Protein Interaction Network Analysis to Test the Synaptic Hypothesis of Autism

蛋白质相互作用网络分析检验自闭症突触假说

• 批准号: 8616138
• 负责人: Stephen Edward Paucha Smith
• 金额: $ 9万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8616138
• 关键词: Achievement; Antibodies; Antigen-Presenting Cells; Autistic Disorder; Award; Bicuculline; Biological; Biological Assay; Biological Process; Biology; Brain; Budgets; Cells; Collection; Data; Data Set; Development; Disease; Exposure to; Financial compensation; Flow Cytometry; Gene Expression Profile; Genes; Genome; Glutamates; Goals; Hippocampus (Brain); Human; Imagery; Immunoprecipitation; In Vitro; Individual; Knowledge; Learning; Link; Lymphocyte antigen; Maps; Measurement; Measures; Membrane; Mentors; Methods; Molecular; Molecular Biology; Mouse Strains; Mus; Mutation; Neurons; Neurotransmitter Receptor; Pathway Analysis; Pathway interactions; Phase; Physiological; Property; Protein-Protein Interaction Map; Proteins; Proteomics; Reporting; Research; Risk Factors; Sampling; Scientist; Signal Transduction; Social isolation; Students; Synapses; System; T-Lymphocyte; Technology; Testing; Tetrodotoxin; Training; Translating; career development; experience; immunogenic; immunological synapse; in vivo; induced pluripotent stem cell; link protein; new technology; new therapeutic target; novel; novel therapeutics; programs; protein complex; protein protein interaction; public health relevance; response; risk variant; signal processing; tool; trafficking; yeast two hybrid system

Project Summary/Abstract "Evaluating the synaptic hypothesis of autism using protein-protein interaction network analysis" Protein-protein interaction (PPI) networks of a cell are thought to represent a system with emergent network properties, integrating signals for a variety of inputs into coordinated responses. A deeper understanding of protein complexes at the synapse will enhance our knowledge of normal brain function and may highlight new therapeutic targets for diseases, including autism. The applicant has recently developed a novel technology, multiplex immunoprecipitation measured by flow cytometry (mIP-FCM) that allows the rapid, simultaneous, quantitative measurement of hundreds of protein-protein interactions (PPIs) from small amounts of biological samples. This proposal involves translating mIP-FCM technology to target synaptic proteins relevant to autism, and investigating the quantitative changes in protein complexes that occur under physiological and disease-associated conditions. The observation of the dynamic activity of protein complexes at the mouse and human glutamate synapse will contribute to a greater understanding of normal brain function, and may highlight pathological molecular mechanisms of autism-associated genes. The applicant proposes three specific aims: 1) Generate a multiplex IP-FCM assay targeted to synaptic proteins, to be completed during year 1 of the K99 portion of the award; 2) Map synaptic PPIs in mice carrying autism-associated mutations. This portion will be completed in years 2-5, during the K99/R00 transition. 3) Map synaptic PPIs in human iPS neurons derived from autistic and typically developing individuals, to be completed during years 3-5 of the R00 phase of the award. The overall goal of this proposal is to define specific changes in the PPI network of the glutamatergic synapse associated with different autism risk factors, and to investigate the extent to which there is convergence of autism risk genes into specific common molecular pathways at the synapse. During the K99 phase of the award, the applicant will 1) Develop the mIP-FCM assay targeted to the glutamatergic synapse, and learn appropriate statistical and visualization methods for the large datasets; 2) Learn to manage a research team, including management of technical support staff and students, overseeing budgets, integrating data from multiple individual projects, and acting as a mentor of junior scientists. The applicant has assembled a group of experienced co-mentors to help with this career development training. Overall, this training will facilitate the achievement of the applicant's long-term goal to develop an internationally-renowned independent research program focused on the molecular mechanisms of autism risk factors.

项目总结/文摘