A living enzymatic adenosine switch couples metabolic rescue to IL-12-driven antitumor immunity

Abstract Cancer cells remodel their metabolic environment to disable infiltrating immune cells. Adenosine is a principal mediator of this immunosuppression, impairing the metabolism and effector functions of T and natural killer (NK) cells. Blocking adenosine receptors or production within a redundant signaling network leaves the metabolite itself intact and has shown limited clinical efficacy. Adenosine deaminase (ADA) instead converts adenosine into inosine, a metabolite that can fuel T cell metabolism. We engineered non-pathogenic, tumor-tropic Escherichia coli K-12 DH5α to surface-display ADA, confining this metabolic switch to the tumor, and co-displayed interleukin-12 (IL-12) to couple metabolic rescue to local immune stimulation. Bacteria-displayed ADA alone restored metabolic and proliferative fitness in adenosine-suppressed lymphocytes to baseline, whereas incorporation of IL-12 coupled this rescue to enhanced effector function. Across multiple preclinical models of hematologic and solid malignancies, the dual-function bacteria were well tolerated, enhanced tumor control, prolonged survival, and induced durable protective immunity against melanoma rechallenge. The bacteria were enriched at tumor sites, reactivated quiescent immune compartments in leukemia, enhanced CAR-NK cell tumor trafficking and activity in pancreatic cancer, and outperformed recombinant proteins in breast cancer. These findings establish a modular living immunotherapeutic strategy that couples a local extracellular metabolic switch with localized cytokine delivery.

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Publication Details

Journal
Research Square
Published
2026-10-29
DOI
https://doi.org/10.21203/rs.3.rs-11368155/v1
Primary Topic
Cancer Research and Treatments
Type
preprint
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preprint

A living enzymatic adenosine switch couples metabolic rescue to IL-12-driven antitumor immunity

Jerome Ritz, Roman M. Shapiro, John Koreth, Mounica Vallurupalli et al.
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Research Square
Cancer Research and Treatments
preprint

A living enzymatic adenosine switch couples metabolic rescue to IL-12-driven antitumor immunity

Jerome Ritz, Roman M. Shapiro, John Koreth, Mounica Vallurupalli, Alaa Kassim Ali, Bartosz J. Grzywacz, Catherine J. Wu, Rizwan Romee, Stephanie Sendker, Robert J. Soiffer, Andrew A. Lane, José A. Cancelas, Rémy Duléry, Mohammad Rashidian, Daniel J. DeAngelo, Xingyu Deng, Andreia Maia, Jiahe Li, Mubin Tarannum, Eden Bobilev, Veronica W. Hui, Shaobo Yang, Ashley Hu, Isabella Salinas, Michael James, Kate Marinchev, Fuguo Liu, Maily Nguyen, Castilleja L. Albert, Alice Chen, Jacqueline Garcia, Jenny Ho, Anna Bader
preprint en

Abstract

Abstract Cancer cells remodel their metabolic environment to disable infiltrating immune cells. Adenosine is a principal mediator of this immunosuppression, impairing the metabolism and effector functions of T and natural killer (NK) cells. Blocking adenosine receptors or production within a redundant signaling network leaves the metabolite itself intact and has shown limited clinical efficacy. Adenosine deaminase (ADA) instead converts adenosine into inosine, a metabolite that can fuel T cell metabolism. We engineered non-pathogenic, tumor-tropic Escherichia coli K-12 DH5α to surface-display ADA, confining this metabolic switch to the tumor, and co-displayed interleukin-12 (IL-12) to couple metabolic rescue to local immune stimulation. Bacteria-displayed ADA alone restored metabolic and proliferative fitness in adenosine-suppressed lymphocytes to baseline, whereas incorporation of IL-12 coupled this rescue to enhanced effector function. Across multiple preclinical models of hematologic and solid malignancies, the dual-function bacteria were well tolerated, enhanced tumor control, prolonged survival, and induced durable protective immunity against melanoma rechallenge. The bacteria were enriched at tumor sites, reactivated quiescent immune compartments in leukemia, enhanced CAR-NK cell tumor trafficking and activity in pancreatic cancer, and outperformed recombinant proteins in breast cancer. These findings establish a modular living immunotherapeutic strategy that couples a local extracellular metabolic switch with localized cytokine delivery.

Research Square
Broad Institute (US), Beth Israel Deaconess Medical Center (US), Harvard University (US), University of Michigan (US), Dana-Farber Cancer Institute (US)
Good health and well-being
Cancer Research and Treatments
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