Lung PGD Biomarkers in Organ Donors

Study Overview
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Brief Summary
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PGD is a syndrome characterized by alveolocapillary barrier structural and functional alterations with surfactant inactivation and vascular permeability increase, which cause lung edema, parenchymal infiltrate and progressive hypoxemia. PGD may be enhanced in lung donor. Inflammatory and structural changes may be present in the lungs before organ recovery and/or after organ preservation. The investigators aim to identify the surfactant protein, inflammatory and structural changes in lung donor before and after cold ischemia, and biomarkers to PGD in lung recipients.

Detailed Description
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Primary graft dysfunction (PGD) is responsible of high early mortality in lung transplanted patients. Rationale The evolution of lung transplantation may be complicated by primary graft dysfunction (PGD), a form of acute respiratory distress syndrome caused by ischemia-reperfusion-related phenomena. PGD occurs in 15-50% of cases and is responsible for a significant increase in mortality, duration of assisted ventilation and length of stay in intensive care. It is also an important risk factor for the medium-term development of acute and long-term rejection, of bronchiolitis obliterans syndrome (BOS) - chronic rejection - which drastically reduces the survival of the graft. Surfactant proteins comprising the secretory protein of Clara cells (16-kd Clara cell protein-CC16) and surfactant proteins -A (SP-A), -B (SP-B) and -D (SP- D) are recognized as markers of the permeability of the alveolocapillary barrier. Based on these findings, we postulate that the gene expression of CC16, SP-A, -B and D is altered in pulmonary biopsies performed in donors of patients developing primary graft dysfunction after pulmonary transplantation compared to those performed in the donors of patients free of this syndrome. This study could therefore be a complementary means of objective assessment of lung quality prior to transplantation. Aims and Objectives Describe, in the organ donor, changes in expression of Clara Cell Protein (CC16), surfactant-associated proteins (A, B or D), pro- and anti-inflammatory cytokines in circulating blood and lung tissue during organ recovery. Describe the biological and structural changes after the period of cold ischemia. Establish a correlation between biomarkers in the organ donor and the occurrence of acute graft dysfunction in the lung recipient. Material and method Inclusion Criteria All lung organ donor patients referred to our network and their recipients will be included after obtaining their informed consent. Data Collection In the donor, we will record demographic data (age, sex), history, cause of death, blood gas measurement, chest x-ray protocol, blood biological parameters, duration of brain death if appropriate and hot ischemia time if appropriate and protocol of bacteriological analyzes. In the recipient, we will record the demographic data (age, sex), indication of transplantation, results of right catheterization performed on pre-transplantation, standard intraoperative data, immunosuppression, Blood gas, chest x-ray protocol, filling balance and blood biological parameters at 24, 48 and 72h. The declamping times are recorded. Patients are automatically followed up for the rest of their lives. Iterative biopsies are performed in the first year to detect possible acute rejection. The data will be included in our study. Biological samples In the donor, before the perfusion of the preservation solutions, 18 cc of peripheral blood are taken (dry tube, 9 cc). 1 tube will be stored at -80 ° C., the other will be centrifuged (15 minutes, 10000 / min, 20 ° C.) and then the serum will be stored at -80 ° C. Immediately after lung recovery, a pulmonary biopsy (6 cm2) is performed at the lower lobes. A fragment will be immediately placed in liquid nitrogen and stored at -80 ° C. A second fragment is stored in formalin for 24h and then stored in paraffin blocks. Before implantation, at the end of the preservation period, a new lung biopsy is performed in the lower lobes. Biological analyzes Histological examination and gradation Lung tissues fixed in formalin are stained with hematoxylin-eosin to gradate lung lesions [(1) neutrophil infiltration, (2) airway epithelial cell damage, (3) interstitial edema, (4) Hyaline membrane and (5) hemorrhage]. Inflammation, apoptosis and Surfactant proteins O Tissue mRNA measurements: in real time Quantification PCR (RTQ-PCR) O Tissue peptide measurements: Western Blot - ELISA - MILLIPLEX O Treatment of blood samples and analyzes: ELISA - MILLIPLEX Protein inflammatory cytokines (TNF-alpha, IL-6, IL-8, IL-1, IL33) (IL-10), intercellular adhesion molecules (ICAM-1, VCAM-1), apoptosis (Bax, Bcl2, Caspases) Evaluation of apoptosis: TUNEL - Immunohistochemistry

Official Title
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Role of Lung Surfactant Proteins in Donor Lung to Predict Primary Graft Dysfunction in Lung Recipients

Conditions
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Lung Transplant Failure

Intervention / Treatment
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* Diagnostic Test: lung biomarkers in organ donor for PGD in lung recipient


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: All organ donors and their lung recipients who reached the criteria for lung recovery and transplantation Exclusion Criteria: None

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Lung recipients with PGD<br>in the first 72 hours following lung transplantation, the lung recipients have blood gases and chest X-Ray to assess the occurrence of primary graft dysfunction. Chest X-Ray infiltrate and pathological PaO2/FiO2 ratio describe PGD occurence | Diagnostic Test: lung biomarkers in organ donor for PGD in lung recipient<br>* Lung biopsies in organ donor before and after cold ischemia<br>|
| Lung recipients PGD Free<br>In the first 72 hoours following lung transplantation, if the Cest X-ray is normal, the patient is considered PGD-free | Diagnostic Test: lung biomarkers in organ donor for PGD in lung recipient<br>* Lung biopsies in organ donor before and after cold ischemia<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Lung Biomarkers for primary graft dysfunction in organ donors | The investigators aim te describe lung donor structural and molecular changes in relation of PGD occurrence in recipient. | 5-7 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Primary graft dysfunction, Lung transplantation, Surfactant Protein, inflammation