Stress Granules and the Biology of TDP-43

应激颗粒和 TDP-43 的生物学

基本信息

批准号：
8625475
负责人：
Benjamin L Wolozin
金额：
$ 41.98万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8625475
关键词：
Accounting Alanine Amyotrophic Lateral Sclerosis Aryl Hydrocarbon Receptor Biochemical Biology C9ORF72 Cessation of life Chemicals Clinic Code Cytoplasmic Granules DNA Development Disease Disease Progression Familial Amyotrophic Lateral Sclerosis Functional disorder Funding Gene Expression Gene Proteins Gene Targeting Genes Genetic Genetic Risk Genetic Transcription Genomics Genotype Hematopoiesis Length Libraries Ligands Link Lymphocyte Malignant Neoplasms Measures Mediating Motor Neurons Mutate Mutation National Institute of Environmental Health Sciences Neurites Neurons Organophosphates Outcome Pathology Patients Phenotype Principal Investigator Process Proteins RNA Regulation Reporter Research Research Personnel Research Project Grants Risk Role Sampling Sequence Analysis Source Stress Testing Toxic Environmental Substances Variant activating transcription factor aryl hydrocarbons cohort environmental chemical gene discovery gene environment interaction genetic analysis genetic linkage genetic variant induced pluripotent stem cell inhibitor/antagonist innovation link protein lipid biosynthesis mutant neurotoxic neurotoxicity novel programs protein TDP-43 protein aggregation protein expression public health relevance research study response risk variant small hairpin RNA toxicant

项目摘要

The major objectives of this ViCTER program are to test novel mechanisms of gene-environment interactions. We will test whether environmental toxicants linked to amyotrophic lateral sclerosis (ALS) act in part through genes linked to ALS, including the newly discovered gene, C9ORF72, as well as the gene PON2, which biotransforms environmental toxicants. We will also test whether toxicants might contribute to ALS by acting through the aryl hydrocarbon (AhR) receptor, which is known to mediate toxicant action in other diseases, such as cancers. Hexanucleotide expansions in C9ORF72 account for 30% of cases of familial ALS. Inclusion of samples from ALS patients through our co-investigator, Dr. Rademakers, allows us to generate iPSCs from subjects with these mutations. The AhR is a ligand- activated transcription factor that mediates many of the effects of environmental toxicants in hematopoiesis, lymphocyte development, adipogenesis and cancer. We hypothesize that AhR ligands and other environmental chemicals, many of which are neurotoxic might also increase the risk of ALS by stimulating transcription of genes linked to ALS. The experiments in this consortium could identify novel mechanisms through which environmental toxicants might contribute to ALS. In addition, the availability of novel, non-toxic AhR antagonists, which were generated by Dr. Sherr's team, offers the exciting prospect of potentially inhibiting the expression of genes linked to ALS, a strategy that might reduce or delay the pathophysiology of ALS. Aim 1 will determine the role of AhR and PON2 genetic variants in ALS using the Mayo ALS cohort; Dr. Rademakers will perform in-depth genetic analyses of AhR in ~750 ALS patients and matched controls ascertained at Mayo Clinic to determine whether there are common genetic risk variants and/or rare modifier variants or disease causing variants in AhR. In Aim 2, Dr. Murphy will generate iPSC lines from subjects identified by Dr. Rademakers who have ALS and express mutated C9ORF72 and PON2. In Aim 3, Dr. Wolozin will determine whether genes linked to ALS (e.g., TARDBP, C9ORF72 or PON2) increase the vulnerability and/or pathology in response to ALS-linked environmental toxicants in MN iPSCs. Toxicants linked to ALS will be utilized, including ¿- methyl amino alanine, neurotoxic organophosphates and environmental AhR ligands. In Aim 4, Dr. Sherr will determine whether AhR ligands increase the expression of ALS-linked genes and proteins, and modify the resulting phenotypes. He will use AhR-specific shRNA, novel inhibitors, qPCR and AhR-reporter-transduced iPSCs to characterize AhR expression and activity in MN iPSCs. Thus, in this consortium, we will determine if AhR contributes to ALS through regulation of ALS-associated genes, and whether such activation modifies the pathophysiology of genes or proteins linked to ALS.

ViCTER 计划的主要目标是测试基因-环境的新机制我们将测试环境毒物是否与肌萎缩侧索硬化症（ALS）有关。部分通过与 ALS 相关的基因发挥作用，包括新发现的基因 C9ORF72 以及基因PON2，它可以对环境毒物进行生物转化，我们还将测试毒物是否可能。通过芳基烃 (AhR) 受体发挥作用，导致 ALS，已知该受体可介导 C9ORF72 中的六核苷酸扩展导致了其他疾病的毒性作用。 30% 的家族性 ALS 病例通过我们的共同研究员 Dr. 纳入 ALS 患者的样本。 Rademakers 使我们能够从具有这些突变的受试者中生成 iPSC。 AhR 是一种配体。激活的转录因子介导环境毒物的许多影响我们采用了 AhR 配体来影响造血、淋巴细胞发育、脂肪生成和癌症。和其他环境化学物质，其中许多具有神经毒性，也可能增加 ALS 的风险该联盟的实验可以通过刺激与 ALS 相关的基因转录来识别。环境毒物可能导致 ALS 的新机制。 Sherr 博士团队开发的新型无毒 AhR 拮抗剂的可用性提供了潜在抑制 ALS 相关基因表达的令人兴奋的前景，这一策略可能减少或延缓 ALS 的病理生理学目标 1 将决定 AhR 和 PON2 遗传的作用。 Rademakers 博士将利用 Mayo ALS 队列对 ALS 变异进行深入的基因分析在 Mayo Clinic 确定了约 750 名 ALS 患者和匹配对照的 AhR，以确定是否存在是 AhR In 中常见的遗传风险变异和/或罕见的修饰变异或致病变异。目标 2，Murphy 博士将从 Rademakers 博士确定的患有 ALS 的受试者中生成 iPSC 系并表达突变的 C9ORF72 和 PON2 在目标 3 中，Wolozin 博士将确定基因是否相关。 ALS（例如 TARDBP、C9ORF72 或 PON2）会增加响应的脆弱性和/或病理 MN iPSC 中与 ALS 相关的环境毒物将被利用，包括 ¿ - 甲基氨基丙氨酸、神经毒性有机磷酸酯和环境 AhR 配体在 Aim 4 中，Dr. Sherr 将确定 AhR 配体是否会增加 ALS 相关基因和蛋白质的表达，并修改由此产生的表型。他将使用 AhR 特异性 shRNA、新型抑制剂、qPCR 和 AhR 报告基因转导的 iPSC 来表征 MN iPSC 中的 AhR 表达和活性。联盟，我们将确定 AhR 是否通过调节 ALS 相关基因来促进 ALS，以及这种激活是否会改变与 ALS 相关的基因或蛋白质的病理生理学。