Experimental Bothrops atrox Envenomation: Blood Plasma Proteome Effects after Local Tissue Damage and Perspectives on Thromboinflammation

Abstract

The clinical manifestations of Bothrops atrox envenoming involve local and systemic changes, among which edema requires substantial attention due to its ability to progress to compartmental syndromes and sometimes cause tissue loss and amputations. However, the impact of edema on the poisoned body’s system has not been explored. Thus, the present study aimed to explore the systemic pathological and inflammatory events that are altered by intraplantar injection of B. atrox venom in a mouse model through hematologic, lipidic, and shotgun proteomics analysis. Plasma samples collected showed a greater abundance of proteins related to complement, coagulation, lipid system, platelet and neutrophil degranulation, and pathways related to cell death and ischemic tolerance. Interestingly, some proteins, in particular, Prdx2 (peroxiredoxin 2), Hba (hemoglobin subunit alpha), and F9 (Factor IX), increased according to the amount of venom injected. Our findings support that B. atrox venom activates multiple blood systems that are involved in thromboinflammation, an observation that may have implications for the pathophysiological progression of envenomations. Furthermore, we report for the first time a potential role of Prdx2, Hba, and F9 as potential markers of the severity of edema/inflammation in mice caused by B. atrox.

Challenges and Opportunities in Clinical Diagnostic Routine of Envenomation Using Blood Plasma Proteomics

“…Human envenomation by venomous animals causes changes in plasma protein abundance that are currently monitored by assays that primarily assess changes in hemostasis [ 20 , 21 ]. Additionally, other proteins that are or are not involved in hemostasis have already been identified as potential biomarker candidates but require validation for monitoring clinical complications associated with envenomation [ 67 , 68 , 69 , 70 ]. Furthermore, the possibility of identifying circulating animal toxins in plasma samples from patients after multiple bee stings [ 50 ] and snake bites [ 41 , 71 ] using proteomics tools has already been reported.…”

“…Studies involving mass spectrometry in toxinology have focused on the study of proteins present in animal venoms, converging on (i) the identification of isolated and purified proteins from venoms [ 72 , 73 , 74 ], (ii) the description of the composition of venoms [ 75 , 76 , 77 , 78 , 79 , 80 , 81 ], and (iii) the identification of altered biological processes in tissues after tissue damage caused by snake venom [ 41 , 67 , 68 , 69 , 70 , 82 , 83 ]. MS can assist in the development and tracking of target toxins and improve the sensitivity and specificity of ELISA and Western blot assays by identifying toxins circulating in the blood of the victims.…”

“…The number of species within the same genus can be a critical challenge for studies of biomarkers in envenomation by venomous animals. As observed in experimental studies, different biomarker candidates may be associated with the species that caused the envenomation, requiring the search for candidates that overlap at the genus level [ 67 , 68 , 69 ]. There is an opportunity for high specificity if the biomarker target is not endogenous, for example, specific toxins with markers [ 41 , 50 , 71 ].…”

“…Furthermore, the ability to actually target and separate the biomarkers associated with clinical complications from those that are not could be a potential challenge. One of the limitations encountered so far in studies is that they display a map of the proteome differentially expressed at a single moment during disease progression compared to healthy individuals or individuals who did not present a specific event [ 37 , 67 , 68 , 69 , 70 ]. Single-point analysis (case versus control) in the case of envenomations by venomous animals, which involves obtaining an overview of biomarkers at the moment the patient enters the hospital service, will help to identify possible diagnostic markers.…”

Crosstalk of Inflammation and Coagulation in Bothrops Snakebite Envenoming: Endogenous Signaling Pathways and Pathophysiology

“…In the current study "thromboinflammation" in Bothrops envenoming was considered to be the activation of integrated processes, which includes classical coagulation, clot formation, and immune responses elicited during Bothrops envenoming [30,[33][34][35]. Thus, the thromboinflammation process was recognized as a broad-spectrum effect on envenoming.…”