Administrative supplement to Regulation of mitochondrial DNA homeostasis and neuroinflammation by Fascin

项目总结/文摘

Project Summary/Abstract Alzheimer’s disease (AD), the most common neurodegenerative disorder, affects one in ten people age 65 and older. Due to limited understanding of mechanisms underlying AD pathogenesis, there is no effective treatment for this devastating disease. The goal of this application is to investigate an unexpected role for actin bundling protein Fascin in regulating mitochondrial nucleoid DNA (mtDNA) homeostasis, oxidative phosphorylation (OXPHOS), mitochondrial oxidative stress, neuroinflammation, and neurodegeneration, as well as how dysregulation of these processes contributes to AD pathogenesis. Fascin is an actin bundling protein essential for the cross-linking of actin filaments into compact and rigid bundles. The current paradigm posits that Fascin promotes cell migration and tumor invasion by generating protrusive membrane structures such as filopodia. We recently made the surprising finding that depletion of Fascin disrupts mitochondrial F- actin bundling, which in turn causes abnormal mitochondrial respiratory complex biogenesis and impaired mitochondrial OXPHOS, suggesting a novel role of Fascin in the regulation of mitochondrial function. Mechanistically the mitochondrial dysfunction in Fascin depleted cells was due to increased mtDNA aggregation and leakage. Given that mtDNA can robustly induce inflammasome activation and inflammatory cytokine expression, Fascin deficiency may play an unexpected role in causing neuroinflammation. Importantly, we found that Fascin is cleaved into a 37kDa functionally dominant-negative form in the brains of AD patients and AD mouse models. Virus-mediated expression of Fascin in AD mouse hippocampus mitigated disease symptoms. In addition, Fascin knockout mice we generated showed profound mitochondrial defects and significant loss of neurons in the brain. Based on these preliminary data, we hypothesize that Fascin controls mitochondrial function and mtDNA homeostasis in the brain. Fascin functional deficiency in AD leads to significant mitochondrial defects, neuroinflammation and neurodegeneration. To test the hypothesis, in Aim 1 we will define the role of Fascin in regulating mitochondrial function, mtDNA homeostasis, neuroinflammation and neuronal cell death in the mouse brain in vivo. In Aim 2 we will investigate the functional deficiency of Fascin caused by proteolytic cleavage during the course of AD pathogenesis using brain tissues from AD patients and mouse models, and to elucidate mechanisms underlying how Fascin functional deficiency causes mtDNA leakage, oxidative stress, neuroinflammation, and degeneration. In Aim 3 we will study the effects of transgenic expression of Fascin on alleviating AD pathological phenotypes and disease symptoms in mice. Successful completion of the proposed studies will reveal Fascin’s novel role in regulating mitochondrial function, mtDNA homeostasis, neuroinflammation and neurodegeneration. Investigating Fascin functional deficiency in AD will help understand disease pathogenic mechanisms and facilitate therapeutic development.