Genetic Models to Study Glial Regulation of Angiogenesis

研究血管生成的神经胶质调节的遗传模型

• 批准号: 8845634
• 负责人: Joseph H McCarty
• 金额: $ 19.05万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8845634
• 关键词: Ablation; Address; Adhesions; Basement membrane; Binding; Biochemical; Biochemistry; Biological Assay; Biological Models; Blood - brain barrier anatomy; Blood Vessels; Brain; Cell Adhesion Molecules; Cell Communication; Cell Culture System; Cell Proliferation; Cerebrum; Cessation of life; Collaborations; Complement; Complex; Congenital Abnormality; Cues; Data; Defect; Development; Disease; Embryo; Embryology; Employee Strikes; Endothelial Cells; Event; Experimental Models; Extracellular Matrix; Extracellular Matrix Proteins; Felis catus; Foundations; Gene Deletion; Gene Expression; Gene Expression Profiling; Gene Targeting; Genes; Genetic; Genetic Models; Genetic screening method; Growth Factor; Health; Homeostasis; Human; Integrins; Knock-out; Knockout Mice; LacZ Genes; Ligands; Link; Mediating; Modeling; Molecular; Molecular Genetics; Mus; Mutagenesis; Mutant Strains Mice; Neuraxis; Neuroepithelial Cells; Neuroglia; Organ; Paracrine Communication; Pathology; Pathway interactions; Phenocopy; Phenotype; Primary Cell Cultures; Protein Family; Proteins; RNA Sequence Analysis; Regulation; Regulator Genes; Regulatory Pathway; Reporter; Reporting; Research; Retina; Signal Pathway; Signal Transduction; Spinal Cord; Stroke; Testing; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Vascular Endothelial Cell; Work; Zebrafish; angiogenesis; beta catenin; blood vessel development; cDNA Arrays; cell type; in vivo; in vivo Model; insight; integrin alphavbeta8; interest; knockout gene; mind control; nervous system disorder; neurovascular unit; new therapeutic target; novel; novel therapeutics; premature; receptor; research study; retinal angiogenesis; tool

DESCRIPTION (provided by applicant): Angiogenesis, or the formation of new blood vessels via vascular endothelial cell proliferation and sprouting, is essential for the development of all vertebrate organs. This is particularly relevant in the central nervous system (CNS), comprised of the brain, spinal cord and retina, where angiogenesis is regulated by glial cells that secrete various growth factors and extracellular matrix (ECM) proteins. How endothelial cells properly interpret glial-derived cues, however, remains largely uncharacterized. Using Cre/lox strategies in mice we have discovered the first glial-endothelial cell signaling axis that regulates angiogenesis in the developing CNS. Components of this pathway include alphavbeta8 integrin in glial cells, its ECM-bound latent transforming growth factor beta (TGFbeta) protein ligands in the ECM, and canonical TGFbeta receptors (TGFbetaR2 and Alk5) in endothelial cells. Cell type-specific ablation of any component in this pathway leads to CNS-specific angiogenesis defects and premature death. Interestingly, other groups have reported that genetic deletion of Wnt growth factors in glial cells, or the Wnt signaling effector beta-catenin in endothelial cells, also leads to angiogenesis pathologies that largely phenocopy those in alphavbeta8 integrin and TGFbeta receptor knockout mice. These results, as well as unpublished mechanistic data that we present in this application, strongly support cross talk between integrin-activated TGFbetas and Wnts, and in this project we will analyze these events in the developing brain. In particular, a unique set of experimental tools, including mouse and zebrafish genetic models, will be generated to characterize signaling mechanisms and gene regulatory pathways underlying glial control of angiogenesis. We will complement the in vivo models with signaling experiments in primary brain endothelial cells stimulated with Wnts and/or TGFbetas, and characterize how components of these pathways are functionally interconnected using biochemistry and gene expression profiling. These experiments will not only reveal new and important insights into how Wnts and TGFbetas are involved in glial regulation of angiogenesis, but may identify novel therapeutic mechanisms underlying vascular-related neurological disorders.

描述（由申请人提供）：血管生成，或通过血管内皮细胞增殖和发芽形成新血管，对于所有脊椎动物器官的发展至关重要。这在由大脑，脊髓和视网膜组成的中枢神经系统（CNS）中尤其重要，其中血管生成受神经胶质细胞的调节，这些细胞分泌各种生长因子和细胞外基质（ECM）蛋白。然而，内皮细胞如何正确解释神经胶质的提示在很大程度上没有表征。使用小鼠中的CRE/LOX策略，我们发现了调节发育中CNS中血管生成的第一个神经胶质 - 内皮细胞信号传导轴。该途径的成分包括胶质细胞中的alphavbeta8整合素，其ECM结合的潜在转化生长因子β（TGFBETA）蛋白配体在ECM中以及内皮细胞中的典型TGFBETA受体（TGFBETAR2和ALK5）。该途径中任何成分的细胞类型特异性消融都会导致CNS特异性血管生成缺陷和过早死亡。有趣的是，其他群体报告说，神经胶质细胞中Wnt生长因子的遗传缺失或内皮细胞中Wnt信号传导效应子β-catenin的遗传缺失， 还导致血管生成病理学，在很大程度上表现为α8整合蛋白和TGFBETA受体敲除小鼠的表观复制。这些结果以及我们在本应用程序中提供的未发表的机械数据，强烈支持整合素激活的TGFBETAS和WNT之间的交叉讨论，在该项目中，我们将在发育中的大脑中分析这些事件。特别是，将生成一套独特的实验工具，包括小鼠和斑马鱼遗传模型，以表征信号机制和基因调节途径的基因调节途径。我们将通过WNT和/或TGFBETA刺激的原代脑内皮细胞中的信号实验来补充体内模型，并表征这些途径的组件如何使用生物化学和基因表达分析在功能上互连。这些实验不仅会揭示有关Wnt和TGFBETA如何参与血管生成的神经胶质调节的新的和重要的见解，而且可以鉴定出与血管相关神经系统疾病的新型治疗机制。

项目成果

RICE CRISPR: Rapidly increased cut ends by an exonuclease Cas9 fusion in zebrafish.

• DOI: 10.1002/dvg.23044
• 发表时间: 2017-08
• 期刊: Genesis (New York, N.Y. : 2000)
• 作者: Clements TP;Tandon B;Lintel HA;McCarty JH;Wagner DS
• 通讯作者: Wagner DS