RNA Processing Misregulation in KIF5A ALS

PROJECT SUMMARY Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease that was previously considered mainly a disorder of protein mislocalization. However, recent findings have unveiled a significant association of ALS with RNA misregulation. Among the 53 genes linked to ALS, the gene KIF5A has recently garnered attention. KIF5A, a kinesin motor protein known for its role in organelle transport, has been implicated in ALS through mutations that purportedly lead to a gain-of-function, resulting in a hyperactive state of the protein. My research has utilized machine learning predictions alongside experimental approaches to demonstrate that KIF5A functions as an RNA-binding protein. Moreover, I found ALS-related mutations in KIF5A modify its RNA-binding profile, uncovering a novel aspect of KIF5A's involvement in ALS. My preliminary findings indicate that KIF5A with ALS mutations exhibits a preference for binding to mRNA of heat shock proteins and several other genes associated with ALS. I propose that mutations in KIF5A contribute to motor neuron degeneration by disrupting RNA processing. The proposed research encompasses three objectives: (1) to investigate the impact of KIF5A ALS mutations on the proteotoxic pathway, (2) to explore the interactions between KIF5A and ALS-associated genes and RNA targets, and (3) to evaluate the effects of KIF5A mutations on transcription, RNA localization, and RNA-binding protein (RBP)-RNA interactions using spinal organoid models. Success in this project will lay the groundwork for innovative methods and insights into the misregulation of RNA processing in ALS. My expertise in quantitative, single-molecule imaging, combined with the Yeo lab's profound knowledge in RNA processing, stem cell models and neurodegeneration, positions me uniquely to lead the proposed research. These objectives will establish a foundation for my career as an independent researcher, focusing on the mechanisms of RNA transport in neurodegenerative disorders.