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Human adipose-derived stem cells genetically programmed to induce necroptosis for cancer immunotherapy

Abstracts

Herein, we present human adipose-derived stem cells (ADSCs) inserted with the receptor-interacting protein kinase-3 (RIP3) gene (RP@ADSCs), which induces cell necroptosis, for tumor immunotherapy. Necroptosis has characteristics of both apoptosis, such as programmed cell death, and necrosis, such as swelling and plasma membrane rupture, during which damage-related molecular patterns are released, triggering an immune response. Therefore, necroptosis has the potential to be used as an effective anticancer immunotherapy. RP@ADSCs were programmed to necroptosis after a particular time after being injected in vivo, and various pro-inflammatory cytokines secreted during the stem cell death process stimulated the immune system, showing local and sustained anticancer effects. It was confirmed that RIP3 protein expression increased in ADSCs after RP transfection. RP@ADSCs continued to induce ADSCs death for 7 days, and various pro-inflammatory cytokines were secreted through ADSCs death. The efficacy of RP@ADSCs-mediated immunotherapy was evaluated in mouse models bearing GL-26 (glioblastoma) and K1735 (melanoma), and it was found that RP resulted in an increase in the population of long-term cytotoxic T cells and a decrease in the population of regulatory T cells. This shows that RP@ADSCs have potential and applicability as an excellent anticancer immunotherapy agent in clinical practice.

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Fig. 1: Preparation and characterization of RP.
Fig. 2: In vitro gene transfection effect after treated with RP in ADSC.
Fig. 3: Analyses of RNA-seq in ADSCs and RP@ADSCs.
Fig. 4: Cancer-homing effect of RP@ADSC into K1735 tumor-bearing Balb/c mice.
Fig. 5: In vivo anticancer activity of RP@ADSC with K1735 tumor-bearing Balb/c mice.
Fig. 6: In vivo anticancer activity of RP@ADSCs with GL-26 tumor-bearing Balb/c nu/nu mice.
Fig. 7: In vivo toxicity evaluation.

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Acknowledgements

This research was supported by the Basic Science Research Program (NRF-2022R1A2B5B03001432) of the National Research Foundation of Korea which is funded by the Ministry of Science and ICT (MSIT).

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S. Bak contributed to the investigation, methodology, formal analysis, and writing—original and revised draft. K.S. Kim contributed to the methodology, formal analysis, and writing—original and revised draft. K. Na contributed to the conceptualization, project administration, supervision, funding acquisition, and writing—revised draft.

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Correspondence to Kun Na.

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The Institutional Animal Care and Use Committee of the Catholic University of Korea granted approval for all animal experiments.

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Bak, S., Kim, K.S. & Na, K. Human adipose-derived stem cells genetically programmed to induce necroptosis for cancer immunotherapy. Cancer Gene Ther 31, 995–1006 (2024). https://doi.org/10.1038/s41417-024-00794-4

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