Mesenchymal stromal cells to protect kidney after traumatic hemorrhagic shock

Abstract

Background & Aim Haemorrhagic shock (HS) refers to the inadequate perfusion of tissues due to the imbalance between systemic oxygen delivery and consumption. Blunt or penetrating trauma is the most common cause of HS. The development of multiorgan failure after severe trauma is one of the leading causes of morbidity and late mortality. Among critical organs, the Acute Kidney Injury (AKI) affects up to 50% of the patients and no etiologic treatment is available. Mesenchymal Stromal Cells (MSC) are multipotent cells found in a large number of adult tissues and used in clinical practice as a therapeutic agent for immunomodulation and tissue repair. Many studies have highlighted their immunomodulatory, anti-apoptotic/anti-oxidative stress, pro-angiogenic effect, stimulation of endogenous regenerative process or anti-fibrotic effect by direct contact or by secreting bioactive molecules. Our objective was to evaluate and optimize an early MSC therapy after traumatic hemorrhagic shock (THS) inducing an AKI. Methods, Results & Conclusion We have developed in vivo THS model and validated functional assays on cultures of immune and renal human cells. In vivo, rats were subjected to 1h of HS at a Mean Arterial Pressure of 30mmHg, associated with administration of 1ml plasma from THS animals. Biological and histological analyses are currently performed on D2, D7 and D21. In vitro, a cellular model of renal injury was developed by adding a "shock cocktail" (SC) containing inflammatory cytokines, HMGB1, histones and H2O2. Several criteria have been studied: Membrane polarization of the mitochondria by the JC1 dye, MitoSOX and CellROX probes to evaluate the mitochondrial and cytoplasmic oxidation state. Aggression of renal cells induced a decrease in the mitochondrial membrane polarization. The CellROX probe also showed an increase in cytoplasmic oxidative stress. We also developed a model of inflammation using THP1 cell line treated with SC. The SC induced an increase in TNFα secretion by THP1. Our main preliminary results indicated that MSCs improved the viability of renal cells, partially restored their mitochondrial membrane potential and decreased their cytoplasmic oxidative stress. In addition, they can exert an anti-inflammatory effect by inhibited the secretion of TNFα by THP1 and increased their secretion of the anti-inflammatory cytokine IL1RA. Finally, the pre-clinical model mimicking a polytrauma could be a formidable tool to evaluate the benefit of MSC therapy, away from shock. Haemorrhagic shock (HS) refers to the inadequate perfusion of tissues due to the imbalance between systemic oxygen delivery and consumption. Blunt or penetrating trauma is the most common cause of HS. The development of multiorgan failure after severe trauma is one of the leading causes of morbidity and late mortality. Among critical organs, the Acute Kidney Injury (AKI) affects up to 50% of the patients and no etiologic treatment is available. Mesenchymal Stromal Cells (MSC) are multipotent cells found in a large number of adult tissues and used in clinical practice as a therapeutic agent for immunomodulation and tissue repair. Many studies have highlighted their immunomodulatory, anti-apoptotic/anti-oxidative stress, pro-angiogenic effect, stimulation of endogenous regenerative process or anti-fibrotic effect by direct contact or by secreting bioactive molecules. Our objective was to evaluate and optimize an early MSC therapy after traumatic hemorrhagic shock (THS) inducing an AKI. We have developed in vivo THS model and validated functional assays on cultures of immune and renal human cells. In vivo, rats were subjected to 1h of HS at a Mean Arterial Pressure of 30mmHg, associated with administration of 1ml plasma from THS animals. Biological and histological analyses are currently performed on D2, D7 and D21. In vitro, a cellular model of renal injury was developed by adding a "shock cocktail" (SC) containing inflammatory cytokines, HMGB1, histones and H2O2. Several criteria have been studied: Membrane polarization of the mitochondria by the JC1 dye, MitoSOX and CellROX probes to evaluate the mitochondrial and cytoplasmic oxidation state. Aggression of renal cells induced a decrease in the mitochondrial membrane polarization. The CellROX probe also showed an increase in cytoplasmic oxidative stress. We also developed a model of inflammation using THP1 cell line treated with SC. The SC induced an increase in TNFα secretion by THP1. Our main preliminary results indicated that MSCs improved the viability of renal cells, partially restored their mitochondrial membrane potential and decreased their cytoplasmic oxidative stress. In addition, they can exert an anti-inflammatory effect by inhibited the secretion of TNFα by THP1 and increased their secretion of the anti-inflammatory cytokine IL1RA. Finally, the pre-clinical model mimicking a polytrauma could be a formidable tool to evaluate the benefit of MSC therapy, away from shock.