Inhalation Delivery of Exosome and microRNA Therapy for Lung Fibrosis

吸入外泌体和 microRNA 治疗肺纤维化

基本信息

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
项目类别：
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

项目摘要

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）是一种最终致命疾病，其唯一的治疗方法是肺移植。 IPF的特征是形成肺中纤维化病变，最终导致疤痕和肺部功能的逐渐丧失。尽管有一些较新的治疗方法，但IPF患者仍然有诊断后的中位生存率仅为3 - 5年。显然，新的治疗方法是需要治疗这种毁灭性疾病。有希望的方法是干细胞疗法。在过去8年，我们的实验室一直在开发肺球体细胞（LSC）作为新的治疗来源正在追求肺部细胞，以及通过LSC治疗IPF患者的LSC治疗的FDA批准。尽管如此，基于干细胞的治疗仍面临一些重要的局限性。活细胞需要在使用前精心保存和处理，并且细胞移植具有某些免疫原性和肿瘤性风险。重要的是，活干细胞不能传递到肺通过吸入，这是向肺部输送治疗剂的最方便和有效途径。最近，我们和其他人对许多成年干细胞进行了小说而令人兴奋的观察主要是通过分泌的再生因素来发挥其有益效果内源修复。在构成本应用基础的初步研究中，我们有发现培养的LSC的分泌物比LSC同样有效，甚至更有效自己在疾病的啮齿动物模型中衰减和解决IPF。寻求活跃 LSC分泌物中的成分，我们发现LSC分泌了大量外泌体（30-150 NM囊泡由多种细胞类型分泌）。我们已经证明了源自LSC的外泌体（LSC-EXO）对受伤的肺有治疗性和再生性，表明这些纳米结构是在IPF的啮齿动物模型中，在很大程度上负责对LSC分泌的重复响应。我们都知道外泌体携带MicroRNA（miR）货物，可以在细胞细胞中起重要作用通信和组织修复，的确我们发现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功效在IPF啮齿动物模型中，肺修复需要的有效剂量的LSC-EXO，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标介导的维修和恢复。我们假设使用LSC-EXO（例如mir-30a）和 Let-7是TGF-BETA信号通路的介体，使用Scrna-Seq WE产生的数据最终将确定这些外泌体中进一步的miR富集是否实现最佳肺修复。从干细胞中得出的无细胞或非生物治疗剂的发展具有革新当前的再生医学实践。