15353-j-veluchamy

1 General introduction and Scope of this Thesis | 19 Infused dose NK-cells Final product characteristics Outcome Escalating doses: 3x10^6 cells/ kg (cohort 1), 10x10^6 cells/kg (cohort 2) and 30x10^6 cells/kg (cohort 3) Mean purity: 74+/-13% CD34+ cell product. 75+/-12% generated CD56+CD3- NK-cells. 0.03+/-0.04% CD3+ cells. 0.16+/-0.21% CD19+ cells. Mean viability: 94% Well tolerated, no GvHD nor toxicity. 4/10 DFS for 55, 47, 17 and 12 months after infusion Repeated dose: level 1: 0.5 x 10^6 NK cells /kg level 2: 1 x 10^6 NK cells /kg level 3: 10x10^6 NK cells /kg Purity: >90% CD56+CD3- cells Well tolerated, no GvHD. 4/13 NE, 4/13 TF- PD, 3/13 CR, 1/13 Cri and 1/13 MLFS Single dose: 1.29-5.53x10^6 cells/ kg Median purity: infused CD3+ cells: 0.65x10^5 cells/kg. Mean viability: 95% Feasible study, moderate toxicity. 9/16 DFS, 7/16 in relapse (3-51 months), 1/16 died of bacterial pneumonia Single dose: 4,3-22,4x10^6 cells/kg Purity: ≥90% CD3-CD56+ cells. CD3+ cells <0.1%. Viability: 82- 100% No GvHD. 3/8 PR, 5/8 no response. Median survival is 12,9 months Single dose: 1X10^6 cells/kg (cohort 1) 1x10^7 cells/kg (cohort 2) Repeated dose: 1x10^6 cells/kg (cohort 3) 3x10^6 cells/kg (cohort 4) 1x10^7cells/kg (cohort 5) 3x10^7 cells/kg (cohort 6) Purity: CD16+/CD56+ cells: 98.13 +/- 1.98%; CD3+ cells: 0.41 +/- 0.43%; CD14+ cells: 0.40 +/- 0.37%; CD19+ cells: 0.15 +/- 0.25%. Fold expansion: 757.5 +/- 232.2. Viability: 92.9 +/- 2.1% No GvHD nor severe toxicities. 8/20 SD, 9/20 PD, 3/20 NE. Median PFS in SD patients: 4 months (2-18 months) Single dose: 3.5-103x10^6 cells/kg Median purity: 98.4% CD56+ cells.0% CD3+CD56- T-cells. 0.31% CD19+ B-cells Well tolerated, no GvHD. 6/29 PR, 14/29 CR, 8/29 no response and 1/29 NE. 4/29 are alive and DFS Single dose: 1x10^6 cells/kg Purity: CD56+ cells 97.17% of which 80% CD56+CD3- cells Serious adverse reactions, no GvHD. 3/7 in CR remained in remission, 1/7 in PR achieved CR, 2/7 relapsed and 1/7 died (6 months follow up). Median OS: 141-910 days Table 2: Summary of allogeneic NK cell clinical trials in a non-transplantation setting Study Malig ncy Clinical Trial design Culture method* Phase I (EudraCT numb r: 2010-0 8988-41) Dolstra et al ref 72 AML (n=10) Conditioning with Cy/F u followed by KIR mismatched UCB-NK-cell infusio Ex vivo expanded, differentiated and activated UCB-NK cells from unrelated donors. Culture duration: 42 days with GM-CSF, G-SCF, IL-6, SCF, Flt3L, TPO, IL-7, IL-2 and IL-15 *CD34+ selected HSPC’s Phase I (NCT01898793) Romee et al (2016) ref 73 AML (n=13) Conditioning with Cy/F u followed by cytokine induced memory-like NK-cell infusion and subsequent IL-2 therapy (every other day, 6x) Ex vivo expanded and activated PBNK-cells from haploidentical donors. Culture duration :12- 16h with IL-15, IL-12 and IL- 18. *CD3 depleted and CD56 selected Phase I (NCT00799799) Curti et al (2016) ref 74 AML (n=16) Conditioning with Cy/ Flu followed b KIR ligand mismatched NK-cell infus on; IL-2 therapy (3x weekly for 2 weeks) PBNK-cells from haploidentical donors. *CD3 deplete and CD56 selected Phase II (NCT00526292) Shaffer et al (2016) ref 75 AML (n=6) and MDS (n=2) Conditioning with Cy/Flu followed by HLA-mismatched NK-cell infusion; IL-2 therapy (6x) starting 1 day before and after NK-cell infusion BNK-cells from haploidentical donors. *CD3 depleted and CD56 selected Phase I (NCT 1212341) Yang et al (2016) ref 76 Lymphoma (n=2) and solid tumor (n=18) KIR ligand mismatched NK cell infusion Ex vivo e panded and activated PBNK-cells from unrelated donors. Culture duration: 14 days with irradiated auto- PBMCs, OKT3 and IL-2 Phase I (NKAML: NCT00697671) Pilot study (NKHEM: NCT00187096) Rubnitz et al (2015) ref 77 Relaps Leukemia post HSCT (n=15) Refractory/relap ed leukemia (no prior HSCT) (n=14) Conditioning with Clo/E /Cy followed by KIR matched or mismatched NK-cell infusion; IL-2 therapy (6x) starting 1 day before and after NK-cell infusion Ex vivo expanded PBNK-cells fr m haploidentical donors. Culture duration :>12h. *CD3 depleted and CD56 selected Phase I (EudracT number: 2005-006087-62) Kottaridis et al (2015) ref 78 AML (n=7) Conditioning with Flu and TBI followed by haploidentical tumor primed NK-cell nfusion Ex vivo expa ded and activated PBNK-cell from haploidentical donors. Culture duration: o/n with CTV-1 lysate and cryopreserved for infusion. *Only CD56 selected