Metabolic characterization of directly reprogrammed renal tubular epithelial cells (iRECs)

Sci Rep. 2018 Mar 1;8(1):3878. doi: 10.1038/s41598-018-22073-7.

Abstract

Fibroblasts can be directly reprogrammed to induced renal tubular epithelial cells (iRECs) using four transcription factors. These engineered cells may be used for disease modeling, cell replacement therapy or drug and toxicity testing. Direct reprogramming induces drastic changes in the transcriptional landscape, protein expression, morphological and functional properties of cells. However, how the metabolome is changed by reprogramming and to what degree it resembles the target cell type remains unknown. Using untargeted gas chromatography-mass spectrometry (GC-MS) and targeted liquid chromatography-MS, we characterized the metabolome of mouse embryonic fibroblasts (MEFs), iRECs, mIMCD-3 cells, and whole kidneys. Metabolic fingerprinting can distinguish each cell type reliably, revealing iRECs are most similar to mIMCD-3 cells and clearly separate from MEFs used for reprogramming. Treatment with the cytotoxic drug cisplatin induced typical changes in the metabolic profile of iRECs commonly occurring in acute renal injury. Interestingly, metabolites in the medium of iRECs, but not of mIMCD-3 cells or fibroblast could distinguish treated and non-treated cells by cluster analysis. In conclusion, direct reprogramming of fibroblasts into renal tubular epithelial cells strongly influences the metabolome of engineered cells, suggesting that metabolic profiling may aid in establishing iRECs as in vitro models for nephrotoxicity testing in the future.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / physiology
  • Cellular Reprogramming / physiology
  • Cluster Analysis
  • Epithelial Cells / metabolism*
  • Fibroblasts / metabolism
  • Gas Chromatography-Mass Spectrometry / methods
  • Kidney Tubules / metabolism*
  • Metabolome / genetics
  • Metabolomics
  • Mice
  • Mice, Inbred C57BL
  • Transcription Factors / metabolism

Substances

  • Transcription Factors