Elucidation of the developmental role of Jakmip1, and autism-susceptibility gene

阐明 Jakmip1 和自闭症易感基因的发育作用

• 批准号: 8011720
• 负责人: Jamee Mae Bomar
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8011720
• 关键词: 15q; 3' Untranslated Regions; Accounting; Adult; Affect; Antibodies; Attention; Autistic Disorder; Behavior Therapy; Binding; Biological; Brain; Brain region; Candidate Disease Gene; Cell Line; Cerebellum; Child; Development; Developmental Process; Disease; Early Diagnosis; Etiology; FMRP; Fragile X Syndrome; GABA Receptor; Gene Expression Profile; Genes; Genetic; Goals; Grant; Human; Immunohistochemistry; Inherited; Janus kinase; Kinesin; Light; Messenger RNA; Microarray Analysis; Microtubules; Morphology; Mus; Mutate; Nature; Nervous System Physiology; Neuraxis; Neurites; Neurodevelopmental Disorder; Neurons; Pathology; Pathway interactions; Patients; Pharmacologic Substance; Population; Process; Protein Isoforms; Proteins; Public Health; RNA; Regulation; Research; Role; Societies; Staging; Structure; Susceptibility Gene; Synapses; System; Techniques; Time; Tissues; Transcript; Translations; Western Blotting; autism spectrum disorder; autistic children; base; design; genome-wide; nerve stem cell; neuronal cell body; receptor; relating to nervous system; research study; small hairpin RNA; synaptic function; synaptogenesis; trafficking

DESCRIPTION (provided by applicant): A major challenge in studying the genetic basis of autism is that many genes are associated with the disorder, but each only accounts for a fraction of the cases. To develop treatments that will help a wide range of autistic subjects, we must identify and understand those genes that are commonly dysregulated. To this end, our group looked at shared genome-wide expression changes across two groups of autistic patients with known genetic causes vs. healthy controls. Janus kinase and microtubule-interacting protein 1, JAKMIP1, was one of the top hits from this screen and was validated in a separate group of autistic patients with unknown causes. JAKMIP1 is an important autism-candidate gene for three reasons. First, it is expressed predominantly in neurons. Secondly, JAKMIP1 regulates the expression and possible transport of GABA receptors and GABAergic systems are known to be abnormal in the autisms. Lastly, JAKMIP1 is downstream of genes that monogenetically cause autism. An important aspect of JAKMIP1 is that is contains four brain-specific isoforms, A, B, C and D, that are conserved between mouse and human and have distinct functional domains. My project aims to understand how irregular Jakmip1 isoform levels could disrupt conserved developmental processes, contributing to autism pathology. Aim 1A will determine which isoforms are expressed at the protein and transcript level during development and throughout adulthood. I will collect protein and mRNA from the cortex and cerebellum, two brain regions affected in autism, throughout development and will determine expression changes using western blotting and quantitative real time PCR. Aim 1B will elucidate the distribution of Jakmip1's isoforms in the developing and adult brain and in which neuronal subtypes these isoforms are located by using immunohistochemistry and isoform-specific antibodies. Aim 2A will ascertain the transcriptome-wide impact of silencing Jakmip1's isoforms in differentiated mouse neural progenitors using shRNA and microarray technology. Lastly, Aim 2B will assess neurite morphological changes and perturbations in the population make-up of dendritically-localized mRNA resulting from decreasing Jakmip1 isoform expression in differentiated mouse neural progenitors pre and post stimulation. I will carry out Aim 2B by employing a recent technique (Poon et al. 2006) to isolate and separate dendritic, axonal, and glial processes from the cell body. This study will shed light on the mechanism by which aberrant JAKMIP1 levels could result in autism and may further implicate JAKMIP1 as an autism-susceptibility gene. These discoveries will aid in pre-symptomatic diagnosis and early behavioral intervention for autistic children as well as the design of pharmaceuticals.

描述（由申请人提供）：研究自闭症遗传基础的一个主要挑战是许多基因与该疾病相关，但每个基因只占病例的一小部分。为了开发出能够帮助广泛的自闭症患者的治疗方法，我们必须识别并了解那些通常失调的基因。为此，我们的小组研究了两组已知遗传原因的自闭症患者与健康对照组之间共有的全基因组表达变化。 Janus 激酶和微管相互作用蛋白 1（JAKMIP1）是本次筛选中最热门的蛋白之一，并在另一组原因不明的自闭症患者中得到了验证。 JAKMIP1 是一个重要的自闭症候选基因，原因有以下三个。首先，它主要在神经元中表达。其次，JAKMIP1 调节 GABA 受体的表达和可能的转运，并且已知自闭症患者的 GABA 能系统存在异常。最后，JAKMIP1 是单基因导致自闭症的基因的下游。 JAKMIP1 的一个重要方面是它包含四种大脑特异性亚型：A、B、C 和 D，它们在小鼠和人类之间是保守的，并且具有不同的功能域。我的项目旨在了解不规则的 Jakmip1 同工型水平如何破坏保守的发育过程，从而导致自闭症病理学。目标 1A 将确定哪些亚型在发育期间和整个成年期在蛋白质和转录水平上表达。我将在整个发育过程中从皮层和小脑（受自闭症影响的两个大脑区域）收集蛋白质和 mRNA，并使用蛋白质印迹和定量实时 PCR 确定表达变化。目标 1B 将通过使用免疫组织化学和异构体特异性抗体来阐明 Jakmip1 异构体在发育中和成人大脑中的分布，以及这些异构体位于哪些神经元亚型中。目标 2A 将使用 shRNA 和微阵列技术确定在分化的小鼠神经祖细胞中沉默 Jakmip1 亚型的转录组范围内的影响。最后，Aim 2B 将评估刺激前后分化小鼠神经祖细胞中 Jakmip1 同工型表达减少导致的神经突形态变化和树突状定位 mRNA 群体构成的扰动。我将通过采用最新技术（Poon 等人，2006）来实现目标 2B，从细胞体中分离和分离树突、轴突和神经胶质过程。这项研究将揭示异常 JAKMIP1 水平可能导致自闭症的机制，并可能进一步表明 JAKMIP1 作为自闭症易感基因。这些发现将有助于自闭症儿童的症状前诊断和早期行为干预以及药物设计。