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Molecular Medicine

, Volume 21, Issue 1, pp 749–757 | Cite as

The HIV Protease Inhibitor Saquinavir Inhibits HMGBl-Driven Inflammation by Targeting the Interaction of Cathepsin V with TLR4/MyD88

  • John P. Pribis
  • Yousef Al-Abed
  • Huan Yang
  • Domokos Gero
  • Hongbo Xu
  • Marcelo F. Montenegro
  • Eileen M. Bauer
  • Sodam Kim
  • Sangeeta S. Chavan
  • Changchun Cai
  • Tunliang Li
  • Petra Szoleczky
  • Csaba Szabo
  • Kevin J. Tracey
  • Timothy R. Billiar
Research Article

Abstract

Extracellular high-mobility group box 1 (HMGB1) (disulfide form), via activation of toll-like receptor 4 (TLR4)-dependent signaling, is a strong driver of pathologic inflammation in both acute and chronic conditions. Identification of selective inhibitors of HMGB1-TLR4 signaling could offer novel therapies that selectively target proximal endogenous activators of inflammation. A cell-based screening strategy led us to identify first generation HIV-protease inhibitors (PI) as potential inhibitors of HMGB1-TLR4 driven cytokine production. Here we report that the first-generation HIV-PI saquinavir (SQV), as well as a newly identified mammalian protease inhibitor STO33438 (334), potently block disulfide HMGB1-induced TLR4 activation, as assayed by the production of TNF-α by human monocyte-derived macrophages (THP-1). We further report on the identification of mammalian cathepsin V, a protease, as a novel target of these inhibitors. Cellular as well as recombinant protein studies show that the mechanism of action involves a direct interaction between cathepsin V with TLR4 and its adaptor protein MyD88. Treatment with SQV, 334 or the known cathepsin inhibitor SID26681509 (SID) significantly improved survival in murine models of sepsis and reduced liver damage following warm liver ischemia/reperfusion (I/R) models, both characterized by strong HMGB1-TLR4 driven pathology. The current study demonstrates a novel role for cathepsin V in TLR4 signaling and implicates cathepsin V as a novel target for first-generation HIV-PI compounds. The identification of cathepsin V as a target to block HMGB1-TLR4-driven inflammation could allow for a rapid transition of the discovery from the bench to the bedside. Disulfide HMGB1 drives pathologic inflammation in many models by activating signaling through TLR4. Cell-based screening identified the mammalian protease cathepsin V as a novel therapeutic target to inhibit TLR4-mediated inflammation induced by extracellular HMGB1 (disulfide form). We identified two protease inhibitors (Pls) that block cathepsin V and thereby inhibit disulfide HMGB1-induced TLR4 activation: saquinavir (SQV), a firstgeneration PI targeting viral HIV protease and STO33438 (334), targeting mammalian proteases. We discovered that cathepsin V binds TLR4 under basal and HMGB1-stimulated conditions, but dissociates in the presence of SQV over time. Thus cathepsin V is a novel target for first-generation HIV PIs and represents a potential therapeutic target of pathologic inflammation.

Notes

Acknowledgments

We would like to thank Patricia Loughran for providing the histology and Rick Shapiro for technical advice on experiments. This work was supported by grants to TRB P50GM053789 and GM098446 to Huan Yang.

Supplementary material

10020_2015_2101749_MOESM1_ESM.pdf (3.5 mb)
Supplementary material, approximately 3.50 MB.

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Authors and Affiliations

  • John P. Pribis
    • 1
    • 2
  • Yousef Al-Abed
    • 3
    • 4
  • Huan Yang
    • 3
  • Domokos Gero
    • 5
  • Hongbo Xu
    • 1
    • 6
  • Marcelo F. Montenegro
    • 1
  • Eileen M. Bauer
    • 1
  • Sodam Kim
    • 1
  • Sangeeta S. Chavan
    • 3
  • Changchun Cai
    • 1
  • Tunliang Li
    • 1
    • 6
  • Petra Szoleczky
    • 5
  • Csaba Szabo
    • 5
  • Kevin J. Tracey
    • 3
  • Timothy R. Billiar
    • 1
  1. 1.Department of SurgeryF1281 Presbyterian University Hospital, University of PittsburghPittsburghUSA
  2. 2.Integrative Molecular and Biomedical Sciences Graduate ProgramBaylor College of MedicineHoustonUSA
  3. 3.Laboratory of Biomedical ScienceThe Feinstein Institute for Medical ResearchManhassetUSA
  4. 4.Department of Medicinal ChemistryThe Feinstein Institute for Medical ResearchManhassetUSA
  5. 5.Department of AnesthesiologyUniversity of Texas Medical BranchGalvestonUSA
  6. 6.The Third Xiangya HospitalCentral South UniversityHunanChina

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