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Expansion of KRAS hotspot mutations reactive T cells from human pancreatic tumors using autologous T cells as the antigen-presenting cells

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Abstract

Adoptive cell therapy (ACT) with expanded tumor-infiltrating lymphocytes (TIL) or TCR gene-modified T cells (TCR-T) that recognize mutant KRAS neo-antigens can mediate tumor regression in patients with advanced pancreatic ductal adenocarcinoma (PDAC) (Tran et al in N Engl J Med, 375:2255–2262, 2016; Leidner et al in N Engl J Med, 386:2112–2119, 2022). The mutant KRAS-targeted ACT holds great potential to achieve durable clinical responses for PDAC, which has had no meaningful improvement over 40 years. However, the wide application of mutant KRAS-centric ACT is currently limited by the rarity of TIL that recognize the mutant KRAS. In addition, PDAC is generally recognized as a poorly immunogenic tumor, and TILs in PDAC are less abundant than in immunogenic tumors such as melanoma. To increase the success rate of TIL production, we adopted a well-utilized K562-based artificial APC (aAPC) that expresses 4-1BBL as the costimulatory molecules to enhance the TIL production from PDCA. However, stimulation with K562-based aAPC led to a rapid loss of specificity to mutant KRAS. To selectively expand neo-antigen-specific T cells, particularly mKRAS, from the TILs, we used tandem mini gene-modified autologous T cells (TMG-T) as the novel aAPC. Using this modified IVS protocol, we successfully generated TIL cultures specifically reactive to mKRAS (G12V). We believe that autologous TMG-T cells provide a reliable source of autologous APC to expand a rare population of neoantigen-specific T cells in TILs.

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Data availability

The data supporting this study's findings are available from the corresponding author, GY, upon reasonable request.

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Acknowledgements

The authors would like to thank Drs. Eric Tran and Rom Leidner of Earle A. Chiles Research Institute for helpful discussions on topics related to this work. This work is supported by the Jiangsu Clinical Medical Center (innovation platform) construction project (No. YXZXA2016006, to XW) and Shanghai Pujiang Program (No.20PJ1417400, to GY).

Funding

It is sponsored by Jiangsu Clinical Medical Center (innovation platform) construction project YXZXA2016006 and Shanghai Pujiang Program 20PJ1417400.

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Authors and Affiliations

Authors

Contributions

SW, XZ, XZ, and MW are joint first authors. XW and GY are PIs of research projects and study guarantors. XW, HH, and GY contributed to the study's concept and design. SW, XZ, XZ, CG, XJ, ML, RS, DZ, AY, and YF collected samples, performed the experiments, and analyzed the data. GY and HH drafted the manuscript. XW and BAF contributed to interpreting the results and critical revision of the manuscript and approved the final version. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Hong-Ming Hu, Guangjie Yu or Xinbo Wang.

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Competing interests

Xiaohui Zhang, Xuemei Zou, Maorong Wen, Chi Gan, Xiaochun Jiang, and Guangjie Yu are full-time employees of ImmuXell ltd. They have filed patent applications related to the TCR technology and could potentially receive licensing royalties. All other authors have no conflict of interest to declare. Hong-Ming Hu and Bernard A. Fox is the founder of UbiVAC, and UbiVAC is a potential licensee of technology developed in this study.

Conflict of interest

Xiaohui Zhang, Xuemei Zou, Maorong Wen, Chi Gan, Xiaochun Jiang, and Guangjie Yu are full-time employees of ImmuXell ltd. They have filed patent applications related to the TCR technology and could potentially receive licensing royalties. All other authors have no conflict of interest to declare. Hong-Ming Hu and Bernard A. Fox is the founder of UbiVAC, and UbiVAC is a potential licensee of technology developed in this study.

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This study was approved by the Ethics Committee of Jinling Hospital (Nanjing, China), reference number 2020NJKY-020–01.

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Wang, S., Zhang, X., Zou, X. et al. Expansion of KRAS hotspot mutations reactive T cells from human pancreatic tumors using autologous T cells as the antigen-presenting cells. Cancer Immunol Immunother 72, 1301–1313 (2023). https://doi.org/10.1007/s00262-022-03335-w

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