ORCID

0000-0002-3828-9562

Date of Award

5-9-2024

Author's School

Graduate School of Arts and Sciences

Author's Department

Chemistry

Degree Name

Doctor of Philosophy (PhD)

Degree Type

Dissertation

Abstract

Rare genetic mutations in proteins expressed in cardiomyocytes lead to proteotoxic cardiomyopathies which are characterized by widespread protein-aggregate pathology. However, the mechanistic link between proteotoxicity and protein-aggregates in the more common ischemic cardiomyopathy remains unclear. Our study revealed the mis-localization of desmin, actin, and α-actinin to protein-aggregates in the myocardium of patients with ischemic cardiomyopathy and in mouse hearts undergoing post-myocardial infarction ventricular remodeling. These findings mimic observations in autosomal-dominant cardiomyopathy resulting from the R120G mutation in the chaperone protein CRYAB where desmin is mis-localized into the aggregates. Importantly, we observed increased phosphorylation of CRYAB at serine-59 (pS59-CRYAB) and its accumulation in the NP40-insoluble aggregate-rich fraction. Further investigation showed that CRYAB undergoes phase separation, and mimicking phosphorylation at serine-59 with an aspartate substitution reduced the fluidity of CRYAB condensates, promoting protein-aggregate formation and increased cell death. Conversely, substituting serine-59 with alanine, by preventing phosphorylation, restored the fluidity of CRYAB condensates and reduced protein aggregation. Moreover, introducing a phospho-deficient S59A mutation, but not the phospho-mimetic S59D mutation, at the CRYAB locus in mice rescued adverse left ventricular remodeling after ischemia reperfusion injury. Treatment with 25-Hydroxycholesterol attenuated serine-59 phosphorylation, increasing the fluidity of R120G-CRYAB condensates, and ameliorating post-myocardial infarction cardiac dysfunction. Experimental interventions aimed at preventing aggregate formation or promoting disaggregation exacerbated post-myocardial infarction cardiomyopathy, highlighting the importance of pS59CRYAB sequestration in aggregates to attenuate disease progression. These findings suggest that modulating CRYAB phase separation could offer a promising approach to mitigating ischemic cardiomyopathy.

Language

English (en)

Chair and Committee

Abhinav Diwan

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