Single-cell-based evidence for GLS1 inhibitor as a bona fide senolytic agent in vivo

Abstract

A plethora of senolytic compounds and technologies have been identified. However, the efficacy of these treatments in eliminating senescent cells and the suppression of chronic inflammation remains to be substantiated in vivo. Here, we employ single-cell RNA sequencing and find the selective elimination of highly inflammatory fibroblasts, endothelial cells in the lung, and proximal tubule cells in the kidney from the GLS1 inhibitor BPTES-treated aged mice. The eliminated fibroblasts share typical phenotypes with in vitro human senescent cells. These cells predominantly express Dpp4 (CD26) and Cadm3, showing activated IFN signaling and lysosomal membrane damage. Cells eliminated by BPTES in lungs exhibit transcriptomes similar to those eliminated in p16-DTR mice, where DTR expression is limited in p16-expressing cells. BPTES treatment results in the T cell population shift from cytotoxic to a protective state, suggesting the suppression of age-related chronic inflammation. CellChat analysis revealed that multiple cytokine signals are transmitted from inflammatory fibroblasts and proximal tubular cells to immune cells in lungs and kidneys. These results provide evidence that GLS1 inhibitor functions as a bona fide senolytic drug to eliminate inflammatory cells, including a subset of senescent cells, and suppresses age-related chronic inflammation.