Novel low‐avidity glypican‐3 specific CARTs resist exhaustion and mediate durable antitumor effects against HCC
Background and Aims Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low‐affinity monoclonal antibo...
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Published in | Hepatology (Baltimore, Md.) Vol. 76; no. 2; pp. 330 - 344 |
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Main Authors | , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
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United States
Wolters Kluwer Health, Inc
01.08.2022
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Abstract | Background and Aims
Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low‐affinity monoclonal antibodies (mAbs) and low‐avidity CARTs for HCC and to test the hypothesis that low‐avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects.
Methods and Results
New human glypican‐3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3‐specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high‐affinity mAb, but with approximately 17‐fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low‐avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high‐avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5‐fold more of 8F8‐BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor‐infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs.
Conclusion
The low‐avidity 8F8‐BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high‐avidity CARTs. |
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AbstractList | Background and AimsChimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low‐affinity monoclonal antibodies (mAbs) and low‐avidity CARTs for HCC and to test the hypothesis that low‐avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects.Methods and ResultsNew human glypican‐3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3‐specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high‐affinity mAb, but with approximately 17‐fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low‐avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high‐avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5‐fold more of 8F8‐BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor‐infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs.ConclusionThe low‐avidity 8F8‐BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high‐avidity CARTs. Background and Aims Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low‐affinity monoclonal antibodies (mAbs) and low‐avidity CARTs for HCC and to test the hypothesis that low‐avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects. Methods and Results New human glypican‐3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3‐specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high‐affinity mAb, but with approximately 17‐fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low‐avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high‐avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5‐fold more of 8F8‐BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor‐infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs. Conclusion The low‐avidity 8F8‐BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high‐avidity CARTs. Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low-affinity monoclonal antibodies (mAbs) and low-avidity CARTs for HCC and to test the hypothesis that low-avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects. New human glypican-3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3-specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high-affinity mAb, but with approximately 17-fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low-avidity 8F8 CARTs killed both hGPC3 and hGPC3 HCC tumor cells to the same extent as high-avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5-fold more of 8F8-BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor-infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs. The low-avidity 8F8-BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high-avidity CARTs. Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low-affinity monoclonal antibodies (mAbs) and low-avidity CARTs for HCC and to test the hypothesis that low-avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects.BACKGROUND AND AIMSChimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low-affinity monoclonal antibodies (mAbs) and low-avidity CARTs for HCC and to test the hypothesis that low-avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects.New human glypican-3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3-specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high-affinity mAb, but with approximately 17-fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low-avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high-avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5-fold more of 8F8-BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor-infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs.METHODS AND RESULTSNew human glypican-3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3-specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high-affinity mAb, but with approximately 17-fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low-avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high-avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5-fold more of 8F8-BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor-infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs.The low-avidity 8F8-BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high-avidity CARTs.CONCLUSIONThe low-avidity 8F8-BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high-avidity CARTs. |
Author | Hussein, Mohamed S. He, Yukai Jiang, Xiaotao Mao, Rui Kehinde‐Ige, Mercy Sadek, Ramses Qiu, Xiangguo Peng, Yibing Caraballo Galva, Leidy D. Brody, Pierce Zhang, Huajun He, Aiwu Ruth Shi, Huidong |
AuthorAffiliation | 4 Lombardi Cancer Center, Georgetown University, Washington, District of Columbia, USA 3 Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Manitoba, Canada 1 Georgia Cancer Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA 2 Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA |
AuthorAffiliation_xml | – name: 2 Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA – name: 4 Lombardi Cancer Center, Georgetown University, Washington, District of Columbia, USA – name: 3 Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Manitoba, Canada – name: 1 Georgia Cancer Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA |
Author_xml | – sequence: 1 givenname: Leidy D. orcidid: 0000-0003-4263-8419 surname: Caraballo Galva fullname: Caraballo Galva, Leidy D. organization: Augusta University – sequence: 2 givenname: Xiaotao orcidid: 0000-0001-7571-1593 surname: Jiang fullname: Jiang, Xiaotao organization: Augusta University – sequence: 3 givenname: Mohamed S. orcidid: 0000-0001-9277-713X surname: Hussein fullname: Hussein, Mohamed S. organization: Augusta University – sequence: 4 givenname: Huajun surname: Zhang fullname: Zhang, Huajun organization: University of Manitoba – sequence: 5 givenname: Rui surname: Mao fullname: Mao, Rui organization: Augusta University – sequence: 6 givenname: Pierce surname: Brody fullname: Brody, Pierce organization: Augusta University – sequence: 7 givenname: Yibing surname: Peng fullname: Peng, Yibing organization: Augusta University – sequence: 8 givenname: Aiwu Ruth surname: He fullname: He, Aiwu Ruth organization: Georgetown University – sequence: 9 givenname: Mercy surname: Kehinde‐Ige fullname: Kehinde‐Ige, Mercy organization: Augusta University – sequence: 10 givenname: Ramses surname: Sadek fullname: Sadek, Ramses organization: Augusta University – sequence: 11 givenname: Xiangguo surname: Qiu fullname: Qiu, Xiangguo organization: University of Manitoba – sequence: 12 givenname: Huidong orcidid: 0000-0003-1137-3390 surname: Shi fullname: Shi, Huidong organization: Augusta University – sequence: 13 givenname: Yukai orcidid: 0000-0001-7636-5549 surname: He fullname: He, Yukai email: yhe@augusta.edu organization: Augusta University |
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Notes | Funding information Supported by Augusta University start‐up fund Leidy D. Caraballo Galva, Xiaotao Jiang, Mohamed S. Hussein, and Huajun Zhang contributed equally to this work. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Present address Xiaotao Jiang, Department of Immunology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China AUTHOR CONTRIBUTIONS Huajun Zhang, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China YH conceive the hypothesis and is in charge of the overall experimental design, experiments, and data analysis and summary. YH, XJ, LDCG, MSH, and HZ designed and conducted the majority of the experiments, analyzed and summarized the data. HZ and XQ did the affinity study. ARH conducted IHC staining on liver tissues and analyzed data. RM, PB, and YP prepared the CAR lentivectors and helped conducted the in vivo animal studies. MK-I and HS performed the RNAseq analysis. RS did the statistical analysis. YH, XJ, LDCG, MSH, and HZ wrote the manuscript. |
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and normal myeloid lineage without HSC toxicity publication-title: Leukemia doi: 10.1038/leu.2016.35 |
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Snippet | Background and Aims
Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers.... Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function... Background and AimsChimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers.... |
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SubjectTerms | Affinity Antigens Antitumor activity Apoptosis Avidity Blood cancer Chimeric antigen receptors Epitopes Hematological diseases Heparan sulfate proteoglycans Hepatology Lymphocytes T Monoclonal antibodies Solid tumors Tumor cells Tumor-infiltrating lymphocytes Tumors Xenografts |
Title | Novel low‐avidity glypican‐3 specific CARTs resist exhaustion and mediate durable antitumor effects against HCC |
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