Supplement to Serotonin Signaling in Mitral Valve Homeostasis, Maintenance and Restoration

补充二尖瓣稳态、维护和恢复中的血清素信号传导

• 批准号: 10852158
• 负责人: Giovanni Ferrari
• 金额: $ 59.5万
• 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/10852158
• 关键词: Age; Agonist; Animal Model; Antidepressive Agents; Award; Bioreactors; Calcium Channel; Carcinoid Tumor; Cardiac; Cell Culture Techniques; Cells; Chromaffin Cells; Clinical Research; Collagen; Data; Diet; Disease; Echocardiography; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Feasibility Studies; Fibrosis; Fluoxetine; Gene Expression Profile; Genes; Grant; Heart Valve Diseases; Homeostasis; Human; Informatics; Intestinal Secretions; Intestines; Investigation; Laboratories; Literature; Maintenance; Mechanical Stimulation; Mechanics; Medicine; Mental Depression; Meta-Analysis; Mitral Valve; Mitral Valve Insufficiency; Mitral Valve Prolapse; Modeling; Morphology; Mus; Nobel Prize; Operative Surgical Procedures; Paper; Parents; Pathway interactions; Patients; Periodicity; Pharmaceutical Preparations; Piezo 1 ion channel; Piezo 2 ion channel; Production; Productivity; Quantitative Reverse Transcriptase PCR; Receptor Signaling; Reporting; Research; Sampling; Selective Serotonin Reuptake Inhibitor; Serotonin; Sheep; Signal Transduction; Small Interfering RNA; Stretching; System; Tissues; Transforming Growth Factor beta; Tryptophan 5-monooxygenase; United States National Institutes of Health; Up-Regulation; Ventricular; Ventricular Remodeling; cell type; experimental study; inhibitor; innovation; interest; interstitial cell; mouse model; novel; programs; receptor; restoration; reuptake; serotonin receptor; serotonin transporter; single-cell RNA sequencing; transcriptome sequencing

Abstract/Summary This one-year Supplement project will investigate the novel relationship of Piezo1, a mechanical sensing channel never studied previously concerning mitral valve prolapse (MVP) or mitral regurgitation (MR) to serotonin (5HT) mechanisms that contribute to the progression of MR. We previously demonstrated that mitral valve interstitial cell (MVIC) serotonin transporter (SERT) activity was inhibited by either the diet drug, Fenfluoramine or Fluoxetine, a selective 5HT reuptake inhibitor (SSRI), thus resulting in diminished 5HT clearance and enhanced 5HT receptor (HTR) activity. In addition, we showed in clinical studies of MR surgical patients that SSRI use was significantly associated with MR surgery at a younger age, and qRTPCR studies of human MR leaflets, compared to normal human mitral valve (MV) leaflets showed diminished SERT activity in MR. Piezo1 and increased 5HT secretion: Prior research has reported that intestinal Piezo1 activation results in increased 5HT, stimulating our research interest. Feasibility studies presented in this proposal demonstrate that Piezo1 agonists increase HTR signaling in MVIC. Hypothesis: MR progression results from increased HTR2B signaling due to diminished SERT expression and increased 5HT secretion, caused by activation of the mechanically sensitive Ca++ channel, Piezo1. Specific Aim 1: To study the effects of Piezo1 activation and inhibition on 5HT mechanisms in human MVIC, and in a mouse model of Fluoxetine induced MR. We will use the specific Piezo1 agonist, Yoda1, to model mechanical stimulation of Piezo1 in MVIC, and we will also study the effects of inhibition of Piezo1, HTR2B, and tryptophan hydroxylase-1 (TPH1) to reduce HTR signaling. MVIC endpoints include differences in extracellular matrix (ECM) production, expression of 5HT related genes, collagens 1&3, and Piezo1 and 2. Studies of our Fluoxetine mouse mitral valvulopathy model will investigate mitigation of the valvulopathy with both a HTR2B inhibitor and the Piezo1 inhibitor, Dooku1, with endpoints including: Ventricular remodeling per echocardiography, ventricular and valve morphology, and qRTPCR studies of 5HT related genes, and Piezo1&2. Specific Aim 2: To study human and sheep mitral valve (MV) leaflets, from both MR (human only) and normal MV (sheep and human) in cycle stretch studies with our mechanical bioreactor system. We will compare static and cyclic-stretch conditions, with exposure to inhibitors of Piezo1, HTR2B, and TPH1 compared to untreated, assessing differences in expression of ECM proteins, 5HT related genes and Piezo1 and 2. RNA sequencing studies will be carried out on bioreactor samples before and after mechanical stimulation with informatics analyses to identify gene expression patterns and pathways, compared to our human MV data. The proposed studies for this Supplement are responsive to NOT-HL-23-078 because they address the underlying basic mechanisms of fibrosis responsible for progression of MVP to MR, and utilize a relevant animal model.

摘要/总结 这个为期一年的补充项目将研究 Piezo1（一种机械传感）的新颖关系 以前从未研究过关于二尖瓣脱垂 (MVP) 或二尖瓣反流 (MR) 血清素的通道 (5HT) 促进 MR 进展的机制。我们之前证明了二尖瓣 间质细胞 (MVIC) 血清素转运蛋白 (SERT) 活性受到节食药物芬氟明的抑制 或氟西汀，一种选择性 5HT 再摄取抑制剂 (SSRI)，从而导致 5HT 清除率降低， 增强 5HT 受体 (HTR) 活性。此外，我们在 MR 手术患者的临床研究中表明， SSRI 的使用与年轻时的 MR 手术显着相关，并且人类 MR 的 qRTPCR 研究 与正常人二尖瓣 (MV) 小叶相比，MR 中 SERT 活性减弱。 Piezo1 和增加 5HT 分泌：先前的研究报道肠道 Piezo1 激活结果 5HT的增加，激发了我们的研究兴趣。本提案中提出的可行性研究表明 Piezo1 激动剂可增加 MVIC 中的 HTR 信号传导。 假设：MR 进展是由于 SERT 表达减少导致 HTR2B 信号传导增加所致 机械敏感 Ca++ 通道 Piezo1 的激活导致 5HT 分泌增加。 具体目标1：研究Piezo1激活和抑制对人体5HT机制的影响 MVIC，以及氟西汀诱导的 MR 小鼠模型。我们将使用特定的 Piezo1 激动剂 Yoda1 来建模 MVIC 中 Piezo1 的机械刺激，我们还将研究抑制 Piezo1、HTR2B 和 色氨酸羟化酶-1 (TPH1) 可减少 HTR 信号传导。 MVIC 终点包括细胞外的差异 基质 (ECM) 产生、5HT 相关基因、胶原蛋白 1 和 3 以及 Piezo1 和 2 的表达。我们的研究 氟西汀小鼠二尖瓣病模型将研究 HTR2B 对瓣膜病的缓解作用 抑制剂和 Piezo1 抑制剂 Dooku1，终点包括： 超声心动图、心室和瓣膜形态以及 5HT 相关基因和 Piezo1&2 的 qRTPCR 研究。 具体目标 2：研究人类和绵羊二尖瓣 (MV) 瓣叶，来自 MR（仅限人类）和 使用我们的机械生物反应器系统进行循环拉伸研究中的正常 MV（绵羊和人类）。我们会比较 静态和循环拉伸条件下，与暴露于 Piezo1、HTR2B 和 TPH1 抑制剂相比 未经处理，评估 ECM 蛋白、5HT 相关基因以及 Piezo1 和 2 表达的差异。RNA 将在机械刺激之前和之后对生物反应器样品进行测序研究 与我们的人类 MV 数据相比，进行信息学分析以确定基因表达模式和途径。 本增补的拟议研究响应 NOT-HL-23-078，因为它们解决了 导致 MVP 进展为 MR 的纤维化的基本机制，并利用相关动物 模型。