Biomarqueurs et rejet chronique pulmonaire: Résultats de l Initiative de la Cohorte de Greffe Pulmonaire (COLT)
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1 Biomarqueurs et rejet chronique pulmonaire: Résultats de l Initiative de la Cohorte de Greffe Pulmonaire (COLT) Antoine Magnan, Adrien Tissot, Aurore Foureau, Maxim Durand, Carole Brosseau, PierreJoseph Royer, Eugénie Durand, Richard Danger, Sophie Brouard Université de Nantes, CHU de Nantes, l institut du thorax, Service de pneumologie, INSERM 187 / CNRS 6291, INSERM UMR 164
2 CLAD : mécanismes Royer PJ et al., transplantation 216
3 COLT Research of early predictive factors of CLAD Towards 4P medicine? Inclusion Transplantation Chronic rejection V V1 V2 V3 5 years samples
4 COLT C Knoop M Stern S Mussot O Brugière (Bichat), V Boussaud (HEGP) R Kessler A Magnan A Tissot C Benden L Nicod JD Aubert JF Mornex C Dromer C Pison M Dahan M ReynaudGaubert
5 COLT / SysCLAD Story 28: LT program VLM / AGL 29: PHRC National INSERM R Pays de Loire ABM Novartis, Sanofi, Astellas : Inclusions: 184 Transplanted: 1451 Recherche Biomédicale Recherche Non Interventionnelle
6 Dysfonction Chronique du Greffon: Classification of adjudicated patients after 3 years post transplant Nov 217: 1 st adjudications at 5 years: 25 patients «stable» at 3 years
7 Recipient s Immune cells Donor s Bronchial epithelial cells (BEC) MMP9 Story Airway epithelial cells exposed to allogeneic T cells produce MMP9 through a CCL2/CCR2 pathway: implications for chronic lung allograft dysfunction. Modification of BEC profile Pain M, Royer PJ, AJT 216
8 MMP9 (ng/ml) Airway epithelial cells exposed to allogeneic T cells produce MMP9 through a CCL2/CCR2 pathway: implications for chronic lung allograft dysfunction. A ZO1 ECadherin B + TGF Without TGF 1 With TGF 1.2 ZO1 ECadherin Relative expression (2 CT).9 Relative expression (2 CT).9 Fibronectin TL + + Monocytes Actin. TL Monocytes TL Monocytes Relative expression (2 CT) Fibronectin MMP9 MMP9 8 *** *** 6 *** 6 * TL Monocytes Relative expression (2 CT) C 5 * 4 * Without TGF With TGF TL Monocytes TL Monocytes Pain M, Royer PJ, AJT 216
9 MMP9 (pg/ml) ng/ml Sensitivity % Airway epithelial cells exposed to allogeneic T cells produce MMP9 through a CCL2/CCR2 pathway: implications for chronic lung allograft dysfunction. A 12 * B 1 C 1 BOS 9 Stable RAS Area:.8561 p< VCLAD Stable BOS %specificity A cutoff value of 139 ng/ml of MMP9 allowed the prediction of BOS at 6 months with 94% sensitivity and 73% specificity Pain M, Royer PJ, AJT
10 Regulatory T cells in transplantation Regulation of inflammation : asthma, autoimmune diseases, Growing interest in studying T regs in organ transplantation : operational tolerance, acute rejection, clinical trials. No consensus about the link between Tregs and BOS occurrence.
11 % Tregs/CD4 CD25 among T Cells 1 to 6 months after TP % Tregs/CD8+ among T Cells 1 to 6 months after TP CD25 % Tregs among CD4 T Cells % Tregs among CD4 T Cells 1 to 6 months after TP Count Regulatory T cells profile 4 Regulatory CD4 T Cells 8 *** 4.5 ** Before Tp 1 to 6 mth after Tp 6 mth before CLAD CLAD BOS 1 BOS CD4 4.1 ***.2 * FoxP BOS. BOS
12 % mtregs among CD4 T Cells 1 to 6 months after TP CD45RA MFI CD25 of Tregs 1 to 6 months after TP MFI CD39 of Tregs 1 to 6 months after TP MFI PD1 of Tregs 1 to 6 months after TP MFI CD15s of Tregs 1 to 6 months after TP Regulatory T cells profile 8 CD25 CD39 PD1 CD15s BOS BOS BOS BOS * FoxP3.5. BOS 1. FoxP3 CD4+ Tregs 1.1 ntregs FoxP3low CD45RA+ 1.2 nontregs FoxP3low CD45RA 1.3 mtregs FoxP3hi CD45RA
13 Sensitivity% BOS free survival (%) Conclusion Increase of Tregs proportion with a memory phenotype early after TP for patients who will declare a BOS in the five years % Specificity% ** 4 2 Log rank p =, Monts post TP Tregs among CD4 < 2,8 % Tregs among CD4 > 2,8 % New potential biomarker of the BOS occurrence, which could help to manage CLAD after lung transplantation in the next decades.
14 Circulating CD9+ B cells: biomarker of longterm BOSfree survival after Lung Transplantation (94/1) Carole Brosseau 1,2,5,6, Maxim Durand 1,2,3, Eugénie Durand 1,2, Richard Danger 1,2, Jennifer Loy 5,6, Aurore Foureau 5,6,, Mélanie Chesneau 1,2, Philippe Lacoste 5,6, PierreJoseph Royer 5,6, Adrien Tissot 1,2,3,5,6, Antoine Roux 5, Martine Reynaud Gaubert 6, Romain Kessler 7, Sacha Mussot 8, Claire Dromer 9, Olivier Brugière 1, Jean François Mornex 11, Romain Guillemain 12, Marcel Dahan 13, Christiane Knoop 14, Christophe Pison 15, Laurent Nicod 16, Antoine Magnan 5,6$, Sophie Brouard 1,2,4$ & COLT and SysCLAD consortia*.
15 % of plamsa cells % of transitional cells % of CD19+ % B cells of CD19+ B cells % of CD19+ cells % of CD19+ B cells % of memory cells % of naive cells SSCA SSCA CD24 CD27 A Transitional Memory Lymphocyte Doublet & dead cells exclusion CD19+ Plas ma cells Naive B FSCA CD19 CD38 B Lymphocytes CD19 Memory mémoires cells Naive naives cells Legend 2 6 Legend CD CD19 plasmo Plasma cells Transitional 2 transi cells 8 15 *** 6 CD19 Legend 1 2 Legend
16 % of T1/T2 among CD19+ cells % of T3 among CD19+ cells % of CD27+ nonconventional among CD19+ cells % of transitional CD9+ among CD19+ cells % of T1/T2 among transitional cells % of T3 among transitional cells % of CD27+ nonconventional among transitional cells % of CD9+ among transitional cells % of CD19+ % B cells of CD19+ B cells CD24 CD27 IgD % of CD9+ cells % of CD19+ % B cells of CD19+ B cells CD19 B Transitional CD38 T1/T2 T2 dans transi T2 dans CD19 CD27+ nonconvention al 8 BOS 6 rejet CD22 T3 T1 dans transi T1 dans CD T3 T1/T2 IgM CD27+ nonconventional T3 dans transi T3 dans CD19 ** *** *** * CD9 CD19 CD19 ** CD19 CD9 dans CD9+ transi transi CD9 dans 19 *** Legend BOS Legend rejet BOS BOS + rejet rejet Legend
17 Blood Gene Expression Predicts Bronchiolitis Obliterans Syndrome Appearance After Lung Transplantation Richard Danger*, PierreJoseph Royer*, Damien Reboulleau, Eugénie Durand, Jennifer Loy, Adrien Tissot, Philippe Lacoste, Antoine Roux, Martine ReynaudGaubert, Carine Gomez, Romain Kessler, Sacha Mussot, Claire Dromer, Olivier Brugière, JeanFrançois Mornex, Romain Guillemain, Marcel Dahan, Christiane Knoop, Karine Botturi, Christophe Pison, Angela Koutsokera, Laurent P. Nicod, Sophie Brouard*, Antoine Magnan* and the COLT and SysCLAD Consortia
18 Blood gene expression associated with BOS Identification of differential genes (tstatistic from R limma package; p.value <5% and fold change >1.5) Enrichment of downregulated and immunerelated genes in PRED versus comparison Gene ontology analysis: number of genes within the enriched ontology
19 Prediction of BOS: independent validation Validation of genes associated with BOS appearance individual qpcr samples from new patients: n= 13 & 11 PRED independent validation Stable during time POU2AF1: POU class 2 associating factor 1 BLK: B lymphoid tyrosine kinase TCL1A: Tcell leukaemia/lymphoma 1A
20 Prediction of BOS: independent validation Validation of genes associated with BOS appearance individual qpcr samples from new patients: n= 13 & 11 PRED independent validation ROC curves: AUC=.83 AUC=.77 AUC=.78 Excellent discriminative ability Prediction of BOS appearance 3 genes to predict BOS POU2AF1: POU class 2 associating factor 1 BLK: B lymphoid tyrosine kinase TCL1A: Tcell leukaemia/lymphoma 1A
21 Blood Gene Expression Predicts Bronchiolitis Obliterans Syndrome Appearance After Lung Transplantation Conclusion Identification of 3 genes as predictive biomarkers of BOS Whole blood and qpcr: noninvasive and compatible with clinical settings Suggests a role of B cells in BOS mechanisms patent: EP work submitted Validation of these 3 genes in a large prospective cohort
22 Conclusions: Biomarqueurs issus de COLT et SysCLAD MMP9 Treg B transitionnels CD9 + A valider en pratique clinique Score composite à construire Pou2AF1, BLK, TCL1A
23
24 Thank you for your attention CRTI INSERM UMR 164 Sophie BROUARD Carole BROSSEAU Maxime DURAND Richard DANGER Institut du thorax INSERM UMR 187 Antoine MAGNAN Philippe LACOSTE Adrien TISSOT Carole BROSSEAU Eugénie DURAND Jennifer LOY PierreJoseph ROYER Aurore FOUREAU COLT Consortium Flow Cytometry Platform
25 Remerciements PHRC 29
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