AML ImmunoPET: 64Cu-DOTA-anti-CD33 PET-CT imaging of AML in Xenograft mouse model

Madabushi, S. S.; Brooks, J. L.; Zuro, D.; Kumar, B.; Sanchez, J.; Echavarria Parra, L.; Orellana, M.; Vishwasrao, P.; Nair, I.; Chea, J.; Poku, K.; Bowles, N.; Miller, A.; Ebner, T.; Molnar, J.; Rosenthal, J.; Vallera, D. A.; Wong, J. Y. C.; Stein, A. S.; Colcher, D.; Shively, J.; Yazaki, P.; Hui, S. K.

doi:10.1101/635078

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bioRxiv · 10.1101/635078

AML ImmunoPET: 64Cu-DOTA-anti-CD33 PET-CT imaging of AML in Xenograft mouse model

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Abstract

PurposeAcute myeloid leukemia (AML) is a highly aggressive form of leukemia that results in poor survival outcomes. Currently, diagnosis and prognosis are based on invasive single-point bone marrow biopsies (iliac crest). There is currently no AML-specific non-invasive imaging method to assess disease in the whole body, including in extramedullary organs, representing an unmet clinical need. About 85-90% of human myeloid leukemia cells express CD33 (an accepted biomarker for AML) cell surface receptors, highlighting CD33 as an ideal candidate for AML immunoPET.\n\nExperimental DesignIn this study, we evaluated if 64Cu-DOTA-anti-CD33 murine monoclonal antibody (mAb) can be used for ImmunoPET-CT imaging of AML in preclinical model. For translational purpose, a humanized anti-CD33 antibody was produced, and after confirming its anti-CD33 PET-CT imaging ability we further explored its therapeutic potential.\n\nResults64Cu-DOTA-anti-CD33 based PET-CT imaging detected CD33+ AML in mice with high sensitivity (95.65%) and specificity (100%). In the entire skeletal system, CD33+ PET activity was significantly high in the bones of AML bearing mice over non-leukemic control mice; femur (p<0.00001), tibia (p=0.0001), humerus (p=0.0014), and lumber spine (p<0.00001). Interestingly, the imaging showed CD33+ PET activity prevelant in epiphysis/diaphysis of the bones, indicating spatial heterogeneity. Anti-CD33 therapy using newly developed humanized anti-CD33 mAb as an ADC (p=0.02) and 225Ac-anti-CD33-RIT (p<0.00001), significantly reduced disease burden over that of respective controls.\n\nConclusionWe have successfully developed a novel anti-CD33 immunoPET-CT based non-invasive imaging tool for diagnosis of AML and its spatial distribution, which indicates a preferential skeletal niche.\n\nTranslational RelevanceAcute myeloid leukemia (AML) exhibits extensive heterogeneity in therapeutic response, even in patients sharing similar characteristics, indicating the inadequacy of currently used biopsy-based prognostic tools. There is a critical need for non-invasive quantitative imaging modality specific to AML, which is able to assess this disease in the whole body, including in extramedullary organs and to longitudinally monitor treatment response. CD33, an accepted AML biomarker is expressed on more than 85-90% of blast cells and Mylotarg (anti-CD33-ADC) is FDA approved immunotherapy for AML. In this preclinical study using anti-human CD33 mAb (64Cu-DOTA-anti-CD33), a CD33 PET-CT imaging modality was developed that detected AML with high sensitivity and specificity. The approach also indicated spatial heterogeneity in leukemic disease, dependent on disease burden, which warrants caution in analyzing results from single point bone marrow biopsies. Therefore, this imaging modality can be used to non-invasively detect AML in humans and also monitor treatment response, an unmet clinical need.

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Madabushi, S. S., Brooks, J. L., Zuro, D., Kumar, B., Sanchez, J., Echavarria Parra, L., Orellana, M., Vishwasrao, P., Nair, I., Chea, J., Poku, K., Bowles, N., Miller, A., Ebner, T., Molnar, J., Rosenthal, J., Vallera, D. A., Wong, J. Y. C., Stein, A. S., Colcher, D., Shively, J., Yazaki, P., Hui, S. K.. 2019-05-10. AML ImmunoPET: 64Cu-DOTA-anti-CD33 PET-CT imaging of AML in Xenograft mouse model. https://doi.org/10.1101/635078

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