Signatures of the Systemic Effects of a Snake Venom and Antivenom: Multiomics Profiling of the Kidney Pathology

Abstract

Animal venoms comprise many toxins that work in concert to break apart the robust homeostatic systems of prey organisms. Conversely, prey organisms actively antagonize each step of envenoming, which displays a complex kinetics involving important changes at molecular, cell, tissue, and organism levels. In this study we explored the mammalian host response to envenoming using proteomics/N-terminomics and phosphoproteomics approaches to evaluate the in vivo effects of Bothrops jararaca venom in the mouse kidney after injection in the thigh muscle (1.6 mg/kg), mimicking a snakebite, and the impact of anti-Bothrops antivenom injected 1 h later (1.6 mg/kg; i.v. tail). For proteomics/N-terminomics, proteins were TMT-labeled, to allow for specific (tryptic) and semi-specific searches of MS/MS spectra to assess both global proteome and degradome. We quantified >7000 proteins, and prominent changes were observed in the kidney tissue, where protein differential abundance was identified after 3, 6, and 24 h, including markers of acute-phase response and injury. Likewise, the N-terminomic analysis revealed a significant impact of venom progressing from 3 h to 24 h, resulting in dysregulated proteolysis and indicating the activation of host proteases. The protease fingerprint matched legumain and cathepsin profiles. Venom toxins also promoted alteration in the dynamics of phosphorylation, with the activation of kinases. Under the conditions tested, antivenom administration (i) did not reduce the number of differentially abundant proteins and inflammation markers, (ii) partially attenuated the generation of proteolytic products in envenomed animals, and (iii) directly perturbed the phosphorylation signaling in control animals. Taken together, our findings underscore for the first time the mouse renal response to a protease-rich venom, revealed by the dynamic alteration in protein abundance, protease targets, and phosphorylation events, providing new facets of snake venom and antivenom systemic effects, which are important for the development of new therapies.

Keywords: Bothrops jararaca; N-terminomics; mouse kidney; phosphoproteomics; proteomics; snake venom.