Exosome treatment-induced mechanisms in chronic wound beds - Resubmission - 1

慢性伤口床中外泌体治疗诱导的机制 - 重新提交 - 1

基本信息

批准号：
10350276
负责人：
Piul Sanjana Rabbani
金额：
$ 16.2万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
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：
2022-03-15 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10350276
关键词：
Acceleration Acute Address Affect Animals Anti-Inflammatory Agents Biochemical Process Biological Assay Blood Vessels Bone Marrow Cell Culture Techniques Cell physiology Cells Chronic Chronic Disease Clinical Coculture Techniques Communication Communities Data Dermal Diabetic mouse Disease Dose Endothelial Cells Epitopes Event Excision Gene Proteins Goals Granulation Tissue Growth Human IL6 gene Immune Impaired healing Impaired wound healing In Vitro Inflammation Inflammatory Knowledge Literature Mass Spectrum Analysis Mediating Mediator of activation protein Medical Mentorship Microvascular Dysfunction Modeling Molecular Mus Non-Insulin-Dependent Diabetes Mellitus Pathologic Pathway interactions Patients Phase Phenotype Physiological Plasminogen Activator Inhibitor 1 Play Proteins Proteomics Publishing RNA Research Research Design Resolution Role STAT3 gene Safety Signal Pathway Source Stromal Cells Testing Therapeutic Effect Time Tissues Training Transcript Translational Research Type 2 diabetic Vascular Endothelial Growth Factors Vesicle Wild Type Mouse Wound models acute wound angiogenesis career career development cell motility cell type chronic wound cost design diabetic diabetic ulcer diabetic wound healing educational atmosphere effective therapy epithelial wound evidence base exosome healing immunoreactivity inhibitor intercellular communication macrophage monocyte nanosized nanovesicle neovascularization overexpression pre-clinical preclinical study rate of change rational design response skin wound social spatiotemporal therapy design tool transcriptomics uptake wound wound bed wound closure wound healing wound treatment

项目摘要

PROJECT SUMMARY Chronic wounds present a costly social and medical dilemma, particularly in patients with type II diabetes, but there are no effective treatments. These wounds are characterized by chronic inflammation, severe microvascular complications, and therefore lack expansion of the granulation tissue and proliferation necessary to heal the wound. Recently, exosome administration has emerged as a potent therapy for promoting wound healing but the mechanisms underlying the therapeutic effect are mostly unknown. Exosomes are secreted membranous nanovesicles that can be isolated from cell culture of multipotent stromal cells. In a preclinical type II diabetic model, a single local administration of exosomes was very effective and reduced time to closure to nearly that of wild type mice. We found extensive neovascularization in granulation tissue of exosome-treated wounds and presence of large numbers of macrophages immunoreactive for arginase1, typically indicative of a pro-healing phenotype. Exosomes from multipotent stromal cells carry plasminogen activator inhibitor-1 (PAI-1) among other angiogenesis-associated proteins. When we applied a PAI-1 inhibitor simultaneously with exosomes, the beneficial effect of exosome treatment was partially mitigated. Exosomes can have multiple signaling pathway targets, and PAI-1 is a major component. Our long term objective is to understand the molecular mechanisms we need to address to alter the chronic wound state, and actually promote wound closure. An approach like this is necessary to understand the rationale and safety of exosome treatment. In this study, we are investigating the hypothesis that PAI-1 in bone marrow MPSC exosomes mediates adaptive effects on macrophages and ECs in chronic wound beds, thereby promoting expansion of granulation tissue with effective changes in wound healing trajectory. In Aim 1, we will determine whether overexpression or loss of PAI-1 in exosomes affects the typically delayed diabetic wound healing. We will assess spatiotemporal distribution of macrophages and endothelial cells. In Aim 2, we will analyze the changes in wound bed macrophages and endothelial cells at a single cell resolution, using an integrated epitope and transcriptomics approach. Additionally, in in vitro culture with inflammatory conditions, we will determine molecular changes in macrophages and microvascular endothelial cells, downstream of exosome uptake. The results will demonstrate the mechanisms orchestrating the efficacy of exosome treatment in normalizing inflammation and promoting diabetic wound closure. We will perform the study under the mentorship of Dr. Bruce Cronstein, Dr. Ann Marie Schmidt, and Dr. Thorsten Kirsch, and gain expertise in modeling chronic diseases in preclinical study designs, hyper-inflammatory disease states, macrophage assays, and exosome-mediated mechanisms and cellular communication in a chronic cutaneous wound bed. The collaborative and scientific learning environment at NYU will provide the optimal setting to gain expertise in rational design of evidence-based translational research. This training period will provide the scientific and career development rubric for an independent career in research.

项目概要慢性伤口带来了代价高昂的社会和医疗困境，特别是对于 II 型糖尿病患者，但没有有效的治疗方法。这些伤口的特点是慢性炎症、严重的微血管并发症，因此缺乏必要的肉芽组织扩张和增殖治愈伤口。最近，外泌体给药已成为促进伤口愈合的有效疗法。治愈，但治疗效果背后的机制大多未知。外泌体被分泌可以从多能基质细胞的细胞培养物中分离出膜纳米囊泡。在临床前类型 II 型糖尿病模型中，单次局部给予外泌体非常有效，并且缩短了闭合时间与野生型小鼠接近。我们发现外泌体处理的肉芽组织中有广泛的新生血管形成伤口和存在大量对精氨酸酶1具有免疫反应性的巨噬细胞，通常表明存在促愈合表型。来自多能基质细胞的外泌体携带纤溶酶原激活剂抑制剂-1 (PAI-1) 其他血管生成相关蛋白。当我们同时应用 PAI-1 抑制剂时外泌体治疗的有益效果部分减弱。外泌体可以有多个信号通路靶点，PAI-1是主要成分。我们的长期目标是了解我们需要解决分子机制来改变慢性伤口状态，并实际上促进伤口关闭。像这样的方法对于了解外泌体治疗的基本原理和安全性是必要的。在本研究中，我们正在研究骨髓 MPSC 外泌体中的 PAI-1 介导的假设对慢性创面床巨噬细胞和内皮细胞的适应性作用，从而促进肉芽扩张有效改变伤口愈合轨迹的组织。在目标 1 中，我们将确定是否过度表达或外泌体中 PAI-1 的丢失会影响典型的糖尿病伤口愈合延迟。我们将评估时空巨噬细胞和内皮细胞的分布。在目标 2 中，我们将分析伤口床的变化使用集成表位和转录组学以单细胞分辨率分析巨噬细胞和内皮细胞方法。此外，在具有炎症条件的体外培养中，我们将确定巨噬细胞和微血管内皮细胞，外泌体摄取的下游。结果将证明协调外泌体治疗在使炎症正常化和促进糖尿病伤口闭合。我们将在 Bruce Cronstein 博士、Ann Marie 博士的指导下进行这项研究 Schmidt 和 Thorsten Kirsch 博士获得了临床前研究设计中慢性病建模方面的专业知识，高炎症性疾病状态、巨噬细胞测定以及外泌体介导的机制和细胞慢性皮肤伤口床中的通讯。纽约大学的协作和科学学习环境将为获得循证转化研究合理设计方面的专业知识提供最佳环境。这培训期将为独立的研究生涯提供科学和职业发展标准。