Off-the-shelf invariant NKT cells expressing anti-PSCA CAR and IL-15 promote pancreatic cancer regression in mice
Zhenyu Dai, … , Michael A. Caligiuri, Jianhua Yu
Zhenyu Dai, … , Michael A. Caligiuri, Jianhua Yu
Published April 15, 2025
Citation Information: J Clin Invest. 2025;135(8):e179014. https://doi.org/10.1172/JCI179014.
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Research Article Immunology Oncology

Off-the-shelf invariant NKT cells expressing anti-PSCA CAR and IL-15 promote pancreatic cancer regression in mice

Abstract

Pancreatic ductal adenocarcinoma cancer (PDAC) continues to pose a significant health burden, with a 5-year survival rate of only 10%. Prostate stem cell antigen (PSCA) is highly expressed on the surface of tumor cells of most PDAC patients, with minimum expression in most normal tissues. Here, we generated cryopreserved, off-the-shelf, allogeneic PSCA chimeric antigen receptor (CAR) invariant NKT (iNKT) cells using human peripheral blood mononuclear cells as a cell source. In multiple in vitro and in vivo PDAC models, freshly manufactured PSCA CAR_sIL-15 iNKT cells and frozen-thawed, off-the-shelf PSCA CAR_sIL-15 iNKT cells demonstrate comparable efficacies, and both show remarkable suppression of PSCA-positive and gemcitabine-resistant PDAC. Importantly, off-the-shelf cryopreserved PSCA CAR_sIL-15 iNKT cells show equivalent efficacy when compared with PSCA CAR T cells using the same PSCA CAR and in the same PDAC model; however, PSCA CAR_sIL-15 iNKT cells do not appear to induce systemic toxicity or graft-versus-host disease, thus allowing for multiple infusions to control recurrent disease. Collectively, our study suggests that PSCA CAR_sIL-15 iNKT cells merit clinical investigation for PDAC patients exhibiting positive PSCA expression. The therapy could be given as a single agent or in combination with established therapeutic modalities for PDAC.

Authors

Zhenyu Dai, Zheng Zhu, Zhiyao Li, Lei Tian, Kun-Yu Teng, Hanyu Chen, Li-Shu Wang, Jianying Zhang, Laleh Melstrom, Michael A. Caligiuri, Jianhua Yu

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Figure 7

Cryopreserved and fresh PSCA CAR_sIL-15 iNKT cells exhibit comparable antitumor activity.

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Cryopreserved and fresh PSCA CAR_sIL-15 iNKT cells exhibit comparable an...
(A) In vitro cytotoxicity of off-the-shelf sIL-15 iNKT cells and off-the-shelf PSCA CAR_sIL-15 iNKT cells against PSCA+ PDAC cell lines, measured by RTCA at an E:T ratio 1:1. Experiments were repeated with 3 donors. (B) In vivo assessment of Capan-1 tumor burden was monitored by BMI during treatment with off-the-shelf cryopreserved sIL-15 iNKT cells and off-the-shelf cryopreserved PSCA CAR_sIL-15 iNKT cells until week 7. (C) Graphical summary of mouse tumor burden during treatment shown in B. Data are displayed as mean ± SD until day 30. ***P < 0.001 (2-way ANOVA). n = 6 for the untreated and sIL-15 groups. n = 7 for the PSCA CAR_sIL-15 group. (D) Overall Kaplan-Meier survival curve as the result of treatment shown in B. ***P < 0.001 (log-rank test). n = 6 for the untreated and the sIL-15 groups. n = 7 for the PSCA CAR_sIL-15 group. (E) NSG mice were injected with 5 × 105 Capan-1-luc cells on day 1. On day 7, mice received fresh PSCA CAR_sIL-15 iNKT cells or frozen PSCA CAR_sIL-15 iNKT cells (i.p. 4 × 106 and i.v. 2 × 106 /mouse, respectively). Tumor burden was monitored by BLI until day 49. (F) Overall Kaplan-Meier survival curve resulting from E. n = 6 per group. One mouse receiving fresh PSCA CAR_sIL-15 iNKT cells died accidentally during imaging. (G) NSG mice were injected with 5 × 105 Capan-1-luc cells on day 1. On day 7, mice were injected with off-the-shelf cryopreserved PSCA CAR-sIL-15 iNKT cells (i.p. 4 × 106 plus i.v. 2 × 106/mouse). Blood, bone marrow, lung, liver, pancreas, kidney, and spleen were harvested on days 1, 7, 14, and 21 after PSCA CAR_sIL-15 iNKT cell injection. PSCA CAR_sIL-15 iNKT cell persistence was detected by flow cytometry using hCD45+ (n = 3 or 4 mice per time point).