Functional Dissection of Alzheimer's Disease Networks in Drosophila: from Association to Causal Modulators of Age-Dependent Neurodegeration

果蝇阿尔茨海默病网络的功能剖析：从年龄依赖性神经变性的关联到因果调节因子

• 批准号: 10228292
• 负责人: Juan Botas
• 金额: $ 13.67万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228292
• 关键词: Adult; Affect; Age; Alleles; Alzheimer disease screening; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Animal Model; Antibodies; Automobile Driving; Autopsy; Biochemical; Bioinformatics; Biological Assay; Brain; Cells; Collection; Complement; Confocal Microscopy; Consensus; Cultured Cells; Data; Data Analyses; Disease; Disease Progression; Dissection; Drosophila genus; Fluorescence; Gene Expression; Gene Expression Alteration; Gene Proteins; Genes; Genetic Screening; Genotype; Glutamates; Goals; Grant; Human; Immunoblotting; Immunofluorescence Immunologic; Individual; Investigation; Knowledge Portal; Maintenance; Mammalian Cell; Medicine; Messenger RNA; Modeling; Mus; NUAK1 gene; Neurodegenerative Disorders; Neuroglia; Neurons; Parents; Pathogenesis; Pathologic; Pathway interactions; Population; Progress Reports; Reagent; Regulator Genes; Sampling; Synapses; Testing; Therapeutic; Time; Tissues; Validation; abeta accumulation; age related; base; case control; causal variant; cell type; cholinergic neuron; differential expression; dopaminergic neuron; experimental study; gene discovery; genetic manipulation; improved; innovation; nerve stem cell; parent grant; protective factors; small hairpin RNA; tau Proteins; tau aggregation; transcriptome sequencing; transcriptomics

The goal of this supplemental project is to extend the bioinformatic analyses and experimental validation of AMP-AD target networks, which are the focus of the parent grant. First (AIM1), we will generate single-cell longitudinal transcriptomic profiles of Drosophila AD models expressing human tau and secreted amyloid-β. These data will allow to distinguish gene expression profiles from distinct neuronal populations as well as different glia subtypes. These studies will also strongly complement ongoing AMP-AD single-cell gene expression profiles from human postmortem tissue. We will include rigorous experimental controls for tau / amyloid-β expression as well as longitudinal sampling to dissect out the specific contributions of age and AD pathologic species on cell-type specific gene expression changes in the brain. Initially we will include 2 time points (early and late) during disease progression. During the first year of the proposed supplement (Y4 of the parent R01 grant) we will generate Drosophila of the appropriate genotypes, extract mRNA, perform scRNAseq, and begin analyses of the results. During the second year of the supplement (Y5 of the parent R01 grant) we will complete analysis of the data, integrating with our findings from whole brain RNAseq and performing cross-species comparisons with AMP-AD scRNAseq. We will also perform independent experiments to confirm the most promising results from scRNAseq, such as immunofluorescence confocal microscopy, taking advantage of available antibodies, and reagents. Selected candidate causal drivers will also be manipulated using cell-type specific drivers, including glia and/or glutamatergic, GABAergic, cholinergic and dopaminergic neurons within Drosophila CNS to examine requirements for brain maintenance and/or function. Second (AIM2), we will extend experimental validation of computationally predicted AD causal genes from Drosophila to mammalian cells. All causal drivers identified in the tau Drosophila screen, will be tested in cultured mouse and human neural progenitor cells using shRNAs. Specifically, we will assess the impact of causal drivers on tau protein levels given the key role of tau accumulation AD pathogenesis. Our data shows that a subset of the identified tau modifier genes (e.g., Hippo pathway components, Nuak1) lower tau protein levels in Drosophila. Thus, we hypothesize that a subset of causal drivers identified in the ongoing AMP-AD screen modulate tau accumulation and this subset may be especially interesting from a therapeutic standpoint. During the first year of the proposed supplement (Y4 of the parent R01 grant) we will test the Hippo pathway genes in Neuro2A and human neural precursor cells plus all 44 tau modifier genes for their ability to modulate tau protein levels in the Drosophila brain using immunoblotting and Homogeneous Time Resolved Fluorescence approaches. During the second year of the supplement (Y5 of the parent R01 grant) we will test in Neuro2A and human neural precursor cells all additional genes lowering tau levels in Drosophila brains. Third (AIM 3), all project data will be made available via the Synapse/AMP-AD Knowledge Portal.

本补充项目的目的是扩展生物信息学分析和实验验证

项目成果

Integrated proteomics reveals brain-based cerebrospinal fluid biomarkers in asymptomatic and symptomatic Alzheimer's disease.

• DOI: 10.1126/sciadv.aaz9360
• 发表时间: 2020-10
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Higginbotham L;Ping L;Dammer EB;Duong DM;Zhou M;Gearing M;Hurst C;Glass JD;Factor SA;Johnson ECB;Hajjar I;Lah JJ;Levey AI;Seyfried NT
• 通讯作者: Seyfried NT

Targeted Quantification of Detergent-Insoluble RNA-Binding Proteins in Human Brain Reveals Stage and Disease Specific Co-aggregation in Alzheimer's Disease.

• DOI: 10.3389/fnmol.2021.623659
• 发表时间: 2021
• 期刊: Frontiers in molecular neuroscience
• 影响因子: 4.8
• 作者: Guo Q;Dammer EB;Zhou M;Kundinger SR;Gearing M;Lah JJ;Levey AI;Shulman JM;Seyfried NT
• 通讯作者: Seyfried NT

Global quantitative analysis of the human brain proteome and phosphoproteome in Alzheimer's disease.

• DOI: 10.1038/s41597-020-00650-8
• 发表时间: 2020-09-28
• 期刊: Scientific data
• 影响因子: 9.8
• 作者: Ping L;Kundinger SR;Duong DM;Yin L;Gearing M;Lah JJ;Levey AI;Seyfried NT
• 通讯作者: Seyfried NT

Insights into the changes in the proteome of Alzheimer disease elucidated by a meta-analysis.

• DOI: 10.1038/s41597-021-01090-8
• 发表时间: 2021-12-03
• 期刊: Scientific data
• 影响因子: 9.8
• 作者: Haytural H;Benfeitas R;Schedin-Weiss S;Bereczki E;Rezeli M;Unwin RD;Wang X;Dammer EB;Johnson ECB;Seyfried NT;Winblad B;Tijms BM;Visser PJ;Frykman S;Tjernberg LO
• 通讯作者: Tjernberg LO

TBK1 interacts with tau and enhances neurodegeneration in tauopathy.

• DOI: 10.1016/j.jbc.2021.100760
• 发表时间: 2021-01
• 期刊: The Journal of biological chemistry
• 作者: Abreha MH;Ojelade S;Dammer EB;McEachin ZT;Duong DM;Gearing M;Bassell GJ;Lah JJ;Levey AI;Shulman JM;Seyfried NT
• 通讯作者: Seyfried NT