Single-Nucleus RNA Sequencing Reveals Loss of Distal Convoluted Tubule 1 Renal Tubules in HIV Viral Protein R Transgenic Mice

Am J Pathol. 2024 Oct;194(10):1844-1856. doi: 10.1016/j.ajpath.2024.06.006. Epub 2024 Jul 18.

Abstract

Although hyponatremia and salt wasting are common in patients with HIV/AIDS, the understanding of their contributing factors is limited. HIV viral protein R (Vpr) contributes to HIV-associated nephropathy. To investigate the effects of Vpr on the distal tubules and on the expression level of the Slc12a3 gene, encoding the sodium-chloride cotransporter (which is responsible for sodium reabsorption in distal nephron segments), single-nucleus RNA sequencing was performed on kidney cortices from three wild-type (WT) and three Vpr transgenic (Vpr Tg) mice. The percentage of distal convoluted tubule (DCT) cells was significantly lower in Vpr Tg mice compared with WT mice (P < 0.05); in Vpr Tg mice, Slc12a3 expression was not significantly different in DCT cells. The Pvalb+ DCT1 subcluster had fewer cells in Vpr Tg mice compared with those in WT mice (P < 0.01). Immunohistochemistry revealed fewer Slc12a3+Pvalb+ DCT1 segments in Vpr Tg mice. Differential gene expression analysis between Vpr Tg and WT samples in the DCT cluster showed down-regulation of the Ier3 gene, which is an inhibitor of apoptosis. The in vitro knockdown of Ier3 by siRNA transfection induced apoptosis in mouse DCT cells. These observations suggest that the salt-wasting effect of Vpr in Vpr Tg mice is likely mediated by Ier3 down-regulation in DCT1 cells and loss of Slc12a3+Pvalb+ DCT1 segments.

MeSH terms

  • AIDS-Associated Nephropathy / genetics
  • AIDS-Associated Nephropathy / metabolism
  • AIDS-Associated Nephropathy / pathology
  • Animals
  • Kidney Tubules, Distal* / metabolism
  • Kidney Tubules, Distal* / pathology
  • Mice
  • Mice, Transgenic*
  • Sequence Analysis, RNA*
  • Solute Carrier Family 12, Member 3 / genetics
  • Solute Carrier Family 12, Member 3 / metabolism
  • vpr Gene Products, Human Immunodeficiency Virus / genetics
  • vpr Gene Products, Human Immunodeficiency Virus / metabolism

Substances

  • Solute Carrier Family 12, Member 3
  • Slc12a3 protein, mouse
  • vpr Gene Products, Human Immunodeficiency Virus