bims-proarb Biomed News
on Proteostasis in aging and regenerative biology
Issue of 2022–01–02
ten papers selected by
Rich Giadone, Harvard University



  1. IUBMB Life. 2022 Jan;74(1): 41-52
      The tight regulation of proteostasis is essential for physiological cellular function. Mammalian cells possess a network of mechanisms that ensure proteome integrity under normal or stress conditions. The proteasome, being the major cellular proteolytic machinery, is central to proteostasis maintenance in response to distinct intracellular and extracellular conditions. The proteasomes are multisubunit protease complexes that selectively catalyze the degradation of short-lived regulatory proteins and damaged peptides. Different forms of the proteasome complexes comprising of different subunits and attached regulators directly affect the substrate selectivity and degradation. Thus, the proteasome participates in the turnover of a multitude of factors that control key processes that affect the cellular state, such as adaptation to environmental cues, growth, development, metabolism, signaling, senescence, pluripotency, differentiation, and immunity. Aberrations on its function are related to normal processes like aging and pathological conditions such as neurodegeneration and cancer. The past few years of research have highlighted that proteasome abundance, activity, assembly, and localization are subject to a dynamic transcriptional control that secures the continuous adaptation of the proteasome to internal or external stimuli. This review focuses on the factors and signaling pathways that are involved in the regulation of the mammalian proteasome at the transcriptional level. A comprehensive understanding of proteasome regulation has critical implications on disease prevention and treatment.
    DOI:  https://doi.org/10.1002/iub.2586
  2. FEBS J. 2021 Dec 30.
      Biological aging is the main driver of age-associated chronic diseases. In 2014, the United States National Institute of Aging (NIA) sponsored a meeting between several investigators in the field of aging biology, who identified 7 biological pillars of aging and a consensus review, "Geroscience: Linking Aging to Chronic Disease," was published. The pillars of aging demonstrated the conservation of aging pathways in diverse model organisms and thus, represent a useful framework with which to study human aging. In this present review, we revisit the 7 pillars of aging from the perspective of exercise and discuss how regular physical exercise can modulate these pillars to stave off age-related chronic diseases and maintain functional capacity.
    Keywords:  aging; disruption in proteostasis; dysregulated stress response; epigenetic drift; inflammaging; macromolecular damage; metabolic dysregulation; stem cell exhaustion
    DOI:  https://doi.org/10.1111/febs.16337
  3. Exp Eye Res. 2021 Dec 27. pii: S0014-4835(21)00471-1. [Epub ahead of print] 108905
      The BALB.NCT-Cpoxnct is a mutant mouse model for hereditary cataracts. We previously uncovered that the primary cause of the cataracts of BALB.NCT-Cpoxnct is a mutation in the coproporphyrinogen oxidase (Cpox) gene. Because of the mutation, excessive coproporphyrin is accumulated in the BALB.NCT-Cpoxnct lens. In this study, we analyzed the changes in transcriptome and proteins in the lenses of 4- and 12-week-old BALB.NCT-Cpoxnct to further elucidate the molecular etiology of cataracts in this mouse strain. Transcriptome analysis revealed that endoplasmic reticulum (ER) stress was increased in the BALB.NCT-Cpoxnct lens that induced persistent activation of the PERK signaling pathway of the ER stress response. Also, levels of crystallin transcripts and proteins were reduced in the BALB.NCT-Cpoxnct lens. Analysis of proteins disclosed aggregation of crystallins and keratins prior to the manifestation of cataracts in 4-week-old BALB.NCT-Cpoxnct mice. At 12 weeks of age, insoluble crystallins were accumulated in the cataractous BALB.NCT-Cpoxnct lens. Overall, our data suggest the following sequence of events in the BALB.NCT-Cpoxnct lens: accumulated coproporphyrin induces the aggregation of proteins including crystallins. Aggregated proteins increase ER stress that, in turn, leads to the repression of global translation of proteins including crystallins. The decline in the molecular chaperone crystallin aggravates aggregation and insolubilization of proteins. This vicious cycle would eventually lead to cataracts in BALB.NCT-Cpoxnct.
    Keywords:  Cataract; Coproporphyrin; Crystallin; Endoplasmic reticulum stress; Mouse; Protein aggregation; Transcriptomics
    DOI:  https://doi.org/10.1016/j.exer.2021.108905
  4. Alzheimers Dement. 2021 Dec;17 Suppl 2 e058727
       BACKGROUND: Impaired proteostasis is associated with normal aging and is accelerated in neurodegeneration. This impairment may lead to the toxic protein accumulation. In a subset of frontotemporal dementia (FTD) cases, mutations in the microtubule-associated protein tau (MAPT) that alter the relative levels of specific tau isoforms are sufficient to cause tau inclusions in neurons and astroglia and neurodegeneration without the presence of mutated protein (e.g. MAPTIVS10+16). However, the pathogenic events triggered by the expression of the alternatively spliced tau remain poorly understood.
    METHOD: To determine whether altered tau splicing induced from MAPT IVS10+16 mutations is sufficient to alter proteostasis in neurons and glia, we used human induced pluripotent stem cell (iPSC)-derived neurons and astrocytes from patients carrying the MAPT IVS10+16 mutation and CRISPR/Cas9, isogenic corrected controls.
    RESULT: We found that neurons from MAPT IVS10+16 carriers exhibited significantly higher levels of 4 repeat tau, deficits in lysosomal trafficking, and reduced lysosomal acidity relative to isogenic-control neurons. Additionally, MAPTIVS10+16 was sufficient to reduce genes associated with lysosomal biogenesis (regulated by TFEB). Interestingly, mutant astrocytes also produce elevated 4 repeat tau levels and exhibit several key hallmarks of cellular aging. Mutant astrocytes were larger in size with enlarged nuclei compared to isogenic controls. In addition to this hypertrophy phenotype, mutant astrocytes exhibited significantly elevated levels of senescence genes. Furthermore, markers of proteostasis were altered in mutant astrocytes: MAPT IVS10+16 carriers exhibited an increase in acidic lysosomes compared to isogenic-control astrocytes, and TFEB-regulated genes were upregulated MAPT IVS10+16 astrocytes.
    CONCLUSION: Our findings suggest that altered tau splicing induced by the MAPT IVS10+16 mutation is sufficient to cause aging signatures, including hypertrophy, senescence, and altered proteostasis in a cell-type specific manner.
    DOI:  https://doi.org/10.1002/alz.058727
  5. Alzheimers Dement. 2021 Dec;17 Suppl 2 e058308
       BACKGROUND: Mutations in microtubule-associated protein tau (MAPT) gene are causative of Frontotemporal Dementia (FTD). Many of the features associated with the development of tau pathology, e.g high levels of tau phosphorylation, are also present in early development. iPSC-neurons have gene expression signatures similar to fetal neurons, and iPSC-neurons with MAPT mutations do not develop tau aggregates. We hypothesise that iPSC-neurons are resistant to developing tau aggregates due to high activity levels of the proteostasis network during early development. To test this hypothesis, we investigated the levels of proteasome subunits in developing iPSC derived neurons and measured the proteasome activity of iPSC-neurons throughout development. To test whether manipulation of proteostasis in iPSC-neurons could lead to tau pathology we treated cells with a proteasome inhibitor, and measured changes in total and phosphorylated tau, proteasome subunits and autophagy regulators.
    METHODS: Human cortical neurons were derived from isogenic iPSCs with the following MAPT genotypes: WT, 10+16 monoallelic, 10+16 biallelic and 10+16/P301S biallelic. RNA and protein analysis of proteasome subunits was performed by qPCR and Western blot at during neuronal development (DIV 0, 10, 30 and 100) and after proteasome inhibition treatments.
    RESULTS: We show that proteasome activity decreases during the differentiation of iPSCs to cortical neurons, accompanied by a reduction in levels of the proteasome regulatory subunits Rpt6 and Rpn6. Proteasome inhibition in WT neurons does not lead to a significant change in total or phosphorylated tau levels but resulted in an increase in the autophagy associated protein BAG3, together with an induction of tau cleaved by caspase-3 at Asp421.
    CONCLUSION: Proteasome activity decreases during the differentiation of iPSCs into neurons, which may be due to a reduction in in proteasome regulators. Our results suggest that proteasome inhibition causes an increase in tau cleavage which may be preferentially cleared through the autophagy pathway. The ability of these 'fetal-like' neurons to adapt and upregulate their protein clearance system may be enhanced compared to FTD brain tissue. In ongoing work we are investigating proteasome expression and activity in FTD post-mortem tissue as well as the localisation and interactions of tau and the proteasome after proteasome inhibition treatments.
    DOI:  https://doi.org/10.1002/alz.058308
  6. Front Mol Biosci. 2021 ;8 787260
      The Hsp90 molecular chaperone, along with a set of approximately 50 cochaperones, mediates the folding and activation of hundreds of cellular proteins in an ATP-dependent cycle. Cochaperones differ in how they interact with Hsp90 and their ability to modulate ATPase activity of Hsp90. Cochaperones often compete for the same binding site on Hsp90, and changes in levels of cochaperone expression that occur during neurodegeneration, cancer, or aging may result in altered Hsp90-cochaperone complexes and client activity. This review summarizes information about loss-of-function mutations of individual cochaperones and discusses the overall association of cochaperone alterations with a broad range of diseases. Cochaperone mutations result in ciliary or muscle defects, neurological development or degeneration disorders, and other disorders. In many cases, diseases were linked to defects in established cochaperone-client interactions. A better understanding of the functional consequences of defective cochaperones will provide new insights into their functions and may lead to specialized approaches to modulate Hsp90 functions and treat some of these human disorders.
    Keywords:  Aha1; CS domain; FKBP; FNIP1; chaperonopathy; tetratricopeptide repeat
    DOI:  https://doi.org/10.3389/fmolb.2021.787260
  7. Alzheimers Dement. 2021 Dec;17 Suppl 2 e058717
       BACKGROUND: In Alzheimer's disease (AD), the accumulation of the microtubule associated protein tau is linked with neuronal loss, but the major cell death pathway remains to be identified. Pathogenic tau promotes endoplasmic reticulum (ER) stress and subsequent unfolded protein response (UPR) activation. However, the contribution of ER stress to tau-mediated toxicity in neurons is unknown. A previous study demonstrated that mice lacking CHOP/Ddit3, an important regulator of ER stress-induced cell death, were protected from ER stress induced neuronal loss. We hypothesized that silencing CHOP would prevent tau toxicity through the ER stress pathway and reveal the contribution of tau toxicity that is mediated through this pathway.
    METHOD: To investigate this, we generated a mouse-targeting CHOP/Ddit3 shRNA AAV9 that also expresses GFP by a separate promoter. We injected shCHOP AAV9 or a shScrambled control AAV9 in the brains of 8-month-old male and female rTg4510 tau transgenic mice and harvested them at 12-months of age, when ER stress is activated, and neuronal loss is occurring in these mice. We performed behavioral studies using the Y-Maze, Open field, and 2-day Radial-Arm water maze task with a reversal paradigm. Unbiased stereology was used to determine neuronal health and effect of CHOP shRNA on CHOP levels was evaluated by immunofluorescence.
    RESULT: AAV9 CHOP shRNA did not alter spatial learning and memory in tau transgenic mice. Evaluation of the tissue revealed a modest, but non-significant increase in neuronal loss from CHOP knock down, while the DG shows a non-significant decrease in neuronal loss. Tissue volume was unchanged in both regions. Very surprising to us, and opposite of what was expected, we found that in these regions of interest the CHOP levels increased in the animal expressing the CHOP.
    CONCLUSION: Overall, this suggests that shCHOP AAV9 does not alter tau-modified behaviors such as learning and memory particularly at this timepoint. Our results also suggest that there may be a compensatory increase in CHOP signaling in neighboring cells, suggesting that the activation of the UPR in the brains of these mice may be a protective mechanism.
    DOI:  https://doi.org/10.1002/alz.058717
  8. Alzheimers Dement. 2021 Dec;17 Suppl 2 e058498
       BACKGROUND: Molecular chaperones have protective functions in neurodegeneration by preventing misfolding of aggregation-prone proteins. These functions have been mostly studied in neurons. However, members of the family of chaperones known as small heat shock proteins (sHSPs), including HSPB1, CRYAB, and HSPB8, are specifically expressed in glial cells rather than in neurons, and their levels increase in astrocytes in human post-mortem Alzheimer's disease (AD) brain. While traditionally considered intracellular proteins, novel extracellular functions have been attributed to chaperones in cancer or the immune system. Interestingly, HSPB1 is also found in senile plaques suggesting these can also be extracellular in the brain. We hypothesize that sHSP can be extracellular and aim to investigate whether sHSPs are secreted from astrocytes in Alzheimer's disease brain.
    METHOD: We used human post-mortem brain sections from Alzheimer's disease patients to investigate changes in extracellular/intracellular localization of HSPB1. To further explore sHSPs secretion, we used primary mouse astrocytes. Reactive astrocytes were induced by TNFα and IL-1α, mimicking the local environment in the diseased brain. We investigated changes in astrocyte cell lysates and the astrocyte conditioned media by Western blot. To investigate the secretion mechanisms, we also use size exclusion chromatography (SEC) and nanoparticle tracking analysis (NTA) RESULT: Although sHSPs are intracellular chaperones, Our analysis of AD patients post-mortem brain sections suggested that extracellular HSPB1 might increase around astrocytes that are close to amyloid plaques. In primary mouse astrocytes, we also found an increase in sHSPs in the extracellular media when astrocytes become reactive upon treatment with TNFα and IL-1α. We further investigated their mechanism of secretion by fractionating the astrocyte conditioned media using SEC. NTA and proteinase K protection assays show that the sHSPs were not present in extracellular vesicle fractions, but rather free in the media.
    CONCLUSION: Our study suggests that HSPB1 is secreted from astrocytes near to plaques in AD brain, as well as from reactive astrocytes in culture, and this secretion is not dependent on extracellular vesicles. The mechanisms behind how sHSPs are secreted and their extracellular functions still need to be explored.
    DOI:  https://doi.org/10.1002/alz.058498
  9. Alzheimers Dement. 2021 Dec;17 Suppl 2 e058572
       BACKGROUND: Astrocytes are vital in the onset and progression of Alzheimer's disease (AD). Accumulation of reactive astrocytes, together with tau phosphorylation, correlates very strongly with cognitive decline. Molecular chaperones are essential for maintaining protein homeostasis. One family of chaperones are the small heat shock proteins (sHSPs), which include HSPB1 and CRYAB. Expression of these sHSPs seems to be restricted to glial cells, and levels are found to increase in those astrocytes found in AD brains. The role that sHSP play in astrocytes in AD is still not known. Emerging evidence suggests vital interplay between different cell types during neurodegenerative diseases. We therefore aim to investigate the non-cell autonomous role of astrocytic sHSPs in AD.
    METHOD: We are using primary mouse neurons and organotypic brain slice cultures in conjunction with the recombinant adeno-associated viral system (rAAV). These systems will allow us to replicate the tau pathology found in AD, investigate neuron-astrocyte interactions and determine whether either can be altered by overexpression of our sHSPs. Using immunofluorescence, confocal microscopy and western blotting we are characterising expression and localization of sHSPs in post-mortem human AD brain tissue and brain slice cultures.
    RESULT: Our data shows that HSPB1 is specifically expressed in astrocytes in both human brain tissue and slice cultures. Interestingly, HSPB1 levels in the human brain increase in GFAP-positive astrocytes surrounding amyloid plaques. When organotypic brain slices are treated with cytokines or Aβ oligomers, reactive astrocytes are induced and levels of HSPB1 and CRYAB are also increased, similar to what we observe in human AD brain.
    CONCLUSION: Our results in human AD brain highlight the importance of astrocytic HSPB1 in AD, and we provide evidence to suggest not only that organotypic brain slices are a good model to replicate and study the function of sHSPs in AD, but that HSPB1 and CRYAB play an important role in the response to AD-relevant pathology. Using rAAVs, we aim to manipulate astrocytic sHSPs levels in organotypic brain slices to investigate whether astrocytic sHSPs could modulate neuronal health and tau pathology in a non-cell autonomous manner.
    DOI:  https://doi.org/10.1002/alz.058572
  10. Biochim Biophys Acta Gen Subj. 2021 Dec 28. pii: S0304-4165(21)00238-5. [Epub ahead of print] 130079
      This mini-review will cover the various chemical biology approaches employed to prepare and modulate ubiquitin chains and the ubiquitin-proteasome system. Emphasis will be given to the biochemistry and chemical biology of poly-ubiquitin chain preparation as a tool to elucidate its roles in biological systems as well as the hijacking of the ubiquitin proteasome system using heterobifunctional compounds to induce intracellular ubiquitination.
    Keywords:  Chemical biology; Protein synthesis; Ubiquitin
    DOI:  https://doi.org/10.1016/j.bbagen.2021.130079