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Proteome and proteostasis network disruptions accompany proteostasis alterations, leading to accumulation of protein aggregates, characteristic of several age-related diseases. Previous work in Caenorhabditis elegans and zebrafish unveiled asymptomatic protein aggregation (ARPA), characterized by generalized increased accumulation of insoluble proteins through aging. However, the consequences of protein homeostasis imbalances in the context of healthy aging in mammals remains mostly unexplored. To elucidate if accumulation of insoluble proteins also occurs through aging in mammals, C57BL/6 mice with different ages corresponding to young (6 months old), adult (13 months old), and old stages (18 and 24 months old) were used. Detergent-insoluble fractions were isolated from total protein extracts of tissues, followed by characterization of both total and detergent-insoluble protein profiles. Our results show tissue-specific proteome alterations through aging. For example, the insoluble fractions increase through aging in brain-derived tissues, but muscular tissues do not display significant alterations until 18 months old. We are now performing SWATH-MS analysis to identify differential proteome signatures of aging and which proteins are more prone to aggregate to further elucidate the functions and biological processes affected by ARPA. published

Aging is characterized by a gradual decline in overall organismal fitness across the lifespan, and greatly enhances the risk for chronic diseases, as neuropathological disorders, diabetes, cancer and cardiovascular diseases. One of the hallmarks of aging is the loss of proteostasis, a regulated process responsible for maintaining the cellular proteome. When proteostasis is altered, proteome and proteostasis network disruptions occur, leading to accumulation of protein aggregates, characteristic of several age-related diseases. Previous work performed in Caenorhabditis elegans and zebrafish unveiled asymptomatic protein aggregation (ARPA), characterized by generalized increased accumulation of insoluble proteins through aging in healthy animals. We hypothesize that proteome imbalances occur with aging in mammals and they correlate with tissue aging and susceptibility for the development of age-related disease. The main goal of this work is to perform a global proteomic characterization of age-dependent protein aggregation in mice. C57BL/6 mice with different ages (6, 13, 18, 24 and 29 months) were used and the detergent-insoluble fractions isolated from total protein extracts of different tissues. Protein profiles were characterized by automated capillary electrophoresis separation using the LabChip GX. Our results suggest that proteome alterations occur during aging in mammals, and a tissue-specific insoluble protein signature across the lifespan is observed. For the cortex, the identification of the proteins more prone to aggregate during aging was performed by mass spectrometry and characterized to establish the functions and biological processes affected by ARPA. This will allow identifying novel aging biomarkers that may be relevant for disease onset and progression. published

Proteostasis is a highly regulated process involving the proteostasis network responsible for maintaining the cellular proteome. When proteostasis is altered, proteome and proteostasis network disruptions occur, leading to accumulation of protein aggregates, characteristic of several age-related diseases. Previous work performed in Caenorhabditis elegans and zebrafish unveiled asymptomatic protein aggregation (ARPA), characterized by generalized increased accumulation of insoluble proteins through aging in healthy animals. Whether ARPA is a causative process in aging and contributes to age-related diseases is not clear and to date the consequences of protein homeostasis imbalances in the context of normal aging in mammals remains largely unexplored. We hypothesize that accumulation of insoluble proteins occurs through aging in mammals and that it contributes to activate the stress response and disrupt protein synthesis rate and protein quality control mechanisms. To elucidate if ARPA occurs along the lifespan of mice, C57BL/6 mice with different ages (1, 6, 13, 18, 24 and 29 months) were used and the detergent-insoluble fractions were isolated from total protein extracts of different tissues. The total and detergent-insoluble protein profiles were characterized by automated capillary electrophoresis separation using the LabChip GX equipment (PerkinElmer). Our preliminary results show that there is an increase in insoluble proteins until at least 18 months old in particular tissues in mice. Identification of the proteins more prone to aggregate during aging will be performed by mass spectrometry and characterized to establish the functions and biological processes affected by ARPA. This will allow identifying novel aging biomarkers that may be relevant for disease onset and progression. published

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