Therapeutic ECM Resorption in Cellular Systems and Precision Cut Lung Slices.

细胞系统中的治疗性 ECM 吸收和精密切割肺切片。

基本信息

批准号：
10318078
负责人：
Daniel J. Tschumperlin
金额：
$ 61.97万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318078
关键词：
Address Age Alveolar Animal Model Appearance Architecture Biological Biological Assay Biological Models Biological Response Modifier Therapy Cell Communication Cells Cellular Assay Characteristics Chemicals Cicatrix Collagen Cyclic AMP Data Deposition Disease Environment Epigenetic Process Extracellular Matrix Failure Fibrillar Collagen Fibroblasts Fibrosis Gases Goals Human Impairment In Vitro Individual Lung Mediating Mediator of activation protein Metabolic Modeling Molecular Natural regeneration Organoids Participant Pathogenesis Pathway interactions Patients Pharmacology Physiological Process Pulmonary Fibrosis Research Signal Transduction Slice Stimulus Structure of parenchyma of lung System Testing Therapeutic Therapeutic Intervention Tissues Variant cell type crosslink fibrotic lung high throughput screening idiopathic pulmonary fibrosis insight lung repair macrophage method development miniaturize novel repaired therapeutic evaluation tissue repair tool ventilation

项目摘要

Project Summary Idiopathic Pulmonary Fibrosis (IPF) is characterized by progressive replacement of functional alveolar gas exchange tissue with collagen rich scar. The accumulation, crosslinking and stiffening of this matrix are defining features of the disease, and definitive barriers to effective repair or regeneration. Emerging evidence indicates that fibrotic scar remains highly resorbable under appropriate conditions], a process that appears to be impaired or absent in humans with IPF. The conditions and pathways that promote fibroblasts (and other cell types) to resorb collagen rich scar are largely unknown. Development of methods and approaches to address this critical gap in understanding is the focus of this U01 proposal. We propose to develop model systems in which ECM deposition and resorption can be efficiently studied in both primary cultured lung fibroblasts as well as precision cut lung slices (PCLS). Our preliminary data show that under appropriate stimuli, IPF-derived human lung fibroblasts can be prompted to degrade and resorb fibrillar collagen. We hypothesize that under appropriate stimuli fibroblasts can be stimulated to not only resorb collagen rich ECM in vitro, but also resorb scar-associated ECM in the lungs from patients with IPF. We propose to develop and leverage novel in vitro tools to test this hypothesis, with the goal of identifying biological pathways and therapeutic interventions that mediate physiologic collagen resorption. We propose to pursue these goals through two aims. In the first aim we will develop culture systems allowing us to identify the signals that promote ECM resorption by lung fibroblasts and delineate the molecular mechanisms of collagen resorption. We will validate these assays for high-throughput discovery and perform a focused screen as proof of concept of the value of this approach. We will also compare the innate ECM deposition and degradation characteristics of IPF and control fibroblasts and fibroblast subsets. In the second aim we will develop ex vivo lung tissue assays to test therapeutic modulation and mechanisms of clearance of scar-associated IPF ECM. We will characterize baseline and stimulus evoked collagenolytic activity in control and IPF lung tissue, and define the association of this activity with lung cell types and histopathological appearance of the tissue. We will also test candidate hits identified in aim 1 for their capacity to increase targeted fibrillar collagen degradation in the native IPF ECM environment. Together the proposed studies will establish robust models of ECM deposition and resorption in primary human IPF fibroblasts and ex vivo lung slices. This platform will open new avenues for identifying signals and mechanisms that shift fibroblasts in IPF toward a matrix resorbing state, generating new opportunities to develop advanced therapeutics for IPF.

项目概要特发性肺纤维化 (IPF) 的特点是功能性肺泡气体的逐渐替代用富含胶原蛋白的疤痕交换组织。该基质的积累、交联和硬化是确定疾病的特征，以及有效修复或再生的明确障碍。新出现的证据表明纤维化疤痕在适当的条件下仍然具有高度可吸收性]，这一过程似乎患有 IPF 的人会受损或缺失。促进成纤维细胞（和其他细胞类型）来吸收富含胶原蛋白的疤痕在很大程度上是未知的。开发方法和途径解决这一理解上的关键差距是 U01 提案的重点。我们建议开发模型可以在原代培养肺中有效研究 ECM 沉积和吸收的系统成纤维细胞以及精密切割肺切片（PCLS）。我们的初步数据表明，在适当的情况下在刺激下，IPF来源的人肺成纤维细胞可以促使纤维状胶原蛋白降解和再吸收。我们假设在适当的刺激下，成纤维细胞不仅可以被刺激以吸收富含胶原蛋白的 ECM 体外，而且还能吸收 IPF 患者肺部中与疤痕相关的 ECM。我们建议开发和利用新颖的体外工具来检验这一假设，目的是确定生物途径和介导生理性胶原吸收的治疗干预。我们建议追求这些目标通过两个目标。在第一个目标中，我们将开发文化系统，使我们能够识别以下信号：促进肺成纤维细胞 ECM 吸收并描绘胶原吸收的分子机制。我们将验证这些检测的高通量发现，并进行聚焦筛选作为概念证明这种方法的价值。我们还将比较先天 ECM 沉积和降解特性 IPF 和对照成纤维细胞和成纤维细胞亚群。在第二个目标中，我们将开发离体肺组织测试疤痕相关 IPF ECM 的治疗调节和清除机制的测定。我们将表征对照和 IPF 肺组织中的基线和刺激诱发的胶原蛋白溶解活性，并定义这种活性与肺细胞类型和组织的组织病理学外观的关联。我们也会测试目标 1 中确定的候选药物能够增加目标纤维状胶原蛋白的降解原生 IPF ECM 环境。拟议的研究将共同建立强大的 ECM 沉积模型以及原代人 IPF 成纤维细胞和离体肺切片中的吸收。该平台将开辟新途径用于识别将 IPF 中的成纤维细胞转变为基质吸收状态的信号和机制，产生开发 IPF 先进疗法的新机遇。