Stress Granules and the Biology of TDP-43

应激颗粒和 TDP-43 的生物学

• 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
• 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

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.

该ViCTER计划的主要目标是测试基因-环境的新机制， 交互.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相关的基因起作用，包括新发现的基因C9 ORF 72，以及 基因PON 2，它可以生物转化环境毒物。我们还将测试有毒物质是否可能 通过芳香烃（AhR）受体起作用而导致ALS，已知芳香烃（AhR）受体介导 在其他疾病中的毒性作用，如癌症。C9 ORF 72中的六核苷酸扩增解释了 30%的家族性ALS病例。通过我们的合作研究者， Rademakers允许我们从具有这些突变的受试者中产生iPSC。AhR是一种配体- 一种激活的转录因子，介导环境毒物的许多影响， 造血、淋巴细胞发育、脂肪生成和癌症。我们假设AhR配体 以及其他环境化学物质，其中许多是神经毒性的，也可能增加ALS的风险。 通过刺激与ALS相关的基因的转录。这个联盟的实验可以识别 环境毒物可能导致ALS的新机制。此外该 由Sherr博士的团队开发的新型无毒AhR拮抗剂的可用性， 一个令人兴奋的前景，可能抑制基因的表达与ALS，一个战略， 减少或延缓ALS的病理生理学。目的1确定AhR和PON 2基因的作用， 使用马约ALS队列研究ALS的变异; Rademakers博士将对 在马约诊所确定的约750例ALS患者和匹配对照中的AhR，以确定是否存在 是AhR中常见的遗传风险变体和/或罕见的修饰变体或致病变体。在 目标2，Murphy博士将从Rademakers博士确定的患有ALS的受试者中产生iPSC系 并表达突变的C9 ORF 72和PON 2。在目标3中，Wolozin博士将确定基因是否与 至ALS（例如，TARDBP、C9 ORF 72或P0 N2）增加了响应于以下的易感性和/或病理学： MN iPSC中ALS相关的环境毒物。将使用与ALS相关的毒物，包括： 甲基氨基丙氨酸、神经毒性有机磷酸酯和环境AhR配体。在目标4中，博士。 Sherr将确定AhR配体是否增加ALS相关基因和蛋白质的表达， 并改变所产生的表型。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以表征MN iPSC中的AhR表达和活性。所以针对本 财团，我们将确定是否AhR有助于ALS通过调节ALS相关基因， 以及这种激活是否改变了与ALS相关的基因或蛋白质的病理生理学。