Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22
Chimeric antigen receptor (CAR) T cell therapies targeting B cell-restricted antigens CD19, CD20, or CD22 can produce potent clinical responses for some B cell malignancies, but relapse remains common. Camelid single-domain antibodies (sdAbs or nanobodies) are smaller, simpler, and easier to recombi...
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Elsevier
2024-03-01
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| Series: | Molecular Therapy: Oncology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2950329924000171 |
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| author | Scott McComb Mehdi Arbabi-Ghahroudi Kevin A. Hay Brian A. Keller Sharlene Faulkes Michael Rutherford Tina Nguyen Alex Shepherd Cunle Wu Anne Marcil Annie Aubry Greg Hussack Devanand M. Pinto Shannon Ryan Shalini Raphael Henk van Faassen Ahmed Zafer Qin Zhu Susanne Maclean Anindita Chattopadhyay Komal Gurnani Rénald Gilbert Christine Gadoury Umar Iqbal Dorothy Fatehi Anna Jezierski Jez Huang Robert A. Pon Mhairi Sigrist Robert A. Holt Brad H. Nelson Harold Atkins Natasha Kekre Eric Yung John Webb Julie S. Nielsen Risini D. Weeratna |
| author_facet | Scott McComb Mehdi Arbabi-Ghahroudi Kevin A. Hay Brian A. Keller Sharlene Faulkes Michael Rutherford Tina Nguyen Alex Shepherd Cunle Wu Anne Marcil Annie Aubry Greg Hussack Devanand M. Pinto Shannon Ryan Shalini Raphael Henk van Faassen Ahmed Zafer Qin Zhu Susanne Maclean Anindita Chattopadhyay Komal Gurnani Rénald Gilbert Christine Gadoury Umar Iqbal Dorothy Fatehi Anna Jezierski Jez Huang Robert A. Pon Mhairi Sigrist Robert A. Holt Brad H. Nelson Harold Atkins Natasha Kekre Eric Yung John Webb Julie S. Nielsen Risini D. Weeratna |
| author_sort | Scott McComb |
| collection | DOAJ |
| description | Chimeric antigen receptor (CAR) T cell therapies targeting B cell-restricted antigens CD19, CD20, or CD22 can produce potent clinical responses for some B cell malignancies, but relapse remains common. Camelid single-domain antibodies (sdAbs or nanobodies) are smaller, simpler, and easier to recombine than single-chain variable fragments (scFvs) used in most CARs, but fewer sdAb-CARs have been reported. Thus, we sought to identify a therapeutically active sdAb-CAR targeting human CD22. Immunization of an adult Llama glama with CD22 protein, sdAb-cDNA library construction, and phage panning yielded >20 sdAbs with diverse epitope and binding properties. Expressing CD22-sdAb-CAR in Jurkat cells drove varying CD22-specific reactivity not correlated with antibody affinity. Changing CD28- to CD8-transmembrane design increased CAR persistence and expression in vitro. CD22-sdAb-CAR candidates showed similar CD22-dependent CAR-T expansion in vitro, although only membrane-proximal epitope targeting CD22-sdAb-CARs activated direct cytolytic killing and extended survival in a lymphoma xenograft model. Based on enhanced survival in blinded xenograft studies, a lead CD22sdCAR-T was selected, achieving comparable complete responses to a benchmark short linker m971-scFv CAR-T in high-dose experiments. Finally, immunohistochemistry and flow cytometry confirm tissue and cellular-level specificity of the lead CD22-sdAb. This presents a complete report on preclinical development of a novel CD22sdCAR therapeutic. |
| format | Article |
| id | doaj-art-2b9541fc680c4b00a851c3c1cf34b814 |
| institution | Kabale University |
| issn | 2950-3299 |
| language | English |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Oncology |
| spelling | doaj-art-2b9541fc680c4b00a851c3c1cf34b8142024-11-24T04:15:30ZengElsevierMolecular Therapy: Oncology2950-32992024-03-01321200775Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22Scott McComb0Mehdi Arbabi-Ghahroudi1Kevin A. Hay2Brian A. Keller3Sharlene Faulkes4Michael Rutherford5Tina Nguyen6Alex Shepherd7Cunle Wu8Anne Marcil9Annie Aubry10Greg Hussack11Devanand M. Pinto12Shannon Ryan13Shalini Raphael14Henk van Faassen15Ahmed Zafer16Qin Zhu17Susanne Maclean18Anindita Chattopadhyay19Komal Gurnani20Rénald Gilbert21Christine Gadoury22Umar Iqbal23Dorothy Fatehi24Anna Jezierski25Jez Huang26Robert A. Pon27Mhairi Sigrist28Robert A. Holt29Brad H. Nelson30Harold Atkins31Natasha Kekre32Eric Yung33John Webb34Julie S. Nielsen35Risini D. Weeratna36Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada; Centre for Infection, Immunity, and Inflammation, University of Ottawa, Ottawa, ON, Canada; Corresponding author: Scott McComb, Human Health Therapeutics Research Centre, National Research Council, Canada.Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, CanadaTerry Fox Laboratory, British Columbia Cancer Research Institute, Vancouver, BC, Canada; Division of Hematology, Faculty of Medicine, University of British Columbia, Vancouver, BC, CanadaDivision of Anatomical Pathology, The Ottawa Hospital/University of Ottawa, Ottawa, ON, Canada; University of Ottawa Faculty of Medicine, Ottawa, ON, CanadaDepartment of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON, CanadaDivision of Anatomical Pathology, The Ottawa Hospital/University of Ottawa, Ottawa, ON, Canada; Division of Hematopathology and Transfusion Medicine, The Ottawa Hospital/University of Ottawa, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biology, Concordia University, Montréal, QC, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaTerry Fox Laboratory, British Columbia Cancer Research Institute, Vancouver, BC, CanadaDepartment of Medical Genetics, University of British Columbia, Vancouver, BC, Canada; Canada’s Michael Smith Genome Sciences Centre, Vancouver, BC, Canada; Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, BC, CanadaDeeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, Canada; Department of Medical Genetics, University of British Columbia, Vancouver, BC, CanadaDivision of Hematology, Department of Medicine, The Ottawa Hospital, Ottawa, ON, CanadaDivision of Hematology, Department of Medicine, The Ottawa Hospital, Ottawa, ON, Canada; Clinical Epidemiology Program, Ottawa Hospital Research Institute, Ottawa, ON, CanadaCanada’s Michael Smith Genome Sciences Centre, Vancouver, BC, CanadaDeeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, CanadaDeeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, CanadaHuman Health Therapeutics Research Centre, National Research Council, Ottawa, ON, CanadaChimeric antigen receptor (CAR) T cell therapies targeting B cell-restricted antigens CD19, CD20, or CD22 can produce potent clinical responses for some B cell malignancies, but relapse remains common. Camelid single-domain antibodies (sdAbs or nanobodies) are smaller, simpler, and easier to recombine than single-chain variable fragments (scFvs) used in most CARs, but fewer sdAb-CARs have been reported. Thus, we sought to identify a therapeutically active sdAb-CAR targeting human CD22. Immunization of an adult Llama glama with CD22 protein, sdAb-cDNA library construction, and phage panning yielded >20 sdAbs with diverse epitope and binding properties. Expressing CD22-sdAb-CAR in Jurkat cells drove varying CD22-specific reactivity not correlated with antibody affinity. Changing CD28- to CD8-transmembrane design increased CAR persistence and expression in vitro. CD22-sdAb-CAR candidates showed similar CD22-dependent CAR-T expansion in vitro, although only membrane-proximal epitope targeting CD22-sdAb-CARs activated direct cytolytic killing and extended survival in a lymphoma xenograft model. Based on enhanced survival in blinded xenograft studies, a lead CD22sdCAR-T was selected, achieving comparable complete responses to a benchmark short linker m971-scFv CAR-T in high-dose experiments. Finally, immunohistochemistry and flow cytometry confirm tissue and cellular-level specificity of the lead CD22-sdAb. This presents a complete report on preclinical development of a novel CD22sdCAR therapeutic.http://www.sciencedirect.com/science/article/pii/S2950329924000171MT: Regular Issuechimeric antigen receptorsCD22CAR-Tnanobodysingle-domain antibody |
| spellingShingle | Scott McComb Mehdi Arbabi-Ghahroudi Kevin A. Hay Brian A. Keller Sharlene Faulkes Michael Rutherford Tina Nguyen Alex Shepherd Cunle Wu Anne Marcil Annie Aubry Greg Hussack Devanand M. Pinto Shannon Ryan Shalini Raphael Henk van Faassen Ahmed Zafer Qin Zhu Susanne Maclean Anindita Chattopadhyay Komal Gurnani Rénald Gilbert Christine Gadoury Umar Iqbal Dorothy Fatehi Anna Jezierski Jez Huang Robert A. Pon Mhairi Sigrist Robert A. Holt Brad H. Nelson Harold Atkins Natasha Kekre Eric Yung John Webb Julie S. Nielsen Risini D. Weeratna Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 Molecular Therapy: Oncology MT: Regular Issue chimeric antigen receptors CD22 CAR-T nanobody single-domain antibody |
| title | Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 |
| title_full | Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 |
| title_fullStr | Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 |
| title_full_unstemmed | Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 |
| title_short | Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22 |
| title_sort | discovery and preclinical development of a therapeutically active nanobody based chimeric antigen receptor targeting human cd22 |
| topic | MT: Regular Issue chimeric antigen receptors CD22 CAR-T nanobody single-domain antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2950329924000171 |
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