Senescent cells inhibit muscle differentiation via the SASP-lipid 15d-PGJ₂mediated modification and control of HRas

Senescent cells, which are characterized by multiple features such as increased expression of Senescence-Associated β-galactosidase activity (SA β-gal) and cell cycle inhibitors such as p21 or p16, accumulate with tissue damage and dysregulate tissue homeostasis. In the context of skeletal muscle, it is known that agents used for chemotherapy such as Doxorubicin cause buildup of senescent cells, leading to the inhibition of tissue regeneration. Senescent cells influence the neighboring cells via numerous secreted factors which form the senescence-associated secreted phenotype (SASP). Lipids are emerging as a key component of SASP that can control tissue homeostasis. Arachidonic acid-derived lipids have been shown to accumulate within senescent cells, specifically 15d-PGJ₂, which is an electrophilic lipid produced by the non-enzymatic dehydration of the prostaglandin PGD₂. In this study, we show that 15d-PGJ₂is also released by Doxorubicin-induced senescent cells as a SASP factor. Treatment of skeletal muscle myoblasts with the conditioned medium from these senescent cells inhibits myoblast fusion during differentiation. Inhibition of L-PTGDS, the enzyme that synthesizes PGD₂, diminishes the release of 15d-PGJ₂by senescent cells and restores muscle differentiation. We further show that this lipid post-translationally modifies Cys184 of HRas in skeletal muscle cells, causing a reduction in the localization of HRas to the Golgi, increased HRas binding to RAF RBD, and activation of cellular MAPK-Erk signaling (but not the Akt signaling). Mutating C184 of HRas prevents the ability of 15d- PGJ₂to inhibit the differentiation of muscle cells and control the activity of HRas. This work shows that 15d-PGJ₂released from senescent cells could be targeted to restore muscle homeostasis after chemotherapy.