Investigating the Effect of Rapamycin on Impaired Proteostasis in Aged Skeletal Muscle After Disuse Atrophy

Older individuals undergo loss of muscle mass and strength, which can be compounded during bed rest and extended hospital stays. Adult and aged individuals lose muscle mass at similar rates during periods of disuse; however, only aged skeletal muscle fails to fully recover once activity resumes. Current efforts to facilitate skeletal muscle recovery are aimed at increasing myofibrillar protein synthesis by stimulating mTORC1(mammalian target of rapamycin complex 1), a major regulator of protein synthesis. However, we showed that skeletal muscle in aged rats already has higher rates of myofibrillar protein synthesis and mTORC1 activity than adult rats. We also showed that aged skeletal muscle has an accumulation of insoluble ubiquitinated proteins compared to adult muscle, indicating impaired proteostasis and possibly impaired autophagy degradation. We hypothesized that mTORC1 inhibition via rapamycin administration would reduce protein synthesis and the accumulation of p62 aggregates, facilitating recovery in aged skeletal muscle after disuse. Methods: Adult (10 months) and old (26 months) OKC HET rats were hindlimb unloaded (HU) for 14-days to induce atrophy, followed by subsequent 14 days of reloading (RE) through normal ambulation. Animals were fed either a control or rapamycin-supplemented (42 ppm) diet. To determine bulk protein synthesis, rats were labeled with deuterium oxide (D 2 O) during weight bearing (WB), HU, and RE, (n=4-6 per time point per group). Finally, to assess the presence of autophagy related protein aggregates we immuno stained for Sequestosome 1/p62, an autophagy cargo receptor protein. Results: Our data showed that myofibrillar protein synthesis rates were elevated in the old muscle compared to the adult ( k = 0.029 1/day ±0.002 vs. 0.039 1/day ±0.002, p<0.0001), and that protein synthesis rates were lower in the old rapamycin cohort (k = 1/day ±0.002 vs. 1/day ±0.003, p<0.0001). Furthermore, p62 puncta was three-fold more abundant (p<0.01) in aged skeletal muscle, compared to the adult muscle. Although the sample size is small, when comparing p62 puncta in the old rapamycin group to the old controls, there was no difference between the cohorts. Conclusion: We show that mTORC1 inhibition reduces protein synthesis in aged skeletal muscle. We also show that aged muscle has more p62 aggregates compared to adult skeletal muscle, indicating that autophagy related targets are not being degraded effciently with age. Further analyses will assess the effects of mTORC1 inhibition on other proteostatic mechanisms in atrophied skeletal muscle of aged rats. NIA Training Grant: 5T32AG052363-04VA I01 BX005592 APS SURF. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.