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，为身体的每个器官注入氧气和营养，运输免疫细胞并与保护身体的免疫细胞相互作用。在成人中，维持完整的血管内皮受到严格的稳态控制，以防止水肿或出血。伤口或组织缺氧可导致血管生成和血管重塑。血管稳态过程高度复杂受到监管并涉及许多分子参与者的协同行动。在疾病条件下，协调这些分子过程可能会出错。对于罕见的孟德尔疾病尤其如此，这些疾病是由以下原因引起的：该机制的关键组成部分发生突变，例如遗传性出血性毛细血管扩张症（HHT），是由 ENG、ACVRL1 或 SMAD4 功能缺失突变引起的。了解分子调节血管稳态的基础对于许多疾病至关重要，包括易感性和从急性肺损伤和 COVID-19 中恢复。在这里，我们将研究蛋白质酪氨酸的作用磷酸酶非受体 14 型 (PTPN14) 作为血液和淋巴调节的关键参与者血管稳态。我们之前表明 PTPN14 基因内的遗传变异与 HHT 患者的肺动静脉畸形 (AVM) 和人类遗传学研究表明其在其中发挥着作用 PTPN14 在淋巴管发育和稳态中的作用。 PTPN14 是 YAP 信号传导的拮抗剂，我们已表明它支持 ALK1(ACVRL1)/SMAD4 信号。我们已经在 PTPN14 基因与 PTPN14 表达以及 HHT 中肺 AVM 的存在相关，表明 PTPN14 表达水平影响 AVM 发生率。我们还发现了两种罕见的非与 AVM 分离的同义 PTPN14 SNP，我们还将确定它们如何影响 PTPN14 SMAD4 和 YAP/TAZ 的功能和分子相互作用。我们将使用人体工程微血管在流动条件下研究 PTPN14 敲低或突变的影响，有或没有 ENG 或 AVRL1 敲低对内皮细胞、大小、增殖、迁移、血流对齐和血管的影响不同流动条件下的渗透率。最后，我们将使用 Cre 介导的 Ptpn14-loxp 等位基因，生成在内部，研究因基因缺失而导致的血管和淋巴畸形的发展体内内皮细胞或实质细胞中的 Ptpn14，并检查 PTPN14 如何与 BMP9- 相互作用内皮糖蛋白-ALK1 信号通路调节体内 AVM 的形成。我们将产生他莫昔芬诱导剂细胞类型特异性 Ptpn14-/- 并研究其如何影响发育性血管生成、病理学受伤的角膜以及成人肺、皮肤、肝脏、肠道和大脑的血管床中的血管生成。我们也会研究 Ptpn14DiEC 对 Eng+/-、EngDiEC 表型的影响，以确定这些基因如何在体内。将使用我们的新方法评估肺部血流和潜在的动静脉畸形通过 S10 资助获得的 Quantum GX2 微型 CT 成像仪。