title: Genetic engineering of chimeric antigen receptors using lamprey derived variable lymphocyte receptors.
abstract: Chimeric antigen receptors (CARs) are used to redirect effector cell specificity to selected cell surface antigens. Using CARs, antitumor activity can be initiated in patients with no prior tumor specific immunity. Although CARs have shown promising clinical results, the technology remains limited by the availability of specific cognate cell target antigens. To increase the repertoire of targetable tumor cell antigens we utilized the immune system of the sea lamprey to generate directed variable lymphocyte receptors (VLRs). VLRs serve as membrane bound and soluble immune effectors analogous but not homologous to immunoglobulins. They have a fundamentally different structure than immunoglobulin (Ig)-based antibodies while still demonstrating high degrees of specificity and affinity. To test the functionality of VLRs as the antigen recognition domain of CARs, two VLR-CARs were created. One contained a VLR specific for a murine B cell leukemia and the other contained a VLR specific for the human T cell surface antigen, CD5. The CAR design consisted of the VLR sequence, myc-epitope tag, CD28 transmembrane domain, and intracellular CD3ζ signaling domain. We demonstrate proof of concept, including gene transfer, biosynthesis, cell surface localization, and effector cell activation for multiple VLR-CAR designs. Therefore, VLRs provide an alternative means of CAR-based cancer recognition.
authors.affiliation: Department of Molecular and Systems Pharmacology, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Moot
authors.affiliation: Department of Pediatrics, Aflac Cancer and Blood Disorders Center, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Sunil S
authors.initials: SS
authors.lastname: Raikar
authors.affiliation: Department of Molecular and Systems Pharmacology, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Fleischer
authors.affiliation: Department of Pediatrics, Aflac Cancer and Blood Disorders Center, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Querrey
authors.affiliation: Department of Pediatrics, Aflac Cancer and Blood Disorders Center, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Tylawsky
authors.affiliation: Department of Pathology and Laboratory Medicine, Emory University School of Medicine , Atlanta, Georgia, USA.
authors.forename: Hirotomo
authors.initials: H
authors.lastname: Nakahara
authors.affiliation: Department of Molecular and Systems Pharmacology, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine, Atlanta, Georgia, USA; Department of Pediatrics, Aflac Cancer and Blood Disorders Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Doering
authors.affiliation: Department of Molecular and Systems Pharmacology, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine, Atlanta, Georgia, USA; Department of Pediatrics, Aflac Cancer and Blood Disorders Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: H Trent
authors.initials: HT
authors.lastname: Spencer