Kurly's Role in Left-Right Patterning and Congenital Heart Disease

Kurly 在左右模式和先天性心脏病中的作用

• 批准号: 7749380
• 负责人: KIMBERLY M JAFFE
• 金额: $ 5.01万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7749380
• 关键词: Affect; Affinity Chromatography; Animal Model; Anterior; Area; Biological Assay; Biological Models; Cell Polarity; Child; Chromosomes, Human, Pair 21; Cilia; Congenital Heart Defects; Defect; Development; Dorsal; Down Syndrome; Embryo; Embryology; Embryonic Development; Event; Gene Expression; General Population; Genes; Genetic; Handedness; Heart; Human; Human Chromosomes; Immunofluorescence Immunologic; Immunohistochemistry; Incidence; Injection of therapeutic agent; Investigation; Knowledge; Laboratories; Left; Life; Location; Maps; Mutation; Organ; Organism; Orthologous Gene; Pathway interactions; Patients; Pattern; Population; Positioning Attribute; Proteins; Research; Role; Signal Pathway; Site; Societies; Staging; Techniques; Vertebrates; Zebrafish; base; cardiogenesis; congenital heart disorder; gene function; genetic analysis; genetic manipulation; insight; mutant; novel; research study

DESCRIPTION (provided by applicant): Vertebrates have internal asymmetries along the left-right axis, including asymmetric placement of the organs. Correct placement of organs, such as the heart, is critical for proper embryonic development and survival. Due to improper gene expression as many as one in 5,000 children are born with left-right patterning defects which often manifest as congenital heart disease. Understanding the genetic pathways involved in establishing proper left-right patterning in the embryo is vital to understanding how these defects may arise. A novel zebrafish mutant, Kurly, affects left-right patterning and subsequent heart development. Interestingly, the novel human ortholog of Kurly maps to a gene located on human chromosome 21 within the Down Syndrome Critical Region. Genes located in this area are perhaps responsible for the high incidence of congenital heart defects in Down Syndrome patients. While 0.8% of the general population is affected by congenital heart defects, 40-50% of those affected by Down Syndrome have congenital heart defects. By understanding how Kurly functions in zebrafish, valuable insight will be gained into how left-right patterning defects and subsequent congenital heart defects may arise. The zebrafish is an excellent model organism due to its external development, transparency, and genetic manipulations that can be made. Also, as many genes involved in left-right patterning have conserved roles in humans, experiments performed in zebrafish and knowledge gained from these experiments are clearly applicable to the greater good of society. To investigate Kurly's effects on left- right patterning and heart development, I will take multiple approaches. I will first use immunohistochemistry and fluorescent protein tagging to examine the location and sites of function for Kurly. I will then explore how Kurly affects primary cilia and cell polarity during these early patterning events by injection assays and immunofluorescence. I will finally identify other proteins that interact with Kurly, through tandem affinity purification, during these stages of development in order to gain insight into significant pathways which may require Kurly function. Relevance: This research greatly benefits mankind through an in-depth analysis of a gene that, when misregulated, leads to left-right internal organ patterning defects as well as congenital heart defects.

描述（由申请人提供）：脊椎动物沿着左右轴具有内部不对称性，包括器官的不对称放置。心脏等器官的正确放置对于胚胎的正常发育和存活至关重要。由于基因表达不当，多达五千分之一的儿童出生时患有左右模式缺陷，通常表现为先天性心脏病。了解在胚胎中建立正确的左右模式所涉及的遗传途径对于了解这些缺陷如何出现至关重要。一种新的斑马鱼突变体 Kurly 会影响左右图案和随后的心脏发育。有趣的是，Kurly 的新型人类直系同源物映射到位于唐氏综合症关键区域内的人类 21 号染色体上的一个基因。位于该区域的基因可能是唐氏综合症患者先天性心脏病高发的原因。虽然总人口中有 0.8% 患有先天性心脏缺陷，但唐氏综合症患者中 40-50% 患有先天性心脏缺陷。通过了解 Kurly 在斑马鱼中的功能，我们将获得关于左右图案缺陷和随后的先天性心脏缺陷如何出现的宝贵见解。斑马鱼因其外部发育、透明度和可进行遗传操作而成为一种优秀的模式生物。此外，由于许多涉及左右模式的基因在人类中具有保守的作用，因此在斑马鱼中进行的实验以及从这些实验中获得的知识显然适用于社会的更大利益。为了研究 Kurly 对左右模式和心脏发育的影响，我将采取多种方法。我将首先使用免疫组织化学和荧光蛋白标记来检查 Kurly 的位置和功能位点。然后，我将通过注射测定和免疫荧光探索 Kurly 如何在这些早期模式形成事件中影响初级纤毛和细胞极性。我最终将在这些开发阶段通过串联亲和纯化来鉴定与 Kurly 相互作用的其他蛋白质，以便深入了解可能需要 Kurly 功能的重要途径。相关性：这项研究通过对基因的深入分析极大地造福了人类，该基因如果调节不当，会导致左右内脏器官模式缺陷以及先天性心脏缺陷。