Circulating cells as tools to study vascular pathobiology of HHT

循环细胞作为研究 HHT 血管病理学的工具

• 批准号: 8825101
• 负责人: ROSEMARY J AKHURST
• 金额: $ 55.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8825101
• 关键词: ACVRL1 gene; Affect; Anastomosis - action; Anemia; Angiogenesis Inhibitors; Arteriovenous malformation; Automobile Driving; Biological; Biological Markers; Biological Process; Biology; Blood; Blood Cells; Blood Vessels; Bone Marrow; Brain; Cell Count; Cell Proliferation; Cell Survival; Cells; Cellular biology; Characteristics; Clinical; Color; Communities; Data; Development; Disease; ENG gene; Endothelial Cells; Epistaxis; Family; Gap Junctions; Gene Expression; Genes; Genetic; Genomic Instability; Hemorrhage; Hereditary Disease; Hereditary hemorrhagic telangiectasia; Home environment; Human; Human Genetics; In Vitro; Individual; Inflammation; Investigation; Knowledge; Lesion; Liver; Lung; Messenger RNA; MicroRNAs; Molecular; Mus; Mutation; NOTCH1 gene; Organ; Pathology; Pathway interactions; Patients; Penetrance; Population; Population Heterogeneity; Prognostic Marker; Rare Diseases; Recruitment Activity; Recurrence; Risk; Severities; Severity of illness; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Stem cells; Stroke; Syndrome; Telangiectasis; Testing; Thalidomide; Vascular Diseases; Visceral; angiogenesis; bevacizumab; cardiovascular disorder risk; cell type; clinically relevant; high risk; human tissue; in vitro testing; induced pluripotent stem cell; insight; knock-down; malformation; monitor peptide receptor; monocyte; mouse model; prognostic; prognostic value; public health relevance; receptor; repaired; response; self-renewal; tool; transcriptomics

DESCRIPTION (provided by applicant): Hereditary Hemorrhagic Telangiectasia (HHT) is a rare disease (affecting ~1/5, 000), caused predominantly by mutations in ENG or ACVRL1 aka ALK1 encoding TGF�MP receptors. Patients develop vascular malformations, including small mucocutaneous telangiectases and/or large visceral arteriovenous malformations (AVMs) in multiple organs. The penetrance and expressivity of clinical manifestations is highly variable between patients, even within a family. The rationale for the project is that investigation of the biological and molecular characteristics of cells circulating within blood, especially, but not onl, circulating endothelial progenitor cells (cEPCs), will provide important new knowledge about basic cellular and molecular mechanisms of HHT pathobiology, and contribute to development of prognostic HHT markers. The overarching hypotheses to be tested are: a) the cellular makeup of circulating bone marrow-derived cell populations is perturbed in the blood of HHT patients; b) there is an elevation of cEPC numbers in HHT that contributes to and correlates with disease pathology including telangiectases and AVMs; c) gene expression networks are perturbed in HHT cEPCs to alter their biological functions, and d) HHT genetic modifiers regulate cEPC numbers and biology. The extent to which cellular and molecular perturbations correlate with disease type (HHT1 versus HHT2) and disease severity will be tested, and molecular mechanisms investigated using cultured endothelial colony forming cells (ECFCs) from HHT bloods. Aim 1) multi-color FACS analysis will be utilized to assess perturbation in circulating cell numbers in HHT patients and HHT mouse models, focusing on cEPCs and monocytes. Aim 2) ECFCs will be isolated from blood of HHT patients and controls, and utilized to investigate alterations in clinically-relevant parameters of in vitro angiogenesis, and biologicl effects of siRNA knock down of HHT modifier genes. A bank of cEPC-derived iPS cells will be generated for use by the HHT community. Aim 3) Blood cell mRNA and miRNA transcriptomic profiles will be compared between wild type, Eng+/- and Alk1+/- mice. Blood gene expression networks will be generated from existing transcriptomic data available from genetically heterogeneous mouse (>100) and human (>100) populations. Finally, new transcriptomic data will be generated from human HHT1, HHT2 and control blood, to test the postulate that gene expression networks are rewired within HHT cEPCs and/or monocytes to alter cellular parameters that can be molecularly probed in Aim 2 in order to provide mechanistic insight into HHT disease mechanisms.

遗传性出血性毛细血管扩张症（HHT）是一种罕见的疾病（影响约1/5000），主要由ENG或ACVRL 1（又名ALK 1）编码TGF β MP受体突变引起。患者发生血管畸形，包括多个器官中的小粘膜皮肤毛细血管扩张和/或大内脏动静脉畸形（AVM）。临床表现的特异性和表现力在患者之间是高度可变的，即使在一个家庭中也是如此。该项目的基本原理是，研究血液中循环的细胞，特别是但不仅仅是循环内皮祖细胞（cEPCs）的生物学和分子特征，将提供关于HHT病理生物学的基本细胞和分子机制的重要新知识，并有助于发展预后HHT标志物。待检验的首要假设为：a）HHT患者血液中循环骨髓源性细胞群的细胞组成受到干扰; B）HHT中cEPC数量升高，导致疾病病理学（包括毛细血管扩张酶和AVM）并与其相关; c）HHT cEPC中的基因表达网络受到干扰以改变其生物学功能，以及d）HHT遗传修饰剂调节cEPC数量和生物学。将测试细胞和分子扰动与疾病类型（HHT 1与HHT 2）和疾病严重程度相关的程度，并使用来自HHT血液的培养内皮集落形成细胞（ECFC）研究分子机制。目的1）多色FACS分析将用于评估HHT患者和HHT小鼠模型中循环细胞数量的扰动，重点是cEPC和单核细胞。目的2）从HHT患者和正常人的血液中分离ECFC，并用于研究体外血管生成的临床相关参数的改变，以及siRNA敲低HHT修饰基因的生物学效应。将产生cEPC衍生的iPS细胞库，供HHT社区使用。目的3）比较野生型、Eng+/-和Alk 1 +/-小鼠的血细胞mRNA和miRNA转录组学谱。血液基因表达网络将从遗传异质性小鼠（>100）和人类（>100）群体的现有转录组学数据中生成。最后，将从人HHT 1、HHT 2和对照血液中生成新的转录组数据，以测试基因表达网络在HHT cEPCs和/或单核细胞内重新连接以改变可以在Aim 2中进行分子探测的细胞参数的假设，以便提供对HHT疾病机制的机制性见解。