Apical proximal tubule fatty acid uptake-generated ceramides cause endoplasmic reticulum stress from altered membrane fluidity.

Circulating fatty acids (FAs) are constitutively taken up by basolateral kidney proximal tubule transporters and are the preferred metabolic substrate. In many chronic kidney diseases, the damaged glomerular filtration barrier permits passage of albumin-bound FAs, which are reabsorbed by apical FA transport protein-2 (FATP2). Bilateral FA uptake leads to lipotoxicity and progressive renal function decline, but the relative apical versus basolateral contribution and intracellular mechanisms are not established. Apical or bilateral (but not basolateral) palmitate incubation with human proximal tubule cells stimulated endoplasmic reticulum (ER) stress gene expression, ER stress pathway activation, and ER fragmentation. Apical or bilateral palmitate was associated with reduced lipid droplets and decreased expression of ER-localized lipid droplet biogenesis transcripts. Inhibition of lipid droplet formation also precipitated ER stress, suggesting diminished sequestration of FA metabolites as the cause. Indeed, C16:0 ceramide was increased in bilateral palmitate-treated cells and in kidneys from mice that phenocopy progressive diabetic kidney disease. Ceramide synthesis inhibition abrogated ER stress, and transfection with C16:0 ceramide decreased ER membrane fluidity and caused ER stress. We conclude that aberrant filtration and uptake of FAs by apical FATP2 exceeded the capacity for lipid droplet incorporation and led to cytotoxicity from ceramide-induced ER lipid bilayer stress.