C-KIT SIGNALING IN COLLATERALS REMODELING

侧枝循环重塑中的 C-KIT 信号传导

• 批准号: 10565856
• 负责人: Roberta Marques Lassance Soares
• 金额: $ 12.64万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-17 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10565856
• 关键词: Alternative Therapies; American; Amputation; Anatomy; Arteries; Biology; Blood; Blood Vessels; Blood Volume; Blood capillaries; Blood flow; Bypass; Cell Proliferation; Coronary; Defect; Disease; Ensure; FBXO32 gene; Failure; Functional disorder; GKLF protein; Gangrene; Goals; Growth; Hematopoietic; Hindlimb; Histology; Human; Hypoxia; Impairment; Intervention; Ischemia; Knock-in; Knockout Mice; Limb structure; Lower Extremity; Mediating; Membrane; Modeling; Molecular; Morbidity - disease rate; Mus; Mutant Strains Mice; Mutation; Operative Surgical Procedures; Outcome Study; Pain; Pathway interactions; Patients; Periodicity; Peripheral arterial disease; Phase; Phenotype; Physiologic pulse; Physiological; Play; Procedures; Process; Protein Kinase C; Proteomics; Proto-Oncogene Protein c-kit; Receptor Protein-Tyrosine Kinases; Recovery; Regulation; Rest; Risk; Role; Secondary to; Signal Transduction; Smooth Muscle Myocytes; Stenosis; Tamoxifen; Testing; Tissues; Training; Transgenic Model; Transgenic Organisms; Ubiquitination; United States; Vascular remodeling; Vascularization; angiogenesis; artery occlusion; cell motility; comparison control; conditional knockout; critical limb Ischemia; effective therapy; experimental study; gain of function; imaging modality; improved; limb ischemia; loss of function; microCT; mutant; new therapeutic target; novel; novel therapeutics; patient population; pharmacologic; prevent; reconstitution; restoration; transcriptome sequencing; vascular stress

TITLE: C-KIT SIGNALING IN COLLATERALS REMODELING Critical limb ischemia (CLI) is the most advanced form of peripheral arterial disease. At the moment, endovascular procedures and bypass surgeries are the only effective treatments for limb revascularization. However, a significant proportion of CLI patients are not good candidates for these interventions, requiring primary amputation as the main treatment option. Therefore, an effective pharmacological strategy that prevents CLI and its devastating consequences represents a much- needed alternative for this patient population. Arteriogenesis is a physiological compensatory process in which pre-existing collaterals enlarge and serve as natural bypasses to severe occlusion of arteries. We have recently identified that the c-Kit receptor tyrosine kinase plays a key role in the remodeling of collaterals during arteriogenesis. We found that defective c-Kit function compromises blood flow recovery after hindlimb ischemia, which was not resolved with hematopoietic reconstitution of c-Kit activity. We have confirmed defective arteriogenesis in c-Kit mutant mice compared to controls. Mechanistically, we advanced our understanding of how the c-Kit/Kruppel-like factor 4 (KLF4) pathway orchestrates collateral remodeling. Therefore, our central hypothesis is that c-Kit/KLF-4 signaling controls the recovery of the smooth muscle cell (SMC) contractile phenotype at the maturation phase of arteriogenesis to prevent excessive remodeling and narrowing of collaterals. We propose two Specific Aims (SA) to test our hypothesis. In SA1, we will demonstrate the involvement of vascular c-Kit signaling in arteriogenesis. We will investigate whether inactivation or activation of c-Kit specifically in SMCs will lead to dysfunction or optimal arteriogenesis in unique transgenic models of loss- and gain- of-function of c-Kit, respectively. We will also identify molecular mechanisms that can be altered by the loss- or gain-of-function on SMC c-Kit. In SA2, we will dissect the molecular pathways upstream and downstream of c-Kit in SMCs. First, we will investigate whether cyclic strain dictates the oscillatory expression of the c-Kit receptor by activating protein kinase C, leading to c-Kit suppression. Next, we will determine whether c-Kit inhibits KLF4 through FBXO32-mediated ubiquitination, thereby restoring the contractile SMC phenotype and preventing defective remodeling. RELEVANCE: Approximately 80,000 major lower-extremity amputations occur in the United States every year due to CLI. In this study, we will identify novel therapeutic targets to improve arteriogenesis and prevent or alleviate the devastating effects of CLI. Moreover, we will identify the molecular mechanism that dictates phenotypic switching in SMCs, which may be applicable to the understanding and treatment of other vascular occlusive diseases.

标题：C-KIT信号在侧链重塑中的作用 严重肢体缺血（CLI）是外周动脉疾病的最高级形式。目前， 血管内手术和旁路手术是肢体血管畸形的唯一有效治疗方法。 血运重建然而，很大一部分CLI患者并不是这些治疗的良好候选者。 主要的治疗选择是截肢。因此，有效的 预防CLI及其破坏性后果的药理学策略代表了一个非常- 这一患者群体需要替代品。动脉形成是一种生理性代偿过程 其中预先存在的侧支扩大并作为动脉严重闭塞的自然旁路。 我们最近发现，c-Kit受体酪氨酸激酶在重构中起着关键作用， 在动脉形成过程中的侧枝。我们发现有缺陷的c-Kit功能会损害血流 后肢缺血后恢复，c-Kit造血重建未解决 活动与对照组相比，我们已经证实c-Kit突变小鼠的动脉生成缺陷。 从机制上讲，我们进一步了解了c-Kit/Kruppel样因子4（KLF 4）通路是如何在细胞内表达的。 协调侧支重塑因此，我们的中心假设是，c-Kit/KLF-4信号转导 控制平滑肌细胞（SMC）收缩表型在成熟期的恢复， 动脉生成以防止过度重塑和侧支狭窄。我们提出两个具体的 目的（SA）来检验我们的假设。在SA 1中，我们将证明血管c-Kit的参与， 动脉生成中的信号传导。我们将研究是否c-Kit的失活或激活，特别是在 SMC将导致功能障碍或最佳动脉生成在独特的转基因模型的损失和获得- c-Kit的功能。我们还将确定可以被改变的分子机制， SMC c-Kit功能丧失或获得。在SA 2中，我们将剖析上游的分子通路， SMC中c-Kit的下游。首先，我们将调查是否周期应变支配振荡 通过激活蛋白激酶C来抑制c-Kit受体的表达，导致c-Kit抑制。接下来我们 将确定c-Kit是否通过FBXO 32介导的遍在化抑制KLF 4，从而恢复 收缩性SMC表型和防止缺陷性重塑。 相关性：在美国，大约有80，000例下肢截肢手术 每年，由于CLI。在这项研究中，我们将确定新的治疗靶点，以改善动脉生成 预防或减轻CLI的破坏性影响。此外，我们将识别分子 决定SMC中表型转换的机制，这可能适用于理解 以及其他血管闭塞性疾病的治疗。

项目成果

c-Kit expression in smooth muscle cells reduces atherosclerosis burden in hyperlipidemic mice.

• DOI: 10.1016/j.atherosclerosis.2021.03.004
• 发表时间: 2021-05
• 期刊: Atherosclerosis
• 影响因子: 5.3
• 作者: Zigmond ZM;Song L;Martinez L;Lassance-Soares RM;Velazquez OC;Vazquez-Padron RI
• 通讯作者: Vazquez-Padron RI

Ischemic-Trained Monocytes Improve Arteriogenesis in a Mouse Model of Hindlimb Ischemia.

• DOI: 10.1161/atvbaha.121.317197
• 发表时间: 2022-03
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Falero-Diaz G;Barboza CA;Pires F;Fanchin M;Ling J;Zigmond ZM;Griswold AJ;Martinez L;Vazquez-Padron RI;Velazquez OC;Lassance-Soares RM
• 通讯作者: Lassance-Soares RM