Epigenetic and Transcriptional Regulation of Angiogenic Regulator CD36 and Transformation of Capillaries into Small Arteries

血管生成调节因子 CD36 的表观遗传和转录调控以及毛细血管向小动脉的转化

基本信息

批准号：
9752936
负责人：
BIN Ren
金额：
$ 37.18万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2021-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9752936
关键词：
Actins Adult Affinity Chromatography Angiogenic Switch Area Arteries Binding Biological Biological Assay Biology Blood Circulation Blood Vessels Blood capillaries CD36 gene Capillary Endothelial Cell Cardiovascular Diseases Cardiovascular system Cells Clinical Clinical Trials Confocal Microscopy Development Diet Dimensions Down-Regulation Endothelial Cells Endothelium Evaluation Event FOXO1A gene Failure Gene Expression Genes Genetic Genetic Transcription Goals Growth Growth Factor HDAC7 histone deacetylase Heart Heart Diseases Hindlimb Histones Impairment In Situ Ischemia KDR gene Knowledge Lead Ligation Lipids Mediating Mediator of activation protein Messenger RNA Modeling Modification Molecular Molecular Biology Mus Myocardial Ischemia Obesity Organ Pathogenesis Pathway interactions Peripheral Vascular Diseases Play Process Recovery Regulation Resolution Ribosomes Role SIRT1 gene Series Signal Pathway Signal Transduction System Testing Therapeutic Thrombosis Time Tissues Transcriptional Regulation Transgenic Organisms Translating Treatment Efficacy Untranslated RNA Vascular Diseases Vascular Endothelial Growth Factors Work Zebrafish angiogenesis arteriole base coronary artery occlusion effective therapy epigenetic regulation experimental study feeding gene repression imaging modality in vivo innovation insight lysophosphatidic acid matrigel molecular imaging mouse model next generation novel novel strategies novel therapeutic intervention promoter protein protein interaction recruit repaired single molecule single-cell RNA sequencing therapeutic target tissue repair tool trafficking transcriptome transcriptome sequencing transdifferentiation

项目摘要

Project Summary Arteriolar differentiation and formation of small arteries from very tiny blood vessels (capillary arterialization) are fundamental processes underlying ischemic organ recovery from a noxious insult. Despite its obvious clinical and biological importance, little is known about the molecular mechanisms of these events and their regulation. We have recently discovered that transcriptional repression of CD36 is involved in a proangiogenic and proarteriogenic reprogramming of capillary endothelial cells (EC) or microvascular EC (MVECs), and is implicated in capillary arterialization. Specifically, we have shown that LPA/PKD-1 signaling-mediated CD36 transcriptional repression plays a key role in promoting arteriogenic gene expression in MVECs, and microvascular remodeling in vivo. Moreover, EC-specific deletion of pkd-1 showed impaired recovery from ischemic insult. These unexpected findings led to the hypothesis that CD36 gene repression via LPA/PKD-1 signaling axis reprograms MVECs to differentiate into arteriolar ECs and promotes capillary arterialization. To test this hypothesis, we have established two- and three-dimensional MVEC culture systems, and in vivo Matrigel assays. More importantly, we have established hindlimb ischemia models in unique cd36 gene deficient mice with the EC-specific translating ribosome affinity purification (TRAP), and EC-specific pkd-1 deficient mice or EC-specific pkd-1 deficient TRAP mice. These novel TRAP transgenic lines have targeted gene deficiency and carry an EC-specific EGFP-tag that allows us to directly purify ribosome-bound mRNA from ECs in vivo for next generation RNA-sequencing, facilitating probes of EC-specific transcriptome and ribosome binding of noncoding RNAs. The availability of this mouse model provides us for the first time with the ability to devise experiments to gain new and fundamental insights into biology of adult arteriolar growth under ischemic conditions. Using these tools,, we aim to 1) prove that in MVECs the LPA/PKD-1-CD36 signaling axis is essential to regulate MVEC reprogramming and arteriolar differentiation; 2) test the hypothesis that CD36 transcriptional repression via the LPA/PKD-1-FoxO1 signaling axis is a critical component of a genetic reprogramming switch to promote capillary arterialization (de novo arteriogenesis). Together with assays using a series of molecular biology and imaging methods single cell RNA-sequencing as well as zebrafish models, we expect to accomplish these aims and provide a comprehensive evaluation of arteriolar differentiation and capillary arterialization. This will illuminate poorly explored and poorly understood aspects of vascular biology. This proposal focuses on de novo arteriogenesis in adult because of its remarkably practical significance. Getting a greater understanding of capillary arterialization in adult tissues will provide important insights into finding novel and effective therapeutic targets against ischemic heart and vascular diseases.

项目概要小动脉分化和由非常微小的血管形成小动脉（毛细血管动脉化）是缺血性器官从有害损伤中恢复的基本过程。尽管其显而易见临床和生物学的重要性，人们对这些事件的分子机制及其影响知之甚少。规定。我们最近发现 CD36 的转录抑制参与促血管生成毛细血管内皮细胞 (EC) 或微血管 EC (MVEC) 的促动脉重编程，并且是与毛细血管动脉化有关。具体来说，我们已经证明 LPA/PKD-1 信号介导的 CD36 转录抑制在促进 MVEC 中动脉生成基因表达中发挥关键作用，并且体内微血管重塑。此外，EC特异性删除pkd-1显示出恢复受损缺血性损伤。这些意外的发现导致了这样的假设：CD36 基因通过 LPA/PKD-1 进行抑制信号轴重新编程 MVEC 以分化为小动脉 EC，并促进毛细血管动脉化。到为了验证这一假设，我们建立了二维和三维 MVEC 培养系统，并在体内基质胶测定。更重要的是，我们建立了独特的cd36基因的后肢缺血模型具有 EC 特异性翻译核糖体亲和纯化 (TRAP) 和 EC 特异性 pkd-1 的缺陷小鼠缺陷小鼠或 EC 特异性 pkd-1 缺陷 TRAP 小鼠。这些新颖的 TRAP 转基因系已靶向基因缺陷并携带 EC 特异性 EGFP 标签，使我们能够直接纯化核糖体结合的 mRNA 来自体内 EC 的下一代 RNA 测序，促进 EC 特异性转录组的探针和非编码 RNA 的核糖体结合。该鼠标模型的上市首次为我们提供了设计实验以获得对成人小动脉生长生物学的新的和基本的见解的能力缺血条件下。使用这些工具，我们的目标是 1) 证明在 MVEC 中 LPA/PKD-1-CD36 信号轴对于调节 MVEC 重编程和小动脉分化至关重要； 2）检验假设通过 LPA/PKD-1-FoxO1 信号轴的 CD36 转录抑制是基因重编程开关促进毛细血管动脉化（从头动脉生成）。连同使用一系列分子生物学和成像方法、单细胞 RNA 测序以及斑马鱼模型，我们期望实现这些目标并提供小动脉的综合评估分化和毛细血管动脉化。这将阐明尚未探索和理解的方面血管生物学。该提案的重点是成人动脉新生，因为它非常实用意义。更好地了解成人组织中的毛细血管动脉化将提供重要的帮助寻找针对缺血性心脏和血管疾病的新颖有效的治疗靶点的见解。