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: 8021828
• 负责人: KIMBERLY M JAFFE
• 金额: $ 2.38万
• 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 2011-09-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8021828
• 关键词: 21q22.1; 21q22.3; 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）：脊椎动物沿左右轴沿着具有内部不对称性，包括器官的不对称放置。器官的正确放置，如心脏，对胚胎的正常发育和存活至关重要。由于基因表达不当，每5,000名儿童中就有一名患有左右图案缺陷，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理解这些缺陷是如何产生的至关重要。一种新的斑马鱼突变体Kurly影响左右模式和随后的心脏发育。有趣的是，Kurly的新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号染色体上的基因。位于这一区域的基因可能是唐氏综合症患者先天性心脏缺陷发病率高的原因。虽然一般人口的0.8%受到先天性心脏缺陷的影响，但受唐氏综合症影响的人中有40-50%患有先天性心脏缺陷。通过了解Kurly如何在斑马鱼中发挥作用，将获得关于左右图案缺陷和随后的先天性心脏缺陷如何产生的有价值的见解。斑马鱼是一个很好的模式生物，因为它的外部发展，透明度和遗传操作，可以进行。此外，由于参与左右模式的许多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，我将采用多种方法。我将首先使用免疫组织化学和荧光蛋白标记来检查Kurly的位置和功能部位。然后，我将探讨如何库利影响初级纤毛和细胞极性在这些早期图案的注射测定和免疫荧光事件。最后，我将通过串联亲和纯化，在这些发展阶段确定与Kurly相互作用的其他蛋白质，以深入了解可能需要Kurly功能的重要途径。相关性：这项研究通过对一种基因的深入分析极大地造福了人类，这种基因在失调时会导致左右内脏模式缺陷以及先天性心脏缺陷。