Abstract: SA-PO322
Intracellular Acetyl-CoA Levels Are Pivotal for Nephron Progenitor Cell Fate Decisions
Session Information
- Development, Organoids, Vascularized Kidneys, Nephrons, and More
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Tortelote, Giovane G., Tulane University School of Medicine, New Orleans, Louisiana, United States
- Diniz, Fabiola, Tulane University School of Medicine, New Orleans, Louisiana, United States
- Hilliard, Sylvia, Tulane University School of Medicine, New Orleans, Louisiana, United States
- El-Dahr, Samir S., Tulane University School of Medicine, New Orleans, Louisiana, United States
Group or Team Name
- Tortelote Lab.
Background
Low nephron endowment at birth correlates with the development of hypertension in later life. Nephron endowment at birth is governed by a delicate balance between nephron progenitor cells (NPC) self-renewal and differentiation. Previous studies showed that inhibition of glycolysis pushes NPC to differentiation and causes progenitor pool depletion.
Methods
Organ culture experiments: E12.5 embryonic kidneys cultured for 24h-48h in the presence of a vehicle or several pharmacological agents: the glycolysis inhibitor drug, YN1; the mitochondrial pyruvate carrier 1 blocker, UK5099; sodium acetate or canonical Wnt signaling activator (CHIR99021). Immunofluorescence was performed to characterize the observed phenotypes. Bulk. RNA seq. and Proteomics: NPCs were extracted by MACS (passage 0), cultured for 24h with the vehicle, CHIR, and YN1, and then processed for proteomics and bulk RNA sequencing. Data Analysis. Differential gene expression, pathway analysis, and GO enrichment analysis were performed in R.
Results
Here, we showed a direct relationship between glucose metabolism and NPC fate decisions. First, E12.5 embryonic kidneys cultured in the presence of YN1 showed reduced extracellular acidification rate (ECAR), and increased differentiation rate. Second, E12.5 embryonic kidneys stimulated with UK5099 showed an increased ECAR, reduced ATP production from oxidative phosphorylation, augmented NPC pool, and reduced differentiation. We compared the proteome and transcriptome of E13.5 NPC stimulated with either a CHIR or YN1 to dissect the molecular mechanism underlying these phenomena. Decreased cholesterol biosynthesis emerged as a top pathway targeted by both drugs. Inhibition of cholesterol synthesis stimulated NPC differentiation at the cost of cap mesenchyme depletion, which, in turn, phenocopied YN1 treatment. Acetyl-CoA links cholesterol production and glucose metabolism. Sodium-acetate is an alternative source of acetyl-CoA. Sodium-acetate supplementation increased cytosolic acetyl-CoA levels and cap mesenchyme size, without hindering NPC differentiation. Co-treatment with exogenous sodium-acetate abrogated the adverse effects of glycolysis inhibition.
Conclusion
This study identifies acetyl-CoA as a key metabolite for kidney development and sodium-acetate as a potential exogenous substrate to promote kidney growth and development
Funding
- NIDDK Support