Metabolic Reprogramming in Renal Tubular Cells in Acute Kidney Injury Following Severe Trauma

Study Overview
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Brief Summary
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Severe trauma remains the leading cause of death in people under 50, and is associated with high morbidity, including severe disability, with a substantial socio-economic impact. Secondary to trauma, multiple mechanisms (inflammatory, ischemic, oxidative, etc.) setting in rapidly, leads to organ failure, one of the three first cause of death. Vascular damage, with vasoplegia, renal damage, with acute kidney injury (AKI), and pulmonary damage, with acute respiratory distress syndrome (ARDS), are the most frequently observed but all organs can be affected whatever the type of trauma. For these reasons, identifying the pathophysiological pathways involved in organ failure induced by severe trauma is a major step towards limiting the morbidity and mortality induced by trauma, and proposing therapies to prevent them. Because of the variability of lesions in these patients, and the multiplicity of pathways activated, the mechanisms involved and their causality with organ failure following severe trauma, are still poorly understood. Given their frequency and importance in terms of morbidity and mortality, the investigators decided to take a particular interest in the mechanisms leading to renal and pulmonary injury. The investigators' hypothesis is that the study of urinary and blood markers not performed as part of clinical routine would provide a better understanding of the pathophysiological mechanisms leading to organ failure secondary to severe trauma, and more specifically to renal and pulmonary injuries. With TRAUMATEC study, the investigators will explore mechanisms leading to AKI and ARDS through blood and urine samples of 60 severe trauma patients sampled over the first 48 hours after ICU admission and a reference of 20 healthy volunteers.

Detailed Description
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The investigators plan to include 60 patients over 18 years old with severe trauma, defined with an ISS≥9 and 20 healthy volunteers 18 years old as a reference group. Blood and urine samples will be collected at ICU arrival, 12-, 24- and 48- hours after ICU admission. Specific dosages will then be realized on blood and urines to study metabolic and hormonal pathway leading to AKI and ARDS. The primary objective of the study is to explore the association between renal metabolic changes and renal function impairment following severe trauma. Secondary objectives are (1) to explore mitochondrial changes observed at the renal cellular level, on in vitro renal culture cells after exposure to trauma patient serum (2) to explore the association between plasma metabolic changes and renal and pulmonary function impairment following severe trauma (3) to explore the association between hormonal metabolic changes and renal and pulmonary function impairment following severe trauma (4) to explore the association between red blood cell-induced oxidative stress and renal function impairment following severe trauma (5) to explore the association between changes in the hemoglobin recycling (chelation) system and impaired renal function following severe trauma (6) to explore renal tubular damage secondary to severe trauma (7) to explore the pathophysiological mechanisms associated with pulmonary damage following severe trauma (8) to describe mortality at day 30.

Official Title
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Metabolic Reprogramming in Renal Tubular Cells in Acute Kidney Injury Following Severe Trauma

Conditions
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Multi-organ Failure After Severe Trauma

Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Trauma patients : Adult patients (age ≥ 18 years) Patient admitted for suspected severe trauma (1 Vittel criteria) Injury Severity Score ≥ 9 Health insurance Written consent obtained from the patient or trusted support person / family member / close friend, or inclusion in an emergency situation and written consent obtained from the patient (trusted support person / family member / close friend if necessary) as soon as possible (article L1122-1-2 of the CSP). Healthy volunteers : Adult patients (≥ 18 years) Affiliated with health insurance Written informed consent́ Patient respecting matching Exclusion Criteria: Trauma patients : Pregnant patient Minor patient Adult under guardianship, curatorship or safeguard of justice Patient under Aide Médicale d'État Chronic renal failure on dialysis Chronic respiratory disease Patient with chronic cardiac insufficiency Systemic inflammatory disease Healthy volunteers : Pregnant patient Minor patient Adult under guardianship, curatorship or safeguard of justice Patient under Aide Médicale d'État Patient with chronic renal failure on dialysis Chronic respiratory disease Patient with chronic cardiac insufficiency Systemic inflammatory disease

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| polytrauma patients<br>polytrauma patients | |
| healthy volunteers<br>healthy volunteers | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Profile of urinary metabolite concentrations measured by mass spectometry | Metabolomic study of patients urine according to AKI and compared to healthy volunteers measured by mass spectometry | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| mitochondrial enzymatic activities of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to serum from healthy volunteers | In vitro mitochondrial function of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to healthy volunteer serum assessed by enzymatic activities by Seahorse XFe96 analyzer | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| mitochondrial membrane potential of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to serum from healthy volunteers | In vitro mitochondrial function of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to healthy volunteer serum assessed by measurement of mitochondrial membrane potential by fluorescence | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| mitochondrial Adenosine TriPhosphate (ATP) content of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to serum from healthy volunteers | In vitro mitochondrial function of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to healthy volunteer serum assessed by measurement of ATP content by spectrofluorimetry | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| expression levels of mitochondrial of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to serum from healthy volunteers | In vitro mitochondrial function of cultured Human Kidney 2 (HK2) kidney cells exposed to patient serum and exposed to healthy volunteer serum assessed by measurement of expression levels of mitochondrial by Western-blot | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Plasma metabolite concentration profile measured by mass spectrometry | Metabolomic study of patients plasma measured by mass spectrometry according to AKI and ARDS and compared to healthy volunteers | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| advanced glycation end products (AGEs) produced by red blood cells | Measurement of oxidative stress produced by red blood cells assessed by advanced glycation end products (AGEs) | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| reactive oxygen derivatives produced by red blood cells | Measurement of oxidative stress produced by red blood cells assessed by reactive oxygen derivatives | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| erythrocyte NO production produced by red blood cells | Measurement of oxidative stress produced by red blood cells assessed by erythrocyte NO production | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| haptoglobin level | Hemoglobin recycling system assessed by haptoglobin | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| haptoglobin-hemoglobin complexes level | Hemoglobin recycling system assessed by haptoglobin-hemoglobin complexes | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| free hemoglobin level | Hemoglobin recycling system assessed by free hemoglobin | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| monocyte CD163 receptor from peripheral blood mononuclear cell (PBMC) isolation level | Hemoglobin recycling system assessed by monocyte CD163 receptor from peripheral blood mononuclear cell (PBMC) isolation | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Heme oxygenase-1 (HO-1) enzyme catabolizing heme level | Hemoglobin recycling system assessed by Heme oxygenase-1 (HO-1) enzyme catabolizing heme | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| renin-angiotensin-aldosterone (RASS) profile | Measurement of RASS | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Antidiuretic hormone (ADH) profile | Measurement of ADH | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| cortisol profile | Measurement of cortisol | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| leptine profile | Measurement of leptine | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Intensity of renal tubular injury | Renal tubular injury assessed by standard urinary markers: neutrophil gelatinase-associated lipocalin (NGAL), Kidney injury molecule 1 (KIM-1), IGFB-7, tissue inhibitor of metalloproteinases-2 (TIMP-2), cystatin C | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| hypoxemia level | Pulmonary injury during hospitalization in intensive care evaluated by hypoxemia assessed by the PaO2/FiO2 ratio | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| use of mechanical ventilation | Pulmonary injury during hospitalization in intensive care evaluated by use of mechanical ventilation | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Radiographic Assessment of Lung Edema (RALE) score | Pulmonary injury during hospitalization in intensive care evaluated by Radiographic Assessment of Lung Edema (RALE) score | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| alveolar epithelial lesions | Pulmonary injury during hospitalization in intensive care evaluated by evaluation of alveolar epithelial lesions by circulating soluble Receptor for Advanced Glycation Endproducts (sRAGE) assay. | On admission, at 12 hours, 24 hours and 48 hours of hospital admission | 
| Death | Vital status at day 30 | Day 30 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Severe trauma, hemorrhagic shock, Acute kidney Injury, Acute respiratory distress syndrome, Multi-organ failure