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Retrograde transmission of pathogenic TDP-43 to the CNS

致病性 TDP-43 逆行传播至中枢神经系统

基本信息

批准号：
10561606
负责人：
Eileen Marie Lynch
金额：
$ 7.4万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10561606
关键词：
Alzheimer&apos s Disease Amyloid Amyloid beta-Protein Amyotrophic Lateral Sclerosis Atrophic Behavior Biological Assay Biopsy Biosensor Brain Brain region Cell Line Cell Nucleus Cells Cellular Stress Central Nervous System Coculture Techniques Collaborations Competence Cytoplasm Cytoplasmic Granules Dementia Development Diagnosis Disease Environment Fluorescence Resonance Energy Transfer Frontotemporal Dementia Future Hindlimb Human In Vitro Induced pluripotent stem cell derived neurons Inflammation Injections Intramuscular Intramuscular Injections Link Methods Monitor Motor Mus Muscle Mutate Mutation Myoblasts Myopathy Natural regeneration Necrosis Nerve Degeneration Neurodegenerative Disorders Neuromuscular Diseases Neurons Nuclear Localization Signal Parkinson Disease Pathogenicity Pathologic Pathology Patients Phosphorylation Proteins Qualifying RNA-Binding Proteins Recombinants Research Research Personnel Resources Skeletal Muscle Spinal Cord Stress Structure Symptoms System TDP-43 aggregation Testing Time Training Wild Type Mouse alpha synuclein cognitive change enhancing factor experimental study improved in vivo misfolded protein motor impairment mouse model multisystem proteinopathy muscle form neurotransmission novel diagnostics novel therapeutics protein TDP-43 protein aggregation sensor stress granule superoxide dismutase 1 tau aggregation timeline transmission process

项目摘要

Project Summary/Abstract TDP-43 is a common protein found aggregated in multiple neurodegenerative diseases and myopathies. TDP- 43 is a ubiquitously expressed RNA-binding protein that is primarily localized in the nucleus but relocates to the cytoplasm under conditions of cellular stress, forming stress granules in neurons and myo-granules in skeletal muscle. Normally TDP-43 returns to the nucleus when the stress is resolved, but through unknown disease mechanisms misfolded TDP-43 remains in the cytoplasm as phosphorylated insoluble aggregates. Studies have shown that TDP-43 forms amyloid-like oligomers which are capable of self-templated conversion in which the misfolded protein causes its native counterpart to become misfolded, leading to propagation of the aggregate pathology. The misfolded aggregation-prone proteins, referred to as proteopathic seeds, are capable of cell-to-cell transmission through connected neuronal networks. In some diseases such as multisystem proteinopathy, patients have TDP-43 aggregation in both skeletal muscle and neurons. In these cases, myopathy symptoms begin several years prior to the onset of dementia. Therefore, the central hypothesis behind this proposal is that TDP-43 is capable of self-templated conversion in skeletal muscle and transmission to the central nervous system (CNS) to cause neurodegeneration. In order to test this, TDP-43 isolated from human skeletal muscle myo-granules will first be tested for the ability to undergo self-templated conversion in vitro using a thioflavin assay and a TDP-43 FRET sensor line. Then, the isolated TDP-43 or recombinant TDP-43 seeds will be injected into the hindlimb muscles of wild type mice and mice expressing TDP-43 with a mutated nuclear localization signal which causes increased cytoplasmic TDP- 43 expression. Mice will be monitored for motor or cognitive changes and sacrificed at specific time points following proteopathic seed injection. Skeletal muscle will be examined for signs of local transmission and pathology to confirm whether seeds can propagate in the environment of a myofiber. Additionally, the spinal cord and brain will be examined for signs of TDP-43 pathology to determine whether intramuscular injected TDP-43 proteopathic seeds can lead to transmission of TDP-43 proteinopathy to the CNS. To confirm muscle- to-neuron transmission of TDP-43 seeds on a shorter timeline, primary mouse myoblasts expressing fluorescently-tagged TDP-43 will be treated with proteopathic seeds and then co-cultured with primary mouse neurons. The neurons will be monitored for fluorescent TDP-43, indicating seeding. If successful, the proposed experiments will significantly improve the understanding of TDP-43 proteinopathy and whether it can spread from the periphery to the CNS. This has the potential to greatly impact the diagnosis methods and treatment options for neurodegenerative diseases. The successful completion of these aims as well as the training plan provided will prepare the applicant to become an independent investigator in the field of protein aggregation and neuromuscular disease.

项目总结/文摘