Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects
Abstract One of the major hurdles that has hindered the success of chimeric antigen receptor (CAR) T cell therapies against solid tumors is on-target off-tumor (OTOT) toxicity due to sharing of the same epitopes on normal tissues. To elevate the safety profile of CAR-T cells, an affinity/avidity fin...
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-03-01
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| Series: | Molecular Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12943-024-01952-w |
| _version_ | 1827316191040897024 |
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| author | Yufei Wang Alicia Buck Brandon Piel Luann Zerefa Nithyassree Murugan Christian D. Coherd Andras G. Miklosi Haraman Johal Ricardo Nunes Bastos Kun Huang Miriam Ficial Yasmin Nabil Laimon Sabina Signoretti Zhou Zhong Song-My Hoang Gabriella M. Kastrunes Marion Grimaud Atef Fayed Hsien-Chi Yuan Quang-De Nguyen Tran Thai Elena V. Ivanova Cloud P. Paweletz Ming-Ru Wu Toni K. Choueiri Jon O. Wee Gordon J. Freeman David A. Barbie Wayne A. Marasco |
| author_facet | Yufei Wang Alicia Buck Brandon Piel Luann Zerefa Nithyassree Murugan Christian D. Coherd Andras G. Miklosi Haraman Johal Ricardo Nunes Bastos Kun Huang Miriam Ficial Yasmin Nabil Laimon Sabina Signoretti Zhou Zhong Song-My Hoang Gabriella M. Kastrunes Marion Grimaud Atef Fayed Hsien-Chi Yuan Quang-De Nguyen Tran Thai Elena V. Ivanova Cloud P. Paweletz Ming-Ru Wu Toni K. Choueiri Jon O. Wee Gordon J. Freeman David A. Barbie Wayne A. Marasco |
| author_sort | Yufei Wang |
| collection | DOAJ |
| description | Abstract One of the major hurdles that has hindered the success of chimeric antigen receptor (CAR) T cell therapies against solid tumors is on-target off-tumor (OTOT) toxicity due to sharing of the same epitopes on normal tissues. To elevate the safety profile of CAR-T cells, an affinity/avidity fine-tuned CAR was designed enabling CAR-T cell activation only in the presence of a highly expressed tumor associated antigen (TAA) but not when recognizing the same antigen at a physiological level on healthy cells. Using direct stochastic optical reconstruction microscopy (dSTORM) which provides single-molecule resolution, and flow cytometry, we identified high carbonic anhydrase IX (CAIX) density on clear cell renal cell carcinoma (ccRCC) patient samples and low-density expression on healthy bile duct tissues. A Tet-On doxycycline-inducible CAIX expressing cell line was established to mimic various CAIX densities, providing coverage from CAIX-high skrc-59 tumor cells to CAIX-low MMNK-1 cholangiocytes. Assessing the killing of CAR-T cells, we demonstrated that low-affinity/high-avidity fine-tuned G9 CAR-T has a wider therapeutic window compared to high-affinity/high-avidity G250 that was used in the first anti-CAIX CAR-T clinical trial but displayed serious OTOT effects. To assess the therapeutic effect of G9 on patient samples, we generated ccRCC patient derived organotypic tumor spheroid (PDOTS) ex vivo cultures and demonstrated that G9 CAR-T cells exhibited superior efficacy, migration and cytokine release in these miniature tumors. Moreover, in an RCC orthotopic mouse model, G9 CAR-T cells showed enhanced tumor control compared to G250. In summary, G9 has successfully mitigated OTOT side effects and in doing so has made CAIX a druggable immunotherapeutic target. |
| first_indexed | 2024-04-24T23:09:26Z |
| format | Article |
| id | doaj.art-90de263ab4864245afd1b4be5c061876 |
| institution | Directory Open Access Journal |
| issn | 1476-4598 |
| language | English |
| last_indexed | 2024-04-24T23:09:26Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Cancer |
| spelling | doaj.art-90de263ab4864245afd1b4be5c0618762024-03-17T12:17:32ZengBMCMolecular Cancer1476-45982024-03-0123111610.1186/s12943-024-01952-wAffinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effectsYufei Wang0Alicia Buck1Brandon Piel2Luann Zerefa3Nithyassree Murugan4Christian D. Coherd5Andras G. Miklosi6Haraman Johal7Ricardo Nunes Bastos8Kun Huang9Miriam Ficial10Yasmin Nabil Laimon11Sabina Signoretti12Zhou Zhong13Song-My Hoang14Gabriella M. Kastrunes15Marion Grimaud16Atef Fayed17Hsien-Chi Yuan18Quang-De Nguyen19Tran Thai20Elena V. Ivanova21Cloud P. Paweletz22Ming-Ru Wu23Toni K. Choueiri24Jon O. Wee25Gordon J. Freeman26David A. Barbie27Wayne A. Marasco28Department of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Medical Oncology, Dana-Farber Cancer InstituteDepartment of Medical Oncology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteONI (Oxford Nanoimaging Ltd.)ONI (Oxford Nanoimaging Ltd.)ONI (Oxford Nanoimaging Ltd.)Molecular Imaging Core, Dana-Farber Cancer InstituteDepartment of Pathology, Brigham and Women’s HospitalDepartment of Pathology, Brigham and Women’s HospitalHarvard Medical SchoolLUMICKSLUMICKSDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteLurie Family Imaging Center, Center for Biomedical Imaging in Oncology, Dana-Farber Cancer InstituteDepartment of Medical Oncology, Dana-Farber Cancer InstituteDepartment of Medical Oncology, Dana-Farber Cancer InstituteDepartment of Medical Oncology, Dana-Farber Cancer InstituteDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteHarvard Medical SchoolLowe Center for Thoracic Oncology, Dana-Farber Cancer InstituteHarvard Medical SchoolHarvard Medical SchoolDepartment of Cancer Immunology and Virology, Dana-Farber Cancer InstituteAbstract One of the major hurdles that has hindered the success of chimeric antigen receptor (CAR) T cell therapies against solid tumors is on-target off-tumor (OTOT) toxicity due to sharing of the same epitopes on normal tissues. To elevate the safety profile of CAR-T cells, an affinity/avidity fine-tuned CAR was designed enabling CAR-T cell activation only in the presence of a highly expressed tumor associated antigen (TAA) but not when recognizing the same antigen at a physiological level on healthy cells. Using direct stochastic optical reconstruction microscopy (dSTORM) which provides single-molecule resolution, and flow cytometry, we identified high carbonic anhydrase IX (CAIX) density on clear cell renal cell carcinoma (ccRCC) patient samples and low-density expression on healthy bile duct tissues. A Tet-On doxycycline-inducible CAIX expressing cell line was established to mimic various CAIX densities, providing coverage from CAIX-high skrc-59 tumor cells to CAIX-low MMNK-1 cholangiocytes. Assessing the killing of CAR-T cells, we demonstrated that low-affinity/high-avidity fine-tuned G9 CAR-T has a wider therapeutic window compared to high-affinity/high-avidity G250 that was used in the first anti-CAIX CAR-T clinical trial but displayed serious OTOT effects. To assess the therapeutic effect of G9 on patient samples, we generated ccRCC patient derived organotypic tumor spheroid (PDOTS) ex vivo cultures and demonstrated that G9 CAR-T cells exhibited superior efficacy, migration and cytokine release in these miniature tumors. Moreover, in an RCC orthotopic mouse model, G9 CAR-T cells showed enhanced tumor control compared to G250. In summary, G9 has successfully mitigated OTOT side effects and in doing so has made CAIX a druggable immunotherapeutic target.https://doi.org/10.1186/s12943-024-01952-wChimeric antigen receptor (CAR) TAffinity/avidity fine-tunedClear cell renal cell carcinoma (ccRCC)Carbonic anhydrase IX (CAIX)Direct stochastic optical reconstruction microscopy (dSTORM)On-target off-tumor (OTOT) toxicity |
| spellingShingle | Yufei Wang Alicia Buck Brandon Piel Luann Zerefa Nithyassree Murugan Christian D. Coherd Andras G. Miklosi Haraman Johal Ricardo Nunes Bastos Kun Huang Miriam Ficial Yasmin Nabil Laimon Sabina Signoretti Zhou Zhong Song-My Hoang Gabriella M. Kastrunes Marion Grimaud Atef Fayed Hsien-Chi Yuan Quang-De Nguyen Tran Thai Elena V. Ivanova Cloud P. Paweletz Ming-Ru Wu Toni K. Choueiri Jon O. Wee Gordon J. Freeman David A. Barbie Wayne A. Marasco Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects Molecular Cancer Chimeric antigen receptor (CAR) T Affinity/avidity fine-tuned Clear cell renal cell carcinoma (ccRCC) Carbonic anhydrase IX (CAIX) Direct stochastic optical reconstruction microscopy (dSTORM) On-target off-tumor (OTOT) toxicity |
| title | Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects |
| title_full | Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects |
| title_fullStr | Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects |
| title_full_unstemmed | Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects |
| title_short | Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects |
| title_sort | affinity fine tuning anti caix car t cells mitigate on target off tumor side effects |
| topic | Chimeric antigen receptor (CAR) T Affinity/avidity fine-tuned Clear cell renal cell carcinoma (ccRCC) Carbonic anhydrase IX (CAIX) Direct stochastic optical reconstruction microscopy (dSTORM) On-target off-tumor (OTOT) toxicity |
| url | https://doi.org/10.1186/s12943-024-01952-w |
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