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: 10581593
• 负责人: Giovanni Ferrari
• 金额: $ 72.18万
• 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 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581593
• 关键词: Adult; Affect; Agonist; Alpha Granule; Anabolism; Antidepressive Agents; Area; Aspirin; BMP4; Blood; Blood Platelets; Bone Marrow; Bone Marrow Ablation; Bone Marrow Transplantation; CD34 gene; Canis familiaris; Carcinoid Tumor; Cells; Childhood; Chronic; Clinical Pathology; Cytoplasmic Granules; Diet; Disease; Disease Progression; Down-Regulation; Endothelial Cells; Endothelium; Event; Experimental Designs; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Feasibility Studies; Flow Cytometry; Functional disorder; Gene Expression Profile; HTR2A gene; Heart Valve Diseases; Heart Valves; Homeostasis; Homing; Human; In Vitro; Maintenance; Measurement; Mediating; Mesenchymal; Mitral Valve; Mitral Valve Insufficiency; Mitral Valve Prolapse; Modeling; Multivariate Analysis; Mus; Neurotransmitters; Operative Surgical Procedures; Pathogenesis; Pathologic; Patients; Pharmaceutical Preparations; Phenotype; Platelet Activation; Population; Preventive treatment; Production; Proteomics; Protocols documentation; Quantitative Reverse Transcriptase PCR; Receptor Signaling; Research; Role; Serotonin; Signal Transduction; Specimen; Therapeutic; Thick; Tissues; Transforming Growth Factor beta; Tryptophan; Tryptophan 5-monooxygenase; United States National Institutes of Health; Up-Regulation; antagonist; aortic valve; coronary fibrosis; experimental study; human old age (65+); human subject; inhibitor; interstitial cell; migration; personalized approach; progenitor; programs; receptor; repaired; restoration; reuptake; serotonin receptor; serotonin transporter; single cell sequencing; stem cells; therapeutic target; transcriptome; transcriptomics

Degenerative mitral regurgitation (MR) due to myxomatous valve disease affects millions. The only treatment is either valve repair or replacement. This competing renewal will advance understanding of the role of the neurotransmitter, serotonin (5HT) in MR. Heart valve disease has been associated with both 5HT-secreting carcinoid tumors and medications, such as the diet drug, Dexfenfluoramine. Based on our progress in the 1ST program period, we will investigate the hypothesis that 5HT-mechanisms involving the serotonin transporter (SERT), and 5HT receptor (HTR) signaling, both in platelets and mitral valve interstitial cells (MVIC) of bone marrow origin, contribute to the pathophysiology MR, and represent promising therapeutic targets. Specific Aim 1: To dissect 5HT-mediated mechanisms that influence pathologic remodeling of MV leaflets in human subjects and mice Subaim 1A: To characterize 5HT-mediated MV remodeling in MR compared to normal mitral valve (MV) leaflets and human-derived MVICs. SERT-antagonists, and HTR inhibitor studies will dissect unique 5HT related transcriptome and cellular phenotypes associated with MR progression in vitro and ex vivo under TGFβ, BMP4, and 5HT stimulation. Endpoints will be transcriptomic, extracellular matrix (ECM) remodeling in human MVIC and tissue from MR compared to normal leaflets exposed to 5HT-related mechanistic interrogation. Subaim 1B: To investigate the role of SERT in the pathogenesis of MR: The SERT deleted mouse (SERT-/-) spontaneously develops a valvulopathy affecting the mitral and aortic valves. Mechanistic studies will compare SERT-/- to SERT+/+ . Endpoints will include valve thickness, myocardial fibrosis, and changes in the MV transcriptome, to understand disease progression in this SERT-related valvulopathy. Specific Aim 2: To investigate the role of 5HT in the pathogenesis of MR involving platelet mechanisms and bone marrow derived blood outgrowth endothelial cells (BOEC) that express both CD34 and HTR2B. Subaim 2A: It is noteworthy that MR results in a chronic state of platelet activation, since platelets are the primary carriers of 5HT in blood, releasing 5HT upon activation. We will study the role of platelets in 5HT related MR pathophysiology in human subjects, canines with MR, and mice. Endpoints will include 5HT levels, platelet activation markers, and differences in TGF-b1 levels resulting from platelet alpha granule release. Subaim 2B: To investigate the role of bone marrow derived cells in the pathophysiology of 5HT mechanisms that affect MR. BOEC will be cultivated from the study groups in Subaim 2A. The experimental designs will use flow cytometry for cell characterization, qRT-PCR studies to assess gene expression patterns, and proteomic approaches to assess HTR activity, and measurements of ECM production in vitro. It is expected that the mechanistic results of these studies will lead to therapeutic directions for MR that target platelet derived 5HT and leverage a personalized approach using BOEC to model the pathophysiology.

由于粘液瓣疾病引起的退化性二尖瓣反流（MR）会影响数百万。唯一的治疗方法是阀门修复或更换。这种竞争的更新将提高人们对神经递质5-羟色胺（5HT）在MR中的作用的理解。心脏瓣膜疾病与5HT分泌的类癌和药物（例如饮食药物，右氟胺）都相关。基于我们在第一个计划期间的进步，我们将研究以下假设：在血小板和二尖瓣间质细胞（MVIC）中，涉及5-羟色胺转运蛋白（SERT）和5HT受体（HTR）信号的5HT机制，有助于病理生理学MR MR，并促进了promistectic的目标。具体目的1：剖析5HT介导的机制，这些机制会影响人类受试者和小鼠MICE subaim 1A中MV小叶的病理重塑：与正常二尖瓣（MV）小叶和人类衍生的MVics相比，MR中5HT介导的MV重塑的表征。 SERT抗逆转肌师和HTR抑制剂研究将在TGFβ，BMP4和5HT刺激下剖析与MR进展相关的5HT相关转录组和细胞表型。与暴露于5HT与5HT相关的机械询问的正常小叶相比，终点将是人类MVIC和MR组织中的细胞外基质（ECM）重塑。 Subaim 1B：研究SERT在MR：SERT删除的小鼠（SERT - / - ）的作用上，在赞助的瓣膜病上会发展出影响二尖瓣和主动脉瓣的瓣膜病。机械研究将将SERT - / - 与SERT+/+进行比较。终点将包括瓣膜厚度，心肌纤维化以及MV转录组的变化，以了解这种与SERT相关的瓣膜病中的疾病进展。具体目的2：研究5HT在涉及血小板机制和骨髓的MR发病机理中的作用，衍生出表达CD34和HTR2B的血液生长内皮细胞（BOEC）。 Subaim 2a：值得注意的是，MR会导致血小板激活的慢性状态，因为血小板是血液中5HT的主要载体，在激活后释放了5HT。我们将研究血小板在与5HT相关的MR病理生理学中的作用在人类受试者，犬类和小鼠中的作用。终点将包括5HT水平，血小板激活标记以及由血小板α颗粒释放引起的TGF-B1水平的差异。 Subaim 2b：研究骨髓衍生细胞在影响MR的5HT机制的病理生理学中的作用。 BOEC将从Subaim 2a的研究小组中培养。实验设计将使用流式细胞仪进行细胞表征，QRT-PCR研究来评估基因表达模式以及评估HTR活性的蛋白质组学方法，并在体外测量ECM产生。预计这些研究的机械结果将导致MR的治疗方向，该MR靶向血小板衍生5HT，并利用BOEC使用BOEC来建模病理生理学的个性化方法。