Serotonin Signaling in Mitral Valve Homeostasis, Maintenance and Restoration

二尖瓣稳态、维护和恢复中的血清素信号传导

• 批准号: 9080961
• 负责人: Giovanni Ferrari
• 金额: $ 70.05万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9080961
• 关键词: Affect; Age; Age-Years; Agreement; Animal Model; Bioinformatics; Biological Assay; Biomechanics; Canis familiaris; Cardiac Surgery procedures; Cell Proliferation; Cells; Clinical; Data; Databases; Disease; Disease Progression; Dose; Event; Excretory function; Extracellular Matrix; Feasibility Studies; Frequencies; Functional disorder; Genetic; Genetic Polymorphism; Genotype; Growth; Heart Valves; Homeostasis; Human; Image; In Vitro; Institutional Review Boards; Intervention; Investigation; Length; Maintenance; Mediating; Mitral Valve; Mitral Valve Insufficiency; Mitral Valve Prolapse; Modeling; Mus; Neurotransmitters; Operative Surgical Procedures; Oxidative Stress; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Process; Production; Promoter Regions; Ptosis; Publishing; Reactive Oxygen Species; Receptor Signaling; Recurrence; Registries; Research; Risk; Role; Serotonin; Signal Transduction; Small Interfering RNA; Specimen; Testing; Therapeutic; Tissues; Variant; Work; base; connective tissue growth factor; decorin; disorder control; high risk; interstitial cell; operation; promoter; public health relevance; receptor; receptor expression; receptor-mediated signaling; reconstruction; repaired; research study; response; restoration; serotonin receptor; serotonin transporter; transcription factor

 DESCRIPTION (provided by applicant): Myxomatous mitral valve (MV) disease (MMVD) affects over 140 million patients worldwide. This application is concerned with maintaining and restoring MV homeostasis in MMVD patients through investigations of serotonin (5HT) mechanisms that affect MMVD pathophysiology. 5HT is a neurotransmitter, which if present at increased levels can cause valvulopathies. Our research indicates that progression of MMVD is mediated by 5HT signaling via 5HT receptors (5HTR) 2A and 2B and by a specific 5HT transporter (SERT) polymorphism in the promoter region (LL-SERT, the full-length promoter) with a Mendelian distribution. Thus, MMVD appears to be a disorder with increased 5HTR responsiveness due to augmented levels of 5HTRs, and is exacerbated by maximal expression of SERT due to the LL-SERT genotype. Downstream processing post-SERT of 5HT results in increased reactive oxygen species (ROS) and TGF-β1 signaling. Specific Aim 1: To determine the impact of LL-SERT polymorphism in MMVD progression and recurrence after surgical repair and 5HT responsiveness. 1. a) Hypothesis: Patients with LL-SERT polymorphism are at higher risk of more rapid progression of MMVD, requiring surgery at a younger age; furthermore post-surgery this genotype with MMVD may have an increased risk for re-operation due to continuing 5HT-enhanced mechanisms. We will conduct genetic and bioinformatics analyses to determine occurrence, progression rate, and clinical recurrence of MVD patients based on SERT polymorphisms, SERT SNP variations, and imaging studies. 1. b) Hypothesis: VIC derived from patients with LL-SERT polymorphism are more responsive to 5HT stimulation resulting in MVICs activation and tissue remodeling. LL-, LS-, and SS-derived VICs and MV tissues will be isolated from MMVD patients and controls and exposed to 5HT stimulation. Endpoints will be MVICs phenotype, ROS production, and ECM remodeling. Specific Aim 2: To investigate the mechanisms of 5HTR signaling in human derived MV cells from all 3 SERT genotypes, focusing on TGF-β1 related pathways. 2. a) Hypothesis: Suppression of 5HTR2A & 2B expression in human derived MV cells and tissues controls cell phenotype and ECM. This subaim will use established siRNA constructs specific for 5HTR2A/2B to investigate their effects over a 5HT dose range, with endpoints including cell proliferation and ECM remodeling. This approach will also study SERT genotypes, SS, LL, and LS separately. 2. b) Hypothesis: Downstream TGF-β1 events related to 5HTR signaling mediate the progression of MMVD. These studies will investigate the effect of 5HTR-TGF-β1 interactions including studies of TGF- β1-receptor expression, TGF-β1 excretion, canonical and non-canonical signaling and the effects of TGF-β1 arising from 5HTR activity on ROS and ECM.

 描述（由应用提供）：粒细胞护孔瓣（MV）疾病（MMVD）影响全球超过1.4亿例患者。该应用涉及通过研究影响MMVD病理生理学的5-羟色胺（5HT）机制来维持和恢复MMVD患者的MV稳态。 5HT是一种神经递质，如果存在以增加水平，可能会导致瓣膜病。我们的研究表明，MMVD的进展是由5HT接收器（5HTR）2A和2B的5HT信号传导介导的，以及特定的5HT转运蛋白（SERT）多态性在启动子区域（LL-SERT，全长启动子）中具有Mendelian分布。这是MMVD似乎是一种疾病，由于5HTR水平增加而导致5HTR响应性增加，并且由于LL-SERT基因型而导致的SERT最大表达而加剧。 5HT的下游处理后，导致活性氧（ROS）和TGF-β1信号传导增加。具体目的1：确定LL-Sert多态性在手术修复和5HT反应性后MMVD进展和复发中的影响。 1。a）假设：LL-SERT多态性患者的MMVD进展更快的风险更高，需要在年轻时进行手术；此外，手术后这种具有MMVD的基因型可能会因继续5HT增强机制而导致重新手术的风险增加。我们将根据SERT多态性，SERT SNP变化和成像研究来确定MVD患者的发生，进展率和临床复发的发生，进展率和临床复发。 1。b）假设：来自LL-Sert多态性患者的VIC对5HT刺激的反应更大，导致MVICS激活和组织重塑。 LL-，LS-和SS衍生的VIC和MV组织将从MMVD患者和对照组中分离出来，并暴露于5HT刺激。终点将是MVICS表型，ROS产生和ECM重塑。具体目的2：研究来自所有3种SERT基因型的人类衍生的MV细胞中5HTR信号的机制，重点是TGF-β1相关途径。 2。a）假设：在人衍生的MV细胞和组织中对5HTR2A和2B表达的抑制控制细胞表型和ECM。该Subaim将使用针对5HTR2A/2B的已建立的siRNA构建体来研究其在5HT剂量范围内的作用，其中包括细胞增殖和ECM重塑，包括细胞增殖和ECM重塑。这种方法还将分别研究SERT基因型，SS，LL和LS。 2。b）假设：与5HTR信号有关的下游TGF-β1事件介导MMVD的进展。这些研究将研究5HTR-TGF-β1相互作用的作用，包括对TGF-β1受体表达，TGF-β1极端，规范和非典型信号传导的研究，以及由5HTR活性对ROS和ECM产生的TGF-β1的影响。