Molecular Therapy: Oncology (Mar 2024)
Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22
- 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
Affiliations
- Scott McComb
- 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, 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.
- Mehdi Arbabi-Ghahroudi
- 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, Canada
- Kevin A. Hay
- Terry Fox Laboratory, British Columbia Cancer Research Institute, Vancouver, BC, Canada; Division of Hematology, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada
- Brian A. Keller
- Division of Anatomical Pathology, The Ottawa Hospital/University of Ottawa, Ottawa, ON, Canada; University of Ottawa Faculty of Medicine, Ottawa, ON, Canada
- Sharlene Faulkes
- Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada
- Michael Rutherford
- Division 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, Canada
- Tina Nguyen
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Alex Shepherd
- 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, Canada
- Cunle Wu
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada; Department of Biology, Concordia University, Montréal, QC, Canada
- Anne Marcil
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Annie Aubry
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Greg Hussack
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Devanand M. Pinto
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Shannon Ryan
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Shalini Raphael
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Henk van Faassen
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Ahmed Zafer
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Qin Zhu
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Susanne Maclean
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Anindita Chattopadhyay
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Komal Gurnani
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Rénald Gilbert
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Christine Gadoury
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Umar Iqbal
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Dorothy Fatehi
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Anna Jezierski
- 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, Canada
- Jez Huang
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Robert A. Pon
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Mhairi Sigrist
- Terry Fox Laboratory, British Columbia Cancer Research Institute, Vancouver, BC, Canada
- Robert A. Holt
- Department 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, Canada
- Brad H. Nelson
- Deeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, Canada; Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada
- Harold Atkins
- Division of Hematology, Department of Medicine, The Ottawa Hospital, Ottawa, ON, Canada
- Natasha Kekre
- Division of Hematology, Department of Medicine, The Ottawa Hospital, Ottawa, ON, Canada; Clinical Epidemiology Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada
- Eric Yung
- Canada’s Michael Smith Genome Sciences Centre, Vancouver, BC, Canada
- John Webb
- Deeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, Canada
- Julie S. Nielsen
- Deeley Research Centre, British Columbia Cancer Research Institute, Victoria, BC, Canada
- Risini D. Weeratna
- Human Health Therapeutics Research Centre, National Research Council, Ottawa, ON, Canada
- Journal volume & issue
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Vol. 32,
no. 1
p. 200775
Abstract
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.
