Identification of Novel Loci Interacting with the Kallmann Syndrome Gene Kal-1

与卡尔曼综合征基因 Kal-1 相互作用的新位点的鉴定

• 批准号: 8006683
• 负责人: Carlos Antonio Diaz-Balzac
• 金额: $ 4.14万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8006683
• 关键词: Accounting; Affect; Anosmia; Axon; Caenorhabditis elegans; Candidate Disease Gene; Cell Adhesion Molecules; Code; Defect; Diagnosis; Disease; Extracellular Matrix; FGF8 gene; FGFR1 gene; Female; Genes; Genetic; Genetic Epistasis; Goals; Hereditary Disease; Human; Hypogonadism; Infertility; Inherited; Kallmann Syndrome; Klinefelter' s Syndrome; Mediating; Mutate; Mutation; Nervous system structure; Neurons; Phenotype; Process; RNA Interference; Role; Single Nucleotide Polymorphism Map; Site; Smell Perception; Transgenic Organisms; gain of function; genome sequencing; hyposmia; knock-down; male; migration; mutant; nervous system development; novel; protein function; public health relevance; research study

DESCRIPTION (provided by applicant): Kallmann Syndrome is a hereditary condition characterized by anosmia (the inability to smell) and hypogonadotropic hypogonadism resulting in infertility. This disorder affects 1 in 10,000 males and 1 in 40,000 females, but may be under diagnosed due to mild cases of hypogonadism or hyposmia. To date, five genes associated with KS have been identified, namely, KAL1, FGFR1, FGF8, PROKR2, and PROK2; though these only account for approximately 30% of all KS cases. The goal of this project is to identify and characterize novel genes that genetically interact with kal-1, a gene that codes for a cell adhesion protein of the extracellular matrix. We propose to accomplish this using a modifier screen of a kal-1 gain of function axon branching phenotype in C. elegans. A pilot screen of this phenotype has been used successfully by our group to identify several novel loci that genetically interact with KAL-1, both in worms and humans. The newly isolated mutations will be molecularly identified through single nucleotide polymorphism mapping and whole genome sequencing approaches. RNAi mediated knock down experiments and transgenic rescue experiments will be performed to further corroborate the identity of the mutation. The identified genes will then be molecularly and genetically characterized by three complementary approaches to gain a deeper understanding of how kal-1 acts in concert with these genes on the development of the nervous system and the role of the extracellular matrix in this process. First, I will perform a detailed neuroanatomical and phenotypic analysis of the modifier mutants with a focus on the nervous system. Second, their site of expression and the sub cellular localization will be determined, which will give important clues to the function of the protein. Third, double mutant and epistasis analyses will be performed in order to place the new mutations within a known genetic context. In the end, as more genes that interact with KAL-1 are identified, we will have a better understanding of their function and how their disruption in humans results in Kallmann Syndrome. PUBLIC HEALTH RELEVANCE: Kallmann Syndrome (KS) is a genetic disease with neuronal targeting and migration defects that manifest in the inability to smell and infertility. We are using a genetic approach in C. elegans to identify genes that interact with the KS gene kal-1 in order to gain a deeper understanding of its function during the development of the nervous system. The identified genes are candidate disease genes to be mutated in molecularly unexplained cases of Kallmann Syndrome.

描述（由申请人提供）：卡尔曼综合征是一种遗传性疾病，其特征是嗅觉缺失（无法嗅觉）和低促性腺激素性功能减退，导致不育。这种疾病影响1/10，000的男性和1/40，000的女性，但可能由于性腺功能减退或嗅觉减退的轻度病例而被诊断。到目前为止，已经确定了与KS相关的五个基因，即KAL 1，FGFR 1，FGF 8，PROKR 2和PROK 2;尽管这些基因仅占所有KS病例的约30%。该项目的目标是识别和表征与kal-1基因相互作用的新基因，kal-1是编码细胞外基质的细胞粘附蛋白的基因。我们建议使用一个kal-1的功能轴突分支表型的增益在C的修饰筛选来实现这一点。优美的我们的研究小组已经成功地使用了这种表型的试验性筛选来鉴定几个与KAL-1在蠕虫和人类中发生遗传相互作用的新基因座。新分离的突变将通过单核苷酸多态性作图和全基因组测序方法进行分子鉴定。将进行RNAi介导的敲除实验和转基因拯救实验以进一步证实突变的身份。然后将通过三种互补方法对所识别的基因进行分子和遗传特征分析，以更深入地了解kal-1如何与这些基因一起作用于神经系统的发育以及细胞外基质在这一过程中的作用。首先，我将进行详细的神经解剖学和表型分析的修饰突变体的神经系统为重点。其次，确定它们的表达位点和亚细胞定位，这将为蛋白质的功能提供重要线索。第三，将进行双突变和上位性分析，以便将新突变置于已知的遗传背景中。最后，随着更多与KAL-1相互作用的基因被发现，我们将更好地了解它们的功能以及它们在人类中的破坏如何导致卡尔曼综合征。 公共卫生相关性：卡尔曼综合征（KS）是一种遗传性疾病，具有神经元靶向和迁移缺陷，表现为无法嗅觉和不育。我们在C中使用遗传方法。elegans来鉴定与KS基因kal-1相互作用的基因，以便更深入地了解其在神经系统发育过程中的功能。所鉴定的基因是在分子上无法解释的卡尔曼综合征病例中待突变的候选疾病基因。