New roles of endothelial regrowth in ischemic tissue recovery and regeneration

内皮再生在缺血组织恢复和再生中的新作用

• 批准号: 10596161
• 负责人: Tsutomu Kume
• 金额: $ 56.68万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596161
• 关键词: Affect; Animals; Attenuated; B-Lymphocytes; Blood Vessels; CXCR4 gene; Cardiovascular Diseases; Cells; Data; Defect; Deposition; Disease; Embryo; Endothelial Cells; Endothelium; Extravasation; FOXC1 gene; FOXC2 gene; Genetic Transcription; Goals; Growth; Homeostasis; Impairment; Intestines; Investigation; Ischemia; Ischemic Bowel Disease; Knock-out; Mission; Molecular; Mucous Membrane; Mus; Mutation; Natural regeneration; Paracrine Communication; Participant; Pathologic; Pathway interactions; Patients; Perinatal mortality demographics; Phenotype; Process; Proliferating; Public Health; Recovery; Reperfusion Injury; Reperfusion Therapy; Research; Role; Signal Transduction; Small Intestines; Stromal Cell-Derived Factor 1; Stromal Cells; Tamoxifen; Testing; Therapeutic; Tissues; United States National Institutes of Health; Vascular Endothelial Cell; Vascular Permeabilities; Vascular System; Villus; angiogenesis; autocrine; beta catenin; experimental study; gene function; improved; injury recovery; intestinal injury; lymphatic vessel; mature animal; paracrine; repaired; restoration; role model; single-cell RNA sequencing; stem cell niche; stem cells; tissue regeneration; tissue repair

Project Summary Tissue regeneration and repair is essential for maintaining physiological homeostasis and relies on the precise control of molecular networks that regulate, or are regulated by, the vasculature. Endothelial cells (ECs) present in the blood and lymphatic vessels (i.e., BECs and LECs, respectively) are crucial participants in the vascular- dependent processes that restore damaged tissue, because they control the secretion of paracrine factors from both the vessels themselves and nearby cells. However, the mechanisms by which ECs govern the activity of other cellular components that participate in the recovery of injured tissues have yet to be adequately characterized. The long-term goal of our lab is to elucidate the fundamental processes that regulate blood- and lymphatic-vessel function and to understand how disruption of these mechanisms leads to pathological vascular defects. We have previously shown that in mice, global homozygous knockout mutations of Foxc1 and/or Foxc2 are associated with vascular anomalies; however, the mutations also lead to embryonic or perinatal lethality, so attempts to determine how the two Foxc genes function in adult animals have generally been inconclusive. To overcome this limitation, we generated a line of mice carrying tamoxifen-inducible, endothelial cell (EC)-specific, compound Foxc1;Foxc2 mutations (i.e., EC-Foxc-DKO mice), and the results from preliminary investigations with these animals indicate that the mutations impair regeneration of the small intestine after ischemia- reperfusion (I/R) injury by causing defects in (1) the regrowth of intestinal blood and lymphatic vessels, (2) the formation of subepithelial stromal cells (e.g., telocytes), (3) the expression of CXCL12 and R-spondin3 in intestinal BECs and LECs, respectively and (4) activation of the Wnt/β-catenin pathway in intestinal stem cells (ISCs). CXCL12 is known to regulate angiogenesis, while R-spondin3 protects mice from vascular leakage, and the two factors cooperatively stimulate canonical Wnt/β-catenin signaling, which subsequently regulates the proliferation of ISCs. Thus, our central hypothesis is that the transcriptional activity of Foxc1/c2 in BECs and LECs contributes to vascular repair and intestinal regeneration by regulating CXCL12 and R-spondin3 signaling. We will test our central hypothesis by pursuing the two Specific Aims: (1) To determine whether Foxc1 and Foxc2 are required for repair of the intestinal vasculature during recovery from intestinal injury, (2) To determine the mechanisms by which Foxc1 and Foxc2 regulate blood vessel recovery and intestinal regeneration, and (3) To determine whether Foxc1 and Foxc2 regulate lymphatic vessel recovery and intestinal regeneration. In summary, the experiments described in this proposal will provide crucial information about how Foxc1/c2 expression in vascular ECs contributes to intestinal repair and regeneration. Furthermore, since vascular deficiencies contribute to a variety of ischemic disorders, our findings may have important implications for other ischemic conditions that are associated with impairments in tissue regeneration, such as cardiovascular disease.

项目概要 组织再生和修复对于维持生理稳态至关重要，并依赖于精确的 控制调节血管系统或受血管系统调节的分子网络。 血管和淋巴管中的细胞（分别是 BEC 和 LEC）是血管生成的重要参与者。 恢复受损组织的依赖过程，因为它们控制旁分泌因子的分泌 然而，ECs 控制血管活性的机制。 参与受损组织恢复的其他细胞成分尚未得到充分的了解 我们实验室的长期目标是阐明调节血液和血液的基本过程。 淋巴管功能并了解这些机制的破坏如何导致病理性血管 我们之前已经证明，在小鼠中，Foxc1 和/或 Foxc2 存在整体纯合敲除突变。 与血管异常有关；然而，突变也会导致胚胎或围产期死亡，因此 确定这两个 Foxc 基因如何在成年动物中发挥作用的尝试通常还没有定论。 克服了这一限制，我们培育了一系列携带他莫昔芬诱导的、内皮细胞（EC）特异性的小鼠， 化合物 Foxc1;Foxc2 突变（即 EC-Foxc-DKO 小鼠），以及初步研究的结果 这些动物表明突变会损害缺血后小肠的再生 再灌注 (I/R) 损伤，导致 (1) 肠道血管和淋巴管的再生缺陷，(2) 上皮下基质细胞（例如远程细胞）的形成，（3）CXCL12 和 R-spondin3 的表达 分别是肠道 BEC 和 LEC，以及 (4) 肠道干细胞中 Wnt/β-catenin 通路的激活 (ISC) 已知 CXCL12 可以调节血管生成，而 R-spondin3 可以保护小鼠免受血管渗漏，并且 这两个因素协同刺激经典的 Wnt/β-catenin 信号传导，随后调节 因此，我们的中心假设是 Foxc1/c2 在 BEC 中的转录活性和 LEC 通过调节 CXCL12 和 R-spondin3 信号传导促进血管修复和肠道再生。 我们将通过追求两个具体目标来检验我们的中心假设：（1）确定 Foxc1 和 Foxc2 是否 在肠道损伤恢复期间修复肠道血管系统所需，（2）确定 Foxc1 和 Foxc2 调节血管恢复和肠道再生的机制，以及 (3) 确定 Foxc1 和 Foxc2 是否调节淋巴管恢复和肠道再生。 本提案中描述的实验将提供有关 Foxc1/c2 如何在 此外，由于血管缺陷，血管内皮细胞有助于肠道修复和再生。 导致多种缺血性疾病，我们的发现可能对其他缺血性疾病具有重要意义 与组织再生受损相关的疾病，例如心血管疾病。