Extracellular protein aggregation is inextricably linked to human neurodegenerative diseases such as Alzheimer's disease, Creutzfeldt-Jakob disease and the transthyretin amyloidoses. The importance of protein aggregation in these disorders has led to significant experimental effort focused on characterizing the cellular pathways that regulate extracellular protein homeostasis (or proteostasis). Two primary determinants in defining extracellular proteostasis identified through these efforts are the efficiency of endoplasmic reticulum (ER) quality control pathways and the spectrum and activity of extracellular chaperones. A primary function of the ER is to facilitate the proper folding of proteins for trafficking to downstream environments of the secretory pathway such as the extracellular environment, while preventing the secretion of destabilized, misfolding-prone proteins. Through this so-called quality control mechanism, the ER indirectly influences extracellular proteostasis by reducing the extracellular population of misfolding prone proteins available for concentration- dependent aggregation. Alternatively, extracellular chaperones such as clusterin directly influence extracellular proteostasis by binding misfolding prone proteins, preventing their aggregation. While the combined activity of ER quality control pathways and extracellular chaperones efficiently regulate extracellular proteostasis under normal conditions, imbalances in ER quality control efficiency induced by environmental, genetic or aging- related insults can lead to increased secretion of aggregation-prone proteins that challenge extracellular chaperoning capacity. To confront this challenge, cells activate the unfolded protein response (UPR) - a stress-responsive signaling pathway that translationally and transcriptionally remodels ER quality control pathways. Thus, UPR activation restores ER function and attenuates the secretion of aggregation-prone proteins. Despite the importance of the UPR in regulating proteostasis in the ER and downstream environments of the secretory pathway, no link between UPR activation and extracellular chaperoning capacity has been established. We hypothesize that UPR activation, in response to stress, increases secretion of extracellular chaperones to prevent the aberrant, extracellular aggregation of misfolding prone proteins. Herein, we identify ERdj3 (DNAJB11) as a previously uncharacterized, UPR-regulated extracellular chaperone. We propose to evaluate the capacity for ERdj3 to attenuate pathologic protein aggregation in the extracellular environment, characterize the biological pathways responsible for stress-induced ERdj3 secretion, and identify the subset of secreted proteins that interacts with ERdj3 during normal physiology and in response to stress. Experimental success herein will provide proof-of-principle of a direct, functional role for the UPR in regulating extracellular proteostasis capacit, catalyzing research to both identify other UPR-regulated extracellular proteostasis factors and explore the potential for inducing the UPR-dependent increase in extracellular chaperone capacity to treat aggregation-associated degenerative disorders.

Public Health Relevance

The extracellular misfolding and aggregation of proteins is inextricably linked to numerous human neurodegenerative diseases including Alzheimer's disease, Creutzfeldt-Jakob disease and the transthyretin amyloidoses. While the contributions of intracellular protein folding pathways to this extracellular protein misfolding/aggregation are well studied, the biological pathways that maintain the stability of the secreted proteome in the extracellular environment remain poorly characterized. Herein, we propose to explore how the unfolded protein response - the stress-responsive signaling pathway that regulates protein homeostasis throughout the secretory pathway - alters the composition and activity of the extracellular chaperone network, revealing novel proteins and pathways that can in principle be therapeutically manipulated to attenuate disease-associated extracellular protein aggregation.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Exploratory/Developmental Grants (R21)
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Cellular and Molecular Biology of Neurodegeneration Study Section (CMND)
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Corriveau, Roderick A
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Scripps Research Institute
La Jolla
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Cooley, Christina B; Ryno, Lisa M; Plate, Lars et al. (2014) Unfolded protein response activation reduces secretion and extracellular aggregation of amyloidogenic immunoglobulin light chain. Proc Natl Acad Sci U S A 111:13046-51
Chen, John J; Genereux, Joseph C; Qu, Song et al. (2014) ATF6 activation reduces the secretion and extracellular aggregation of destabilized variants of an amyloidogenic protein. Chem Biol 21:1564-74