ASN's Mission

ASN leads the fight to prevent, treat, and cure kidney diseases throughout the world by educating health professionals and scientists, advancing research and innovation, communicating new knowledge, and advocating for the highest quality care for patients.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005


The Latest on Twitter

Kidney Week

Abstract: FR-PO951

Protective Effect of Hydroxychloroquine on Cultured Mouse Podocytes Expressing the HIV Accessory Protein Vpr

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology


  • Kajiyama, Hiroshi, Saitama Medical University, Iruma, Saitama, Japan
  • Kopp, Jeffrey B., NIDDK, NIH, Bethesda, Maryland, United States
  • Mimura, Toshihide, Saitama Medical University, Iruma, Saitama, Japan

Studies in HIV-transgenic mice have implicated the HIV accessory protein R (Vpr) in podocyte injury, culminating in HIV-associated nephropathy. Clinical studies indicate that hydroxychloroquine reduces kidney damage in lupus. In this study, we tested protective effects of hydroxychloroquine on cultured mouse podocytes expressing Vpr.


Stably-transfected mouse podocytes bearing three transgenes, expressing Vpr (thermosensitive SV40 T antigen, podocin-promoter-rtTA and tet-responsive element-Vpr) or control AI podocytes expressing two transgenes (thermosensitive SV40 T and podocin-promoter-rtTA) were grown at 33°C and differentiated at 37°C. Differentiated podocytes were plated on day 1, hydroxychloroquine at concentrations ranging from 0.63 to 80 μg/mL was added to podocyte cultures on day 3, and 1 μg/mL doxycycline (DOX) was added day 4). Cell death was observed by phase-contrast microscopy on day 6 and 9, and total cell number and dead cell number were counted in each condition to obtain the percentage of dead cells.


AI control and Vpr-expressing podocytes tolerated hydroxychloroquine concentrations of 10 μg/mL of hydroxychloroquine or less but died at higher concentrations. Vpr podocytes died 48 h after 1 μg/mL DOX was added, likely due to induced expression of Vpr. DOX-treated Vpr podocytes were protected from cell death with 24 h pretreatment of 0.63, 1.25, 2.5, 5 and 10 μg/mL hydroxychloroquine (H), in a dose-dependent manner (H0: 30.2%, H0.63: 25.0%, H1.25: 20.8%, H2.5: 4.3%, H5: 0%, H10: 0%). DOX-untreated Vpr podocytes without hydroxychloroquine also underwent cell death on day 9 of 37°C culture due to leaky expression of Vpr, which was partly inhibited by 5 and 10 μg/mL of hydroxychloroquine pretreatment (H0: 56.3%, H5: 32.0%, H10: 2.6%).


In cultured mouse podocytes expressing Vpr, hydroxychloroquine showed protective effects at up to 10 μg/mL. Hydroxychloroquine has diverse effects on mammalian cells, including increasing lysosomal pH, which in turn alters protein processing such as glycosylation. Hydroxychloroquine also alters Toll-like receptor signaling. The effects of hydroxychloroquine on Vpr-induced podocyte injury deserve further study.