Insulin-Dependent Regulation of mTORC2-Akt-FoxO Suppresses TLR4 Signaling in Human Leukocytes: Relevance to Type 2 Diabetes

Diabetes. 2016 Aug;65(8):2224-34. doi: 10.2337/db16-0027. Epub 2016 May 10.

Abstract

Leukocyte signaling in patients with systemic insulin resistance is largely unexplored. We recently discovered the presence of multiple Toll-like receptor 4 (TLR4) signaling intermediates in leukocytes from patients with type 2 diabetes or acute insulin resistance associated with cardiopulmonary bypass surgery. We extend this work to show that in addition to matrix metalloproteinase 9, hypoxia-inducible factor 1α, and cleaved AMPKα, patient leukocytes also express IRS-1 phosphorylated on Ser(312), Akt phosphorylated on Thr(308), and elevated TLR4 expression. Similar signaling intermediates were detected in leukocytes and neutrophils treated with lipopolysaccharide (LPS), a ligand of TLR4, in vitro. In contrast, insulin, but not LPS, induced mammalian target of rapamycin complex 2 (mTORC2)-dependent phosphorylation of Akt on Ser(473) and FoxO1/O3a on Thr(24/32) in leukocytes and neutrophils. Insulin suppressed LPS-induced responses in a dose- and time-dependent manner. AS1842856, a FoxO1 inhibitor, also suppressed TLR4 signaling. We propose that insulin is a homeostatic regulator of leukocyte responses to LPS/TLR4 and that the signaling intermediates expressed in leukocytes of patients with type 2 diabetes indicate TLR4 signaling dominance and deficient insulin signaling. The data suggest that insulin suppresses LPS/TLR4 signals in leukocytes through the mTORC2-Akt-FoxO signaling axis. Better understanding of leukocyte signaling in patients with type 2 diabetes may shed new light on disease causation and progression.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Aged
  • Diabetes Mellitus, Type 2 / metabolism*
  • Female
  • Forkhead Transcription Factors / metabolism*
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Insulin / metabolism*
  • Leukocytes / drug effects
  • Leukocytes / metabolism*
  • Lipopolysaccharides / pharmacology
  • Male
  • Matrix Metalloproteinase 9 / metabolism
  • Mechanistic Target of Rapamycin Complex 2
  • Middle Aged
  • Multiprotein Complexes / metabolism*
  • Neutrophils / drug effects
  • Neutrophils / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Signal Transduction / drug effects
  • TOR Serine-Threonine Kinases / metabolism*
  • Toll-Like Receptor 4 / metabolism*

Substances

  • Forkhead Transcription Factors
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Insulin
  • Lipopolysaccharides
  • Multiprotein Complexes
  • TLR4 protein, human
  • Toll-Like Receptor 4
  • Mechanistic Target of Rapamycin Complex 2
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • AMP-Activated Protein Kinases
  • Matrix Metalloproteinase 9