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Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer.

Sakamoto N, Ishikawa T, Kokura S, Okayama T, Oka K, Ideno M, Sakai F, Kato A, Tanabe M, Enoki T, Mineno J, Naito Y, Itoh Y, Yoshikawa T - J Transl Med (2015)

Bottom Line: Total cell population had a median expansion of 586-fold (range 95-1102), with a significantly pure (90.96 %) NK cell population.This NK cell therapy was very well tolerated with no severe adverse events.We successfully generated large numbers of activated NK cells from small quantities of blood without prior purification of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. jrsaka@koto.kpu-m.ac.jp.

ABSTRACT

Background: NK cells can destroy tumor cells without prior sensitization or immunization. Tumors often lose expression of MHC molecules and/or antigens. However, NK cells can lyse tumor cells in a non-MHC-restricted manner and independent of the expression of tumor-associated antigens. NK cells are therefore considered ideal for adoptive cancer immunotherapy; however the difficulty of obtaining large numbers of fully functional NK cells that are safe to administer deters its clinical use. This phase I clinical trial seeks to address this obstacle by first developing a novel system that expands large numbers of highly activated clinical grade NK cells, and second, determining if these cells are safe in a mono-treatment so they can be combined with other reagents in the next round of clinical trials.

Methods: Patients with unresectable, locally advanced and/or metastatic digestive cancer who did not succeed with standard therapy were enrolled. NK cells were expanded ex vivo by stimulating PBMCs with OK432, IL-2, and modified FN-CH296 induced T cells. Patients were administered autologous natural killer cell three times weekly via intravenous infusions in a dose-escalating manner (dose 0.5 × 10(9), 1.0 × 10(9), 2.0 × 10(9) cells/injection, three patients/one cohort).

Results: Total cell population had a median expansion of 586-fold (range 95-1102), with a significantly pure (90.96 %) NK cell population. Consequently, NK cells were expanded to approximately 4720-fold (range 1372-14,116) with cells being highly lytic in vitro and strongly expressing functional markers such as NKG2D and CD16. This NK cell therapy was very well tolerated with no severe adverse events. Although no clinical responses were observed, cytotoxicity of peripheral blood was elevated approximately twofolds up to 4 weeks post the last transfer.

Conclusion: We successfully generated large numbers of activated NK cells from small quantities of blood without prior purification of the cells. We also determined that the expanded cells were safe to administer in a monotherapy and are suitable for the next round of clinical trials where their efficacy will be tested combined with other reagents.

Trial registration: UMIN UMIN000007527.

No MeSH data available.


Related in: MedlinePlus

Treatment protocol. PBMCs were separated to prepare RN-T as stimulator cells. One week later, PBMCs were again separated similarly, and re-suspended in culture medium supplemented with heat-inactivated autologous plasma, IL-2 and OK-432. RN-T cells were added to the same flask or culture bag on day 0 and day 7. On days 21–22, the cultured cells were harvested and administered to the patients immediately. Expanded NK cells were intravenously injected for 60 min on days 0, 7, 14 in a dose-escalating manner (dose 0.5 × 109, 1 × 109, 2 × 109 cells/injection, three patients/one cohort). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the last administration of cultured cells. Blood samplings for immune monitoring were done just before the 1st and 3rd administration and 4 weeks after the 3rd administration. PBMCs peripheral blood mononuclear cells, RN-T cells RetroNectin®-induced T cells
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Fig1: Treatment protocol. PBMCs were separated to prepare RN-T as stimulator cells. One week later, PBMCs were again separated similarly, and re-suspended in culture medium supplemented with heat-inactivated autologous plasma, IL-2 and OK-432. RN-T cells were added to the same flask or culture bag on day 0 and day 7. On days 21–22, the cultured cells were harvested and administered to the patients immediately. Expanded NK cells were intravenously injected for 60 min on days 0, 7, 14 in a dose-escalating manner (dose 0.5 × 109, 1 × 109, 2 × 109 cells/injection, three patients/one cohort). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the last administration of cultured cells. Blood samplings for immune monitoring were done just before the 1st and 3rd administration and 4 weeks after the 3rd administration. PBMCs peripheral blood mononuclear cells, RN-T cells RetroNectin®-induced T cells

Mentions: The eligibility criteria for transferred cells was as follows: ① The cultured cell viability was more than 80 %. ② The mean purity value of the three cultured cells was more than 50 %. ③ The number of transferred cells was 70–130 % of the number that was set in each cohort. The patients were divided into three cohorts of three to four patients each: Cohort 1, 0.5 × 109 cells per dose; Cohort 2, 1.0 × 109 cells per dose; and Cohort 3, 2.0 × 109 cells per dose. Expanded NK cells that passed quality tests were intravenously injected for 60 min on days 0, 7, 14 (Fig. 1). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the third cell infusion. DLT was defined as grade ≥3 for any adverse event related to the administration of cultured cells. If no DLT was observed in the previous cohort, another cohort was treated at the next higher dose. There was no intra-patient dose escalation in this study.Fig. 1


Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer.

Sakamoto N, Ishikawa T, Kokura S, Okayama T, Oka K, Ideno M, Sakai F, Kato A, Tanabe M, Enoki T, Mineno J, Naito Y, Itoh Y, Yoshikawa T - J Transl Med (2015)

Treatment protocol. PBMCs were separated to prepare RN-T as stimulator cells. One week later, PBMCs were again separated similarly, and re-suspended in culture medium supplemented with heat-inactivated autologous plasma, IL-2 and OK-432. RN-T cells were added to the same flask or culture bag on day 0 and day 7. On days 21–22, the cultured cells were harvested and administered to the patients immediately. Expanded NK cells were intravenously injected for 60 min on days 0, 7, 14 in a dose-escalating manner (dose 0.5 × 109, 1 × 109, 2 × 109 cells/injection, three patients/one cohort). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the last administration of cultured cells. Blood samplings for immune monitoring were done just before the 1st and 3rd administration and 4 weeks after the 3rd administration. PBMCs peripheral blood mononuclear cells, RN-T cells RetroNectin®-induced T cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4548900&req=5

Fig1: Treatment protocol. PBMCs were separated to prepare RN-T as stimulator cells. One week later, PBMCs were again separated similarly, and re-suspended in culture medium supplemented with heat-inactivated autologous plasma, IL-2 and OK-432. RN-T cells were added to the same flask or culture bag on day 0 and day 7. On days 21–22, the cultured cells were harvested and administered to the patients immediately. Expanded NK cells were intravenously injected for 60 min on days 0, 7, 14 in a dose-escalating manner (dose 0.5 × 109, 1 × 109, 2 × 109 cells/injection, three patients/one cohort). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the last administration of cultured cells. Blood samplings for immune monitoring were done just before the 1st and 3rd administration and 4 weeks after the 3rd administration. PBMCs peripheral blood mononuclear cells, RN-T cells RetroNectin®-induced T cells
Mentions: The eligibility criteria for transferred cells was as follows: ① The cultured cell viability was more than 80 %. ② The mean purity value of the three cultured cells was more than 50 %. ③ The number of transferred cells was 70–130 % of the number that was set in each cohort. The patients were divided into three cohorts of three to four patients each: Cohort 1, 0.5 × 109 cells per dose; Cohort 2, 1.0 × 109 cells per dose; and Cohort 3, 2.0 × 109 cells per dose. Expanded NK cells that passed quality tests were intravenously injected for 60 min on days 0, 7, 14 (Fig. 1). We investigated the dose-limiting toxicity (DLT) occurring over a 28-day period after the third cell infusion. DLT was defined as grade ≥3 for any adverse event related to the administration of cultured cells. If no DLT was observed in the previous cohort, another cohort was treated at the next higher dose. There was no intra-patient dose escalation in this study.Fig. 1

Bottom Line: Total cell population had a median expansion of 586-fold (range 95-1102), with a significantly pure (90.96 %) NK cell population.This NK cell therapy was very well tolerated with no severe adverse events.We successfully generated large numbers of activated NK cells from small quantities of blood without prior purification of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan. jrsaka@koto.kpu-m.ac.jp.

ABSTRACT

Background: NK cells can destroy tumor cells without prior sensitization or immunization. Tumors often lose expression of MHC molecules and/or antigens. However, NK cells can lyse tumor cells in a non-MHC-restricted manner and independent of the expression of tumor-associated antigens. NK cells are therefore considered ideal for adoptive cancer immunotherapy; however the difficulty of obtaining large numbers of fully functional NK cells that are safe to administer deters its clinical use. This phase I clinical trial seeks to address this obstacle by first developing a novel system that expands large numbers of highly activated clinical grade NK cells, and second, determining if these cells are safe in a mono-treatment so they can be combined with other reagents in the next round of clinical trials.

Methods: Patients with unresectable, locally advanced and/or metastatic digestive cancer who did not succeed with standard therapy were enrolled. NK cells were expanded ex vivo by stimulating PBMCs with OK432, IL-2, and modified FN-CH296 induced T cells. Patients were administered autologous natural killer cell three times weekly via intravenous infusions in a dose-escalating manner (dose 0.5 × 10(9), 1.0 × 10(9), 2.0 × 10(9) cells/injection, three patients/one cohort).

Results: Total cell population had a median expansion of 586-fold (range 95-1102), with a significantly pure (90.96 %) NK cell population. Consequently, NK cells were expanded to approximately 4720-fold (range 1372-14,116) with cells being highly lytic in vitro and strongly expressing functional markers such as NKG2D and CD16. This NK cell therapy was very well tolerated with no severe adverse events. Although no clinical responses were observed, cytotoxicity of peripheral blood was elevated approximately twofolds up to 4 weeks post the last transfer.

Conclusion: We successfully generated large numbers of activated NK cells from small quantities of blood without prior purification of the cells. We also determined that the expanded cells were safe to administer in a monotherapy and are suitable for the next round of clinical trials where their efficacy will be tested combined with other reagents.

Trial registration: UMIN UMIN000007527.

No MeSH data available.


Related in: MedlinePlus