Date of Award

5-2017

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Program

Biomedical Sciences

Track

Neuroscience

Research Advisor

Francesca-Fang Liao, Ph.D.

Committee

Ioannis Dragatsis, Ph.D. Michael P. McDonald, Ph.D. Edwards A. Park, Ph.D. Junmin Peng, Ph.D.

Keywords

Neurodegenerative diseases, Protein degradation, Proteinopathies, Stress response

Abstract

Cellular protein homeostasis is achieved by a delicate network of molecular chaperones and various proteolytic processes such as ubiquitin–proteasome system (UPS) to avoid a build-up of misfolded protein aggregates. The latter is a common denominator of neurodegeneration. Neurons are found to be particularly vulnerable to toxic stress from aggregation-prone proteins such as α-synuclein. Induction of heat-shock proteins (HSPs), such as through activated heat shock transcription factor 1 (HSF1) via Hsp90 inhibition, is being investigated as a therapeutic option for proteinopathic diseases. HSF1 is a master stress-protective transcription factor which activates genes encoding protein chaperones (e.g. iHsp70) and anti-apoptotic proteins. However, whether and how HSF1 is dysregulated during neurodegeneration has not been studied. Here, we discover aberrant HSF1 degradation by aggregated α-synuclein (or α-synuclein-induced proteotoxic stress) in transfected neuroblastoma cells. HSF1 dysregulation via α-synuclein was confirmed by in vivo assessment of mouse and in situ studies of human specimens with α-synucleinopathy. We demonstrate that elevated NEDD4 is implicated as the responsible ubiquitin E3 ligase for HSF1 degradation through UPS. Furthermore, pharmacologically induced SIRT1-mediated deacetylation can attenuate aberrant NEDD4-mediated HSF1 degradation. Indeed, we define the acetylation status of the Lys 80 residue located in the DNA-binding domain of HSF1 as a critical factor in modulating HSF1 protein stability in addition to its previously identified role in the transcriptional activity. Together with the finding that preserving HSF1 can alleviate α-synuclein toxicity, the first part of the study strongly suggests that aberrant HSF1 degradation is a key neurodegenerative mechanism underlying α-synucleinopathy. Chronic activation of another cellular stress response, unfolded protein response in ER, has been implicated in tauopathy including Alzheimer’s disease (AD). The unfolded protein response (UPR) in the endoplasmic reticulum (ER) and the cytoplasmic heat stress response are two major stress response systems necessary for maintaining proteostasis for cellular health. Failure of either of these systems, such as in sustained UPR activation or in insufficient heat shock response activation, can lead to the development of neurodegeneration. Alleviation of ER stress and enhancement of heat shock response through heat shock factor 1 (HSF1) activation have previously been considered as attractive potential therapeutic targets for AD—a prevalent and devastating tauopathy. The second part of the study concentrates on our attempts to understand the interplay of the two aforementioned systems and their cooperative role in AD. We provide compelling in vitro and in vivo evidence that strongly suggests an auto-propagating interplay of UPR activation and HSF1 degradation being a common pathogenic feature in both human AD and tau transgenic mouse AD models. We identify aging-associated AD-like neuropathological changes in the hippocampus of HSF1 heterozygous knock-out mice. We speculate that HSF1 loss as an early (earliest) event which constitutes a mechanistic connection between ER stress and tau hyperphosphorylation in tau pathology. Finally, we demonstrate that aged mice lacking HSF1 gene exhibited deficits in hippocampal-dependent functions including short-term working memory, spatial learning and long-term memory. All together, our work supports a previously underappreciated importance of this master stress regulator HSF1 in neuronal functions and in maintaining brain homeostasis.

ORCID

http://orcid.org/0000-0002-9297-2848

DOI

10.21007/etd.cghs.2017.0432

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