Roles of cellular senescence in driving bone marrow adiposity in radiation-and aging-associated bone loss

Abstract

Osteoporosis is associated with an increase in marrow adipocytes, collectively termed bone marrow adipose tissue (BMAT). An increase in BMAT is linked with decline of mesenchymal progenitors that give rise to osteoblasts which are responsible for bone accrual. Oxidative stress-induced reactive oxygen species, DNA damage, apoptosis and cellular senescence have been associated with reduced osteoprogenitors in a reciprocal fashion to BMAT; however, a direct (causal) link between cellular senescence and BMAT is still elusive. Accumulation of senescent cells occur in naturally aged and in focally radiated bone tissue, but despite amelioration of age- and radiation-associated bone loss after senescent cell clearance, molecular events that precede BMAT accrual are largely unknown. Using a mouse model here we show by RNA-Sequencing data that BMAT-related genes were the most upregulated gene subset in radiated bones. Using focal radiation as a model to understand age-associated changes in bone, we performed a longitudinal assessment of cellular senescence and BMAT. Using qRT-PCR, RNA in situ hybridization and histological assessment of telomere dysfunction as a marker of senescence, we observed increased p21 transcripts in bone lining cells, osteocytes and bone marrow cells, and elevated dysfunctional telomeres in osteocytes starting from day 1 post-radiation, without the presence of BMAT. BMAT was significantly elevated in radiated bones at day 7, confirming the qRT-PCR data in which most BMAT-related genes were elevated by day 7, and the trend continued until day 42 post-radiation. Similarly, elevation in BMAT-related genes was observed in aged bones. The senolytic cocktail of Dasatinib (D) plus Quercetin (Q) - D+Q, which clears senescent cells, reduced BMAT in aged and radiated bones. MicroRNAs (miRs) linked with senescence marker p21 were downregulated in radiated- and aged-bones, while miR-27a, a miR that is associated with increased BMAT, was elevated both in radiated- and aged-bones. D+Q downregulated miR-27a in radiated bones at 42 days post-radiation. Overall, our study provides evidence that BMAT occurrence in oxidatively stressed bone environments, such as radiation and aging, is induced following a common pathway and is dependent on the presence of senescent cells.