Title : Optimized Yamanaka factors and TERT gene therapy modulates senescence-associated inflammatory phenotype in human fibroblasts
Abstract:
Cellular senescence, a hallmark of aging, is closely linked to chronic inflammation and dysregulation of innate immune responses through the Senescence-Associated Secretory Phenotype (SASP). The Yamanaka factors (OSKM) can rejuvenate aged cells but pose oncogenic risks, primarily due to c-Myc. As an alternative, we evaluated a safer partial reprogramming approach using optimized factors (Oct4, Sox2, Klf4 – OSK) combined with telomerase (TERT) gene therapy in human MRC-5 lung fibroblasts at late passages (P60–P70). Cells were transfected with plasmids expressing OSK and/or TERT. Gene expression analysis revealed that co-expression of OSK and TERT significantly upregulated youthful markers (Nanog, LAP2A) while downregulating senescence-associated genes (p16, p21, ZSCAN4, ATF3, BTG2, MMP13). Importantly, the pro-inflammatory cytokine IL-6, a key SASP component linked to innate immunity, was markedly reduced. Western blot confirmed corresponding protein level changes. Functional assays showed enhanced cell viability, altered cell cycle distribution, and decreased SA-β-galactosidase activity. These findings suggest that OSK+TERT combination therapy not only delays cellular aging but also attenuates the inflammatory SASP profile. Given the role of SASP in chronic inflammation and immune dysfunction, this strategy may offer a novel intervention to modulate innate immune responses in age-related conditions. Further in vivo studies are warranted to explore its effects on tissue inflammation and immune homeostasis. This work was supported by the Russian Science Foundation (Grant number 25-74-31017).
