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）：Kallmann综合征是一种遗传性疾病，其特征是厌食（无法闻起来）和性腺功能低下的性腺功能减退，导致不育。这种疾病会影响10,000名男性中的1人和40,000名女性中有1个，但由于轻度性低肿瘤或低体性病例，可能被诊断出来。迄今为止，已经确定了与KS相关的五个基因，即KAL1，FGFR1，FGF8，PROKR2和PROK2；尽管这些仅占所有KS案例的30％。该项目的目的是识别和表征与Kal-1相互作用的新型基因，该基因编码了细胞外基质的细胞粘附蛋白。我们建议使用秀丽隐杆线虫中功能轴突分支表型的KAL-1增益的修饰符屏幕来实现这一目标。我们的小组成功地使用了该表型的试验屏幕，以识别几个在蠕虫和人类中与Kal-1相互作用的新型基因座。新分离的突变将通过单核苷酸多态性映射和整个基因组测序方法来分子鉴定。 RNAi介导的敲除实验和转基因救援实验将进一步证实突变的身份。然后，鉴定的基因将以分子和遗传的特征在于三种互补方法，以更深入地了解Kal-1如何与这些基因共同作用于神经系统的发展以及细胞外基质在此过程中的作用。首先，我将对修饰子突变体进行详细的神经解剖学和表型分析，重点是神经系统。其次，将确定他们的表达部位和亚细胞定位，这将为蛋白质的功能提供重要的线索。第三，将进行双重突变体和上毒分析，以将新突变放置在已知的遗传环境中。最后，随着越来越多的与KAL-1相互作用的基因，我们将更好地理解其功能以及它们在人类中的破坏如何导致Kallmann综合征。 公共卫生相关性：Kallmann综合征（KS）是一种遗传疾病，具有神经元靶向和迁移缺陷，表现为无法闻起来和不育。我们正在使用秀丽隐杆线虫中的遗传方法来识别与KS基因KAL-1相互作用的基因，以便在神经系统的发展过程中更深入地了解其功能。确定的基因是候选疾病基因，在分子无法解释的Kallmann综合征中被突变。