IL10 Regulation of Fibroblast Progenitor Cell Mediated Cardiac Fibrosis

IL10 对成纤维祖细胞介导的心脏纤维化的调节

• 批准号: 9492609
• 负责人: Suresh K Verma
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9492609
• 关键词: Adenoviruses; Attenuated; Blood; Bone Marrow; Bone Marrow Transplantation; Cardiac; Cell physiology; Cells; Data; Development; Disease; Dominant-Negative Mutation; Exhibits; Extracellular Signal Regulated Kinases; Fibroblasts; Fibrosis; Gene Expression; Goals; Heart; Heart Diseases; Heart Injuries; Heart failure; Hematopoietic stem cells; Home environment; Homing; In Vitro; Interleukin-10; Knockout Mice; Length; MAPK3 gene; MEKs; Mediating; Mediator of activation protein; Molecular; Morbidity - disease rate; Mus; Myocardial; Myocardial dysfunction; Myocardium; Myofibroblast; Paracrine Communication; Pathologic; Patients; Phosphotransferases; Population; Process; Proteins; Public Health; Quality of life; Regulation; Reporter; Reporting; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Stem cells; Stimulus; Subfamily lentivirinae; System; Testing; Therapeutic; Translating; Transplantation; United States; base; cardiac repair; coronary fibrosis; exosome; experimental study; heart function; improved; macrophage; monocyte; mortality; novel; novel therapeutics; paracrine; pre-clinical; pressure; prevent; progenitor; protein expression; response; transdifferentiation

Abstract Activated cardiac fibroblasts (myofibroblast; myoFBs) are crucial players in excessive fibrosis, which ultimately results in myocardial stiffness and heart failure development. Thus, regulation of fibroblasts activation and associated fibrosis would be a potential therapeutic strategy in the treatment of cardiac diseases. Previous studies suggest that in addition to resident fibroblasts, bone marrow fibroblast progenitor cells (FPCs) may home to the failing heart and contribute to excessive fibrosis. The overall goal of this application is to determine the role of BM-derived FPCs and their paracrine signaling (via exosomes) on pressure overload (PO)-induced cardiac fibrosis, and to determine whether inhibition of function of this cell population results in the reduction of cardiac fibrosis and remodeling. We have reported that PO-induced cardiac fibrosis is attenuated by systemic IL10 treatment and IL10-knockout mice (IL10 KO) display exaggerated cardiac fibrosis. But the precise mechanism of anti-fibrotic role of IL10 is not well defined. Thus we hypothesized that “IL10 inhibits fibroblast progenitor cell (FPCs) homing and trans-differentiation into myoFB and modifies FPC pro-fibrotic paracrine signaling thereby reducing cardiac fibrosis and improving cardiac function”. We will use WT (some with GFP reporter system), IL10 KO, TGFbr1/2 KO and CCR2KO (RFP reporter system) mice in this study. In SA1, via bone marrow transplantation strategy, we will determine whether FPC contributes in Ang II-induced cardiac fibrosis in WT and IL10 Knock out mice and whether IL10 treatment inhibits it. In specific aim 2 we will identify the TGFb downstream signaling targets [both canonical (Smad2/3), non-canonical (ERK-MAP kinase) and miR associated with fibrosis] and their roles in FPC's trans-differentiation. To identify their role, we will infect/transfect dominant negative adenovirus or kinase-dead construct (Smad/ERK) or antagomir (miRs) with their respective control in WT and IL10KO FPCs. In specific aim 3 we propose to study the paracrine regulation of FPCs on resident cardiac fibroblast activation and exaggerated fibrosis in IL10KO mice. In this aim we will explore whether pro-fibrotic factors secreted by FPC (WT/IL10KO) via exosomes, induce cardiac fibroblast transition to myoFB. To translate this finding in preclinical settings, in specific aim 3 we will investigate whether transplantation of exosomes-derived from ex vivo-modified FPC (for target miRs/proteins) in IL10 KO mice heart inhibits Ang II-induced fibrosis. The significance of this study is to identify the novel mechanism(s) of IL10 in regulation of cardiac fibrosis. Our proposed experiments can potentially identify novel therapeutic strategies for cardiac repair based on manipulation of bone marrow fibroblast progenitor cells signaling after pressure overload-induced cardiac injury.

摘要 活化的心脏成纤维细胞在过度纤维化中起关键作用。 最终导致心肌僵硬和心力衰竭的发展。因此，成纤维细胞的调节 活化和相关的纤维化将是治疗心脏疾病的潜在治疗策略。 疾病。先前的研究表明，除了常驻成纤维细胞，骨髓成纤维细胞 祖细胞(FPC)可能是衰竭心脏的归宿，并导致过度纤维化。总目标 这一应用的目的是确定骨髓来源的FPC及其旁分泌信号的作用(通过 Exosome)对压力超负荷(PO)诱导的心肌纤维化的影响，并确定是否抑制 这种细胞群的功能导致心脏纤维化和重构的减少。我们有 有报道称，系统性IL10治疗和IL10基因敲除可减轻PO诱导的心肌纤维化 小鼠(IL10KO)表现出夸大的心肌纤维化。但IL10抗纤维化作用的确切机制 没有很好地定义。因此，我们假设“IL10抑制成纤维细胞前体细胞(FPC)的归巢和 转分化为myoFB并修饰FPC促纤维化旁分泌信号，从而减少心脏 肝纤维化和改善心功能“。我们将使用WT(一些带有GFP报告系统)，IL10 KO， TGFBR1/2 KO和CCR2KO(RFP报告系统)小鼠。在SA1中，通过骨髓 移植策略，我们将确定FPC是否在Ang II诱导的心脏纤维化中起作用 WT和IL10敲除小鼠，以及IL10处理是否抑制它。在具体目标2中，我们将确定 TGFb下游信号转导靶点[规范的(Smad2/3)、非规范的(ERK-MAPK)和 MIR与纤维化相关]及其在FPC转分化中的作用。为了确定他们的角色，我们将 感染/转染显性负性腺病毒或激酶死亡结构物(Smad/ERK)或反义寡核苷酸(MiRs) 在WT和IL10KO FPC中拥有各自的控制权。在具体目标3中，我们建议研究旁分泌 FPC对IL10KO小鼠心脏成纤维细胞活化和过度纤维化的调节。在……里面 为此，我们将探讨FPC(WT/IL10KO)通过外切体分泌的促纤维化因子是否能诱导 心脏成纤维细胞向myoFB转化。为了将这一发现转化为临床前环境，在特定目标3中，我们 将研究从体外修饰的FPC(用于靶标)获得的外切体的移植 IL10KO小鼠心脏中的MIRS/蛋白质)抑制血管紧张素转换酶II诱导的纤维化。这项研究的意义在于 明确IL10调节心肌纤维化的新机制(S)。我们提议的实验可以 潜在地发现基于骨髓操作的心脏修复的新治疗策略 成纤维细胞前体细胞在压力超负荷致心脏损伤后的信号转导。