Vaccination with a new generation of tumorspecific mrna loaded dendritic cells prolong progression free survival in patients with different types of malignancies Iris Bigalke 25.09.2015 ISCT Europe 2015 Regional Meeting
DC vaccine studies Stage IV melanoma/ prostate cancer Id vs in Pediatric cancers Stage IV Melanoma: in + IL-2 prostate cancer: Id + aldara Stage III melanoma Temodal Transfection of mdc Irradiation and intratumoral idc NHL Glioblastoma Amplified neurosphere mrna + htert/survivin mrna 2000 2001 2002 2006 2007 2009 Stage IV melanoma +Temodal +Transfection of idc +T cell expansion Prostate cancer Autologous mrna +htert/survivin mrna Amplified ovarian cancer stem cell mrna htert/survivin Acute myeloid leukaemia/ Glioblastoma Fast DC Munich collaboration Autologous mrna and /or htert/survivin mrna 2009 2010 2011 2013-2015 Department of Cellular Therapy
Main conclusions in early studies: RNA/DC-vaccine production is feasible RNA/DC-vaccine is well tolerated Immune responses in ~50% of patients Immune responses observed after 3 6 months Intradermal injection intranodal injection Department of Cellular Therapy
Antigen-tailored dendritic cell (DC) vaccines: optimal delivery of 3 signals for T cell activation CCR7 Signal 1 MHC-Peptid---------------------------TCR Zytokine Signal 2 Costimulation-----------CD28 B7- Family (CD80, CD86); CD40 CD40L IL-12 IL-4 IL-10 Signal 3 Th1: IL-2, IFN,TNF Th2: IL-4,IL-5, IL-13
Production of mrna loaded autologous dendritic cells Leukapheresis PBMC Elutriation Monocytes fresh or frozen Gas-Permeable Teflon Bags Differentiation Day 0 GM-CSF IL-4 Maturation Day 2-3 GM-CSF IL-4 IL-1β TNFα IFNγ PGE 2 R848 24 hours Electroporation Mature DCs mrna: 2-3 antigens Recovery Freezing DC-vaccine A. Zobywalski GMP production and analysis in the cleanroom facility of the Department of Cellular Therapy at the Oslo University Hospital
Patient CU020: Phenotype mdcs mdcs htert mdcs survivin CD 274 CD 14 CD 80 CD 83 CD 86 CCR 7 HLA-DR CD 40 Monocytes Immature Dendritic Cells Mature Dendritic Cells
Functionality Signal 3: Culture medium with DCs Layer with CD40L- Mouse Fibroblasts or soluble CD40L in culture medium 786pg 400 300 200 IL-12p70 IL-10 100 0 HD CU012 CU020 CU024 CU028 CU031 CU040 Controls
Study flow chart mdct Further therapy depending on clinical/immunological response CT CT CT Week -2 1 2 3 4 5 6 7 10 11 12 14 15 18 19 22 23 24 Vaccines 4 vaccines DTH Booster vaccines every 4th week Frequent blood sampling for immune response assessment
Treatment of a patient with NSCLC with mrna htert transfected DCs 61 year old patient, Radiosurgery of the 2 brain metastases with Cyberknife Cycles 4+5 chemotherapy Cisplatin/Gemcitabine Obstruction of bronchus Irradiation of primary tumor (60Gy) Cortisone therapy New brain metastasis Radio surgery with Cyberknife Cyberknife treatment of a lung metastasis in 2014 and a brain metastasis in 2015 Cycles 1-3 chemotherapy Cisplatin/Pemetrexed DC-vaccination 07 2011 08 09 10 11 12 2011 02-06 2013 04 2014 01 2015 09 2015 07/2011 Diagnosis Non small cell lung cancer Tumor stage IV T2N2M1 b (lung and brain) Stable Disease Stable Disease
AML patients not eligible for BM transplantation treated with mrna WT-1 and PRAME transfected DCs Age at diagnosis Gender Time of diagnosis Start DC vaccination WT-1 in BM at time of diagnosis Status ObservationtTime since start of vaccination (months) Observation time total (months) CU030 59 f 09/2013 05/2014 3029 CR 16 24 CU031 51 m 08/2013 08/2014 6978 Relapse 08/2015 13 23 Allotransplantation 09/2015 CU033 69 f 05/2014 11/2014 33110 CR 10 16 CU040 73 f 07/2014 04/2015 4027 CR 5 14 CU041 59 f 10/2014 09/2015 3405 CR started 9/2015 11
Events CU030 59 year old patient, AML diagnosed in 9/2013 Not eligible for BM transplantation due to severe side effects to chemotherapy Start of vaccination with DCs loaded with WT-1 and PRAME mrna at the beginning of May 2014 10000 1000 100 50 10 1 3029 WT-1 Start of vaccination 68 90 89 55 83 100 132 133 153 4000 2000 400 200 BM 60 40 20 0 CD8 WT-1 CD8 PRAME CU030 CD8 CD8 htert CD8 survivin Baseline w5 w9 w13 w45 w53 w61 w69
Events CU031 51 year old patient, male AML diagnosed in 8/2013 Not eligible for BM transplantation Start of vaccination with DCs loaded with WT-1 and PRAME mrna at the end of August 2014 WT-1 CU031 CD8 10000 1000 100 50 10 1 6978 Start of vaccination 45 48 45 51 47 1036 3034 BM 800 600 400 200 60 40 20 0 WT-1 PRAME Baseline w5 w13 w18 w22 w29 w33 w37 w41
Vaccination with TC mrna transfected DCs in Glioblastomas DC-GSC (7 pat): median PFS = 534 days Standard 11 pat): median PFS = 223 days Vik-Mo et al. 2013 Radio/C Surgery hemo Leukapheresis DC vaccination 02.10.2015 Department of Cellular Therapy
Glioblastoma patients treated with DCs loaded with autologous tumor mrna and mrna htert and survivin Age at diagnosis Gender OP Vaccine 1 Last Boost Status Observation Time from surgery OS CU024 73 f May-13 Jul-13 Apr-15 alive 840 days ongoing CU027* 64 f Sep-13 Dec-13 Aug-15 Pseudorelapse, OP 09/2015 720 days ongoing CU028* 69 m Sep-13 Jan-14 Jan-14 Pseudorelapse 04/2014 and 08/2014, 2x OP CU029 63 m Jan-13 Jan-14 Jun-14 Decision to stop DC vaccination by patient CU035 66 m Oct-14 Feb-15 Jul-15 Progression/pseudorelapse (?) in 07/2015, chemo- and cortisone therapy CU038 65 m Nov-14 Jan-15 Aug-15 Pseudorelapse 06/2015, cortisone therapy 570 days 570 days 330 days 330 days 330 days ongoing 300 days ongoing No autologous tumor RNA was available
Main conclusions from these productions and patients RNA/DC-vaccine production with our new generation DCs is feasible for patients with different types of cancer RNA/DC-vaccine of our new generation DCs is well tolerated DTH responses appear earlier and are more prominent than observed with the standard DCprotocol, indicating specific immune responses Specific CD8 responses could be detected Phase I/II study to treat AML patients has just started in Oslo Randomized studyto treat Glioblastoma patients in preparation Department of Cellular Therapy
RESEARCH GROUPS Dept. of Cellular Therapy DC-Projects Gunnar Kvalheim Iris Bigalke Kirsti Hønnåshagen Marianne Lundby Guri Solum Lisbeth Skoge Grete Andreassen Dagny Merete S. Knudtzon Lene Mowinckel Jens Andreas L. Jørgensen Lena Tjeldhorn Else Marit Inderberg Suso Stein Sæbøe-Larssen Anne-Merete Tryggestad Kristina Anderson Leukapheresis-Team QC-Lab Dept. of Clinical Cancer Research Steinar Aamdal Paal Brunsvig Study coordinators/nurses Helmholtz Center Munich Dolores J. Schendel (now Medigene AG) Julitta Kasten AML-Trial Munich University Hospital Munich, Med. III Marion Subklewe AML-Trial Oslo Yngvar Fløisand Glioblastoma Trial Oslo Iver Langmoen Einar Vik-Mo Prostate-Cancer Trial Oslo Svein Dueland