Extracellular Matrix Turnover Research Articles

Abstract 15246: Sex-Biased Preoperative Circulating MicroRNA Levels in Patients Who Developed Postoperative Heart Failure With Preserved Ejection Fraction

Introduction: Acute postoperative heart failure (PoHF) is encountered in 20-35% of elderly patients after cardiac surgery but predictors are not well defined. Circulating microRNAs (miRNAs) predict HF or postoperative atrial fibrillation, yet, their role in identifying those at risk for PoHF with preserved LV function and gender-related differences is not known. Hypothesis: miRNAs involved in CVD can identify patients who develop PoHF in gender-specific manner. Methods: Preoperative blood samples from patients with preserved LV function undergoing cardiac surgery with no prior history of HF, supraventricular, or ventricular tachycardia was used for RNA isolation. Differences in relative plasma levels of miRNA between PoHF vs. No-PoHF were assessed using qPCR. Univariate and multiple logistic regressions were performed to assess risk factors for PoHF. Results: Out of 68 patients (mean age 68.3±12.4), 13 (19%) developed PoHF (54% males; mean age 64.1±15.1; p=0.23), while 55 (51% males; mean age 69.3y ± 11.6) remained free of PoHF. Although all patients had preserved LV function, those who developed PoHF had lower LVEF, (52 vs 58; P<0.05). No significant differences were observed in the prevalence of hypertension, diabetes, myocardial infarction, heart failure, stroke, COPD, or the use of cardiac medications. Out of 13 miRNAs analyzed, stepwise discriminant analysis showed miR-423 (11-fold), -187 (6-fold), -26a2 (4-fold), and -15b (-1.9-fold) were significantly (p<0.05) altered in patients who developed PoHF. When miRNAs were assessed for gender differences, levels of only four miRNAs [miR-423(P<0.01), -23b2(p<0.05), -15b1(p<0.01), and -29b (p<0.05)] principally involved in TGFβ1-SMAD 2/3 regulated extracellular matrix (ECM) turnover pathways were significantly altered in male but not in female patients who developed PoHF compared to No-PoHF group. Conclusions: The differences in preoperative miRNA expression in pathways involved in ECM turnover in male patients who develop PoHF is suggestive of pathophysiological substrate differences between males and females and identifies a group at risk of PoHF with preserved LV function.

Abstract 14063: Circulating Levels of Collagen Formation and Degradation Biomarkers Are Prognostic for Risk of Hospitalization and Mortality in Patients With HFpEF - Results From the TRAINING-HF Cohort

Introduction: heart failure with preserved ejection fraction (HFpEF) is projected to become the dominant form of heart failure, and currently there are no viable treatment options to reduce mortality. Extracellular matrix turnover plays an important role in the pathology of heart failure. Our aim was to evaluate the association between adverse outcome in patients with HFpEF and circulating levels of novel collagen formation and degradation biomarkers in samples from HFpEF patients. Methods: fifty-nine symptomatic HFpEF patients were included for analysis. Biomarkers of formation of type III, VI, VIII, XXVIII collagens (PRO-C3, PRO-C6, PRO-C8 and PRO-C28) and degradation of type I, III and IV collagens (C1M, C3M and C4M) were assessed in baseline serum by means of competitive ELISA. Three models were generated by logistic regression with forward selection in relation to readmission and mortality, with the aim of assessing prognostic abilities for either outcome. The models were compared head to head using ROC curve analyses. Model 1 (CONF): age, sex, HDL, LDL, systolic blood pressure, diabetes status, BMI and smoking status. Model 2 (FORM): collagen formation biomarkers. Model 3 (DEGR): collagen degradation biomarkers. Results: all three models significantly predicted readmission (CONF: P<0.0001, FORM: P=0.0029, DEGR: P=0.0026). For mortality, CONF and FORM were significantly predictive, while DEGR was borderline significant (CONF: P=0.0097, FORM: P<0.0001, DEGR: P=0.059). ROC curves for mortality showed a significant difference between FORM and CONF (P=0.017), while no significant difference was seen when comparing CONF and DEGR. The CONF model was mainly driven by LDL (P=0.02) in relation to readmission, whereas no CONF variables were significant for mortality. No variables in the FORM or DEGR models were independently significant for readmission, but PRO-C6 and PRO-C8 were significant in the FORM model in relation to mortality (both P=0.028). Conclusion: in stable symptomatic HFpEF patients, turnover of collagen predicts outcome. Moreover, formation of collagens provides significant information on risk, suggesting a potential role for collagen biomarkers in stratifying HFpEF patients at higher risk of adverse outcomes.

Tumor cell MT1-MMP is dispensable for osteosarcoma tumor growth, bone degradation and lung metastasis

The membrane-anchored matrix metalloprotease MT1-MMP is a potent collagenolytic enzyme with a well-established role in extracellular matrix turnover and cellular invasion into collagen-rich tissues. MT1-MMP is highly expressed in various types of cancer and has been demonstrated to be directly involved in several stages of tumor progression, including primary tumor growth, angiogenesis, invasion and metastasis. Osteosarcoma is the most common type of primary bone cancer. This disease is characterized by invasive tumor growth, leading to extensive bone destruction, and metastasis to the lungs. The tumor cells in human osteosarcoma display a strong expression of MT1-MMP, but the role of MT1-MMP in osteosarcoma progression is currently unknown. In this study, we investigated the role of MT1-MMP during various stages of osteosarcoma development. We utilized an optimized orthotopic murine osteosarcoma model and human osteosarcoma cells in which the MT1-MMP gene was knocked out using CRISPR/Cas9. We observed a strong expression of MT1-MMP in wildtype cells of both primary tumors and lung metastases, but, surprisingly, MT1-MMP deficiency did not affect primary tumor growth, bone degradation or the formation and growth of lung metastases. We therefore propose that, unlike findings reported in other cancers, tumor-expressed MT1-MMP is dispensable for all stages of osteosarcoma progression.

Chronic Obstructive Pulmonary Disease

Chronic obstructive pulmonary disease (COPD) is a leading cause of disability and death worldwide. This edition reviews the epidemiology and etiologies of COPD, including the gender effects, racial differences, and more recently identified genetic factors associated with this condition. It details the many pathogenetic mechanisms thought to be associated with this disease state, such as increased airway inflammation and turnover of extracellular matrix. There is a detailed discussion about diagnosis, classification, and the therapeutic options available for both stable disease and acute exacerbations. The recent evidence supporting various treatments, such as vaccinations, inhaled bronchodilators, inhaled corticosteroids, oral corticosteroids, antibiotics, supplemental oxygen, pulmonary rehabilitation, and surgery, is reviewed in depth. There is further evaluation of experimental approaches, such as bronchoscopic lung reduction procedures and the use of extracorporeal carbon dioxide removal for hypercapnic respiratory failure. The many complications associated with COPD are described, acknowledging that evidence continues to suggest that COPD has a significant systemic component associated with increased rates of psychiatric illness, cardiovascular disease, osteoporosis, and skeletal muscle dysfunction along with lung cancer. Overall this text serves as an excellent evidence-based guide to better understand, diagnose, and manage COPD and its array of associated complications. This review contains 6 highly rendered figures, 6 tables, and 241 references Key words: chronic obstructive pulmonary disease (COPD), COPD complications, COPD diagnosis, COPD management, COPD pathophysiology, Global Initiative for Chronic Obstructive Lung Disease (GOLD)

Chronic Obstructive Pulmonary Disease

Chronic obstructive pulmonary disease (COPD) is a leading cause of disability and death worldwide. This edition reviews the epidemiology and etiologies of COPD, including the gender effects, racial differences, and more recently identified genetic factors associated with this condition. It details the many pathogenetic mechanisms thought to be associated with this disease state, such as increased airway inflammation and turnover of extracellular matrix. There is a detailed discussion about diagnosis, classification, and the therapeutic options available for both stable disease and acute exacerbations. The recent evidence supporting various treatments, such as vaccinations, inhaled bronchodilators, inhaled corticosteroids, oral corticosteroids, antibiotics, supplemental oxygen, pulmonary rehabilitation, and surgery, is reviewed in depth. There is further evaluation of experimental approaches, such as bronchoscopic lung reduction procedures and the use of extracorporeal carbon dioxide removal for hypercapnic respiratory failure. The many complications associated with COPD are described, acknowledging that evidence continues to suggest that COPD has a significant systemic component associated with increased rates of psychiatric illness, cardiovascular disease, osteoporosis, and skeletal muscle dysfunction along with lung cancer. Overall this text serves as an excellent evidence-based guide to better understand, diagnose, and manage COPD and its array of associated complications. This review contains 6 highly rendered figures, 6 tables, and 241 references Key words: chronic obstructive pulmonary disease (COPD), COPD complications, COPD diagnosis, COPD management, COPD pathophysiology, Global Initiative for Chronic Obstructive Lung Disease (GOLD)

Altered balance between collagen formation and degradation after successful direct-acting antiviral therapy of chronic hepatitis C.

The effect of direct-acting antiviral (DAA) therapy on extracellular matrix (ECM) turnover, a prominent feature of chronic hepatitis C (CHC), is unknown. ECM protein degradation and formation generate fragments reflecting the tissue turnover balance when quantified in the blood. PRO-C3 and PRO-C4 reflect type III and IV collagen formation; C3M and C4M are degradation markers of type III and IV. We aimed to assess the markers' dynamics with DAA therapy in CHC patients. Plasma PRO-C3, PRO-C4, C3M and C4M were assessed before, during and up till one year after 12-24weeks of DAA therapy in 77 CHC patients with advanced fibrosis (n=14) or cirrhosis (n=63). Liver stiffness was evaluated using transient elastography. PRO-C3, C3M and C4M levels decreased significantly (P<.00001) while PRO-C4 was unchanged (P=.20) during the study period. There was a steep decrease in the PRO-C3/C3M ratio during DAA therapy and follow-up (P<.02). The PRO-C4/C4M ratio was unchanged (P>.27). The dynamics of the collagen markers behaved similarly between patients with advanced fibrosis and cirrhosis. However, the cirrhosis patients had >20% higher levels of C3M, PRO-C4 and C4M at all time points (P<.05). The collagen markers correlated with liver stiffness at baseline and follow-up.Markers of type III and IV collagen formation and degradation decreased during and after successful DAA therapy in CHC patients with advanced liver disease, and associated with disease severity. These results indicate an altered balance between collagen formation and degradation after viral clearance suggesting favourable effects on liver fibrosis.

Collagenases MMP-1, MMP-13, and Tissue Inhibitors TIMP-1, TIMP-2: Their Role in Healthy and Complicated Pregnancy and Potential as Preeclampsia Biomarkers—A Brief Review

Extracellular matrix (ECM) turnover is characterized by a unique balance between matrix metalloproteinases’ degradation activity and their natural inhibition by collagen specific tissue inhibitors. Human uterine ECM is a complex structure, majorly consisting of proteins as fibrillar collagen types I and III, fibronectin, and laminin. Collagenases are enzymes from the matrix metalloproteinases’ family, which are predominantly involved in fibrillar collagen types I and III degradation. They are mainly represented by matrix metalloproteinase-1, -13 (MMP-1, -13), naturally inhibited by tissue inhibitors (TIMP-1, -2). The collagen structure of the uterus has been shown to be impaired in women with preeclampsia. This is a result of MMPs/TIMPs dysregulation interplay. This review article summarizes the actual available research data in the literature about the role of MMP-1, MMP-13 and TIMP-1, and TIMP-2 in collagen types I and III turnover in healthy and complicated pregnancy. Their potential use as circulating markers for diagnosis, prognosis, and monitoring of the development of preeclampsia is discussed as well.

Genetic Susceptibility and Protein Expression of Extracellular Matrix Turnover-Related Genes in Oral Submucous Fibrosis.

Betel quid (BQ) chewing increased the risk of oral cancer and oral submucous fibrosis (OSMF), an oral premalignant disorder (OPMD) with malignant transformation potential. BQ components such as areca nut (AN), trauma by coarse AN fiber, catechin, copper, alkaloids, stimulated reactive oxygen species (ROS), inflammation and cytotoxicity are suggested to be the contributing factors. They may induce tissue inflammation, proliferation of fibroblasts and collagen deposition, myofibroblast differentiation and contraction, collagen cross-links and inhibit collagen phagocytosis, finally leading to the development of OSMF and oral cancer. These events are mediated by BQ components-induced changes of extracellular matrix (ECM) turnover via regulation of TGF-β1, plasminogen activator inhibitor-1 (PAI-1), cystatin, lysyl oxidase (LOX) and tissue inhibitors of metalloproteinases (TIMPs) and metalloproteinases (MMPs). Genetic susceptibility is also involved in these disease processes. Further understanding the molecular mechanisms of BQ-induced OSMF and oral cancer can be helpful for future disease prevention and treatment.

Associations of TIMP-3 Genetic Polymorphisms with EGFR Statuses and Cancer Clinicopathologic Development in Lung Adenocarcinoma Patients.

Lung adenocarcinoma (LADC) is a major subtype of lung cancer, particularly among populations of East Asia. The epidermal growth factor receptor (EGFR) is the most frequently mutated oncogene promoting LADC progression and can serve as a therapeutic target in LADC. The tissue inhibitor of metalloproteinases (TIMP)-3 is a major regulator of extracellular matrix turnover via targeting of matrix metalloproteinases (MMPs), and thus, plays a critical role in tumor development and progression. The purpose of this study was to investigate potential associations among TIMP-3 genetic polymorphisms, EGFR statuses, and cancer clinicopathologic development in patients with LADC. In this study, 277 LADC patients with different EGFR statuses were recruited to dissect the allelic discrimination of TIMP-3 -1296 T>C (rs9619311), TIMP3 249T>C (rs9862), and TIMP3 261C>T (rs11547635) polymorphisms using a TaqMan allelic discrimination assay. Our data showed that compared to those LADC patients with wild-type CC homozygotes of TIMP-3 rs9862, patients harboring TT homozygotes of rs9862 were at a higher risk of developing mutant EGFR (adjusted odds ratio (AOR) = 2.530; 95% confidence interval (CI): 1.230–5.205; p = 0.012), particularly the EGFR L858R point mutation (AOR = 2.975; 95% CI: 1.182–7.488; p = 0.021). Moreover, we observed that TIMP-3 TT homozygotes of rs9862 were correlated with the incidence of EGFR mutations in patients with a smoking habit (p = 0.045). Within male patients harboring a mutant EGFR, TIMP-3 rs9862 T (CT+TT) allele carriers were at higher risk of developing an advanced stage (p = 0.025) and lymph node metastasis (p = 0.043). Further analyses of clinical datasets revealed correlations of TIMP-3 expression with a favorable prognosis in patients with LADC. In conclusion, the data suggest that TIMP-3 rs9862 polymorphisms may contribute to identify subgroups of lung cancer patients at high risk for tumor progression, among carriers of LADC-bearing mutant EGFR.

Prediction of severity and subtype of fibrosing disease using model informed by inflammation and extracellular matrix gene index.

Fibrosis is a chronic disease with heterogeneous clinical presentation, rate of progression, and occurrence of comorbidities. Systemic sclerosis (scleroderma, SSc) is a rare rheumatic autoimmune disease that encompasses several aspects of fibrosis, including highly variable fibrotic manifestation and rate of progression. The development of effective treatments is limited by these variabilities. The fibrotic response is characterized by both chronic inflammation and extracellular remodeling. Therefore, there is a need for improved understanding of which inflammation-related genes contribute to the ongoing turnover of extracellular matrix that accompanies disease. We have developed a multi-tiered method using Naïve Bayes modeling that is capable of predicting level of disease and clinical assessment of patients based on expression of a curated 60-gene panel that profiles inflammation and extracellular matrix production in the fibrotic disease state. Our novel modeling design, incorporating global and parametric-based methods, was highly accurate in distinguishing between severity groups, highlighting the importance of these genes in disease. We refined this gene set to a 12-gene index that can accurately identify SSc patient disease state subsets and informs knowledge of the central regulatory pathways in disease progression.

Emerging Technologies and Platforms for the Immunodetection of Multiple Biochemical Markers in Osteoarthritis Research and Therapy.

Biomarkers, especially biochemical markers, are important in osteoarthritis (OA) research, clinical trials, and drug development and have potential for more extensive use in therapeutic monitoring. However, they have not yet had any significant impact on disease diagnosis and follow-up in a clinical context. Nevertheless, the development of immunoassays for the detection and measurement of biochemical markers in OA research and therapy is an active area of research and development. The evaluation of biochemical markers representing low-grade inflammation or extracellular matrix turnover may permit OA prognosis and expedite the development of personalized treatment tailored to fit particular disease severities. However, currently detection methods have failed to overcome specific hurdles such as low biochemical marker concentrations, patient-specific variation, and limited utility of single biochemical markers for definitive characterization of disease status. These challenges require new and innovative approaches for development of detection and quantification systems that incorporate clinically relevant biochemical marker panels. Emerging platforms and technologies that are already on the way to implementation in routine diagnostics and monitoring of other diseases could potentially serve as good technological and strategic examples for better assessment of OA. State-of-the-art technologies such as advanced multiplex assays, enhanced immunoassays, and biosensors ensure simultaneous screening of a range of biochemical marker targets, the expansion of detection limits, low costs, and rapid analysis. This paper explores the implementation of such technologies in OA research and therapy. Application of novel immunoassay-based technologies may shed light on poorly understood mechanisms in disease pathogenesis and lead to the development of clinically relevant biochemical marker panels. More sensitive and specific biochemical marker immunodetection will complement imaging biomarkers and ensure evidence-based comparisons of intervention efficacy. We discuss the challenges hindering the development, testing, and implementation of new OA biochemical marker assays utilizing emerging multiplexing technologies and biosensors.

Gene Expression Signatures of Synovial Fluid Multipotent Stromal Cells in Advanced Knee Osteoarthritis and Following Knee Joint Distraction.

Osteoarthritis (OA) is the most common musculoskeletal disorder. Although joint replacement remains the standard of care for knee OA patients, knee joint distraction (KJD), which works by temporarily off-loading the joint for 6–8 weeks, is becoming a novel joint-sparing alternative for younger OA sufferers. The biological mechanisms behind KJD structural improvements remain poorly understood but likely involve joint-resident regenerative cells including multipotent stromal cells (MSCs). In this study, we hypothesized that KJD leads to beneficial cartilage-anabolic and anti-catabolic changes in joint-resident MSCs and investigated gene expression profiles of synovial fluid (SF) MSCs following KJD as compared with baseline. To obtain further insights into the effects of local biomechanics on MSCs present in late OA joints, SF MSC gene expression was studied in a separate OA arthroplasty cohort and compared with subchondral bone (SB) MSCs from medial (more loaded) and lateral (less loaded) femoral condyles from the same joints. In OA arthroplasty cohort (n = 12 patients), SF MSCs expressed lower levels of ossification- and hypotrophy-related genes [bone sialoprotein (IBSP), parathyroid hormone 1 receptor (PTH1R), and runt-related transcription factor 2 (RUNX2)] than did SB MSCs. Interestingly, SF MSCs expressed 5- to 50-fold higher levels of transcripts for classical extracellular matrix turnover molecules matrix metalloproteinase 1 (MMP1), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), and tissue inhibitor of metalloproteinase-3 (TIMP3), all (p < 0.05) potentially indicating greater cartilage remodeling ability of OA SF MSCs, compared with SB MSCs. In KJD cohort (n = 9 patients), joint off-loading resulted in sustained, significant increase in SF MSC colonies’ sizes and densities and a notable transcript upregulation of key cartilage core protein aggrecan (ACAN) (weeks 3 and 6), as well as reduction in pro-inflammatory C–C motif chemokine ligand 2 (CCL2) expression (weeks 3 and 6). Additionally, early KJD changes (week 3) were marked by significant increases in MSC chondrogenic commitment markers gremlin 1 (GREM1) and growth differentiation factor 5 (GDF5). In combination, our results reveal distinct transcriptomes on joint-resident MSCs from different biomechanical environments and show that 6-week joint off-loading leads to transcriptional changes in SF MSCs that may be beneficial for cartilage regeneration. Biomechanical factors should be certainly considered in the development of novel MSC-based therapies for OA.

The Janus kinase 1/2 inhibitor baricitinib reduces biomarkers of joint destruction in moderate to severe rheumatoid arthritis

BackgroundTissue released blood-based biomarkers can provide insight into drug mode of action and response. To understand the changes in extracellular matrix turnover, we analyzed biomarkers associated with joint tissue turnover from a phase 3, randomized, placebo-controlled study of baricitinib in patients with active rheumatoid arthritis (RA).MethodsSerum biomarkers associated with synovial inflammation (C1M, C3M, and C4M), cartilage degradation (C2M), bone resorption (CTX-I), and bone formation (osteocalcin) were analyzed at baseline, and weeks 4 and 12, from a subgroup of patients (n = 240) randomized to placebo or 2-mg or 4-mg baricitinib (RA-BUILD, NCT01721057). Mixed-model repeated measure was used to identify biomarkers altered by baricitinib. The relationship between changes in biomarkers and clinical measures was evaluated using correlation analysis.ResultsTreatment arms were well balanced for baseline biomarkers, demographics, and disease activity. At week 4, baricitinib 4-mg significantly reduced C1M from baseline by 21% compared to placebo (p < 0.01); suppression was sustained at week 12 (27%, p < 0.001). Baricitinib 4-mg reduced C3M and C4M at week 4 by 14% and 12% compared to placebo, respectively (p < 0.001); they remained reduced by 16% and 11% at week 12 (p < 0.001). In a pooled analysis including all treatment arms, patients with the largest reduction (upper 25% quartile) in C1M, C3M, and C4M by week 12 had significantly greater clinical improvement in the Simplified Disease Activity Index at week 12 compared to patients with the smallest reduction (lowest 25% quartile).ConclusionBaricitinib treatment resulted in reduced circulating biomarkers associated with joint tissue destruction as well as concomitant RA clinical improvement.Trial registrationClinicalTrials.gov NCT01721057; date of registration: November 1, 2012

ERBB2 drives YAP activation and EMT-like processes during cardiac regeneration.

Cardiomyocyte loss after injury results in adverse remodelling and fibrosis, inevitably leading to heart failure. The ERBB2-Neuregulin and Hippo-YAP signalling pathways are key mediators of heart regeneration, yet the crosstalk between them is unclear. We demonstrate that transient overexpression of activated ERBB2 in cardiomyocytes (OE CMs) promotes cardiac regeneration in a heart failure model. OE CMs present an epithelial-mesenchymal transition (EMT)-like regenerative response manifested by cytoskeletal remodelling, junction dissolution, migration and extracellular matrix turnover. We identified YAP as a critical mediator of ERBB2 signalling. In OE CMs, YAP interacts with nuclear-envelope and cytoskeletal components, reflecting an altered mechanical state elicited by ERBB2. We identified two YAP-activating phosphorylations on S352 and S274 in OE CMs, which peak during metaphase, that are ERK dependent and Hippo independent. Viral overexpression of YAP phospho-mutants dampened the proliferative competence of OE CMs. Together, we reveal a potent ERBB2-mediated YAP mechanotransduction signalling, involving EMT-like characteristics, resulting in robust heart regeneration.

Targeted urine proteomics in lupus nephritis – a meta-analysis

ABSTRACT Background Proteomic approaches are central in biomarker discovery. While mass-spectrometry-based techniques are widely used, novel targeted proteomic platforms have enabled the high-throughput detection of low-abundance proteins in an affinity-based manner. Urine has gained growing attention as an ideal biofluid for monitoring renal disease including lupus nephritis (LN). Methods Pubmed was screened for targeted proteomic studies of LN urine interrogating ≥1000 proteins. Data from the primary studies were combined and a meta-analysis was performed. Shared proteins elevated in active LN across studies were identified, and relevant pathways were elucidated using ingenuity pathway and gene ontology analysis. Urine proteomic data was cross-referenced against renal single-cell RNAseq data from LN kidneys. Results Two high-throughput targeted proteomic platforms with capacity to interrogate ≥1000 proteins have been used to investigate LN urine. Twenty-three urine proteins were significantly elevated in both studies, including 10 chemokines, and proteins implicated in angiogenesis, and extracellular matrix turnover. Of these, Cathepsin S, CXCL10, FasL, ferritin, macrophage migration inhibitory factor (MIF), and resistin were also significantly elevated within LN kidneys. Conclusion Targeted urinary proteomics have uncovered multiple novel biomarkers for LN. Further validation in prospective cohorts and mechanistic studies are warranted to establish their clinical utility.

Modulation of Extracellular Matrix by Scrophularia striata Extract in Vitro: A Potential Antiscarring Agent

Background: Hypertrophic scars are the consequences of the aberration of normal wound healing. To date, therapeutic strategies for abnormal scarring have been unsuccessful. ‎‏The abnormal extracellular‎ matrix is one of the most important contributing factors to ‎hypertrophic scars. ‏Scrophularia striata has been used in Iranian folk medicine for the treatment of burn wounds. ‏The plant extract accelerates wound healing and attenuates scar formation. Objectives: The study was performed to investigate the effects of Scrophularia striata hydroalcoholic extract (SSE) on MMP1, MMP8, fibronectin, collagen type I, and total collagen produced by human skin fibroblasts in the culture medium. Methods: The effects of SSE on the expression of MMP1, MMP8, fibronectin, and collagen type I in human skin fibroblast (HSF) were evaluated using Q-PCR and Western blotting methods. In addition, the effect of SSE on the total collagen content was measured in cultured HSF using Red Sirius Kit. Results: SSE significantly induced the expression of MMP1 and suppressed the production of fibronectin at the mRNA and protein levels. The total collagen content was significantly lower in SSE-treated cells than in untreated cells. SSE did not have any significant effect on MMP8 and collagen type I expression. Conclusions: The results of this study revealed that SSE could modulate the extracellular matrix turnover and had the potential for the prevention and treatment of hypertrophic scars.

Association of endothelin-1 and cell surface adhesion molecules levels in patients with systemic sclerosis

Abstract Objectives Endothelin-1 (ET-1) has been implicated in the pathogenesis of inflammatory and fibrotic diseases, including systemic sclerosis. In addition to modulating vascular tone and extracellular matrix turnover, ET-1 up-regulates cell surface adhesion molecules–intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1).The aim of the study was to evaluate the diagnostic value of the detection of ET-1, VCAM-1 and ICAM-1 in the diagnosis of systemic sclerosis. Methods A total of 30 patients with systemic sclerosis from Vilnius University Hospital Santaros Clinics were included in the study. Serum levels of ICAM-1, VCAM-1 and ET-1 were assessed by enzyme immunoassay. Results ET-1 concentration was associated with VCAM-1 concentration (r=0.687; p&lt;0.001). No associations between ET-1 and ICAM-1 concentrations were detected. Depending on the duration of the disease no significant differences in the concentrations of ET-1, ICAM-1 and VCAM-1 were detected. Conclusions The results of this study indicated that ET-1 and VCAM-1 may be assessed together as markers of inflammation and the identification of patients at high risk for disease progression.

Comprehensive assessment of ECM turnover using serum biomarkers establishes PBC as a high-turnover autoimmune liver disease.

Background & AimsPrimary sclerosing cholangitis (PSC), primary biliary cholangitis (PBC) and autoimmune hepatitis (AIH) are phenotypically distinct autoimmune liver diseases that progress to cirrhosis and liver failure; however, their histological fibrosis distribution differs. We investigated the extracellular matrix (ECM) profiles of patients with PSC, PBC, and AIH to establish whether the diseases display differential patterns of ECM turnover.MethodsSerum samples were retrospectively collected from the UK (test cohort; PSC n = 78; PBC n = 74; AIH n = 58) and Norway (validation cohort; PSC n = 138; PBC n = 28; AIH n = 27). Patients with ulcerative colitis without liver disease (n = 194) served as controls. We assessed specific serological biomarkers of ECM turnover: type III and V collagen formation (PRO-C3, PRO-C5), degradation of type III and IV collagen (C3M, C4M), biglycan (BGM) and citrullinated vimentin (VICM).ResultsMost of the ECM markers showed elevated serum levels in PBC compared with PSC or AIH (p <0.01). PRO-C3 correlated well with liver stiffness and showed the most striking differences between advanced and non-advanced liver disease; several of the other ECM markers were also associated with stage. PRO-C3 and other ECM markers were inversely associated with ursodeoxycholic acid treatment response in PBC and remission in AIH. All ECM remodelling markers were significantly elevated (p <0.05) in patients with PSC, PBC, or AIH compared with ulcerative colitis.ConclusionsIn this first study comparing ECM turnover in autoimmune liver diseases, we found increased ECM turnover in PBC compared with either PSC or AIH. The study indicates that ECM remodelling is different in PSC, PBC, and AIH, suggesting differing opportunities for therapeutic intervention.Lay summaryThe level of scarring is linked to prognosis in autoimmune liver diseases such as primary sclerosing cholangitis, primary biliary cholangitis, and autoimmune hepatitis; hence, the scarring process is a possible target for novel therapy. Investigating the scarring process using highly specific technology, we show that the scarring process is different between the 3 autoimmune liver diseases, and this may have important implications for the development of medical treatment.

MiR-21-5p: A viable therapeutic strategy for regulating intraocular pressure

Lowering intraocular pressure (IOP) is the most effective treatment of glaucoma, however most of the current available glaucoma drugs target a single molecule. MicroRNAs (miRNAs) are noncoding RNAs that target a network of molecules. This study aims to investigate the role of miR-21-5p in regulating IOP and the mechanism of function. miR-21-5p mimics was topically applied to C57/BL6 mouse eyes, which significantly increased miR-21-5p expression in the conventional outflow tissue and reduced IOP by a maximum of 17.77% at 24 h after treatment. The conventional outflow facility measured by ex vivo moue eye perfusion of miR-21-5p was significantly increased by 60.14%. Moreover, miR-21-5p overexpression significantly reduced the transendothelial electrical resistance in porcine angular aqueous plexus cells. Transcriptome analysis and further quantification by Western blot and PCR revealed that SMAD7 and FGF18 might be the downstream target of miR-21-5p in regulating aqueous humor outflow. The predicted functional pathways PTEN/eNOS, RhoB/pMLC and TIMP3/MMP9 were significantly altered after miR-21-5p transfection. Dual luciferase assay verified the direct targets of miR-21-5p. In conclusion, miR-21-5p seems to regulate IOP by modulating multiple genes that are associated with aqueous humor outflow, including genes those regulating cell adhesion, cytoskeletal dynamics and extracellular matrix turnover. Thus, miR-21-5p represents a new therapeutic strategy for glaucoma and a viable alternative to existing multidrug regimens.

Cell signaling model for arterial mechanobiology.

Arterial growth and remodeling at the tissue level is driven by mechanobiological processes at cellular and sub-cellular levels. Although it is widely accepted that cells seek to promote tissue homeostasis in response to biochemical and biomechanical cues-such as increased wall stress in hypertension-the ways by which these cues translate into tissue maintenance, adaptation, or maladaptation are far from understood. In this paper, we present a logic-based computational model for cell signaling within the arterial wall, aiming to predict changes in extracellular matrix turnover and cell phenotype in response to pressure-induced wall stress, flow-induced wall shear stress, and exogenous sources of angiotensin II, with particular interest in mouse models of hypertension. We simulate a number of experiments from the literature at both the cell and tissue level, involving single or combined inputs, and achieve high qualitative agreement in most cases. Additionally, we demonstrate the utility of this modeling approach for simulating alterations (in this case knockdowns) of individual nodes within the signaling network. Continued modeling of cellular signaling will enable improved mechanistic understanding of arterial growth and remodeling in health and disease, and will be crucial when considering potential pharmacological interventions.