Protein tyrosine phosphatase non-receptor 14 in vascular stability and remodeling

蛋白酪氨酸磷酸酶非受体 14 在血管稳定性和重塑中的作用

基本信息

批准号：
10660507
负责人：
ROSEMARY J AKHURST
金额：
$ 72.79万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-15 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660507
关键词：
ACVRL1 gene Acute Lung Injury Adult Affect Alleles Amino Acids Architecture Arteriovenous malformation Binding Biological Assay Blood Vessels Blood flow Brain Breeding COVID-19 Cells Cellular biology Chemicals Co-Immunoprecipitations Collagen Complex Corneal Injury Cytoprotection Development Disease Edema Endoglin Endothelial Cells Endothelium Equilibrium Extracellular Matrix Eye diseases Fibrocartilages Genes Genetic Genetic Variation Genetic study Growth Growth Factor Hemorrhage Hereditary hemorrhagic telangiectasia Homeostasis Human Human Engineering Human Genetics Hypoxia Immune In Vitro Incidence Inflammation Intercellular Junctions Knock-out Knowledge Learning Life Liver Lung Lymphatic Endothelium Lymphedema MADH4 gene Maintenance Malignant Neoplasms Mechanical Stress Mediating Molecular Molecular Biology Mus Mutation Nuclear Nutrient Organ Oxygen Pathologic Pathologic Neovascularization Pathology Pathway interactions Patients Peripheral arterial disease Pharmaceutical Preparations Phenotype Physiological Predisposition Process Proliferating Protein Dephosphorylation Protein Tyrosine Phosphatase Proteins Pulmonary Hypertension Recovery Regenerative Medicine Regulation Reporter Reporting Retina Role S10 grant Signal Pathway Signal Transduction Site-Directed Mutagenesis Skin Syndrome Tamoxifen Tissues Transforming Growth Factor beta Variant Vascular Diseases Vascular Endothelial Cell Vascular Endothelial Growth Factors Vascular Endothelium Vascular Permeabilities Vascular System Vascular remodeling angiogenesis antagonist assault cadherin 5 cell type drug repurposing gene interaction genetic variant human disease imager in vivo insight knock-down loss of function mutation lymphatic development lymphatic malformations lymphatic vessel malformation mechanotransduction microCT migration molecular marker new therapeutic target notch protein prevent quantum rare mendelian disorder receptor response retinal angiogenesis segregation shear stress vascular abnormality vascular bed wound

项目摘要

ABSTRACT The vascular system is critical to life, infusing each organ of the body with oxygen and nutrients, and transporting and interacting with immune cells that protect the body. In the adult, maintenance of an intact vascular endothelium is under strict homeostatic control to prevent edema or hemorrhage. Wounding or tissue hypoxia can result in angiogenesis and vascular remodeling. The process of vascular homeostasis is highly regulated and involves many molecular players acting in concert. Under disease conditions, orchestration of these molecular processes may go awry. This is especially true in rare Mendelian disorders that are caused by mutations in key components of this machinery, such as Hereditary Hemorrhagic Telangiectasia (HHT), which is caused by loss of function mutations in ENG, ACVRL1, or SMAD4. Understanding the molecular underpinnings that regulate vascular homeostasis is critical to many diseases, including susceptibility to, and recovery from, acute lung injury and COVID-19. Here, we will investigate the role of protein tyrosine phosphatase non-receptor, type 14 (PTPN14) as a critical player in regulation of both blood and lymphatic vessel homeostasis. We previously showed that genetic variation within the PTPN14 gene associates with pulmonary arteriovenous malformations (AVMs) in HHT patients, and human genetics studies suggest a role for PTPN14 in lymphatic development and homeostasis. PTPN14 is an antagonist of YAP signaling and we have shown that it supports ALK1(ACVRL1)/SMAD4 signaling. We have identified several cis-eQTL in the PTPN14 gene that associated with PTPN14 expression and with the presence of pulmonary AVM in HHT, suggesting that PTPN14 expression levels influence AVM incidence. We have also identified two rare non- synonymous PTPN14 SNPs that segregate with AVMs and we will also determine how these affect PTPN14 function and molecular interactions with SMAD4 and YAP/TAZ. We will use human engineered microvessels under flow conditions to investigate the effects of PTPN14 knockdown or mutation, with or without ENG or ACVRL1 knockdown, on endothelial cell, size, proliferation, migration, alignment with flow, and vascular permeability under differing flow conditions. Finally, we will use our Cre-mediated Ptpn14-loxp allele, generated in-house, to investigate development of vascular and lymphatic malformations that result from genetic loss of Ptpn14 in endothelial or parenchymal cells in vivo, and examine how PTPN14 interacts with the BMP9- endoglin-ALK1 signaling pathway to modulate formation of AVMs in vivo. We will generate tamoxifen-inducible cell type-specific Ptpn14-/- and investigate how this affects developmental angiogenesis, pathological angiogenesis in wounded cornea, and vascular beds of adult lung, skin, liver, gut and brain. We will also investigate the effects of Ptpn14DiEC on Eng+/-, EngDiEC phenotypes to determine how these genes interact in vivo. Blood flow in the lung and potential arteriovenous malformations will be assessed using our new Quantum GX2 micro-CT imager obtained through an S10 grant.

抽象的血管系统对生命至关重要，将身体的每个器官注入氧气和营养素，以及与保护人体的免疫细胞运输和相互作用。在成年人中，保持完整血管内皮受到严格的稳态控制，以防止水肿或出血。受伤或组织缺氧会导致血管生成和血管重塑。血管稳态的过程很高受监管并涉及许多演唱会演奏的分子玩家。在疾病条件下，编排这些分子过程可能会出错。在罕见的孟德尔疾病中尤其如此该机械的关键组成部分中的突变，例如遗传性出血性毛细血管扩张（HHT）是由ENG，ACVRL1或SMAD4中功能突变丧失引起的。了解分子调节血管稳态的基础对许多疾病至关重要，包括对和从急性肺损伤和共同19中恢复。在这里，我们将研究蛋白酪氨酸的作用磷酸酶非受体，14型（PTPN14）作为血液和淋巴管调节的关键参与者船只体内平衡。我们先前表明，PTPN14基因中的遗传变异与 HHT患者的肺动脉畸形（AVM）和人类遗传学研究表明作用用于PTPN14在淋巴发育和稳态中。 PTPN14是YAP信号的拮抗剂，我们已经表明它支持ALK1（ACVRL1）/SMAD4信号传导。我们已经确定了几个CIS-EQTL PTPN14基因与PTPN14表达以及HHT中肺AVM的存在相关，表明PTPN14表达水平会影响AVM的发生率。我们还确定了两个罕见的非 - 与AVMS分离的同义PTPN14 SNP，我们还将确定这些SNP如何影响PTPN14 功能和分子相互作用与SMAD4和YAP/TAZ。我们将使用人类工程的微血管在流动条件下，以研究PTPN14敲低或突变的影响，有或没有ENG或 ACVRL1敲低，在内皮细胞上，大小，增殖，迁移，与流量排列和血管在不同流动条件下的渗透性。最后，我们将使用我们的CRE介导的PTPN14-LOXP等位基因，生成内部，研究因遗传丧失而导致的血管和淋巴畸形的发展在体内内皮或实质细胞中的PTPN14，并检查PTPN14如何与BMP9-相互作用 Endoglin-Alk1信号传导途径调节体内AVM的形成。我们将产生他莫昔芬诱导细胞类型特异性PTPN14 - / - 并研究这如何影响发育性血管生成，病理学成年肺，皮肤，肝脏，肠道和脑的受伤角膜和血管床的血管生成。我们也会研究PTPN14DIEC对ENG +/-的影响，以确定这些基因在中的相互作用体内。肺中的血流和潜在的动静脉畸形将使用我们的新来评估通过S10赠款获得的量子GX2 Micro-CT成像器。