title: Requirement of clusterin expression for prosurvival autophagy in hypoxic kidney tubular epithelial cells.
abstract: Cellular autophagy is a prosurvival mechanism in the kidney against ischemia-reperfusion injury (IRI), but the molecular pathways that activate the autophagy in ischemic kidneys are not fully understood. Clusterin (CLU) is a chaperone-like protein, and its expression is associated with kidney resistance to IRI. The present study investigated the role of CLU in prosurvival autophagy in the kidney. Renal IRI was induced in mice by clamping renal pedicles at 32°C for 45 min. Hypoxia in renal tubular epithelial cell (TEC) cultures was induced by exposure to a 1% O2 atmosphere. Autophagy was determined by either light chain 3-BII expression with Western blot analysis or light chain 3-green fluorescent protein aggregation with confocal microscopy. Cell apoptosis was determined by flow cytometric analysis. The unfolded protein response was determined by PCR array. Here, we showed that autophagy was significantly activated by IRI in wild-type (WT) but not CLU-deficient kidneys. Similarly, autophagy was activated by hypoxia in human proximal TECs (HKC-8) and WT mouse primary TECs but was impaired in CLU-null TECs. Hypoxia-activated autophagy was CLU dependent and positively correlated with cell survival, and inhibition of autophagy significantly promoted cell death in both HKC-8 and mouse WT/CLU-expressing TECs but not in CLU-null TECs. Further experiments showed that CLU-dependent prosurvival autophagy was associated with activation of the unfolded protein response in hypoxic kidney cells. In conclusion, these data suggest that activation of prosurvival autophagy by hypoxia in kidney cells requires CLU expression and may be a key cytoprotective mechanism of CLU in the protection of the kidney from hypoxia/ischemia-mediated injury.
authors.affiliation: Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada;
authors.forename: Hatem A
authors.initials: HA
authors.lastname: Alnasser
authors.affiliation: Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada; Immunity and Infection Research Centre, Vancouver Coastal Health Research Institute, Vancouver, British Columbia, Canada; and.
authors.forename: Qiunong
authors.initials: Q
authors.lastname: Guan
authors.affiliation: Vancouver Prostate Centre, Vancouver, British Columbia, Canada.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada; Vancouver Prostate Centre, Vancouver, British Columbia, Canada.
authors.forename: Martin E
authors.initials: ME
authors.lastname: Gleave
authors.affiliation: Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada;
authors.forename: Christopher Y C
authors.initials: CY
authors.lastname: Nguan
authors.affiliation: Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada; Immunity and Infection Research Centre, Vancouver Coastal Health Research Institute, Vancouver, British Columbia, Canada; and caigan@mail.ubc.ca.
authors.forename: Caigan
authors.initials: C
authors.lastname: Du
mesh_terms: D000818:Animals; D001343:Autophagy / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D051152:Clusterin / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000378:metabolism* / Q000473:pathology; D007684:Kidney Tubules / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D015427:Reperfusion Injury / Q000378:metabolism* / Q000473:pathology
chemical_list: D051152:Clusterin
keywords: acute kidney injury; autophagy; clusterin; hypoxia; kidney ischemia-reperfusion