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1,2,5, Nesrine Aroua , Fabienne de Toni X , Virginie Feliu ... · established, mice were treated...
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Studying cytarabine resistance through PDX models in acute myeloid leukemia Thomas Farge 1,2,5 , Nesrine Aroua 1,2,5 , Fabienne de Toni 1,2,5 , Virginie Feliu 1,2,5 , Robin Perry 3 , François Vergez 1,2,5 , Estelle Saland 1,2,5 , Marie-Laure Travers 1,2,4,5 , Suzanne Tavitian 4,5 , Olivier Duchamp 5,6 , Gwenn Danet-Desnoyers 3 , Martin Carroll 3 , Christian Récher 1,2,4,5 and Jean-Emmanuel Sarry 1,2,5 1 INSERM, Cancer Research Center of Toulouse, U1037, F-31024 Toulouse, France; 2 University of Toulouse, F-31300 Toulouse, France; 3 Division of Hematology & Oncology, Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; 4 Département d’Hématologie, Centre Hospitalier Universitaire de Toulouse, Institut Universitaire du Cancer Toulouse Oncopole, F-31059 Toulouse, France; 5 Consortium IMODI; 6 Oncodesign, 21000 Dijon The major therapeutic barrier in acute myeloid leukemia (AML) is chemotherapy resistance. AML cells resistant to conventional chemotherapy targeting DNA synthesis are thought to be enriched in quiescent leukemic stem cells (LSCs). In order to better understand chemotherapy resistance in AML, we analyzed the response to cytarabine (AraC) through patient-derived xenograft (PDX) models with 20 primary AML patient specimens from two clinical sites and in the context of a French “Innovative models initiative” (IMODI) program. Introduction Results Materials and Methods Conclusion Peripheral blood cells from 20 AML patient were injected in NSG mice (n=15-20/patients). Engraftment was monitored by flow cytometry in mice peripheral blood. When the AML is established, mice were treated with AraC administered IP for 5 days as a single agent at 60 mg/kg daily. In vivo AraC response in PDX models has been compared to clinicobiological data of their matched patients. Finally, residual leukemic cells (RLCs), surviving after in vivo AraC treatment, have been characterized for their cell surface phenotype, stem cell frequency, cell cycle and metabolic status. Altogether, these results suggest a novel model of AraC chemotherapy resistance uncovering the control of the oxidative and mitochondrial energy metabolism in vivo and the relevance of PDX models for clinical investigations and new preclinical drug assessment. Further studies of the role of immune and stromal microenvironment will be assessed in this model to extend our findings in a more relevant setting - + - + - + - + - + - + - + - + - + - + - + - + - + 0 20 40 60 80 100 ** #Ps1 * #Ps4 #Ps5 n.d. * #Ps6 *** #Ps8 #Ps7 n.s. ** n.s * #Ps9 #Ps10 #Ps11 n.d. #Ps12 ** #Ps13 ** #Ps14 *** #Ps15 Ara-C Induction (Fold) 3 13 5 3 2 2 2 2 3 6 10 6 6 Post AraC apoptosis induction index Post AraC tumor burden reduction index In Vivo response to Ara-C (Reduction fold) Dx Rel 0 10 20 30 40 50 In Vivo PDX response to Ara-C (Fold increase) Dx Rel 0 5 10 15 In Vivo PDX response to Ara-C (Fold increase) M F 0 5 10 15 In Vivo response to Ara-C (Reduction fold) M F 0 10 20 30 40 50 Dx/Rel Gender Fig1 In vivo analysis of the total cell tumor burden reduction at D8 post-Tx in PDX model of AML Failure/CR In Vivo PDX response to Ara-C (Fold increase) Failure CR 0 5 10 15 In Vivo response to Ara-C (Reduction fold) Failure CR 0 10 20 30 40 50 FAB In Vivo PDX response to Ara-C (Fold increase) M1 M2 M4/5 0 5 10 15 In Vivo response to Ara-C (Reduction fold) M1 M2 M4/M5 NOS/MLD 0 10 20 30 40 50 * * In Vivo PDX response to Ara-C (Fold increase) <65y >65y 0 5 10 15 Cytogenetic Risk In Vivo PDX response to Ara-C (Fold increase) Fav In t Defa 0 5 10 15 In Vivo response to Ara-C (Reduction fold) Fav in t Defa Nd 0 10 20 30 40 50 In Vivo response to Ara-C (Reduction fold) <65y >65y 0 10 20 30 40 50 0.053 Age at Dx CD34+ In Vivo PDX response to Ara-C (Fold increase) <20% >20% 0 5 10 15 In Vivo response to Ara-C (Reduction fold) <20% >20% 0 10 20 30 40 50 In Vivo PDX response to Ara-C (Fold increase) <337d >337d 0 5 10 15 In Vivo PDX response to Ara-C (Fold increase) <337d >337d 0 10 20 30 40 50 * Overall survival - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + 0.01 0.1 1 10 100 #53 #262 #325 #973 #1956 Ara-C BM SP Reduction (Fold) 5 n.d. n.d. n.s. * 10 6 0.1 7 8 6 4 3 6 3 9 6 * * *** ** * *** #Ps1 * ** #Ps2 * * #Ps3 * #Ps4 ** * 10 6 3 15 6 #Ps5 ** #Ps6 *** #Ps7 #Ps8 34 3 12 16 5 34 21 45 n.s. *** * * n.s. n.s. n.s. #Ps9 ** n.s. #Ps10 n.s. n.s. #Ps11 * n.s. #Ps12 n.s. n.s. #Ps13 * n.s. #Ps14 * n.s. #Ps15 *** * BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP BM SP 3 112 7 1 134 15 2 9 2 10 0.1 4 1 13 Fig2 Comparative analysis between preclinical PDX data and clinical data (patients informations) Vehicle Ara-C 2.5x 20x 2.5x 20x Vehicle Ara-C 2.5x 20x 2.5x 20x Apoptosis induction in BM after at D8 post-Tx Tumor reduction in bone marrow and spleen after at D8 post-Tx Fig3 In vivo response to cytarabine (AraC) in PDX: Signature phenotype, cell cycle and red/ox status of residual leukemic cells DAPI Vehicle Cortical zone Trabecular zone CD33 PIMO Merge DAPI Ara-C Cortical zone Trabecular zone CD33 PIMO Merge DAPI Vehicle Cortical zone Trabecular zone CD33 PIMO Merge DAPI Ara-C Cortical zone Trabecular zone CD33 PIMO Merge 0 20 40 60 80 n.s. Vehicule Ara-C Ki67 + blasts (%) 0 20 40 60 80 n.s. Vehicule Ara-C Ki67 + blasts (%) - + - + - + - + - + - + - + - + - + - + 0.1 1 10 100 #262 #325 n.s. #973 #Ps1 n.s. Ara-C n.s. #Ps2 n.s. #Ps3 n.s. #Ps4 n.s. #Ps6 n.s. #Ps7 n.s. #Ps8 n.s. * G 0 cells in hCD45 + CD33 + cells (%) - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + - + 0.1 1 10 100 * #262 #325 n.s. Ara-C #973 n.s. #1956 n.d. * #Ps1 * #Ps2 #Ps5 n.d. #Ps6 n.s. #Ps3 n.d. #Ps4 ** #Ps7 n.s. * #Ps8 #Ps9 ** #Ps10 n.s. #Ps12 n.s. #Ps13 n.s. #Ps14 n.s. #Ps15 n.s. hCD34 + CD38 - in viable hCD45 + CD33 + cells (%) AraC treatment of AML cells exhibits an enrichment of the CD34+CD38- phenotype in BM post-Tx (6 out of 12) AraC treatment of AML does not necessarily lead to a cell cycle arrest in BM at D8 post-Tx AraC-resistant AML cells are hypoxic (PIMO+) and have high total ROS content in vivo at D8 post-Tx - + - + - + - + - + - + - + - + - + - + 1 10 100 1000 10000 100000 ** #Ps10 #Ps11 n.s #Ps8 0.057 Ara-C #Ps12 * #Ps13 0.06 #Ps14 ** #Ps15 * #Ps1 * #Ps4 n.s #Ps6 * Total ROS content in viable hCD45 + CD33 + blasts ( MFI) CD34 CD38 Vehicle Ara-C CD34 CD38 Viable hCD45 + CD33 + cells (Millions) AnxV+ % in total hCD45 + CD33 + cells Utilization of cytarabine in patient derived xenograft models of acute myeloid leukemia
Transcript of 1,2,5, Nesrine Aroua , Fabienne de Toni X , Virginie Feliu ... · established, mice were treated...
Studying cytarabine resistance through PDX models in acute myeloid leukemia
Thomas Farge1,2,5, Nesrine Aroua1,2,5, Fabienne de Toni1,2,5, Virginie Feliu1,2,5, Robin Perry3, François Vergez1,2,5, Estelle Saland1,2,5, Marie-Laure Travers1,2,4,5, Suzanne Tavitian4,5, Olivier Duchamp5,6, Gwenn Danet-Desnoyers3, Martin Carroll3, Christian Récher1,2,4,5 and Jean-Emmanuel Sarry1,2,5
1INSERM, Cancer Research Center of Toulouse, U1037, F-31024 Toulouse, France; 2University of Toulouse, F-31300 Toulouse, France; 3Division of Hematology & Oncology, Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; 4Département d’Hématologie, Centre Hospitalier Universitaire de Toulouse, Institut Universitaire du Cancer Toulouse Oncopole, F-31059 Toulouse, France; 5 Consortium IMODI; 6 Oncodesign, 21000 Dijon
The major therapeutic barrier in acute myeloid leukemia (AML) is chemotherapy resistance.AML cells resistant to conventional chemotherapy targeting DNA synthesis are thought to beenriched in quiescent leukemic stem cells (LSCs). In order to better understand chemotherapyresistance in AML, we analyzed the response to cytarabine (AraC) through patient-derivedxenograft (PDX) models with 20 primary AML patient specimens from two clinical sites and inthe context of a French “Innovative models initiative” (IMODI) program.
Introduction
Results
Materials and Methods ConclusionPeripheral blood cells from 20 AML patient were injected in NSG mice (n=15-20/patients).Engraftment was monitored by flow cytometry in mice peripheral blood. When the AML isestablished, mice were treated with AraC administered IP for 5 days as a single agent at 60mg/kg daily. In vivo AraC response in PDX models has been compared to clinicobiological dataof their matched patients. Finally, residual leukemic cells (RLCs), surviving after in vivo AraCtreatment, have been characterized for their cell surface phenotype, stem cell frequency, cellcycle and metabolic status.
Altogether, these results suggest a novel model of AraC chemotherapy resistance uncoveringthe control of the oxidative and mitochondrial energy metabolism in vivo and the relevance ofPDX models for clinical investigations and new preclinical drug assessment. Further studies ofthe role of immune and stromal microenvironment will be assessed in this model to extend ourfindings in a more relevant setting
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Fig1 In vivo analysis of the total cell tumor burden reduction at D8 post-Tx in PDX model of AML
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Fig3 In vivo response to cytarabine (AraC) in PDX: Signature phenotype, cell cycle and red/ox status of residual leukemic cells
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Utilization of cytarabine in patient derived xenograft models of acute myeloid leukemia
