Phosphoproteome and transcription factor activity profiling identify actions of the anti-inflammatory agent UTL-5g in LPS stimulated RAW 264.7 cells including disrupting actin remodeling and STAT-3 activation

  • Eur J Pharmacol. 2017 Sep 15;811:66-73. doi: 10.1016/j.ejphar.2017.05.049.
Nicholas J Carruthers  1 ,  Paul M Stemmer  2 ,  Ben Chen  3 ,  Frederick Valeriote  4 ,  Xiaohua Gao  5 ,  Subhash C Guatam  6 ,  Jiajiu Shaw  7
Affiliations
  • 1. Wayne State University, Institute of Environmental Health Sciences, 540 East Canfield Ave., Room 2105, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 2. Wayne State University, Institute of Environmental Health Sciences, 540 East Canfield Ave., Room 2105, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 3. 21st Century Therapeutics, Inc., 440 Burroughs, Suite 447, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 4. Henry Ford Health System, Internal Medicine, 440 Burroughs, Suite 415, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 5. Henry Ford Health System, Department of Surgery, One Ford Place, Oncology Research Laboratory, 4D, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 6. Henry Ford Health System, Department of Surgery, One Ford Place, Oncology Research Laboratory, 4D, Detroit, MI 48202, United States. Electronic address: [email protected].
  • 7. 21st Century Therapeutics, Inc., 440 Burroughs, Suite 447, Detroit, MI 48202, United States; Henry Ford Health System, Internal Medicine, 440 Burroughs, Suite 415, Detroit, MI 48202, United States. Electronic address: [email protected].
PMID: 28576409 DOI: 10.1016/j.ejphar.2017.05.049
Abstract

UTL-5g is a novel small-molecule TNF-alpha modulator. It reduces cisplatin-induced side effects by protecting kidney, liver, and platelets, thereby increasing tolerance for cisplatin. UTL-5g also reduces radiation-induced acute liver toxicity. The mechanism of action for UTL-5g is not clear at the present time. A phosphoproteomic analysis to a depth of 4943 phosphopeptides and a luminescence-based transcription factor activity assay were used to provide complementary analyses of signaling events that were disrupted by UTL-5g in RAW 264.7 cells. Transcriptional activity downstream of the interferon gamma, IL-6, type 1 Interferon, TGF-β, PKC/CA2+ and the Glucocorticoid Receptor pathways were disrupted by UTL-5g. Phosphoproteomic analysis indicated that hyperphosphorylation of proteins involved in actin remodeling was suppressed by UTL-5g (gene set analysis, FDR < 1%) as was phosphorylation of STAT3, consistent with the IL-6 results in the transcription factor assay. Neither analysis indicated that LPS-induced activation of the NF-kB, cAMP/PKA and JNK signaling pathways were affected by UTL-5g. This global characterization of UTL-5g activity in a macrophage cell line discovered that it disrupts selected aspects of LPS signaling including STAT3 activation and actin remodeling providing new insight on how UTL-5g acts to reduce cisplatin-induced side effects.

Keywords
Anti-inflammatory; LPS; Macrophage; Mass spectrometry; Phosphoproteomics; UTL-5g.
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