Translating OCD gene-association studies into mice to examine SLC1A1 function

将强迫症基因关联研究转化为小鼠以检查 SLC1A1 功能

• 批准号: 8476278
• 负责人: Susanne Elizabeth Ahmari
• 金额: $ 6.75万
• 依托单位: NEW YORK STATE PSYCHIATRIC INSTITUTE DBA RESEARCH FOUNDATION FOR MENTAL HYGIENE, INC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-30 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8476278
• 关键词: Alleles; Animal Model; Animals; Anxiety; Autopsy; Behavior; Behavioral; Biochemical; Biological Assay; Biological Models; Brain; Brain region; Candidate Disease Gene; Cell model; Chronic; Clinical; Code; Corpus striatum structure; Data; Development; Disease; Dissection; EAAT3; Event; Functional disorder; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Glutamate Transporter; Glutamates; Glutathione; Goals; Grant; Grooming; High Pressure Liquid Chromatography; Human; Human Genetics; Immunohistochemistry; In Situ; Institutes; Investigation; Knock-in Mouse; Knowledge; Lead; Link; Measurement; Measures; Mediating; Mental disorders; Modeling; Molecular; Molecular Abnormality; Mus; Neurons; Neurotransmitters; New York; Obsessive-Compulsive Disorder; Outcome Measure; Patients; Prevalence; Process; Proteins; Regulation; Research; Research Personnel; Reversal Learning; Risk Factors; Role; Scientist; Side; Signal Transduction; Simulate; Specificity; Staining method; Stains; Structure; Susceptibility Gene; Symptoms; Synapses; Synaptosomes; System; Technology; Testing; Thalamic structure; Time; Tissues; Transgenic Mice; Translating; Universities; Western Blotting; Work; base; behavior test; brain tissue; clinical phenotype; critical developmental period; disorder risk; early onset; flexibility; gamma-Aminobutyric Acid; in vivo; interest; mouse model; neural circuit; neurochemistry; novel; overexpression; postsynaptic; prepulse inhibition; protein transport; research study; severe mental illness; transmission process

DESCRIPTION (provided by applicant): Support is requested for a 2-year collaborative grant between scientists at the New York State Psychiatric Institute and Vanderbilt University to investigate the pathophysiology underlying OCD. The proposal bridges basic and clinical OCD research by integrating findings from the research team's ongoing human genetic studies into the proposed mouse experiments. The research plan thus capitalizes on the expertise of the team in 1) human OCD genetic studies, 2) development of transgenic mice, 3) biochemical assays, and 4) mouse behavioral analysis. Current understanding of the molecular and cellular abnormalities underlying OCD is limited, in part because post-mortem studies in humans have not been performed. In addition, mouse studies have not yet been convincingly linked to the clinical phenotype and genetic abnormalities seen in OCD patients. To date, the only gene which has been consistently linked to OCD in human genetic studies is SLC1A1, which codes for a protein that transports the neurotransmitter glutamate. In addition, there is evidence from human studies that abnormal regulation of glutamate transmission in striatum is correlated with OCD symptoms. This has led to the hypothesis that abnormal levels of the SLC1A1 glutamate transporter in striatum lead to 1) abnormalities in the glutamate system, 2) changes in brain structure, and 3) OCD symptoms. The proposed R21 will test this hypothesis using novel knock-in mouse technology. In the first aim, the researchers will use an efficient system they have previously developed for manipulating gene expression in mice called the FAST system (Flexible Accelerated STOP TetO-knockin). This will allow them to develop a novel knock-in mouse line called tetO-Slc1a1, which will permit precise regulation of SLC1A1 expression levels in brain regions implicated in OCD. They will then use this mouse line to generate abnormally high levels of the SLC1A1 glutamate transporter specifically in striatum. This will simulate the effect of the version of the gene found most commonly in OCD patients. In the second aim, the mice with abnormally high levels of Slc1a1 in striatum will be characterized by: 1) measuring glutamate system functioning~ 2) examining brain structure~ and 3) testing behavior in OCD-relevant paradigms that measure anxiety and repetitive behaviors. This will provide the first direct test of whether OCD-related dysfunction is caused by abnormal expression of the leading human OCD candidate gene. Completion of this grant will lead to an amenable system for 1) further dissection of the molecular, cellular, and electrophysiologic underpinnings of observed changes~ and 2) determination of whether there is a particular time in development during which the brain is more vulnerable to developing OCD. These studies will lead to a better understanding of how dysfunctional circuits lead to OCD symptoms, which is necessary to guide development of new treatments for this severe mental illness.

描述（由申请人提供）：请求纽约州精神病学研究所和范德比尔特大学的科学家提供为期 2 年的合作资助，以研究强迫症的病理生理学。该提案通过将研究小组正在进行的人类遗传学研究的结果整合到拟议的小鼠实验中，将基础和临床强迫症研究联系起来。因此，该研究计划利用了团队在 1) 人类强迫症遗传学研究、2) 转基因小鼠开发、3) 生化分析和 4) 小鼠行为分析方面的专业知识。目前对强迫症背后的分子和细胞异常的了解是有限的，部分原因是尚未对人类进行尸检研究。此外，小鼠研究尚未令人信服地将其与强迫症患者的临床表型和遗传异常联系起来。迄今为止，在人类遗传学研究中唯一与强迫症相关的基因是 SLC1A1，它编码一种转运神经递质谷氨酸的蛋白质。此外，人体研究证据表明纹状体谷氨酸传输的异常调节与强迫症症状相关。这导致了这样的假设：纹状体中 SLC1A1 谷氨酸转运蛋白的异常水平会导致 1) 谷氨酸系统异常，2) 大脑结构变化，以及 3) 强迫症症状。拟议的 R21 将使用新型敲入小鼠技术来测试这一假设。在第一个目标中，研究人员将使用他们之前开发的一种高效系统，用于操纵小鼠的基因表达，称为 FAST 系统（灵活加速停止 TetO-knockin）。这将使他们能够开发出一种名为 tetO-Slc1a1 的新型敲入小鼠品系，该品系将允许精确调节与 OCD 相关的大脑区域中的 SLC1A1 表达水平。然后，他们将使用该小鼠品系在纹状体中产生异常高水平的 SLC1A1 谷氨酸转运蛋白。这将模拟在强迫症患者中最常见的基因版本的效果。在第二个目标中，纹状体中 Slc1a1 水平异常高的小鼠的特征将是：1）测量谷氨酸系统功能~2）检查大脑结构~3）在测量焦虑和重复行为的强迫症相关范式中测试行为。这将首次直接测试强迫症相关功能障碍是否是由人类强迫症候选基因的异常表达引起的。这项资助的完成将产生一个可靠的系统，用于：1）进一步剖析观察到的变化的分子、细胞和电生理学基础；2）确定发育中是否存在某个特定时间，在此期间大脑更容易患上强迫症。 这些研究将有助于更好地了解功能失调的回路如何导致强迫症症状，这对于指导开发这种严重精神疾病的新疗法是必要的。