iTAG-RNA Isolates Cell-Specific Transcriptional Responses to Environmental Stimuli and Identifies an RNA-Based Endocrine Axis

Cell Rep. 2020 Mar 3;30(9):3183-3194.e4. doi: 10.1016/j.celrep.2020.02.020.

Abstract

Biofluids contain various circulating cell-free RNAs (ccfRNAs). The composition of these ccfRNAs varies among biofluids. They constitute tantalizing biomarker candidates for several pathologies and have been demonstrated to be mediators of cellular communication. Little is known about their function in physiological and developmental settings, and most works are limited to in vitro studies. Here, we develop iTAG-RNA, a method for the unbiased tagging of RNA transcripts in mice in vivo. We use iTAG-RNA to isolate hepatocytes and kidney proximal epithelial cell-specific transcriptional responses to a dietary challenge without interfering with the tissue architecture and to identify multiple hepatocyte-secreted ccfRNAs in plasma. We also identify specific transfer of liver-derived ccfRNAs to adipose tissue and skeletal muscle, where they likely constitute a buffering system to maintain lipid homeostasis under acute high-fat-diet feeding. Our findings directly demonstrate in vivo transfer of RNAs between tissues and highlight its implications for endocrine signaling and homeostasis.

Keywords: RNA tagging, mouse genetics, circulating RNA, 5EU prodrug, epigenetics, RNA biomarker.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / metabolism
  • Animals
  • Cell-Free Nucleic Acids / blood
  • Cellular Reprogramming / genetics
  • Chemical Precipitation
  • Cytochrome P-450 CYP3A / metabolism
  • Deoxyuridine / analogs & derivatives
  • Deoxyuridine / chemistry
  • Deoxyuridine / metabolism
  • Diet, High-Fat
  • Endocrine System / metabolism*
  • Environment*
  • Hepatocytes / metabolism
  • Homeostasis
  • Lipid Metabolism / genetics
  • Liver / metabolism
  • Mass Spectrometry
  • Mice
  • Muscle, Skeletal / metabolism
  • Organ Specificity
  • Prodrugs / chemistry
  • Prodrugs / metabolism
  • RNA / blood
  • RNA / metabolism*
  • Reproducibility of Results
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology
  • Staining and Labeling
  • Transcription, Genetic*

Substances

  • Cell-Free Nucleic Acids
  • Prodrugs
  • Small Molecule Libraries
  • RNA
  • Cytochrome P-450 CYP3A
  • 5-ethynyl-2'-deoxyuridine
  • Deoxyuridine