Immuno-stromal axes regulate fibroblast heterogeneity in tissue fibrosis and regeneration

免疫基质轴调节组织纤维化和再生中成纤维细胞的异质性

• 批准号: 10879203
• 负责人: Daniel Abebayehu
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10879203
• 关键词: Immuno; stromal; axes; regulate; fibroblast

Project Summary Tissue damage due to injury, disease, or congenital defect continues to be a critical clinical obstacle in human health. Regenerative medicine-based therapies are a promising strategy, yet the critical determinants of success are the immune response and fibrosis. Typically, in tissue repair, inflammation is closely controlled before remodeling by activated fibroblasts, whose presence is also tightly regulated. Immunomodulation is a promising strategy to avoid fibrotic outcomes, yet it is not the immune cells that directly participate in tissue remodeling and collagen production. Therefore, it is critical to understand how immune cells coordinate stromal cells to promote tissue repair or fibrosis. In fibrosis, evidence suggests that chronic immune cell infiltration and proinflammatory cytokines accompany fibroblasts poised to promote fibrosis. Those fibroblasts are characterized by aberrant mechanotransduction (i.e. sensing “normal”/soft as stiff/scarred) leading to the overproduction of ECM. We now know that immune cells and fibroblasts exist as phenotypically distinct subpopulations, including those with a molecular signature tied to fibrosis. Whether inflammatory signals elicit distinct fibroblast subpopulations with aberrant mechanotransduction in fibrosis is unknown and is a key objective. Understanding the relationship between inflammation and fibroblast heterogeneity is essential to predict the regenerative potential of therapies. I hypothesize that fibroblast subpopulations exist during fibrosis with aberrant mechanotransduction that is regulated by inflammatory signaling. During the mentored K99 phase of this application, I will identify the immune and fibroblast subpopulations that emerge during tissue regeneration and fibrosis leveraging non-fibrotic/regenerative microporous annealed particle hydrogels (MAP gels) and fibrotic nonporous bulk hydrogels (NP gels). I will determine the mechanotransductive differences across those subpopulations by purifying them with identified markers and measuring ECM production, myofibroblastic differentiation, integrin activation, and focal adhesion signaling. I will identify signaling pathways necessary for the phenotypes unique to fibroblast subpopulations. I will validate these findings by confirming the identified fibroblast subpopulations and signaling pathways in fibrotic clinical implants. I will continue my scientific and professional development in preparation for the independent phase with consistent guidance from my scientific advisory committee, diversity advisory committee, and collaborators. During the independent phase, I will determine mediators unique to regenerative and fibrotic microenvironments using MAP and NP gels. I determine if IL-1 signaling in fibroblasts is necessary for fibrosis by implanting NP gels in fibroblast-specific IL-1RI knockout mice. I will also determine whether IL-1 signaling is sufficient to promote fibrosis using MAP gels conjugated with IL-1?. We pose an innovative hypothesis that seeks to bridge the gap between inflammation and disrupted mechanotransduction distinctive of fibrotic disorders. This proposal will equip me with scientific and professional training that will enable me to launch a successful and diverse research program as an independent investigator.

项目概要 由于损伤、疾病或先天性缺陷造成的组织损伤仍然是临床上的一个关键障碍 基于再生医学的疗法是一种有前途的策略，但也是人类健康的关键决定因素。 成功的是免疫反应和纤维化 通常，在组织修复中，炎症得到严格控制。 在被激活的成纤维细胞重塑之前，其存在也受到严格调节。 避免纤维化结果的有希望的策略，但直接参与组织的不是免疫细胞 因此，了解免疫细胞如何协调基质至关重要。 细胞促进组织修复或纤维化，有证据表明慢性免疫细胞浸润和纤维化。 促炎细胞因子伴随着准备促进纤维化的成纤维细胞。 通过异常的机械传导（即感觉“正常”/柔软为僵硬/伤痕累累）导致过度生产 ECM。我们现在知道免疫细胞和成纤维细胞作为表型不同的亚群存在，包括 那些具有与纤维化相关的分子特征的炎症信号是否引起不同的成纤维细胞。 纤维化中机械转导异常的亚群尚不清楚，但这是一个关键目标。 炎症和成纤维细胞异质性之间的关系对于预测再生至关重要 我急切地指出，纤维化期间存在着异常的成纤维细胞亚群。 在此指导的 K99 阶段，受炎症信号调节的机械转导。 应用程序中，我将识别组织再生过程中出现的免疫和成纤维细胞亚群 利用非纤维化/再生微孔退火颗粒水凝胶（MAP 凝胶）和纤维化的纤维化 我将确定这些无孔块状水凝胶（NP 凝胶）的力传导差异。 通过使用已识别的标记物纯化亚群并测量 ECM 的产生、肌纤维母细胞 我将确定分化、整合素激活和粘着斑信号传导所需的信号通路。 我将通过确认已识别的成纤维细胞亚群独特的表型来验证这些发现。 纤维化临床植入物中的成纤维细胞亚群和信号通路我将继续我的科学和研究。 在我的科学的一致指导下，为独立阶段做准备的专业发展 顾问委员会、多元化顾问委员会和合作者 在独立阶段，我将。 使用 MAP 和 NP 凝胶确定再生和纤维化微环境特有的介质。 通过在成纤维细胞特异性 IL-1RI 敲除中植入 NP 凝胶，确定成纤维细胞中的 IL-1 信号传导是否是纤维化所必需的 我还将使用 MAP 凝胶确定 IL-1 信号传导是否足以促进纤维化。 IL-1？我们提出了一个创新的假设，旨在弥合炎症和破坏之间的差距。 这项建议将为我提供科学和专业的纤维化疾病的机械转导。 培训将使我能够作为独立调查员启动成功且多样化的研究计划。