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.

项目摘要慢性伤口带来了昂贵的社会和医疗困境，特别是在II型糖尿病患者中，但没有有效的治疗方法。这些伤口的特征是慢性炎症，严重微血管并发症，因此缺乏肉芽组织的扩张和必要的增殖治愈伤口。最近，外部给药已成为促进伤口的有效疗法愈合，但是治疗效果的基础机制大多未知。外泌体分泌可以从多能基质细胞的细胞培养物中分离出来的膜纳米膜。在临床前类型 II糖尿病模型，单一的局部外泌体给药非常有效，并且减少了闭合时间几乎是野生型小鼠。我们发现外泌体治疗的肉芽组织中广泛的新血管化伤口和大量巨噬细胞免疫反应性精氨酸酶1，通常表明亲治表型。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）在其他与血管生成相关的蛋白质中。当我们同时应用PAI-1抑制剂时外泌体，外泌体治疗的有益作用得到了部分缓解。外泌体可以有多个信号通路目标，PAI-1是主要组成部分。我们的长期目标是了解分子机制我们需要解决以改变慢性伤口状态，并实际促进伤口关闭。这样的方法是了解外泌体治疗的基本原理和安全性的必要方法。在这项研究中，我们正在研究以下假设：骨髓MPSC外泌体中的PAI-1介导对慢性伤口床中巨噬细胞和EC的适应性影响，从而促进了肉芽的扩张组织具有有效变化伤口愈合轨迹的组织。在AIM 1中，我们将确定是过表达还是外泌体中PAI-1的丧失会影响典型延迟的糖尿病伤口愈合。我们将评估时空巨噬细胞和内皮细胞的分布。在AIM 2中，我们将分析伤口床的变化使用集成的表位和转录组学以单细胞分辨率的巨噬细胞和内皮细胞方法。此外，在炎症条件下的体外培养中，我们将确定分子变化巨噬细胞和微血管内皮细胞，外部摄取的下游。结果将证明策划外泌体治疗在标准化炎症和促进的机制糖尿病伤口闭合。我们将在布鲁斯·克朗斯坦博士的指导下进行研究，安·玛丽博士 Schmidt和Thorsten Kirsch博士，并在临床前研究设计中建模慢性疾病方面获得了专业知识，高炎性疾病状态，巨噬细胞测定和外泌体介导的机制和细胞在慢性皮肤伤口床上进行交流。纽约大学的协作和科学学习环境将提供最佳环境，以获得基于证据的转化研究理性设计的专业知识。这培训期将为研究独立职业提供科学和职业发展标题。