Table 2.
Summary of allogeneic NK cell clinical trials in a non-transplantation setting.
| Study | Malignancy | Clinical Trial design | Culture methoda | Infused dose NK cells | Final product characteristics | Outcome |
|---|---|---|---|---|---|---|
| Phase I (EudraCT number: 2010-018988-41) Dolstra et al. (73) | AML (n = 10) | Conditioning with Cy/Flu followed by KIR-mismatched UCB-NK-cell infusion | 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 aCD34+ selected HSPC’s | Escalating doses: 3 × 106 cells/kg (cohort 1), 10 × 106 cells/kg (cohort 2), and 30 × 106 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 |
| Phase I (NCT01898793) Romee et al. (72) | AML (n = 13) | Conditioning with Cy/Flu followed by cytokine-induced memory-like NK-cell infusion and subsequent IL-2 therapy (every other day, 6×) | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: 12–16 h with IL-15, IL-12, and IL-18. aCD3 depleted and CD56 selected | Repeated dose: level 1: 0.5 × 106 NK cells/kg level 2: 1 × 106 NK cells/kg level 3: 10 × 106 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 |
| Phase I (NCT00799799) Curti et al. (71) | AML (n = 16) | Conditioning with Cy/Flu followed by KIR ligand-mismatched NK-cell infusion; IL-2 therapy (3× weekly for 2 weeks) | PBNK cells from haploidentical donors. aCD3 depleted and CD56 selected | Single dose: 1.29–5.53 × 106 cells/kg | Median purity: infused CD3+ cells: 0.65 × 105 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 |
| Phase II (NCT00526292) Shaffer et al. (70) | AML (n = 6) and MDS (n = 2) | Conditioning with Cy/Flu followed by HLA-mismatched NK-cell infusion; IL-2 therapy (6×) starting 1 day before and after NK-cell infusion | PBNK cells from haploidentical donors. aCD3 depleted and CD56 selected | Single dose: 4.3–22.4 × 106 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 |
| Phase I (NCT01212341) Yang et al. (69) | Lymphoma (n = 2) and solid tumor (n = 18) | KIR ligand-mismatched NK-cell infusion | Ex vivo expanded and activated PBNK cells from unrelated donors. Culture duration: 14 days with irradiated auto-PBMCs, OKT3 and IL-2 | Single dose: 1 × 106 cells/kg (cohort 1) 1 × 107 cells/kg (cohort 2) Repeated dose: 1 × 106 cells/kg (cohort 3) 3 × 106 cells/kg (cohort 4) 1 × 107 cells/kg (cohort 5), and 3 × 107 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) |
| Phase I (NKAML: NCT00697671) Pilot study (NKHEM: NCT00187096) Rubnitz et al. (67) | Relapsed leukemia post HSCT (n = 15) Refractory/relapsed leukemia (no prior HSCT) (n = 14) | Conditioning with Clo/Eto/Cy followed by KIR-matched or -mismatched NK-cell infusion; IL-2 therapy (6×) starting 1 day before and after NK-cell infusion | Ex vivo expanded PBNK cells from haploidentical donors. Culture duration: >12 h. aCD3 depleted and CD56 selected | Single dose: 3.5–103 × 106 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 |
| Phase I (EudracT number: 2005-006087-62) Kottaridis et al. (66) | AML (n = 7) | Conditioning with Flu and TBI followed by haploidentical tumor primed NK-cell infusion | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: o/n with CTV-1 lysate and cryopreserved for infusion. aOnly CD56 selected | Single dose: 1 × 106 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 |
| Phase I (BB-IND-14560) Szmania et al. (65) | MM (n = 8) | Conditioning with Bor (+/−Cy/Flu/Dex) followed by fresh haplo-(n = 6) or cryopreserved auto (n = 2) NK cells | Ex vivo expanded and activated PBNK cells from haploidentical (fresh) and autologous (cryopreserved) donors. Culture: 8–9 days with K562-mb15-41BBL stimulator cells and IL-2 | Single dose: 2 × 107–1 × 108 cells/kg | Median purity: 78% CD3− CD56+ cells. CD3+/CD56– 0.1%. Viability cryopreserved: 94%. Viability fresh: 93%. Recovery cryopreserved: 16%. Recovery fresh: 119% | Feasible and safe. 1/8 PR, 6/8 PD, 1/8 NE, and 3/8 died between days 11 and 98 after NK-cell infusion |
| Phase II (NCT00274846) Bachanova et al. (64) | AML (n = 57) | Conditioning with Cy/Flu; IL2DT in cohort 3 followed by haploidentical NK-cell infusion 1 day later; IL-2 therapy (14×, daily) | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: o/n with IL-2. aCD3 depleted (cohort 1) or CD3 depleted/CD56 selected (cohort 2) or CD3/CD19 depleted (cohort 3) | Single dose: 0.96 ± 0.3 × 107 cells/kg (cohort 1) 0.34 ± 0.05 × 107 cells/kg (cohort 2) 2.6 ± 1.5 × 107 cells/kg (cohort 3) | Purity: NK cells 39 ± 9%, T cells: 0.7% (cohort 1) NK cells 75 ± 6%, T cells: 1.3% (cohort 2) NK cells 54 ± 16%, T cells: 0.3% (cohort 3) | Well tolerated, no GvHD and mild toxicities. 9/42 in remission (1.8–15 months) (cohorts 1 and 2, n = 42). 8/15 in remission (1–32 months) (cohort 3, n = 15). DFS: 5% (cohorts 1 and 2) and 33% in cohort 3 |
| Tonn et al. (43) | Solid tumors/sarcoma (n = 12) Leukemia/lymphoma (n = 2) | Pretreatment with mPred following NK-92 cell infusion | Ex vivo expanded and activated allogeneic NK-92 cells. Culture duration: 100–300 h with IL-2. aNo selection | Repeated doses (2 × 48 h apart): 1 × 109 (cohort 1), 3 × 109 (cohort 2) and 1 × 106 (cohort 3) cells/m2 and additional dose level of 1010 cells/m2 in some patients | Viability: >80%. Fold expansion: 32 | Infusion of 1010 NK-92 cells/m2 were well tolerated. 12/15 PD, 2/15 MR, 1/15 SD for 2 years, OS: 13–801 days |
| Pilot study (NCT00799799) Curti et al. (63) | AML (n = 13) | Conditioning with Cy/Flu followed by KIR ligand-mismatched NK-cell infusion; IL-2 therapy (3× weekly for 2 weeks) | PBNK cells from haploidentical donors. aCD3 depleted and CD56 selected | Single dose: 1.11–5 × 106 CD3− CD56+ cells/kg | Mean viability: 95%. Median purity: 93.5% NK cells. Maximum T-cell dose 105 cells/kg | Feasible and safe, no GvHD. 5/13 active disease: 1/5 CR (6 months), 4/5 died of PD. 3/6 treated in CR are DFS (34, 32, and 18 months), 2/13 in MR in CR (4 and 9 months) |
| Phase II (BB-IND 8847) Geller et al. (33) | Refractory metastatic breast cancer (n = 14) Ovarian cancer (n = 6) | Conditioning with Cy/Flu with or without TBI followed by allogeneic NK-cell infusion; IL-2 therapy (3× weekly for 2 weeks) | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: o/n with IL-2 | Single dose: 8.33 × 106–3.94 × 107 cells/kg | Viability: >70%. Median T cells: 0.11% CD3+ cells | TLS and PLS and limited infusion or IL-2 related toxicities. 1/20 died due to grade 5 toxicity. 4/20 PR, 12/20 SD, and 3/20 PD (between 31 and 109 days) |
| Pilot study Bachanova et al. (62) | B-cell NHL (n = 6) | Conditioning with Cy/Flu and mAb (rituximab, 4×) before and after haplo NK-cell infusion followed by IL-2 therapy (6×, every other day) | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: 8–16 h with IL-2 | Single dose: 21 ± 19 × 106 NK cells/kg | Purity: 43 ± 11% NK cells. 0.16 ± 0.12% T cells | Feasible and safe. 2/6 CR, 2/6 relapsed at 6 months, 2/6 died |
| Pilot study NKAML Rubnitz et al. (61) | AML (n = 10) | Conditioning with Cy/Flu followed by KIR-mismatched NK-cell infusion; IL-2 therapy (6×) starting 1 day before and after NK-cell infusion | PBNK cells from haploidentical donors. aCD3 depleted and CD56 selected | Single dose: 5–81 × 106 cells/kg | Median purity: B-cells 0.097 × 106 cells/kg. T cells 1 × 103 cells/kg | Feasible and safe. 10/10 in remission (569–1,162 days) |
| Phase I (EudraCT number: 2005-005125-58) Iliopoulou et al. (60) | Non-SCLC (n = 16) | Haploidentical NK-cell infusion after chemotherapy | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: 21–23 days with IL-15 followed by 1 h with IL-15 and hydrocortisone. aOnly CD56 selected | Repeated doses (2–4): 0.2–29 × 106 cells/kg per dose | Median purity: (T cells) CD3+ CD56+ CD28− 0.12 × 106 cells/kg. CD56+ CD3 cells 97.9% (after 20 days culture). Fold expansion: 23 | Safe, no GvHD. 2/16 PR, 6/16 SD, 7/16 PD, 1/16 not treated. 1-year OS 56% (9/16), 2-year OS 19% (4/16) |
| Phase I Arai et al. (42) | Metastatic RCC (n = 11) or Malignant Melanoma (n = 1) | NK-92-cell infusion | Ex vivo expanded and activated allogeneic NK 92 cells. Culture duration: 15–17 days with or without IL-2. aNo selection | Repeated doses (3× in cohort): 1 × 108 (cohort 1), 3 × 108 (cohort 2), 1 × 109 (cohort 3), and 3 × 109 (cohort 4) cells/m2 | Fold expansion: 200 over 15–17 days. Viability: ≥80% | Safe and feasible, mild toxicities (1 grade 4, hypoglycemia). 10/12 PD (died between day 101 and 1,059), 1/12 alive (1,450 days) and 1/12 died of bronchopneumonia (day 832) |
| Phase I (BB-IND 8847) Miller et al. (50) | Metastatic Melanoma (n = 10), Metastatic RCC (n = 13), Refractory HL (n = 1), and AML (n = 19) | Conditioning with low Cy/mPred or Flu or high-Cy/Flu followed by NK-cell infusion; IL-2 therapy (14×, daily) | Ex vivo expanded and activated PBNK cells from haploidentical donors. Culture duration: o/n with IL-2 | Escalating doses: low cy/mPred: 1 × 105, 1 × 106, 1 × 107, or 2 × 107 cells/kg (at least three per cohort). Flu or high-Cy/Flu: 2 × 107 cells/kg | Viability: >70%. Purity: NK cells 40 ± 2%. T cells 1.75 ± 0.3 × 105 cells/kg is 0.9 ± 0.1%. Monocytes 25 ± 1.6% and B-cells 19 ± 2% | Feasible and tolerated without toxicities. Low-Cy/mPred: 2/17 with MRCC SD for 20 and 21 months. 4/17 with MM SD for 4–9 months (n = 17) High-Cy/Flu: 5/19 AML pts in CR (n = 19) |
CNS, central nervous system; MPDs, myeloproliferative disorders; LPD, lymphoproliferative disorder; MM, multiple myeloma; MDS, myelodysplastic syndromes; MDN, myelodysplastic neoplasms; MPN, myeloproliferative neoplasms; AML, acute myeloid leukemia; LBLL, lymphoblastic leukemia-lymphoma; ALL, acute lymphoblastic leukemia; NB, neuroblastoma; RMS, rhabdomyosarcoma; CML, chronic myelogenous leukemia; NHL, non-Hodgkin’s lymphoma; MCL, mantle cell lymphoma; DLBCL, diffuse large B cell lymphoma; FL, follicular lymphoma; ALCL, anaplastic large cell lymphoma; HL, Hodgkin’s lymphoma; RCC, renal cell cancer; SCLC, small cell lung cancer; CLL, chronic lymphocytic leukemia; HCC, hepatocellular carcinoma; PNET, primitive neuroectodermal tumor; ACC, adrenal cortical carcinoma; EWS, Ewing sarcoma; DSRCT, desmoplastic small round cell tumor; Cy, cyclophosphamide; Flu, fludarabine; Bor, bortezomib; Dex, dexamethasone; Clo, clofarabine; Eto, etoposide; Cis, cisplatin; Pac, paclitaxel; Doc, docetaxel; Vin, vinorelbine; Gem, gemcitabine; Car, carboplatin; Pem, pemetrexed; TBI, total body irradiation; Tac, tacrolimus; Pred, prednisolone; mPred, methylprednisolone; Thy, thymoglobulin; Vin, vincristine; Adr, adriamycin; Predn, prednisone; Mel, melphalan; OKT3, muromonab-CD3; HSPC, human stem and progenitor cells; aGvHD, acute graft versus host disease; DNKI, donor NK-cell infusion; Stim, stimulated; Unstim, unstimulated; DFS, disease-free survival, CR, complete remission; PR, partial response; MR, minimal response; SD, stable disease; PD, progressive disease; aNK-DLI, donor-derived IL-15/4-1BBL activated NK-cell infusion; TRM, transplant-related mortality; MC, mononuclear cell; TLS, tumor lysis syndrome; PLS, passenger lymphocyte syndrome; NE, not evaluable.
aCulture method displays CD3 depleted PBMC’s, otherwise deviated selection method is mentioned in product characteristics.