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Principal Investigator: Samuel Hickenlooper
Organization: LUMINARY THERAPEUTICS, INC.
Fiscal Year: 2024
Award: $399,256
Funding agency: National Cancer Institute
Abstract
CAR T cell therapies have produced robust remissions in several hematological malignancies. However, they
have not achieved the same success against solid tumors. These treatments fail for multiple reasons, including
antigen escape and poor T cell persistence in the immunosuppressive tumor microenvironment. Additionally,
many patients receiving T cell therapy experience serious, sometimes life-threatening side effects. Here, we
propose a CAR T product – an armored split CAR – that combines 2 innovative strategies specifically tailored to
address the unique challenges presented by solid tumors, as well as an inducible suicide switch to enhance
safety. Moreover, we are developing this product on our proprietary allogeneic γδ T cell platform. These cells
display innate anti-tumor properties and do not mediate GVHD. We use a “Split CAR” approach, wherein two
CAR molecules, each targeting a different antigen, are expressed by a single T cell and dimerize upon antigen
binding. By targeting 2 antigens, we reduce the potential for antigen escape. Each CAR molecule contains either
a CD28 or 4-1BB costimulatory domain which are independently activated to cooperatively enhance cytokine
release, cell survival, and proliferation. The heterodimerized CARs trigger activation through a single shared
CD3ζ domain incorporated on one of the CAR chains. In addition, our engineered T cells express a binding
molecule to sequester and disable the tumor-secreted immunosuppressive cytokine, TGFβ. Lastly, we are
including a tEGFR safety switch so that the clinical product can be rapidly eliminated in vivo in case of severe
side effects. Efficient gene delivery of such a large cassette is made feasible by using the Tc Buster transposon
system, which can stably integrate transgenes as large as 10 kb with an acceptable copy number and a superior
safety profile compared to viral transduction. Transposon-based gene delivery also circumvents the long
manufacturing timelines and bottlenecks that beset viral methods. We are adapting the armored split CAR to
treat epithelial ovarian carcinoma by targeting the tumor antigens mesothelin (MSLN) and chondroitin sulfate
proteoglycan-4 (CSPG4). These antigens are overexpressed in at least 40% of ovarian cancer patients and are
associated with poor outcomes. We will test our armored split CAR in vitro using a co-culture target cell killing
assay with ovarian cancer lines that are single- or double-positive for MSLN and CSPG4. We will test how well
the engineered cells resist exhaustion in a serial killing assay in which T cells are exposed to antigen in co-
culture over ~14 days and multiple passages, then assessed for relevant activation and exhaustion markers. We
will also test the tEGFR safety switch by culturing the cells with the inducer, cetuximab, and measuring selective
lysis of engineered cells. Lastly, we will assay tumor killing by the armored split CAR in a mouse model of ovarian
cancer. These experiments are designed to collectively demonstrate efficacy of our product under stress
conditions and will lead into anticipated Phase II preclinical studies to assess toxicity and safety.
Terms: <Address><Adoptive Cell Transfers><Allogenic><Anti-EGFR Monoclonal Antibody><Anti-Epidermal Growth Factor Receptor Monoclonal Antibody><Antigen Targeting><Antigens><Antimorphic mutation><Assay><Binding><Bioassay><Biological Assay><Biotech><Biotechnology><Bone-Derived Transforming Growth Factor><CAR T cell therapy><CAR T cells><CAR T therapy><CAR modified T cells><CAR-T><CAR-Ts><CD28><CD28 gene><Cancer Patient><Cancers><Cell Body><Cell Line><Cell Survival><Cell Viability><CellLine><Cells><Centrocytic Small-Cell Lymphoma><Cetuximab><Chondroitin 4 Sulfate><Chondroitin Sulfate A><Chondroitin Sulfate Proteoglycan><Clinical><Co-culture><Cocultivation><Coculture><Coculture Techniques><Cultured Cells><Cytolysis><D Cells><DNA cassette><Delta Cell><Dimerization><Disease remission><Dominant Negative><Dominant-Negative Mutant><Dominant-Negative Mutation><Elements><Engineering><Environment><Epithelium><Experimental Designs><Exposure to><Failure><Gamma-delta T cells><Gene Delivery><GvHD><Health><Hematologic Cancer><Hematologic Malignancies><Hematologic Neoplasms><Hematological Malignancies><Hematological Neoplasms><Hematological Tumor><Hematopoietic Cancer><Heterodimerization><Homologous Wasting Disease><Human Resources><Immune mediated therapy><Immunocompromised><Immunocompromised Host><Immunocompromised Patient><Immunologically Directed Therapy><Immunology><Immunosuppressed Host><Immunotherapy><In Vitro><Individual><Investigational Drugs><Investigational New Drugs><Lead><Life><Ligands><Lymphoma, Lymphocytic, Diffuse, Poorly-Differentiated><Lysis><Malignant Hematologic Neoplasm><Malignant Neoplasms><Malignant Ovarian Neoplasm><Malignant Ovarian Tumor><Malignant Tumor><Malignant Tumor of the Ovary><Malignant neoplasm of ovary><Manpower><Mantle Cell Lymphoma><Mantle-Zone Lymphoma><Marketing><Measures><Methods><Mice><Mice Mammals><Milk Growth Factor><Molecular Interaction><Multiple Myeloma><Murine><Mus><New Drug Approvals><Outcome><Ovarian Carcinoma><Ovary Cancer><Ovary Carcinoma><Pathway interactions><Patients><Pb element><Phase><Plasma-Cell Myeloma><Platelet Transforming Growth Factor><Platinum><Platinum Black><Production><Prognosis><Proliferating><Property><Protein Dimerization><Pt element><Publications><Recurrence><Recurrent><Remission><Remission Induction Therapy><Research><Runt Disease><Safety><Scientific Publication><Solid Neoplasm><Solid Tumor><Soluble Mpf/Mesothelin-Related Protein><Somatostatin Cells><Somatostatin Secreting Cell><Source><Strains Cell Lines><Stress><Suicide><Survival Rate><System><T cell based therapeutics><T cell based therapy><T cell directed therapies><T cell targeted therapeutics><T cell therapy><T cells for CAR><T-Cells><T-Lymphocyte><T-cell therapeutics><T-cell transfer therapy><T44><TGF B><TGF-Beta Type II Receptor><TGF-beta><TGF-β><TGFBR2><TGFBR2 gene><TGFbeta><TGFβ><Techniques><Technology><Testing><Therapeutic><Toxic effect><Toxicities><Transforming Growth Factor beta><Transforming Growth Factor-Beta Family Gene><Transgenes><Tumor Antigens><Tumor Cell><Tumor-Associated Antigen><Viral><adoptive T cell transfer><adoptive T-cell therapy><adoptive cell therapy><adoptive cellular therapy><anti-cancer><antigen binding><antigen bound><cancer antigens><cell killing><chemotherapy><chimeric antigen T cell receptor><chimeric antigen receptor (CAR) T cell therapy><chimeric antigen receptor (CAR) T cells><chimeric antigen receptor T cell therapy><chimeric antigen receptor T cells><chimeric antigen receptor T therapy><chimeric antigen receptor fusion protein T-cells><chimeric antigen receptor modified T cells><cost effective><cultured cell line><cytokine><design><designing><engineered T cells><enhancer cassette><exhaustion><experience><experiment><experimental research><experimental study><experiments><expression cassette><fatal attempt><fatal suicide><gene cassette><gene delivery system><genetic cassette><genetically engineered T-cells><graft versus host disease><graft vs host disease><graft vs. host disease><heavy metal Pb><heavy metal lead><immune microenvironment><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunogen><immunosuppressed patient><immunosuppressive microenvironment><immunosuppressive tumor microenvironment><improved><in vivo><innovate><innovation><innovative><integration cassette><intent to die><malignancy><manufacture><mesothelin><metabolic fitness><mouse model><murine model><myeloma><myelomatosis><neoplasm/cancer><neoplastic cell><non-viral gene delivery><nonviral gene delivery><novel><ovarian cancer><overexpress><overexpression><pathway><personnel><pre-clinical study><preclinical study><promoter cassette><reporter cassette><resistance cassette><response><selectable cassette><selection cassette><side effect><stop cassette><success><suicides><therapeutic T-cell platform><thymus derived lymphocyte><timeline><transcription cassette><transcriptional cassette><transforming growth factor-beta type II receptor><transforming growth factor-β type II receptor><transgene><transgene cassette><transgenic T- cells><treatment group><tumor><tumor immune microenvironment><tumor-immune system interactions><tumor-specific antigen><γδ T cells><γδT cells>