Mechanisms for treating ischemic wounds with human adipocyte derived stem cell exosomes

人脂肪细胞来源的干细胞外泌体治疗缺血性伤口的机制

• 批准号: 9349163
• 负责人: DENISE Ratzlaff COOPER
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9349163
• 关键词: Accident and Emergency department; Address; Adipocytes; Adipose tissue; Adult; Aging; Alpha Cell; Arginine; Beds; Binding; Biological; Biological Assay; Burn injury; CRISPR/Cas technology; Caring; Cell Cycle; Cell physiology; Cells; Ceramides; Chronically Ill; Clinical; Collagen; Collection; Deposition; Dermal; Diabetes Mellitus; Endocytosis Pathway; Engineering; Environment; Face; Fibroblasts; Freezing; Gene Expression; Genes; Goals; Home environment; Homeostasis; Hospitals; Human; Hypoxia; Impaired wound healing; In Vitro; Inflammation; Inflammatory Response; Injury; Innovative Therapy; Knock-out; Lipids; Liver; MALAT1 gene; Mediating; Mediator of activation protein; Methods; MicroRNAs; Modeling; Nuclear; Ontology; Patients; Phagocytosis; Population; Populations at Risk; Porifera; Preparation; Proteins; Proteomics; RNA; Rattus; Regenerative Medicine; Research; Rodent Model; Role; Serine; Site; Soldier; Source; Spinal cord injury; Standardization; Stem Cell Factor; Stem Cell Research; Stem cells; Supporting Cell; Testing; Therapeutic; Time; Topical application; Traumatic injury; Ulcer; Untranslated RNA; Veterans; Work; Wound Healing; acrosome stabilizing factor; angiogenesis; bone; cell motility; chronic wound; cohort; cold temperature; derepression; diabetic patient; exosome; experience; extracellular; healing; immunoregulation; improved; in vitro Assay; in vivo; inhibitor/antagonist; injury and repair; intercellular communication; interest; migration; nanovesicle; novel therapeutics; older patient; overexpression; paracrine; pre-clinical; programs; protein expression; regenerative; relating to nervous system; repaired; tissue regeneration; transcriptome sequencing; uptake; wound; wound closure

This research will develop an innovative therapy to promote tissue regeneration and reverse the consequences of recalcitrant wound healing. It is particularly relevant to our Veteran population that suffers the consequences of injury and aging. The $5 billion global market for “advanced wound care” is anticipated to triple in the next ten years. We have a rapidly growing cohort of older veterans with diabetes, a new cohort of wounded warriors with injuries that include non-healing ulcers from spinal cord injury, multiple traumatic injuries and burns. Hence Veteran populations are at risk for a staggering number of chronic wounds. We propose to use a secreted nanovesicle from adipose derived stem cells (human ADSC). Adipose tissue is an abundant and renewable source of stem cells. Human ADSC release trophic factors that stimulate endogenous repair mechanisms in wounds, and they have immunomodulatory effects, responding to the microenvironment. They have been shown to heal wounds. However, hADSC research is flawed by the lack of standardization and delivery methods. Our goal is to harness the promise of stem cell regenerative secreted factors to heal wounds. We have identified a wound healing treatment using conditioned media (CM) collected from hADSC in culture. We recognize that CM contains exosomes secreted by hADSC, and the factors within the exosomes act in a paracrine manner. In addition to lipids and proteins, exosomes contain various species of RNA. One type, long non- coding RNA, is of particular interest in that it is only secreted by stem cells. Once cells start to differentiate to lineages such as bone and adipocytes, it is retained for a nuclear function. The secreted long non-coding RNA that we find in exosomes is MALAT1. It functions in wound healing, and we predict that exosomes rich in MALAT1 will be able to heal a variety of wounds. We hypothesize that MALAT probably derepresses genes for migration, proliferation, and angiogenesis via its interaction with microRNAs. We will examine how exosomes function in vitro in a human dermal fibroblast wound healing assay. We will use a rat ischemic wound model for examining how exosomes function in vivo. Both of these models will let us optimize the number and concentrations of exosomes that allow for wound closure and modulation of angiogenesis. We propose improvements to hADSC that will allow for collection of more exosomes. Exosomes are more stable than stem cells for wound applications. They can be stored in low temperatures, applied topically and be made available to wounds in emergency rooms and for home use. This project is the first step in making hADSC exosomes a clinical reality for wound repair and tissue regeneration.

这项研究将开发一种创新疗法来促进组织再生并逆转 伤口难以愈合的后果。这与我们的退伍军人群体特别相关 遭受受伤和衰老的后果。价值 50 亿美元的“先进技术”全球市场 “伤口护理”预计在未来十年内将增加两倍。我们的老年人群正在快速增长 患有糖尿病的退伍军人，这是一群新的受伤战士，他们的伤势包括无法治愈 脊髓损伤、多处外伤和烧伤引起的溃疡。因此退伍军人人口 面临数量惊人的慢性伤口的风险。我们建议使用分泌的纳米囊泡 来自脂肪干细胞（人 ADSC）。脂肪组织是一种丰富且可再生的 干细胞的来源。人类 ADSC 释放刺激内源修复的营养因子 伤口机制，并且它们具有免疫调节作用，对 微环境。它们已被证明可以治愈伤口。然而，hADSC 研究存在缺陷 由于缺乏标准化和交付方法。我们的目标是利用茎的承诺 细胞再生分泌因子来治愈伤口。我们已经确定了伤口愈合治疗方法 使用从培养物中的 hADSC 收集的条件培养基 (CM)。我们认识到 CM 包含 hADSC 分泌的外泌体，外泌体内的因子以旁分泌方式发挥作用。在 除了脂质和蛋白质外，外泌体还含有多种 RNA。一种，长非 编码RNA的蛋白质特别令人感兴趣，因为它仅由干细胞分泌。一旦细胞开始 分化为骨细胞和脂肪细胞等谱系，保留核功能。这 我们在外泌体中发现的分泌性长非编码RNA是MALAT1。它作用于伤口 我们预测富含 MALAT1 的外泌体将能够治愈各种伤口。 我们假设 MALAT 可能会去抑制迁移、增殖和增殖的基因 通过与 microRNA 的相互作用来促进血管生成。我们将研究外泌体如何在体外发挥作用 人真皮成纤维细胞伤口愈合测定。我们将使用大鼠缺血性伤口模型 检查外泌体在体内的功能。这两种模型都可以让我们优化数量 以及允许伤口闭合和调节血管生成的外泌体浓度。我们 提出对 hADSC 进行改进，以收集更多的外泌体。外泌体是 在伤口应用中比干细胞更稳定。它们可以在低温下保存， 局部使用，可用于急诊室和家庭的伤口。这 该项目是使 hADSC 外泌体成为伤口修复和组织临床现实的第一步 再生。