopathological, biochemical, and immunotoxicity of chronic lead exposure in the spleen of a rat model.Methods The rats were divided into five groups of 10 rats each: group I received normal saline orally as a control group; groups II and III received lead acetate for 4 and 8 weeks, respectively; and groups IV and V received lead acetate and vitamin C for 4 and 8 weeks, respectively. The spleen was excised and processed for light, electron microscopic, histopathological, and biochemical analyses. Quantitative assessments of matrix metalloproteinase-2 (MMP-2), MMP-9, interleukin-2 (IL-2), IL-6, and tumor necrosis factor-alpha gene expressions were performed by real-time PCR.Results The examination of control and vitamin C with lead acetate supplemented groups revealed normal splenic architecture. In contrast, the spleen of lead-intoxicated groups exhibited degenerative changes in the spleen, with a significantly decreased expression of IL-2, glutathione peroxidase, superoxide dismutase, and hemoglobin (P<0.05), with significantly increased proinflammatory cytokine (IL-6 and tumor necrosis factor-alpha) expressions, concomitantly with increased oxidative products (malondialdehyde) and protease enzymes (MMP-2 and MMP-9) in the spleen tissues. The coadministration of vitamin C with lead for 4 weeks markedly resolved these changes.Conclusion This study may specify the efficiency of vitamin C in lead toxicity prevention in the spleen, represented by the reduced splenic harmful changes produced by lead administration.

Abstract PR006: The racial disparity-linked gene CRYβB2 alters cell adhesion and tumor microenvironment to promote proliferation and invasion in triple-negative breast cancer

Abstract Background: Clinical and epidemiological data showed that African American (AA) women suffer higher incidence and mortality rates due to triple-negative breast cancer (TNBC) compared to European American (EA) women. These disparate disease outcomes are attributed to multiple factors including socioeconomic status, access to health care, and earlier disease onset. However, a growing body of evidence also shows the existence of different gene expression signatures and tumor microenvironments between these patient racial groups. We and others identified CRYβB2 as a cancer racial disparity gene that has higher expression in AA patients with breast, colon, and prostate cancers. We previously demonstrated, via in vitro and in vivo studies, that CRYβB2 promotes proliferation, metastasis, and IL-6 signaling. The objective of the current research is to identify the cellular and molecular mechanisms downstream of CRYβB2 leading to its tumor-promoting phenotypes. Methods: CRISPR/Cas9 and lentiviral knock-in gene editing tools were used to create TNBC cell lines with CRYβB2 knockout and overexpression respectively. The functional consequences of CRYβB2 were assessed in vitro using colony formation, tumor spheroid invasion, and collagenase activity assays. Additionally, tumor spheroids were grown and RNA sequencing (RNA-seq) was performed to assess the effect of CRYβB2 on gene expression. Results: Herein, functional assays showed that CRYβB2 promotes proliferation and invasion in vitro in both AA and EA cell lines. RNA-seq and Gene Ontology enrichment analyses revealed that CRYβB2 alters key pathways such as cell adhesion and extracellular matrix (ECM) organization. RT-qPCR validation also showed that CRYβB2 inhibits the expression of a specific class of adhesion molecules known as protocadherins (PCDHs). Downregulation of PCDHs has been shown to enhance tumor progression and is a predictive marker for low survival in many cancer types. Indeed, data mining of publicly available data shows that three of the identified PCDHs (PCDHGB5, PCDHGB6 and PCDHGB7) have significantly lower expression levels in breast cancer compared to normal tissue and their low expression leads to low overall survival. Furthermore, RNA-seq data showed that CRYβB2 promotes the expression of multiple matrix metalloproteinase (MMP) genes. This was functionally validated using collagenase activity assay where CRYβB2 enhanced collagen degradation in vitro. Conclusions: Our findings provide a mechanistic explanation for the role of CRYβB2 in driving disparate clinical outcome among AA and EA TNBC patients. We demonstrate a paradigm where high CRYβB2 expression promotes tumor progression by suppressing cell adhesion molecules and increasing ECM degradation to enhance tumor growth, invasion, and metastasis. Disclaimer: This work was prepared while Jodie Fleming was employed at NCCU. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. Citation Format: Amr A. Waly, London Harper, Jodie M. Fleming, Lindsey M. Costantini. The racial disparity-linked gene CRYβB2 alters cell adhesion and tumor microenvironment to promote proliferation and invasion in triple-negative breast cancer [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PR006.

Bioinformatic analysis of MMP family members in GBM

Glioblastoma Multiforme (GBM) is the most serious type of glioma with the highest tumorigenesis rate. Studies have found that the increased expression levels of matrix metalloproteinases (MMPs) contribute to tumor cell proliferation, anti-apoptosis, disseminated tumor growth, neovascularization, and anti-tumor immune surveillance inhibition. The prognostic value of the MMP gene family in different cancer types is becoming more apparent. As a result, the current study sought to perform a complete bioinformatics analysis to clarify the prognostic value of this family in GBM. Consequently, univariate, multivariate Cox, and time-dependent receiver operating curve analyses were performed on the data. Further, UALCAN, GEPIA2, cBioPortal, GeneMANIA, Enrichr, and TIMER were used for various analyses. We found that the mRNA expressions of MMP1/2/7/9/11/14/15/16/25 were all meaningfully upregulated in GBM relative to normal cases. Additionally, the overexpression of MMP2/10/11/13 and MMP1/9/10/14/19 was substantially linked to shorter overall survival (OS) and recurrence-free survival (RFS), respectively. Likewise, the high levels of MMP21 and MMP23B were correlated with longer OS and RFS, respectively. Our results provide new ideas for the role of MMPs in GBM, which may advance the use of MMPs as prognostic biomarkers in clinical applications.