Cell-free regenerative approach in wound healing

伤口愈合中的无细胞再生方法

• 批准号: 10616469
• 负责人: Niketa A. Patel
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616469
• 关键词: Acceleration; Adult; Affect; Aging; Amputation; Biochemical; Biological; COVID-19; COVID-19 patient; Cell Death; Cells; Chemicals; Chronic; Cicatrix; Clinic; Complement; Data; Dermal; Diabetes Mellitus; Economic Burden; Evaluation; Fibroblasts; Genomics; Goals; Harvest; Homeostasis; Human; Impaired wound healing; In Vitro; Infection; Inflammation; Inflammatory; Injury; Intervention; Ischemia; Joints; Laceration; Learning; Lesion; Measures; Mechanics; Mediating; Molecular Target; Natural regeneration; Operative Surgical Procedures; Outcome; Pathway interactions; Patients; Pharmacodynamics; Physiological; Probability; Process; Proliferating; Psychological Impact; Quality of life; Readiness; Recovery of Function; Regenerative pathway; Research; Research Design; Risk Factors; Rodent; Safety; Severities; Site; Skin; Skin repair; Small Interfering RNA; Smoking; Strategic Planning; Structure; Surgical Wound Infection; Surgical incisions; System; Therapeutic; Tissues; Topical application; Transcriptional Activation; Translating; United States Department of Veterans Affairs; Untranslated RNA; Veterans; Veterans Disability Claims; Wound models; adipose derived stem cell; chronic wound; comorbidity; exosome; experience; experimental study; genome-wide; healing; improved; improved outcome; in vivo; in vivo Model; infection risk; injury recovery; interdisciplinary approach; keratinocyte; knock-down; novel; overexpression; pre-clinical; regenerative approach; repaired; response; skin wound; stem cell exosomes; tissue regeneration; tissue repair; transcriptome sequencing; treatment response; wound; wound closure; wound dressing; wound healing; wound treatment

Wounds, which are breaks in structure and function of skin, are healed through a regulated process initiated with tissue homeostasis followed by repair response involving inflammation, proliferation and remodeling. Wounds may be due to cuts and lacerations or incisional wounds post surgeries. At times, this healing is delayed or slowed resulting in chronic recalcitrant wounds. Risk factors for impaired wound healing include surgical site infection and comorbidities such as aging, smoking, diabetes. Strikingly, ongoing research with COVID19 patients showcased increased severity in wounds and lesions compared to normal patients(1-4). Recalcitrant wounds per se are not life-threatening but often lead to chronic wound related sequelae. The long-term outcome of impaired wound healing may lead to scarring or amputations, and often has a substantial psychological impact. The treatment of recalcitrant wounds and stimulation of healing is a critical need and consistent with priorities outlined by Department of Veterans Affairs (2018-2024 Strategic plan). While multiple advances in wound dressing materials promote healing, a noteworthy gap remains in the interventions available which considerably improve the outcome of injury. Towards this goal, a therapeutic approach was undertaken to harvest the potential of exosomes derived from human adipose stem cells (hASCexo) mediating tissue repair and regeneration. Thus, the overarching hypothesis is that hASCexo enable wound healing and recovery of function by modulating genomic pathways following injury. This project will elucidate the safety and efficacy of hASC exosomes applied topically to repair dermal wounds to move the therapy closer to the clinic. Towards this goal, the proposal will undertake an in vitro evaluation of genomic targets complemented with an in vivo approach to determine response to treatment. These hypotheses will be validated using a multi-disciplinary approach including pre- clinical, physiological, cellular and biochemical experiments. Specific Aim 1: Determine the molecular targets of the lncRNA cargo of hASC exosomes that promote wound healing: Using RNAseq to examine the contents of hASC exosomes, two long noncoding RNAs (lncRNAs) were identified which are highly enriched in the hASCexo and are pivotal in recovery post injury. Human dermal fibroblasts (HDF) and human epidermal keratinocytes (HEK) will be used with siRNA mediated knockdown or over-expression of these lncRNA in hASCexo. Cellular and biochemical outcomes will be measured in in vitro wound models along with elucidation of genomic changes in response to hASCexo treatment. These will be validated ex vivo in human skin explants. Specific Aim 2: Evaluation of response to hASC exosomes’ treatment in an in vivo wound model: The hypothesis is that repair and regeneration of wounds by topical application of hASCexo depends on its pharmacodynamics and efficacy. To elucidate this along with its genomic impact, an in vivo rodent ischemic wound model will be used to determine physiological response of hASCexo treatment to improve healing outcomes. Its efficacy to promote healing in conditions of underlying infection will be evaluated The hASCexo provide a novel, cell-free regenerative approach to accelerate wound closure and to alleviate chronic effects of impaired wound healing. Robust preliminary data supports feasibility and successful implementation of hASC exosomes’ topical treatment in wound healing. The research design is expected to greatly enhance translatability by the “learn and confirm” approach by integrating the in vitro results in the in vivo model. The therapeutic potential of hASCexo will be thoroughly evaluated for safety and scalability such that it may be translated successfully to the treatment of Veterans’ wounds and substantially improve outcome of chronic wound related sequelae.

伤口是皮肤结构和功能的断裂，通过调节过程得到治愈 通过组织体内稳态开始，然后修复反应炎症，增殖和 重塑。伤口可能是由于切割，撕裂或切开的逻辑后手术。有时， 这种愈合延迟或缓慢，导致慢性顽固逻辑。受损的风险因素 伤口愈合包括手术部位感染和合并症，例如衰老，吸烟，糖尿病。 令人惊讶的是，与COVID19患者进行的正在进行的研究表明，伤口的严重程度增加 与正常患者相比，病变（1-4）。顽固的逻辑本身不是威胁生命的，而是经常 导致慢性伤口相关的后遗症。伤口愈合受损的长期结果可能导致 疤痕或截肢，通常会产生重大的心理影响。处理 顽固的逻辑和刺激治愈是一个迫切的需求，并且与概述的优先级一致 由退伍军人事务部（2018-2024战略计划）。而伤口的多次进步 敷料材料促进了康复，在可用的干预措施中仍然存在一个值得注意的差距 大大改善了伤害的结果。朝向这个目标，一种治疗方法是 旨在收获源自人脂肪干细胞的外泌体的潜力（Hascexo） 介导组织修复和再生。那就是总体假设是hascexo启用 伤口愈合和功能通过调节损伤后基因组途径的恢复。这 项目将阐明局部应用HASC外泌体的安全性和效率 伤口使治疗更靠近诊所。为了实现这一目标，该提议将进行 通过体内方法完成的基因组靶标的体外评估，以确定对 治疗。这些假设将使用多学科方法进行验证 临床，物理，细胞和生化实验。特定目标1：确定分子 促进伤口愈合的HASC外泌体的LNCRNA货物的靶标：使用RNASEQ 检查了HASC外泌体的含量，两个长的非编码RNA（LNCRNA）已被鉴定出来 在Hascexo中高度富集，并且在受伤后恢复方面是关键的。人真皮成纤维细胞 （HDF）和人表皮角质形成细胞（HEK）将与siRNA介导的敲低或 这些lncRNA在hascexo中的过表达。将测量细胞和生化结果 体外世界模型以及对Hascexo的响应阐明基因组变化 治疗。这些将在人类皮肤外植体中被验证。特定目标2：评估 在体内伤口模型中对HASC外泌体治疗的反应：假设是修复和 通过局部应用Hascexo的伤口再生取决于其药效学和 功效。为了阐明这一点及其基因组影响，体内啮齿动物缺血性伤口模型将 用于确定Hascexo治疗的身体反应以改善愈合结果。它是 将评估在潜在感染条件下促进愈合的功效 Hascexo提供了一种新型的，无细胞的再生方法来加速伤口闭合 并减轻伤口愈合受损的慢性影响。强大的初步数据支持可行性 并成功实施了HASC外泌体在伤口愈合中的局部治疗方法。研究 预计设计通过“学习和确认”方法可以通过整合来增强可转换性 体外导致体内模型。 Hascexo的治疗潜力将彻底 评估安全性和可伸缩性，使其可以成功地转化为 退伍军人的胜利并大大改善了与慢性伤口相关的后遗症的结果。