Type: Oral
Session: 604. Molecular Pharmacology and Drug Resistance: Myeloid Neoplasms: Novel Approaches to Targeting Epigenetics, Apoptosis and Immune Effectors in Myeloid Malignancies
Hematology Disease Topics & Pathways:
Acute Myeloid Malignancies, Research, AML, Bispecific Antibody Therapy, Translational Research, Diseases, Biological therapies, Treatment Considerations, Myeloid Malignancies
CBX-250 has been developed as a TCR-mimetic (TCRm) bispecific T Cell Engager (TCE) antibody (Ab) that binds to the CG1/HLA-A2*02:01 pMHC complex with high affinity. CBX-250 induces potent T cell mediated killing in vitro of leukemia cell lines with varying levels of target expression at sub-nanomolar EC50 levels and exhibits highly effective tumor control in vivo against multiple AML cell line (CDX) murine models at doses as low as 0.005 mg/kg. Importantly, CBX-250 also demonstrates potent efficacy, reduced tumor burden, and a clear survival benefit vs. a control arm in an aggressive primary AML patient derived (PDX) tumor mouse model that harbors FLT3-ITD, DNMT3a, and NPM1 mutations. Consistent with these results, CBX-250 was able to induce significant levels of IFNg release from several primary AML patient samples co-cultured with donor effector T cells vs. a negative control.
Successful immunotherapeutic approaches for the treatment of AML have been limited due to a lack of tumor specific surface targets (e.g., CD33, CD123) that avoid undesirable toxicities due to their expression on normal hematopoietic cells. Significantly, CBX-250 does not induce T cell activation or IFNg production when co-cultured with HLA-A2 positive healthy donor (HD) neutrophils and displays no toxicity towards normal hematopoietic stem cells (HSC) and progenitor cells in a colony forming unit (CFU) assay, in contrast to a cytarabine-treated positive control. In keeping with these findings, CBX-250 does not induce IFNg or T cell activation when co-cultured with HD CD34+ HSCs and PBMCs, whereas a significant increase in IFNg levels is observed when these stem cells are co-cultured with a CD123 TCE bispecific Ab. Moreover, when HD PBMCs are incubated with increasing concentrations of CBX-250 in the absence of tumor cells for 3 days, no changes are observed in any immune cell subpopulation (T, B cells, monocytes). By comparison, a CD123 TCE Ab induces a dose-dependent reduction in monocytes (CD123+, CD14+) and a concomitant increase in T cells under similar conditions, reflecting on-target T cell-mediated killing by the CD123 TCE Ab that is not observed with CBX-250.
Bystander killing has been recognized as a feature of TCE bispecific Abs that may help to overcome potential resistance mechanisms which may arise due to heterogeneous tumor antigen expression3-6. The ability to kill both CG1/pMHC-positive tumor cells and proximal CG1/pMHC-negative cells (bystanders) would be desirable to prevent escape and outgrowth of target-negative tumor cells during treatment. To investigate CBX-250-mediated bystander killing, we measured cytotoxicity of CG1/pMHC-negative AML cells in vitro when co-cultured with CG1/pMHC-positive tumor cells, effector T cells, and CBX-250 at varying concentrations. Our results demonstrated that CBX-250 was able to induce dose-dependent bystander killing of neighboring CG1/pMHC target-negative leukemia cells that was observed only in the presence of CG1/pMHC positive cells, highlighting the beneficial potency effects of CBX-250 in a tumor microenvironment with heterogenous target antigen expression.
Taken together, these data provide strong preclinical evidence of the potency, specificity, and safety of CBX-250 against a novel tumor antigen that may provide a wider therapeutic index relative to other well characterized AML targets. These findings support further development in the clinic and IND-enabling studies for CBX-250 are currently ongoing.
- 1.Zhang et al CCR 2013
- 2.Alatrash et al Leukemia 2017
- 3.Ross et al PLOS One 2017
- 4.Upadhyay et al Can Disc 2021
- 5.Teijeira et al Theranostics 2022
- 6.Gaspar et al JITC 2023
Disclosures: Lee: Coherus Biosciences: Current equity holder in publicly-traded company; Radionetics Oncology: Consultancy. O'Connor: MatTek Life Sciences: Ended employment in the past 24 months. Mak: Crossbow Therapeutics, Inc.: Current holder of stock options in a privately-held company. Princiotta: EpiVac Therapeutics: Other: Independent Contractor. Wiederschain: Sanofi S.A.: Current equity holder in publicly-traded company, Patents & Royalties.