Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells
Abstract Adoptive cell therapies (ACT) have shown reduced efficacy against solid tumor malignancies compared to hematologic malignancies, partly due to the immunosuppressive nature of the tumor microenvironment (TME). ACT efficacy may be enhanced with pleiotropic cytokines that remodel the TME; howe...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-024-07410-z |
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| author | Mara C. Inniss Sean G. Smith Dan Jun Li Benjamin Primack Dexue Sun Grace Y. Olinger Kerri-Lynn Sheahan Theresa Ross Meghan Langley Violet Young Andres Alvarado Shabnam Davoodi Jiefei Geng Michael Schebesta Michelle L. Ols Jeremy Tchaicha Jan ter Meulen Dhruv K. Sethi |
| author_facet | Mara C. Inniss Sean G. Smith Dan Jun Li Benjamin Primack Dexue Sun Grace Y. Olinger Kerri-Lynn Sheahan Theresa Ross Meghan Langley Violet Young Andres Alvarado Shabnam Davoodi Jiefei Geng Michael Schebesta Michelle L. Ols Jeremy Tchaicha Jan ter Meulen Dhruv K. Sethi |
| author_sort | Mara C. Inniss |
| collection | DOAJ |
| description | Abstract Adoptive cell therapies (ACT) have shown reduced efficacy against solid tumor malignancies compared to hematologic malignancies, partly due to the immunosuppressive nature of the tumor microenvironment (TME). ACT efficacy may be enhanced with pleiotropic cytokines that remodel the TME; however, their expression needs to be tightly controlled to avoid systemic toxicities. Here we show T cells can be armored with membrane-bound cytokines with surface expression regulated using drug-responsive domains (DRDs) developed from the 260-amino acid protein human carbonic anhydrase 2 (CA2). The CA2-DRD can be stabilized in vitro and in vivo with the FDA-approved small-molecule CA2 inhibitor acetazolamide (ACZ). We develop conditional degrons using library-based screening of mutants and show characterization of one DRD using crystallography and molecular dynamics (MD) simulations. Using protein-engineering solutions to increase the valency of DRDs fused to the cargo we have developed “modulation hubs” and show tight regulation of membrane-bound cytokines IL2, IL12, IL15, IL21, IL23, and IFNα in genetically engineered T cells. Finally, CA2-DRD regulated IL12 mediates regulated efficacy in a solid tumor model. Regulation of pleotropic cytokines potentially paves the way to safely use these powerful cytokines in ACT for cancer treatment. |
| format | Article |
| id | doaj-art-f2dec03d1ee9463aad8e9f1b30914350 |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-f2dec03d1ee9463aad8e9f1b309143502025-01-12T12:35:46ZengNature PortfolioCommunications Biology2399-36422025-01-018111610.1038/s42003-024-07410-zCarbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cellsMara C. Inniss0Sean G. Smith1Dan Jun Li2Benjamin Primack3Dexue Sun4Grace Y. Olinger5Kerri-Lynn Sheahan6Theresa Ross7Meghan Langley8Violet Young9Andres Alvarado10Shabnam Davoodi11Jiefei Geng12Michael Schebesta13Michelle L. Ols14Jeremy Tchaicha15Jan ter Meulen16Dhruv K. Sethi17Obsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsObsidian TherapeuticsAbstract Adoptive cell therapies (ACT) have shown reduced efficacy against solid tumor malignancies compared to hematologic malignancies, partly due to the immunosuppressive nature of the tumor microenvironment (TME). ACT efficacy may be enhanced with pleiotropic cytokines that remodel the TME; however, their expression needs to be tightly controlled to avoid systemic toxicities. Here we show T cells can be armored with membrane-bound cytokines with surface expression regulated using drug-responsive domains (DRDs) developed from the 260-amino acid protein human carbonic anhydrase 2 (CA2). The CA2-DRD can be stabilized in vitro and in vivo with the FDA-approved small-molecule CA2 inhibitor acetazolamide (ACZ). We develop conditional degrons using library-based screening of mutants and show characterization of one DRD using crystallography and molecular dynamics (MD) simulations. Using protein-engineering solutions to increase the valency of DRDs fused to the cargo we have developed “modulation hubs” and show tight regulation of membrane-bound cytokines IL2, IL12, IL15, IL21, IL23, and IFNα in genetically engineered T cells. Finally, CA2-DRD regulated IL12 mediates regulated efficacy in a solid tumor model. Regulation of pleotropic cytokines potentially paves the way to safely use these powerful cytokines in ACT for cancer treatment.https://doi.org/10.1038/s42003-024-07410-z |
| spellingShingle | Mara C. Inniss Sean G. Smith Dan Jun Li Benjamin Primack Dexue Sun Grace Y. Olinger Kerri-Lynn Sheahan Theresa Ross Meghan Langley Violet Young Andres Alvarado Shabnam Davoodi Jiefei Geng Michael Schebesta Michelle L. Ols Jeremy Tchaicha Jan ter Meulen Dhruv K. Sethi Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells Communications Biology |
| title | Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells |
| title_full | Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells |
| title_fullStr | Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells |
| title_full_unstemmed | Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells |
| title_short | Carbonic anhydrase 2-derived drug-responsive domain regulates membrane-bound cytokine expression and function in engineered T cells |
| title_sort | carbonic anhydrase 2 derived drug responsive domain regulates membrane bound cytokine expression and function in engineered t cells |
| url | https://doi.org/10.1038/s42003-024-07410-z |
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