Muscle Cell Migration Research Articles

Proteomic biomarkers related to obesity in heart failure with reduced ejection fraction and their associations with outcome.

Heart failure (HF) pathophysiology in patients with obesity may be distinct. To study these features, we identified obesity-related biomarkers from 4210 circulating proteins in patients with HF with reduced ejection fraction (HFrEF) and examined associations of these proteins with HF prognosis and biological mechanisms. In 373 patients with trimonthly blood sampling during a median follow-up of 2.1 (25th-75th percentile: 1.1-2.6) years, we applied an aptamer-based multiplex approach measuring 4210 proteins in baseline samples and the last two samples before study end. Associations between obesity (BMI > 30 kg/m2) and baseline protein levels were analyzed. Subsequently, associations of serially measured obesity-related proteins with biological mechanisms and the primary endpoint (PEP; composite of cardiovascular mortality, HF hospitalization, left ventricular assist device implantation, and heart transplantation) were examined. Obesity was identified in 26% (96/373) of patients. A total of 30% (112/373) experienced a PEP (with obesity: 26% [25/96] vs. without obesity: 31% [87/277]). A total of 141/4210 proteins were linked to obesity, reflecting mechanisms of neuron projection development, cell adhesion, and muscle cell migration. A total of 50/141 proteins were associated with the PEP, of which 12 proteins related to atherosclerosis or hypertrophy provided prognostic information beyond clinical characteristics, N-terminal pro-B-type natriuretic peptide, and high-sensitivity troponin T. Patients with HFrEF and obesityshow distinct proteomic profiles compared to patients with HFrEF without obesity. Obesity-related proteins are independently associated with HF outcome. These proteins carry potential to improve management of obesity-related HF and could be leads for future research.

AEBP1 upregulation contributes to cervical cancer progression by facilitating cell proliferation, migration, and invasion.

Aberrant expression of adipocyte enhancer-binding protein 1 (AEBP1) has been demonstrated to be involved in the tumorigenesis and progression of numerous cancers. This study was aimed to investigate the mechanism of AEBP1 in the development of cervical cancer. The expression of AEBP1 in cervical cancer was assessed by immunohistochemistry. The function of AEBP1 on cell proliferation, migration, and invasion was determined by methyl thiazolyl tetrazolium assay, colony formation, and transwell assay. The activation of related signaling pathway was determined by western blot. The bioinformatics analysis was performed by Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Higher protein expression of AEBP1 was observed in patients with cervical cancer. Overexpressed AEBP1 promoted cell proliferation, migration, and invasion abilities in cervical cancer cells. Moreover, the research manifested that AEBP1 activated the phosphorylation of STAT3. GO and KEGG analysis showed that genes positively related to AEBP1 were highly enriched in functions like epithelial cell proliferation, muscle cell migration, myoblast migration, smooth muscle tissue development, ECM-receptor interaction, transcriptional misregulation in cancer, and proteoglycans in cancer. While genes negatively related to AEBP1 were associated with immunity, including inflammatory response, external-stimulus response, neutrophil, granulocyte, and macrophage chemotaxis. This study suggested that AEBP1 acts as an oncogened and might be a potential therapeutic target for the treatment of cervical cancer.

ATR1 A1166C (rs5186), FII G20210A (rs1799963), FV G1691A (rs6025), FXIII 97G > T (rs11466016) and MTHFR A1298C (rs1801131) polymorphisms and the risk of ST-elevation myocardial infarction in young Mexican individuals.

Previous studies had identified genetic variants associated with Myocardial Infarction, but results are inconclusive. We examined the association between FII G20210A (rs1799963), FV G1691A (rs6025), FXIII 97G > T (rs11466016), ATR1 A1166C (rs5186) and MTHFR A1298C (rs1801131) polymorphisms and ST elevation Myocardial Infarction in young Mexican individuals. We included a total of 350 patients with Myocardial Infarction <45 years old and 350 controls matched by age and gender. The polymorphisms were analyzed by PCR-RFLP using specific restriction enzymes. DNA fragments were separated by electrophoresis in 2% gel of agarose and visualized using SYBR green. The A1166C (p = 0.004) but not FXIII 97G > T (p = 0.19), G20210A (p = 0.32), G1691A (p = No significant) and A1298C (p = 0.21) polymorphisms were associated with increased risk for ST elevation Myocardial Infarction. Moreover, dyslipidemia, hypertension, smoking and family history of atherothrombotic disease were associated. We found that A1166C represented increased risk for ST elevation Myocardial Infarction. However, G20210A, G1691A, 97G > T, and A1298C were not associated. In addition, we had determined that Glu298Asp, PLA1/A2, TAFI Thr325Ile, ACE I/D, AGT M235T and PAI-1 4G/5G polymorphisms represented increased risk in the same group of patients. However, MTHFR C677T, AGT T174M, FV G1691A, TSP-1 N700S, MTHFR C677T and TAFI 174M polymorphisms were no associated. Our results suggest that in young patients with ST Myocardial Infarction, those polymorphisms could contribute to premature endothelial dysfunction, atherothrombosis, vasoconstriction, increased platelet aggregation, muscle cell migration and proliferation. Further studies are required to try to better assess gene-gene and gene-modifiable factors interaction.

Photobiomodulation Using Different Infrared Light Sources Promotes Muscle Precursor Cells Migration and Proliferation

Photobiomodulation (PBM) has demonstrated positive effects on the muscle repair process. The aim of the study was to evaluate the effects of infrared PBM using different light sources—low-level laser (LLL) at 780 nm (40 or 70 mW, 10 J/cm2, 0.4 J) or LED at 850 nm (40 or 70 mW, 0.13 J/cm2, 0.4 J)—and dosimetric parameters on the proliferation and migration of muscle cells. The results showed that LLL 40 mW and 70 mW, with the same radiation exposure, led to an increase in proliferation after 24 h, but no differences at 48 and 72 h. Cells irradiated with LED 70 mW exhibited an increase in proliferation in comparison to the control group and 40mW after 24 and 48 h, but not at 72 h. Moreover, cell migration was greater in comparison to the control after 6 and 24 h, and no differences were found at 12 h when LLL was used with an output power of 70 mW. Furthermore, no differences were found at 6 and 12 h with the 70 mW output power-LED, but an increase was observed in the cell migration after 24 h. In conclusion, PBM using different light sources and dosimetric parameters was able to modulate the proliferation of C2C12 myoblasts, but only PBM at 70 mW was able to modulate the migration of these cells.

Pipersentan: A De Novo Synthetic Endothelin Receptor Antagonist that Inhibits Monocrotaline- and Hypoxia-Induced Pulmonary Hypertension.

Background: Although major advances have been made in the pathogenesis and management of pulmonary arterial hypertension (PAH), the endothelin system is still considered to play a vital role in the pathology of PAH due to its vasoconstrictive action. Endothelin receptor antagonists (ERAs), either as monotherapy or in combination with other drugs, have attracted much attention in the treatment of this lethal disease, and research is continuing. Methods: A novel ERA, pipersentan 5-(1,3-Benzodioxol-5-yl)-6-[2-(5-bromopyrimidin-2-yl)oxyethoxy]-N-(2-methoxyethylsulfamoyl)pyrimidin-4-amine, was recently synthesized and the physicochemical characterizations and the pharmacology both in vitro and in vivo were studied. Results: This orally administered ERA can both competitively and selectively inhibit the binding of endothelin-1 (ET-1) to its receptors with good physicochemical characteristics. Pipersentan efficaciously antagonized the effects of ET-1 on pulmonary artery smooth muscle cell proliferation, migration and calcium mobilization and effectively improved right ventricular hypertrophy and pulmonary arterial pressure in both monocrotaline- and hypoxia-induced pulmonary hypertension (PH) rat models. Conclusions: This profile identifies pipersentan as a new agent for treating ET-1 system activation-related PH.

447-P: C/EBPß-Lin28a Positive Feedback Loop Mediated by C/EBPß Hypomethylation Promotes the Proliferation and Migration of Vascular Smooth Muscle Cells in Restenosis

Background: The main reason for restenosis (RS) after percutaneous transluminal angioplasty is the excessive proliferation and migration of vascular smooth muscle cells (VSMCs) . C/EBPβ plays an important role in the process of vascular remodeling. However, the role of C/EBPβ in RS has not been clarified. Methods: The type 2 diabetes rat models of RS and atherosclerosis (AS) were established to detect the difference in DNA methylation and expression levels of C/EBPβ. Simultaneously, the effect of C/EBPβ on the proliferation and migration of VSMCs was clarified by regulating C/EBPβ and Lin28a in vitro. The effect of decitabine, a demethylating agent, on the expression of C/EBPβ and Lin28a, and the further regulation of VSMCs proliferation and migration was confirmed. Results: C/EBPβ was highly expressed and had a lower methylation level in RS compared with AS. In vitro C/EBPβ overexpression facilitated the proliferation and migration of VSMCs with increased expression of Lin28a. Conversely, C/EBPβ knockdown resulted in opposite effects. Chromatin immunoprecipitation assays further demonstrated that C/EBPβ could directly bind to the promoter of Lin28a. Furthermore, increased expression of C/EBPβ and enhanced proliferation and migration of VSMCs were observed after decitabine treatment. And mechanical stretch promoted the expression of C/EBPβ and Lin28a. Additionally, C/EBPβ expression increased following the overexpression of Lin28a. The reversed result was found in the Lin28a knockdown cells. Conclusion: Our results have demonstrated that C/EBPβ expression is regulated partly by DNA methylation. Meanwhile, as a transcription factor, C/EBPβ promotes the transcription of Lin28a and enhances the proliferation and migration of VSMCs through a positive feedback loop between the two. Our study suggests C/EBPβ-Lin28a axis is a driver of the RS progression, providing a promising therapeutic strategy for RS. Disclosure S.Jiang: n/a. L.Liao: None. J.Dong: None. X.Zhou: n/a. Q.Zhang: None. S.Guo: None. Q.Sheng: None. X.Li: None. R.Zhang: None. C.Xu: None. P.Gong: None. Funding National Natural Science Foundation of China Grants (81800732,81670757,82170824) , Shandong Provincial Medicine and Health Science and Technology Development Program (No.202103060766) and Shandong Provincial Hospital Research incubation Fund (No.2021FY059)

Guanxinping Tablets Inhibit ET-1-Induced Proliferation and Migration of MOVAS by Suppressing Activated PI3K/Akt/NF-κB Signaling Cascade

Background/Aim Abnormal proliferation and migration of vascular smooth muscle cells is one of the main causes of atherosclerosis (AS). Therefore, the suppression of abnormal proliferation and migration of smooth muscle cells are the important means for the prevention and inhibition of AS. The clinical effects of Guanxinping (GXP) tablets and preliminary clinical research on the topic have proved that GXP can effectively treat coronary heart disease, but its underlying mechanism remains unclear. This study aimed to confirm the inhibitory effect of GXP on the abnormal proliferation of mouse aortic vascular smooth muscle (MOVAS) cells and to explore the underlying mechanism. Methods MOVAS cells were divided into two major groups: physiological and pathological groups. In the physiological group, MOVAS cells were directly stimulated with GXP, whereas in the pathological group, the cells were stimulated by endothelin-1 (ET-1) before intervention by GXP. At the same time, atorvastatin calcium, which effectively inhibits the abnormal proliferation of MOVAS cells, was used in the negative control group. CCK8 assay, scratch test, ELISA, Western blotting, and immunofluorescence staining were performed to observe the proliferation and migration of MOVAS cells and the expression levels of related factors after drug intervention in each group. Results In the physiological group, GXP had no significant effect on the proliferation and migration of MOVAS cells and the related factors. In the pathological group, a high dose of GXP reduced the abnormal proliferation and migration of MOVAS cells. Further, it reduced the expression levels of PI3K; inhibited the phosphorylation of Akt (protein kinase B); upregulated IκB-α levels; prevented nuclear factor kappa B (NF-κB) from entering the nucleus; downregulated the expression of interleukin 6 (IL6), IL-1β, and iNOS; and upregulated the ratio of apoptosis-related factor Bax/Bcl-2. There was no significant difference between the high-dose GXP group and the atorvastatin calcium group (negative control group). Conclusion Our findings revealed that GXP was able to inhibit the proliferation and migration of MOVAS cells by regulating the PI3K/Akt/NF-κB pathway.

Atherosclerosis

For the past decade, cardiovascular disease (CVD) has been the leading cause of death and disability worldwide, and in most cases, atherosclerosis is the dominant underlying pathologic process. Atherosclerosis is characterized by subendothelial accumulation of lymphocytes, lipid-filled macrophages, and inflammation-induced migration of vascular smooth muscle cells slowly forming a fibroinflammatory lipid plaque. Over time, the plaque progressively weakens and occludes the vessel; however, with continued inflammation and hemodynamic changes, atheromas can dangerously manifest as ischemia (acute or chronic) or aneurysmal disease. This review discusses the economic impact, historical background, and pathogenesis of atherosclerosis, as well as normal arterial biology, risk factors, and screening. Tables outline physiologic properties of nitric oxide in native vessels, selected cellular and chemical mediators and their role in atherogenesis, current risk factors for peripheral arterial disease (PAD) development, and diagnosis criteria for diseases that confer increased PAD risk. Graphs demonstrate mortality due to PAD, PAD prevalence, and the economic impact of CVD. Illustrations depict normal vessel anatomy, blood flow with plaque at carotid bifurcation, the superficial femoral artery (SFA) with plaque, and the progression of atherosclerotic changes. A flow chart for atherogenesis, photograph of a human carotid endarterectomy specimen, and angiogram showing the classic occluding lesion of the SFA are also provided. This review contains 9 figures, 5 tables, and 91 references.

The zinc transporter ZIP12 regulates monocrotaline-induced proliferation and migration of pulmonary arterial smooth muscle cells via the AKT/ERK signaling pathways

BackgroundThe zinc transporter ZIP12 is a membrane-spanning protein that transports zinc ions into the cytoplasm from the extracellular space. Recent studies demonstrated that upregulation of ZIP12 is involved in elevation of cytosolic free zinc and excessive proliferation of pulmonary arterial smooth muscle cells (PASMCs) induced by hypoxia. However, the expression of ZIP12 and its role in pulmonary arterial hypertension (PAH) induced by monocrotaline (MCT) in rats have not been evaluated previously. The aim of this study was to investigate the effect of ZIP12 on the proliferation and migration of PASMCs and its underlying mechanisms in MCT-induced PAH.MethodsA PAH rat model was generated by intraperitoneal injection of 20 mg/kg MCT twice at one-week intervals. PASMCs were isolated from the pulmonary arteries of rats with MCT-induced PAH or control rats. The expression of ZIP12 and related molecules was detected in the lung tissues and cells. A ZIP12 knockdown lentivirus and an overexpressing lentivirus were constructed and transfected into PASMCs derived from PAH and control rats, respectively. EdU assays, wound healing assays and Western blotting were carried out to explore the function of ZIP12 in PASMCs.ResultsIncreased ZIP12 expression was observed in PASMCs derived from MCT-induced PAH rats. The proliferation and migration of PASMCs from PAH rats were significantly increased compared with those from control rats. These results were corroborated by Western blot analysis of PCNA and cyclin D1. All these effects were significantly reversed by silencing ZIP12. Comparatively, ZIP12 overexpression resulted in the opposite effects as shown in PASMCs from control rats. Furthermore, selective inhibition of AKT phosphorylation by LY294002 abolished the effect of ZIP12 overexpression on enhancing cell proliferation and migration and partially suppressed the increase in ERK1/2 phosphorylation induced by ZIP12 overexpression. However, inhibition of ERK activity by U0126 resulted in partial reversal of this effect and did not influence an increase in AKT phosphorylation induced by ZIP12 overexpression.ConclusionsZIP12 is involved in MCT-induced pulmonary vascular remodeling and enhances the proliferation and migration of PASMCs. The mechanism of these effects was partially mediated by enhancing the AKT/ERK signaling pathways.

Downregulation of lncRNA SNHG16 inhibits vascular smooth muscle cell proliferation and migration in cerebral atherosclerosis by targeting the miR-30c-5p/SDC2 axis.

Atherosclerosis (AS) is the basic lesion underlying the occurrence and development of cerebrovascular diseases. Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a crucial role in AS. We aimed to explore the role of SNHG16 in AS and the molecular mechanism of VSMC involvement in the regulation of AS. The expression levels of SNHG16, miR-30c-5p and SDC2 were detected by qRT-PCR. CCK-8, wound healing and Transwell assays were used to assess ox-LDL-induced VSMC proliferation, migration, and invasion, respectively. Western blot analysis was used to detect SDC2 and MEK/ERK pathway-related protein levels. A dual-luciferase reporter assay confirmed the binding of SNHG16 with miR-30c-5p and miR-30c-5p with SDC2. SNHG16 and SDC2 expression was upregulated in patients with AS and ox-LDL-induced VSMCs, while miR-30c-5p was downregulated. Ox-LDL-induced VSMC proliferation and migration were increased, and the MEK/ERK signalling pathway was activated. MiR-30c-5p was targeted to SNHG16 and SDC2. Downregulating SNHG16 or upregulating miR-30c-5p inhibited ox-LDL-induced VSMC proliferation and migration and inhibited MEK/ERK signalling pathway activation. In contrast, downregulating miR-30c-5p or upregulating SDC2 reversed the effects of downregulating SNHG16 or upregulating miR-30c-5p. Furthermore, downregulating SDC2 inhibited ox-LDL-induced proliferation and migration of VSMCs and inhibited activation of the MEK/ERK signalling pathway, while upregulating lncRNA SNHG16 reversed the effects of downregulating SDC2. Downregulation of SNHG16 inhibited VSMC proliferation and migration in AS by targeting the miR-30c-5p/SDC2 axis. This study provides a possible therapeutic approach to AS.

PDGF regulates guanylate cyclase expression and cGMP signaling in vascular smooth muscle

The nitric oxide-cGMP (NO-cGMP) pathway is of outstanding importance for vascular homeostasis and has multiple beneficial effects in vascular disease. Neointimal hyperplasia after vascular injury is caused by increased proliferation and migration of vascular smooth muscle cells (VSMCs). However, the role of NO-cGMP signaling in human VSMCs in this process is still not fully understood. Here, we investigate the interaction between platelet derived growth factor (PDGF)-signaling, one of the major contributors to neointimal hyperplasia, and the cGMP pathway in vascular smooth muscle, focusing on NO-sensitive soluble guanylyl cyclase (sGC). We show that PDGF reduces sGC expression by activating PI3K and Rac1, which in turn alters Notch ligand signaling. These data are corroborated by gene expression analysis in human atheromas, as well as immunohistological analysis of diseased and injured arteries. Collectively, our data identify the crosstalk between PDGF and NO/sGC signaling pathway in human VSMCs as a potential target to tackle neointimal hyperplasia.

Deletion of Smooth Muscle Lethal Giant Larvae 1 Promotes Neointimal Hyperplasia in Mice.

Vascular smooth muscle cell (VSMC) proliferation and migration contribute to neointimal hyperplasia after injury, which causes vascular remodeling related to arteriosclerosis, hypertension, and restenosis. Lethal giant larvae 1 (LGL1) is a highly conserved protein and plays an important role in cell polarity and tumor suppression. However, whether LGL1 affects neointimal hyperplasia is still unknown. In this study, we used smooth muscle-specific LGL1 knockout (LGL1SMKO) mice generated by cross-breeding LGL1flox/flox mice with α-SMA-Cre mice. LGL1 expression was significantly decreased during both carotid artery ligation in vivo and PDGF-BB stimulation in vitro. LGL1 overexpression inhibited the proliferation and migration of VSMCs. Mechanistically, LGL1 could bind with signal transducer and activator of transcription 3 (STAT3) and promote its degradation via the proteasomal pathway. In the carotid artery ligation animal model, smooth muscle-specific deletion of LGL1 accelerated neointimal hyperplasia, which was attenuated by the STAT3 inhibitor SH-4-54. In conclusion, LGL1 may inhibit neointimal hyperplasia by repressing VSMC proliferation and migration via promoting STAT3 proteasomal degradation.

Cigarette smoke extract induces migration of rat vascular smooth muscle cells through regulation of expressions of MOB2 and LATS1

&#x0D; &#x0D; &#x0D; &#x0D; Purpose: To study the impact of cigarette smoke extract (CSE) on the migratory capacity of rat vascular smooth muscle cells (VSMCs), and the likely mechanism of action involved.&#x0D; Methods: Rat VSMCs were exposed to varying concentrations of CSE. The migration capacity of the cells was determined by Transwell migration assay and scratch test. Protein and RNA expression levels of LATS1 (large tumor suppressor kinase 1) and MOB2 (monopolar spindle-one-binder protein 2) were assayed using immunoblotting and quantitative real-time-polymerase chain reaction (qRT-PCR), respectively.&#x0D; Results: The results showed that within a certain concentration range, the migratory capacity of VSMCs increased as the concentration of CSE increased (p &lt; 0.05). In contrast, the protein and RNA expressions of LATS1 and MOB2 were negatively correlated with CSE concentrations (p &lt; 0.05).&#x0D; Conclusion: CSE promotes abnormal migration of rat VSMCs by inhibiting the expressions of LATS1 and MOB2 associated with Hippo signaling pathway. Thus, smoking may induce vascular remodeling and cardiovascular events via this mechanism.&#x0D; &#x0D; &#x0D; &#x0D;

Abnormalities in esophageal smooth muscle induced by mutations in collagen XIX.

Collagen XIX is a nonfibrillar collagen that localizes in restricted tissues at very low amounts. A previous study on Col19a1 null mice revealed that collagen XIX is involved in esophageal muscle physiology and morphogenesis. Here, we use histological analysis to show that mice with a Col19a1 mutant lacking the NC3 domain and seven collagen triplets display abnormal transition of smooth to striated muscle in the abdominal segment of esophagus, and a widened esophagus with age. With two newly prepared antibodies, we analyzed the expression of collagen XIX in the mouse esophagus and show that collagen XIX colocalizes with α-smooth muscle actin. By immunoelectron microscopy, we confirmed the localization of collagen XIX in esophageal smooth muscle cells. Col19a1 mutant mice contained reduced levels of mutated Col19a1 mRNA. Interestingly, hepatocyte growth factor, which has an important role in esophageal striated muscle development, was reduced in the esophagus of the Col19a1 mutant mice. These findings suggest that collagen XIX may be critical for the function of esophageal smooth muscle cells as a scaffold for anteroposterior migration of esophagus-striated muscle cells.

MiR-3571 modulates the proliferation and migration of vascular smooth muscle cells by targeting claudin 1.

Background and aims: The miRNA-based post-transcription modification has been extensively studied in hypertension. It however remains elusive how miRNA expression is regulated in this pathological process. We hypothesize that hydroxymethylation in the promoter regions tightly controls the levels of key miRNAs, which in turn affects the development of hypertension.Methods: The levels of hydroxymethylation in the promoter regions from thoracic aortic tissues were compared between spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto rats (WKYs), using hydroxymethylcytosine DNA immunoprecipitation (hMeDIP) sequencing. The altered hydroxymethylation level of miR-3571 was confirmed by glucosylation-coupled hydroxymethylation-sensitive qPCR. We further identified claudin 1(CLDN1) as a key target of miR-3571 via bioinformatic prediction (targetscan) and dual-luciferase activity assays. Finally, we analyzed the contribution of miR-3571/CLDN1 axis in the proliferation and migration of vascular smooth muscle cells (VSMCs).Results: The hydroxymethylation level of miR-3571 promoter region in thoracic aortic tissue from spontaneously hypertensive rats was lower than that from normotensive Wistar-Kyoto rats. Accordingly, the expression of miR-3571 was lower during hypertension, with up-regulated CLDN1 protein levels. More importantly, we found that miR3571 overexpression led to phenotypic changes of VSMCs, and inhibited the proliferation and migration of muscle cells via suppressing CLDN1 as well. Our findings further suggested that CLDN1 up-regulation increase the activity of ERK1/2 in VSMCs.Conclusions: Our study suggested that hydroxymethylation in the promoter regions controlled the level of miR-3571 and revealed the important roles of miR-3571 and CLDN1 in VSMCs during the development of hypertension. In addition, our results also indicated that miR-3571/CLDN1 axis regulated the functions of VSMCs via the ERK1/2 pathway. Taken together, our findings support miR-3571 as a novel biomarker for the diagnosis and prevention of hypertension.

GPR143, a L-DOPA receptor, induced migration and proliferation of vascular smooth muscle cells is involved in monocrotaline-induced pulmonary hypertension in rats

GPR143, a L-DOPA receptor, induced migration and proliferation of vascular smooth muscle cells is involved in monocrotaline-induced pulmonary hypertension in rats

Role of MicroRNA-134-5p in Metabolic Syndrome-Associated Pulmonary Hypertension in Heart Failure with Preserved Ejection Fraction

Background: Pulmonary hypertension in heart failure with preserved ejection fraction (PH-HFpEF) is the most common cause of PH worldwide. It is closely linked to risk factors for metabolic syndrome, including obesity and diabetes - factors known to increase proliferation and migration of pulmonary artery smooth muscle cells (PASMCs), leading to pulmonary vascular remodeling. Qualitative studies have shown that patients with progressive vascular abnormalities develop more severe symptoms and suffer frequent hospitalization. However, underlying mechanisms involved in the regulation of pulmonary vascular remodeling in metabolic syndrome-associated PH-HFpEF are still unclear.&#x0D; Aim: We have recently observed decreased levels of the tumor suppressor WW domain-containing oxidoreductase (WWOX), which plays a housekeeping role in repressing cellular proliferation, in PASMCs isolated from rats with experimental PH-HFpEF and human subjects with obesity and diabetes. As microRNAs (miRNAs) have been shown to regulate WWOX expression in cancers, here we aimed at examining the involvement of miRNAs in WWOX-associated pulmonary vasculature regulation in metabolic syndrome-associated PH-HFpEF.&#x0D; Methods and Results: Among miRNAs that have been associated with reduced WWOX expression, including miR -134-5p, -153-3p, -29a-3b, -29b-3p and -187-5p, we found that miR-134-5p was significantly increased in PASMCs of obese and diabetic subjects. To determine the role of miR-134-5p in the regulation of WWOX in the pulmonary vasculature, we applied exogenous miR-134-5p to human PASMCs. Treatment with miR-134-5p decreased WWOX expression, increased PCNA expression (a cell proliferation marker) and enhanced cellular proliferation. Additionally, human PASMCs challenged with high concentration of glucose, palmitic acid and insulin, which mimic hyperglycemic, hyperlipidemic and hyperinsulinemic conditions, exhibited increased miR-134-5p, accompanied by elevated cellular proliferation. &#x0D; Conclusions: These studies suggest that miR-134-5p may have a potential role in metabolic syndrome-associated PH-HFpEF through regulating WWOX in the pulmonary vasculature. These studies identify miR-134-5p as a potential therapeutic target for the treatment of metabolic syndrome-associated PH-HFpEF.&#x0D; This project was funded, in part, with support from the NIH NHLBI Short-Term Training Program in Biomedical Sciences Grant funded, in part by T35HL110854 from the National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Abstract 11489: Oral Rhamnan Sulfate Reduces Vascular Inflammation and Atherosclerotic Plaque Formation

Introduction: The glycocalyx, an interior lining of blood vessels, forms a barrier against inflammation and atherosclerotic risk factors. It is degraded during plaque development, allowing fat deposition in the intima. Rhamnan sulfate (RS), a polysaccharide from green seaweed, resembles GAGs such as heparan sulfate in the glycocalyx. Hypothesis: We hypothesize that RS can replace and rejuvenate the glycocalyx to enhance barrier function and prevent plaque formation. Methods: In vitro, we tested the effects of RS on proliferation, migration and permeability to LDL. We used western blots to measure the efficacy of RS in inhibiting the NF-κb pathway. We also performed atomic force microscopy on endothelial cells to test effects of RS on glycocalyx stiffness and thickness post-inflammation. In vivo, we conducted a study in female and male ApoE -/- mice on a high fat diet for 13 weeks where RS treatment was provided orally (n = 10 per group). Results: Rhamnan sulfate decreased growth factor induced proliferation and migration of endothelial and vascular smooth muscle cells. It increased heparan sulfate coverage and decreased the permeability of endothelial cells to LDL. It also decreased the activation of NF-κb in response to TNF-α treatment while also binding to NF-κb subunits. In ApoE -/- mice, oral consumption of RS decreased cholesterol levels in the plasma by 22.5% in female mice. En face analysis of aorta showed that plaque area decreased by 45.2% in female and 36.4% in male mice. In female mice, lipid deposition and stenosis decreased in the aortic arches and thoracic aortas while only lipid deposition was reduced in the aortic arches of male mice. Inflammatory factors were also decreased in the aortic arches of female mice. In livers of these mice, RS treatment reduced NAFLD activity scores and lipid deposition in females and unsaturated lipid content in males. RNA sequencing showed significant differences in genes related to metabolism, circadian rhythm and lipid regulation between high fat diet and RS groups and male and female mice. Conclusions: Rhamnan sulfate reduces inflammation in endothelial cells and proliferation of vSMCs. It also reduces plaque formation in aorta of ApoE -/- mice.

Abstract 12121: Coronary Disease Association With ADAMTS7 is Due to Protease Activity: Implications for Therapeutic Development

Background: Despite contemporary therapy, coronary artery disease (CAD) remains a leading cause of mortality. Genetic variants at ADAMTS7 have been associated with CAD and the loss of ADAMTS7 is protective for atherosclerosis. ADAMTS7 (a disintegrin and metalloproteinase with thrombospondin motifs 7) is a secreted metalloproteinase and complex proteoglycan, yet the mechanism linking ADAMTS7 to CAD risk remains unresolved. Methods and Results: We established a new purification strategy for full-length mouse ADAMTS7 and demonstrated the loss of activity in the catalytic mutant form of ADAMTS7. To test if the enzymatic activity of ADAMTS7 mediates atherosclerosis, we generated a catalytically inactive mutant mouse allele and compared it to the Adamts7 knockout. Using two models of atherosclerosis, we found that reducing either ADAMTS7 dosage or catalytic function decreased the burden of atherosclerosis. We demonstrate impaired vascular smooth muscle migration in both Adamts7 catalytic mutant and null cells using a lateral migration wound healing assay. Expression of the WT allele rescued the migration phenotype in Adamts7 null cells while expression of the catalytic mutant protein did not. We then characterized a human ADAMTS7 coding variant rs3825807 (Ser214Pro) associated with reduced CAD risk. This variant had a hypomorphic effect on ADAMTS7 secretion and migration of vascular smooth muscle cells, findings consistent with our mouse studies. Conclusions: We demonstrated that loss of ADAMTS7 catalytic function protects against atherosclerosis via phenotype switch of VSMCs and that the atherosclerosis protective effects could be mediated by a loss-of-function coding variant associated with CAD risk. In aggregate, we provide compelling evidence that dosage of ADAMTS7 and catalytic function are responsible for the atherosclerotic phenotype, suggesting that the catalytic domain would be an attractive therapeutic target for CAD.

MIA SH3 Domain ER Export Factor 3 Deficiency Prevents Neointimal Formation by Restoring BAT-Like PVAT and Decreasing VSMC Proliferation and Migration.

Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) and excessive accumulation of dysfunctional PVAT are hallmarks of pathogenesis after angioplasty. Recent genome-wide association studies reveal that single-nucleotide polymorphism (SNP) in MIA3 is associated with atherosclerosis-relevant VSMC phenotypes. However, the role of MIA3 in the vascular remodeling response to injury remains unknown. Here, we found that expression of MIA3 is increased in proliferative VSMCs and knockdown of MIA3 reduces VSMCs proliferation, migration, and inflammation, whereas MIA3 overexpression promoted VSMC migration and proliferation. Moreover, knockdown of MIA3 ameliorates femoral artery wire injury-induced neointimal hyperplasia and increases brown-like perivascular adipocytes. Collectively, the data suggest that MIA3 deficiency prevents neointimal formation by decreasing VSMC proliferation, migration, and inflammation and maintaining BAT-like perivascular adipocytes in PVAT during injury-induced vascular remodeling, which provide a potential therapeutic target for preventing neointimal hyperplasia in proliferative vascular diseases.