Progressive Cellular Senescence Mediates Renal Dysfunction in Ischemic Nephropathy

• Published in: Journal of the American Society of Nephrology Vol. 32; no. 8; pp. 1987 - 2004
• Main Authors: Kim, Seo Rin, Puranik, Amrutesh S., Jiang, Kai, Chen, Xiaojun, Zhu, Xiang-Yang, Taylor, Ian, Khodadadi-Jamayran, Alireza, Lerman, Amir, Hickson, LaTonya J., Childs, Bennett G., Textor, Stephen C., Tchkonia, Tamara, Niewold, Timothy B., Kirkland, James L., Lerman, Lilach O.
• Format: Journal Article
• Language: English
• Published: United States American Society of Nephrology 01.08.2021
• Subjects: Animals; Apoptosis - drug effects; Basic Research; Caspase 8 - metabolism; Cellular Senescence - drug effects; Cellular Senescence - genetics; Cellular Senescence - physiology; Chronic Disease; Cyclin-Dependent Kinase Inhibitor p16 - genetics; Cyclin-Dependent Kinase Inhibitor p16 - metabolism; Cyclin-Dependent Kinase Inhibitor p19 - metabolism; Cyclin-Dependent Kinase Inhibitor p21 - genetics; Dasatinib - pharmacology; Disease Models, Animal; Enzyme Activation - drug effects; Epithelial Cells - physiology; Epithelial-Mesenchymal Transition; Gene Expression; Heparin-binding EGF-like Growth Factor - genetics; Humans; Ischemia - etiology; Ischemia - physiopathology; Kidney - blood supply; Kidney - pathology; Kidney - physiopathology; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Osteopontin - genetics; p21-Activated Kinases - genetics; p21-Activated Kinases - metabolism; Protein Kinase Inhibitors - pharmacology; Renal Artery Obstruction - complications; Renal Insufficiency, Chronic - etiology; Renal Insufficiency, Chronic - pathology; Renal Insufficiency, Chronic - physiopathology; Sequence Analysis, RNA; Single-Cell Analysis; Tacrolimus - analogs & derivatives; Tacrolimus - pharmacology; Up-Regulation; senescence; quercetin; renal artery stenosis; dasatinib; transcriptome
• ISSN: 1046-6673; 1533-3450; 1533-3450
• DOI: 10.1681/ASN.2020091373

Significance Statement Renal artery stenosis (RAS) engenders stenotic-kidney ischemia, dysfunction, and injury, but whether these are mediated by cellular senescence has not been elucidated. INK-ATTAC transgenic mice, high-resolution imaging, and unbiased single-cell RNA sequencing of murine kidneys demonstrated cellular senescence as an important mechanism of progressive injury to epithelial/stromal cells within poststenotic kidneys. Both p16-specific and broad quercetin/dasatinib interventions to blunt senescence improved renal function and structure, underscoring the central role of senescence in the pathogenesis. Furthermore, this mechanism was conserved in human subjects with RAS. These observations reveal new mechanisms that contribute to the pathogenesis of chronic ischemic renal injury, and support the development of senolytic therapy to reduce senescent cell burden and delay renal injury. Background Peripheral vascular diseases may induce chronic ischemia and cellular injury distal to the arterial obstruction. Cellular senescence involves proliferation arrest in response to stress, which can damage neighboring cells. Renal artery stenosis (RAS) induces stenotic-kidney dysfunction and injury, but whether these arise from cellular senescenceand their temporal pattern remain unknown. Methods Chronic renal ischemia was induced in transgenic INK-ATTAC and wild type C57BL/6 mice by unilateral RAS, and kidney function (in vivo micro-MRI) and tissue damage were assessed. Mouse healthy and stenotic kidneys were analyzed using unbiased single-cell RNA-sequencing. To demonstrate translational relevance, cellular senescence was studied in human stenotic kidneys. Results Using intraperitoneal AP20187 injections starting 1, 2, or 4 weeks after RAS, selective clearance of cells highly expressing p16Ink4a attenuated cellular senescence and improved stenotic-kidney function; however, starting treatment immediately after RAS induction was unsuccessful. Broader clearance of senescent cells, using the oral senolytic combination dasatinib and quercetin, in C57BL/6 RAS mice was more effective in clearing cells positive for p21 (Cdkn1a) and alleviating renal dysfunction and damage. Unbiased, single-cell RNA sequencing in freshly dissociated cells from healthy and stenotic mouse kidneys identified stenotic-kidney epithelial cells undergoing both mesenchymal transition and senescence. As in mice, injured human stenotic kidneys exhibited cellular senescence, suggesting this process is conserved. Conclusions Maladaptive tubular cell senescence, involving upregulated p16 (Cdkn2a), p19 (Cdkn2d), and p21 (Cdkn1a) expression, is associated with renal dysfunction and injury in chronic ischemia. These findings support development of senolytic strategies to delay chronic ischemic renal injury.