Inhalation Delivery of Exosome and microRNA Therapy for Lung Fibrosis

吸入外泌体和 microRNA 治疗肺纤维化

• 批准号: 10502817
• 负责人: Phuong-Uyen C Dinh
• 金额: $ 37.07万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10502817
• 关键词: Allogenic; Apoptosis; Attenuated; Autologous; Bleomycin; COVID-19/ARDS; Cell Communication; Cell Therapy; Cell Transplantation; Cell secretion; Cells; Cicatrix; Clinical; Clinical Trials; Connective Tissue; Coupled; Data; Development; Diagnosis; Disease; Disease Progression; Disease model; Dose; Down-Regulation; Electroporation; Face; Fibroblasts; Fibrosis; Future; Gene Expression; Gene Expression Profiling; Genes; Genomic approach; Infiltration; Infusion procedures; Inhalation; Label; Lung; Lung Transplantation; Mediating; Mediator of activation protein; MicroRNAs; Modeling; Molecular; Molecular Target; Nanostructures; Ontology; Pathway Analysis; Pathway interactions; Patients; Play; Population; Process; Pulmonary Fibrosis; Rattus; Recombinant Proteins; Recovery; Regenerative Medicine; Resolution; Risk; Rodent Model; Role; Route; Safety; Severities; Severity of illness; Small Interfering RNA; Source; Survival Rate; TGF Beta Signaling Pathway; Testing; Therapeutic; Therapeutic Effect; Transforming Growth Factor beta; Tumorigenicity; Vesicle; Visualization; Work; adult stem cell; antifibrotic treatment; cell type; cellular targeting; clinical translation; curative treatments; differential expression; engineered exosomes; exosome; extracellular vesicles; fibrotic lung; first-in-human; idiopathic pulmonary fibrosis; immunogenicity; in vivo; inhibitor; knock-down; loss of function; lung injury; lung repair; macrophage; non-genetic; novel; novel therapeutic intervention; preclinical efficacy; preclinical safety; preservation; pulmonary function; regenerative; repaired; response; single cell sequencing; single-cell RNA sequencing; stem cell exosomes; stem cell therapy; stem cells; tissue repair; uptake

PROJECT ABSTRACT Idiopathic pulmonary fibrosis (IPF) is an ultimately fatal disease whose only curative treatment is lung transplant. IPF is characterized by formation of fibrotic lesions in the lung, eventually resulting in scarring and progressive loss of lung function. Despite some newer treatments, IPF patients still have a median survival rate of only 3-5 years once diagnosed. Clearly, new therapeutic approaches are needed to treat this devastating disease. A promising avenue of approach is stem cell therapy. In the past 8 years, our lab has been developing lung spheroid cells (LSCs) as a novel source of therapeutic lung cells, and FDA approval of clinical trials with LSC treatment of patients with IPF are being pursued. Nonetheless, stem cell-based therapy faces several important limitations. Live cells need to be carefully preserved and processed before usage, and cell transplantation carries certain immunogenicity and tumorigenicity risks. Importantly, live stem cells cannot be delivered to the lung via inhalation, which is the most convenient and effective route to deliver therapeutics to the lung. Recently, we and others have made the novel and exciting observation that many adult stem cells exert their beneficial effects mainly through secretion of regenerative factors that go on to promote endogenous repair. In the preliminary studies that form the basis for this application, we have discovered that secretions from cultured LSCs are just as, if not more, effective than the LSCs themselves in attenuating and resolving IPF in rodent models of the disease. In the quest for active components in the LSC secretions, we found that LSCs secrete large numbers of exosomes (30-150 nm vesicles secreted by numerous cell types). We have shown that exosomes derived from LSCs (LSC-Exo) are therapeutic and regenerative to the injured lung, suggesting these nanostructures are largely responsible for the reparative response to LSC secretions in rodent models of IPF. It is known that exosomes carry microRNAs (miRs) cargoes that could play important roles in cell-cell communication and tissue repair, and indeed we found that LSC-Exo are highly enriched with miR- 30a and Let-7. In this proposed study, we plan to determine safety and efficacy as well as medium effective dose of LSC-Exo required for lung repair in rodent models of IPF, to determine the major recipient cells of LSC-Exo in the lung, and determine that the relevant molecular target(s) of exosomal mediated repair and recovery. We hypothesize that key miRs withing LSC-Exo such as miR-30a and Let-7 are mediators of the TGF-beta signaling pathway, using the data produced by scRNA-Seq we will finally determine whether further miR enrichment in these exosomes achieves optimal lung repair. The development of cell-free or non-living therapeutics derived from stem cells has the potential to revolutionize current regenerative medicine practice.

项目摘要 特发性肺纤维化（IPF）是一种最终致命的疾病，唯一的治疗方法是肺治疗 移植。 IPF 的特点是肺部形成纤维化病变，最终导致 疤痕和肺功能进行性丧失。尽管有一些新的治疗方法，IPF 患者仍然患有 一旦确诊，中位生存率仅为3-5年。显然，新的治疗方法 需要治疗这种毁灭性的疾病。干细胞疗法是一种有前景的方法。在 在过去的 8 年里，我们的实验室一直在开发肺球状细胞 (LSC) 作为一种新的治疗来源 肺细胞，FDA 正在寻求批准 LSC 治疗 IPF 患者的临床试验。 尽管如此，基于干细胞的疗法面临着几个重要的限制。活细胞需要 使用前经过精心保存和处理，细胞移植具有一定的 免疫原性和致瘤性风险。重要的是，活干细胞不能被输送到肺部 通过吸入，这是将治疗药物输送到肺部的最方便、最有效的途径。 最近，我们和其他人进行了新颖且令人兴奋的观察，即许多成体干细胞 主要通过分泌再生因子来发挥其有益作用，这些再生因子继续促进 内源性修复。在构成本申请基础的初步研究中，我们有 发现培养的 LSC 的分泌物与 LSC 一样有效，甚至更有效 自身在啮齿动物模型中减轻和解决 IPF 的作用。在追求主动的过程中 通过对LSC分泌物中的成分进行分析，我们发现LSC分泌大量的外泌体（30-150 nm 囊泡由多种细胞类型分泌）。我们已经证明，源自 LSC 的外泌体 （LSC-Exo）对受伤的肺部具有治疗作用和再生作用，表明这些纳米结构是 很大程度上负责 IPF 啮齿动物模型中 LSC 分泌物的修复反应。我们都知道 外泌体携带的 microRNA (miR) 货物可能在细胞间发挥重要作用 通讯和组织修复，事实上我们发现 LSC-Exo 高度富含 miR- 30a 和 Let-7。在这项拟议的研究中，我们计划确定安全性和有效性以及中度 IPF 啮齿动物模型肺修复所需的 LSC-Exo 有效剂量，以确定主要 肺中LSC-Exo的受体细胞，并确定外泌体的相关分子靶点 介导的修复和恢复。我们假设 LSC-Exo 中的关键 miR，例如 miR-30a 和 Let-7 是 TGF-β 信号通路的介质，使用 scRNA-Seq 产生的数据，我们 最终将确定这些外泌体中进一步富集 miR 是否能实现最佳的肺修复。 源自干细胞的无细胞或无生命疗法的开发有可能 彻底改变当前的再生医学实践。