Structural and biochemical studies of patient-extracted amyloid fibrils
- Jiang, Yi Xiao
- Advisor(s): Eisenberg, David S
Abstract
Human neurodegenerative diseases cause the gradual loss of neurons and progressive decline in memory, cognitive ability, movement and behavior. These devastating conditions afflict millions of patients worldwide and their families, however the molecular mechanisms of pathogenesis are not well understood, resulting in the lack of effective treatments and early diagnostic tools. Gross analyses of post-mortem brains using immunohistochemistry has identified abnormal deposits of amyloid proteins as defining features of neurodegenerative diseases. Amyloid proteins, like prions, can template aggregation of monomers into highly ordered, stable fibrils composed of tightly interdigitating β-sheets. Recent advances in cryogenic-electron microscopy (cryo-EM) have enabled the structural determination of amyloid fibrils to near-atomic resolution. These structures provide insight into the formation and propagation of amyloid fibrils, and can guide the design of therapies that prevent, delay, or reverse the aggregation of proteins in neurodegenerative diseases. Moreover, previous work has shown that the structures of brain-extracted fibrils differ from the structures of fibrils assembled in vitro, underscoring the importance of studying patient-derived samples. In my thesis research, I extracted amyloid fibrils from autopsied brains of patients with frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) and determined their structures using cryo-EM. Surprisingly, the fibrils examined were not formed by TDP-43 but by a fragment of transmembrane protein 106B (TMEM106B), revealing a previously unsuspected amyloid protein. In addition, I used a biosensor cell line, which detects proteopathic TDP-43 aggregates from sarkosyl-insoluble brains extracts, to study cohorts of TDP-43 proteinopathy cases including amyotrophic lateral sclerosis (ALS), FTLD-TDP and Alzheimer’s disease (AD). My work expands the insight into the molecular basis of amyloid disorders, and inspires future research to address newly raised questions about TMEM106B and TDP-43 in neurodegenerative diseases.