Converting wound scar into healing with regeneration

通过再生将伤口疤痕转化为愈合

• 批准号: 10359160
• 负责人: Mayumi Ito
• 金额: $ 46.71万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10359160
• 关键词: Address; Adult; Agonist; Binding; Cell Differentiation process; Cells; ChIP-seq; Cicatrix; Cues; Data; Dermal; Dermis; Development; Developmental Biology; Excision; Failure; Fibroblasts; Fibrosis; Future; Genetic; Genetic Transcription; Goals; Grant; Hair follicle structure; Human; Hypersensitivity skin testing; Immunohistochemistry; Injury; Investigation; Lead; Ligands; Mammals; Mediating; Molecular Target; Mus; Myofibroblast; Natural regeneration; Outcome; Pathway interactions; Phenotype; Play; Process; Receptor Signaling; Recombinant Proteins; Regenerative Medicine; Regulation; Regulator Genes; Role; SHH gene; Signal Pathway; Signal Transduction; Site; Skin; Skin Cancer; Skin graft; Testing; Tissues; Transfection; Up-Regulation; Viral; WNT Signaling Pathway; Wound models; Xenograft Model; Xenograft procedure; appendage; base; beta catenin; bone morphogenetic protein receptors; experimental study; gain of function; healing; human tissue; improved; loss of function; novel strategies; prevent; recombinant virus; regeneration potential; regenerative; regenerative biology; repaired; response; single-cell RNA sequencing; skin regeneration; skin wound; smoothened signaling pathway; synergism; tongue papilla; trait; transcriptome sequencing; wound; wound environment; wound healing

Summary The goal of this grant is to understand the role of SHH signaling in regulation of regenerative healing in mammalian skin wounds. Most mammals including humans are deficient in regenerative skin healing. After injury, wound healing typically results in 'repair' (also defined as fibrosis/scarring). This process fails to fully restore the lost tissue to its original form, resulting in “a cellular band-aid”. The inability of mammals to regenerate normal skin is most notably illustrated by the lack of skin appendages including hair follicles in the wound scar. A major hurdle in hair follicle regeneration in skin wounds is the failure to reestablish the hair follicle inductive niche known as the dermal papilla (DP). Here, we show that SHH activation within 'repaired' (scarring) wound dermis induces de novo DP formation leading to HFN. These data clearly demonstrate that myofibroblasts within scarring wounds are not intrinsically incapable of forming the regenerative niche. We hypothesize that scarring cells may possess the regenerative potential but are missing the crucial developmental cue, SHH. To address this hypothesis, first, we will ask if Wnt signaling, which is active in fibrotic wounds, is required for SHH-driven DP formation and resultant HFN in wounds. For this we will perform loss-of-function studies on b-catenin, an essential effector of canonical Wnt signaling, in genetically modified wounds with ectopic SHH activation (Aim1). We will also examine the potential of mature Wnt-active fibroblasts to convert to DP in Hh-activated wounds after healing is complete. These experiments will address the potential plasticity of fully differentiated cells and how induction by the Hh pathway may alter their phenotype (Aim 1). Second, preliminary single cell RNAseq analyses comparing Hh-activated and WT wounds have identified downstream expression of several BMPs following Hh activation. We will examine the role of BMP signaling in mediating the function of Hh signaling through loss- and gain-of-function studies on BMP receptor signaling. We will also examine the ability of immediate Hh regulators Gli1 and 2, to directly bind BMP gene regulatory sites by Chip-Seq analysis (Aim 2). Third, human wounds strongly resemble 'repaired' murine wounds in many traits. We will test whether human wounds can respond to SHH signals to produce DP by introducing viral active-SmoM2 or soluble SHH agonists into xenograft-and-wounding models. Importantly, we will delineate the response of human dermal cells to SHH through single cell RNAseq analyses, and also compare these results with those from murine wounds (Aim3). These combined studies will advance our understanding of the mechanisms required to induce hair follicle regeneration in an otherwise hairless wound environment and will provide essential new information about how human wounds may compare with murine wounds and be manipulated for improved healing.

概括 这笔赠款的目的是了解SHH信号在调节再生治疗中的作用 哺乳动物皮肤伤口。包括人类在内的大多数哺乳动物都缺乏再生皮肤愈合。后 受伤，伤口愈合通常会导致“修复”（也定义为纤维化/疤痕）。这个过程无法完全 将丢失的组织恢复为原始形式，从而导致“细胞创可贴”。哺乳动物无法再生 正常的皮肤是由于缺乏皮肤附属物在内的包括伤口疤痕中的毛发毛的附属物所说明的。一个 皮肤伤口中毛发再生的主要障碍是未能重新建立头发叶片感应的小众 被称为真皮乳头（DP）。 在这里，我们表明“修复”（疤痕）伤口真皮诱导的shh激活dp dp 形成导致HFN。这些数据清楚地表明，疤痕伤口中的肌纤维细胞不是 本质上无法形成再生生态位。我们假设疤痕细胞可能拥有 再生潜力，但缺少关键的发展提示，嘘。解决这个问题 假设，首先，我们将询问SHH驱动的DP是否需要在纤维化伤口中活跃的Wnt信号传导 伤口中的形成和由此导致的HFN。为此，我们将对B-catenin进行功能丧失研究， 在具有生态SHH激活的转基因伤口中，规范Wnt信号的基本效应器 （AIM1）。我们还将检查成熟的Wnt活性成纤维细胞在HH激活中转化为DP的潜力 愈合后的伤口完成。这些实验将解决完全分化的潜在可塑性 细胞以及HH途径诱导如何改变其表型（AIM 1）。其次，初步单细胞 RNASEQ分析比较HH激活和WT伤口的RNASEQ分析已经确定了几种的下游表达 HH激活后的BMP。我们将检查BMP信号在介导HH信号功能中的作用 通过对BMP受体信号传导的丧失和功能性研究。我们还将研究 立即进行HH调节剂GLI1和2，通过CHIP-SEQ分析直接结合BMP基因调节位点（AIM 2）。第三，人的伤口在许多特征中强烈类似于“修复”的鼠伤口。我们将测试是否 人的伤口可以通过引入病毒活性-SMOM2或固体SHH来响应SHH信号以产生DP 激动剂变成Xenographicang and-nounging模型。重要的是，我们将描述人真皮细胞的反应 通过单细胞RNASEQ分析进行SHH，并将这些结果与鼠伤口的结果进行比较 （AIM3）。这些合并的研究将提高我们对诱导头发所需机制的理解 在原本无毛伤口环境中的叶片再生，将提供有关的基本新信息 人的伤口如何与鼠风相比，并被操纵以改善愈合。