SHP2 inhibitor PHPS1 ameliorates acute kidney injury by Erk1/2-STAT3 signaling in a combined murine hemorrhage followed by septic challenge model

Jihong Jiang,Elizabeth W Tindal,Yaping Chen,Chun-Shiang Chung,Baoji Hu,Yunhe Zhang,Alfred Ayala,Jinbao Li

doi:10.1186/s10020-020-00210-1

Abstract

BackgroundHypovolemic shock and septic challenge are two major causes of acute kidney injury (AKI) in the clinic setting. Src homology 2 domain-containing phosphatase 2 (SHP2) is one of the major protein phosphatase tyrosine phosphatase (PTPs), which play a significant role in maintaining immunological homeostasis by regulating many facets of immune cell signaling. In this study, we explored whether SHP2 signaling contributed to development of AKI sequential hemorrhage (Hem) and cecal ligation and puncture (CLP) and whether inactivation of SHP2 through administration of its selective inhibitor, phenylhydrazonopyrazolone sulfonate 1 (PHPS1), attenuated this injury.MethodsMale C57BL/6 mice were subjected to Hem (a “priming” insult) followed by CLP or sham-Hem plus sham-CLP (S/S) as controls. Samples of blood and kidney were harvested at 24 h post CLP. The expression of neutrophil gelatinase-associated lipocalin (NGAL), high mobility group box 1 (HMGB1), caspase3 as well as SHP2:phospho-SHP2, extracellular-regulated kinase (Erk1/2): phospho-Erk1/2, and signal transducer and activator of transcription 3 (STAT3):phospho-STAT3 protein in kidney tissues were detected by Western blotting. The levels of creatinine (Cre) and blood urea nitrogen (BUN) in serum were measured according to the manufacturer’s instructions. Blood inflammatory cytokine/chemokine levels were detected by ELISA.ResultsWe found that indices of kidney injury, including levels of BUN, Cre and NGAL as well as histopathologic changes, were significantly increased after Hem/CLP in comparison with that in the S/S group. Furthermore, Hem/CLP resulted in elevated serum levels of inflammatory cytokines/chemokines, and induced increased levels of HMGB1, SHP2:phospho-SHP2, Erk1/2:phospho-Erk1/2, and STAT3:phospho-STAT3 protein expression in the kidney. Treatment with PHPS1 markedly attenuated these Hem/CLP-induced changes.ConclusionsIn conclusion, our data indicate that SHP2 inhibition attenuates AKI induced by our double-hit/sequential insult model of Hem/CLP and that this protective action may be attributable to its ability to mitigate activation of the Erk1/2 and STAT3 signaling pathway. We believe this is a potentially important finding with clinical implications warranting further investigation.

Highlights

Trauma patients often experience hypovolemic shock and sepsis simultaneously, resulting in severe organ dysfunction (Karasu et al 2019; Spahn et al 2019)
In conclusion, our data indicate that Src homology 2 domain-containing phosphatase 2 (SHP2) inhibition attenuates acute kidney injury (AKI) induced by our double-hit/ sequential insult model of Hem/cecal ligation and puncture (CLP) and that this protective action may be attributable to its ability to mitigate activation of the Extracellular-regulated kinase (Erk1/2) and signal transducer and activator of transcription 3 (STAT3) signaling pathway
Hem/CLP-induced SHP2 activation in the kidney is attenuated by phenylhydrazonopyrazolone sulfonate 1 (PHPS1) treatment PHPS1, a cell-permeable highly selective inhibitor for SHP2, has been shown to inhibit SHP2-dependent cellular signaling and tumor cell colony formation (Hellmuth et al 2008)

Summary

Introduction

Trauma patients often experience hypovolemic shock and sepsis simultaneously, resulting in severe organ dysfunction (Karasu et al 2019; Spahn et al 2019). The kidneys are one of the most commonly affected organs, since they receive 20–25% of the resting cardiac output, despite making up less than 1% of the total body mass. This combination makes them more vulnerable and susceptible to acute injury following blood volume losses and the fluid shifts associated with sepsis. Our prior research as well as that of others has shown that immune coinhibitory receptor molecules play a role in the pathological inflammatory response which occurs in animals after hemorrhagic shock and sepsis (Patil et al 2017; Wakeley et al 2020). We explored whether SHP2 signaling contributed to development of AKI sequential hemorrhage (Hem) and cecal ligation and puncture (CLP) and whether inactivation of SHP2 through administration of its selective inhibitor, phenylhydrazonopyrazolone sulfonate 1 (PHPS1), attenuated this injury

Methods

Results

Conclusion