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Production of an interleukin-10 blocking antibody by genetically engineered macrophages increases cancer cell death in human gastrointestinal tumor slice cultures

Cancer Gene Therapy volume 30, pages 1227–1233 (2023)Cite this article

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Abstract

Although it can promote effector T-cell function, the summative effect of interleukin-10 (IL-10) in the tumor microenvironment (TME) appears to be suppressive; therefore, blocking this critical regulatory cytokine has therapeutic potential to enhance antitumor immune function. As macrophages efficiently localize to the TME, we hypothesized that they could be used as a delivery vehicle for drugs designed to block this pathway. To test our hypothesis, we created and evaluated genetically engineered macrophages (GEMs) that produce an IL-10-blocking antibody (αIL-10). Healthy donor human peripheral blood mononuclear cells were differentiated and transduced with a novel lentivirus (LV) encoding BT-063, a humanized αIL-10 antibody. The efficacy of αIL-10 GEMs was assessed in human gastrointestinal tumor slice culture models developed from resected specimens of pancreatic ductal adenocarcinoma primary tumors and colorectal cancer liver metastases. LV transduction led to sustained production of BT-063 by αIL-10 GEMs for at least 21 days. Transduction did not alter GEM phenotype as evaluated by flow cytometry, but αIL-10 GEMs produced measurable quantities of BT-063 in the TME that was associated with an ~5-fold higher rate of tumor cell apoptosis than control.

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Fig. 1: Lentiviral constructs.
Fig. 2: Monocyte-derived macrophages are efficiently transduced and produce IL-10-neutralizing antibody (BT-063).
Fig. 3: αIL-10 GEMs have similar phenotype to CD19t GEMs and non-transduced macrophages.
Fig. 4: αIL-10 GEMs decrease IL-10 concentrations and increase tumor cell death in human tumor slice culture.

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Data availability

All data relevant to the study are included in the article or uploaded as online Supplementary Information. Source data will be made available upon direct request to the corresponding author.

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Acknowledgements

We acknowledge Dr. James Oh Park, Raymond S. Yeung and Jonathan G. Sham and our patients for providing tumor specimens for this research. We acknowledge the NIH S10 grant (S10 OD016240) funding the Leica SP8X confocal microscope and Dr. Nathaniel Peters at the University of Washington W.M. Keck Microscopy Center for assistance with live image confocal microscopy.

Funding

VGP received funding from the US Army Medical Research Acquisition Activity (USAMRAA; CA150370P2), Seattle Translational Tumor Research, Brotman Baty Institute for Precision Medicine, and Merck Investigator Studies Program. TSK received funding from the UW/Fred Hutchinson Cancer Center Support Grant (CCSG), P30 CA015704, New Investigator Award. This work was also supported by the Washington Research Foundation, Stand Up to Cancer, and the National Institute of Neurological Disorders and Stroke, the National Institutes of Health (R25NS079200 to CIE), and the National Cancer Institute, National Institutes of Health (U54CA193461-03 and R01CA195718-02 to ECH).

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Authors and Affiliations

  1. Department of Surgery, University of Washington School of Medicine, Seattle, WA, USA

    Kevin P. Labadie, Sara K. Daniel, Xiuyun Jiang, Kevin M. Sullivan, Alan F. Utria, Heidi L. Kenerson, Teresa S. Kim & Venu G. Pillarisetty

  2. Ben Towne Center for Childhood Cancer Research, Seattle Children’s Research Institute, Seattle, WA, USA

    Shannon A. Kreuser, Katherine J. Brempelis & Courtney A. Crane

  3. Brotman Baty Institute for Precision Medicine, Seattle, WA, USA

    Venu G. Pillarisetty

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Contributions

VGP and CAC conceived the idea, designed experiments, interpreted data, and provided funding and resources. KPL, SAK and KJB designed and performed experiments and analyzed and interpreted data. SKD, XJ, KMS, AFU and HLK assisted on slice culture experiments, collection and interpretation of data. SAK and KJB generated CAR T cells. KPL and VGP wrote the manuscript with input from SAK, KJB, KMS and CAC. All authors contributed feedback for the final manuscript.

Corresponding author

Correspondence to Venu G. Pillarisetty.

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COMPETING INTERESTS

The authors declare that that they have read and understood the policy on declarations of interest. VGP is a member of the scientific advisory board for TriSalus Life Sciences. He served as a consultant for Merck & Company in 2018, GlaxoSmithKline in 2019, Imvax in 2019, Takeda in 2020, Umoja and Sensei in 2022. He has had research funding from AstraZeneca (completed), Ipsen (completed), Merck (completed), NGM (completed), and OncoResponse (current). The funders had no role in the conceptualization, design, data collection, analysis, decision to publish or preparation of the manuscript. CAC is an inventor on issued patent US20170087185A1 “Genetic engineering of macrophages for immunotherapy”.

Ethical approval

All investigations were performed according to the principles expressed in the Declaration of Helsinki. Samples for slice culture were procured from patients undergoing hepatic or pancreatic resection who provided prior written-informed consent under research protocol (no. 1852) approved by the University of Washington Institutional Review Board.

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Labadie, K.P., Kreuser, S.A., Brempelis, K.J. et al. Production of an interleukin-10 blocking antibody by genetically engineered macrophages increases cancer cell death in human gastrointestinal tumor slice cultures. Cancer Gene Ther 30, 1227–1233 (2023). https://doi.org/10.1038/s41417-023-00632-z

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