Premature Senescence and Telomere Shortening Induced by Oxidative Stress From Oxalate, Calcium Oxalate Monohydrate, and Urine From Patients With Calcium Oxalate Nephrolithiasis

Abstract

Oxidative stress, a well-known cause of stress-induced premature senescence (SIPS), is increased in patients with calcium oxalate (CaOx) kidney stones (KS). Oxalate and calcium oxalate monohydrate (COM) induce oxidative stress in renal tubular cells, but to our knowledge, their effect on SIPS has not yet been examined. Here, we examined whether oxalate, COM, or urine from patients with CaOx KS could induce SIPS and telomere shortening in human kidney (HK)-2 cells, a proximal tubular renal cell line. Urine from age- and sex-matched individuals without stones was used as a control. In sublethal amounts, H2O2, oxalate, COM, and urine from those with KS evoked oxidative stress in HK-2 cells, indicated by increased protein carbonyl content and decreased total antioxidant capacity, but urine from those without stones did not. The proportion of senescent HK-2 cells, as indicated by SA-βgal staining, increased after treatment with H2O2, oxalate, COM, and urine from those with KS. Expression of p16 was higher in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS than it was in cells treated with urine from those without stones and untreated controls. p16 was upregulated in the SA-βgal positive cells. Relative telomere length was shorter in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS than that in cells treated with urine from those without stones and untreated controls. Transcript expression of shelterin components (TRF1, TRF2 and POT1) was decreased in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS, in which case the expression was highest. Urine from those without KS did not significantly alter TRF1, TRF2, and POT1 mRNA expression in HK-2 cells relative to untreated controls. In conclusion, oxalate, COM, and urine from patients with CaOx KS induced SIPS and telomere shortening in renal tubular cells. SIPS induced by a lithogenic milieu may result from upregulation of p16 and downregulation of shelterin components, specifically POT1, and might contribute, at least in part, to the development of CaOx KS.

HydroZitLa inhibits calcium oxalate stone formation in nephrolithic rats and promotes longevity in nematode Caenorhabditis elegans

“…ROS and oxidative stress are well-known inducers of SIPS, and oxidative stress accelerates the shortening of telomeres 57 . We recently demonstrated that oxalate, COM, and urine from patients with CaOx stones induced premature senescence, p16 upregulation and telomere attrition in renal tubular cells through oxidative stress 58 . In this study, we showed that HydroZitLa inhibited telomere attrition, p16 upregulation, and premature senescence in HK-2 cells exposed to H 2 O 2 , oxalate, and COM.…”

Telomere Attrition in Induced Pluripotent Stem Cell-Derived Neurons From ALS/FTD-Related C9ORF72 Repeat Expansion Carriers

“…TRF2 levels were found to be decreased in patients with AD ( Wu et al, 2019 ). In addition, treatment with agents that induce ROS reduces levels of POT1 in HK-2 cells ( Chuenwisad et al, 2021 ). Moreover, the decreases in TRF2 and POT1 levels cause unprotected chromosomes to trigger the DDR.…”

“…We performed telomere length quantification by qPCR as previously described by ( Chuenwisad et al, 2021 ). Briefly, we performed genomic DNA extraction from iPSC, NEP, MNP, and MN using the PureLink Genomic DNA Mini kit (Thermo Fischer Scientific).…”

Cancer treatment-induced NAD+ depletion in premature senescence and late cardiovascular complications

“…RS is accompanied by the shortening of telomeres until a critical length at which senescence is induced, known as the Hayflick limit [ 40 ] . SIPS may or may not be associated with telomere shortening [ 41 – 44 ] . For example, the telomerase-immortalized human foreskin fibroblast (hTERT-BJ1) cells exposed to ultraviolet B light or H 2 O 2 develop SIPS, suggesting that SIPS can be independent of telomerase activity and telomere shortening [ 42 ] .…”