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 Article has an altmetric score of 17

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 6

PSCA CAR_sIL-15 iNKT cells overcome gemcitabine resistance in PDAC.

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PSCA CAR_sIL-15 iNKT cells overcome gemcitabine resistance in PDAC. (A) ...
(A) MFI of PSCA (blue solid histograms) on GR cell lines (Capan-1 GR and MIA Paca-2 GR) compared with parental cell lines (red solid histograms) as measured flow cytometry. (B) Cytotoxicity of gemcitabine measured by RTCA in the presence of different concentrations of gemcitabine on GR and parental PDAC cell lines. Capan-1 GR and MIA Paca-2 GR were not while their parental cell lines were killed by gemcitabine at the concentrations of 1.6 μM and 3.2 μM. (C) Cytotoxicity of sIL-15 iNKT cells and PSCA CAR_sIL-15 iNKT cells against GR PDAC cell lines (Capan-1 GR and MIA Paca-2 GR) and the parental PDAC cell lines at an E:T ratio of 1:2, measured by RTCA assay. PSCA CAR_sIL-15 iNKT cells maintained their potent killing ability against Capan-1 GR and MIA PaCa-2 GR cells compared with the parental cells. (A–C) Experiments were repeated 3 times or with 3 different donors. Expression of T cell activation markers CD69 (D) and CD25 (E) on sIL-15 iNKT cells and PSCA CAR_sIL-15 iNKT cells following a 24-hour coincubation with Capan-1 GR, Capan-1, MIA PaCa-2 GR, or MIA PaCa-2 (gated on EGFR+ cells). Data are represented as mean ± SD (n = 3). There were comparable levels of CD69 and CD25 expression in PSCA CAR_sIL-15 iNKT cells when they were coincubated with the GR cells and the parental cells. (F) The parent cell lines Capan-1_luc and Capan-1 GR_luc were injected i.p. (2 × 105 cells/mouse). Three days later, tumor engraftment was confirmed by BLI and visually displayed in vivo weekly up to 8 weeks. The fold changes of BLI for each treatment group were measured. n = 4 for the untreated group. n = 5 for the sIL-15 group. n = 6 for the PSCA CAR_sIL-15 group. (G) Overall Kaplan-Meier survival curve. log-rank test (n = 4 for the untreated group. n = 5 for the sIL-15 group. n = 6 for the PSCA CAR_sIL-15 group.). (H) The growth of the tumor was monitored by BLI imaging until week 8. A single dose of PSCA CAR_sIL-15 iNKT cells still significantly suppressed Capan-1 GR tumor progression, and the antitumor effect of CAR_sIL-15 iNKT cells was not different when compared with the Capan-1 mouse tumor.