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A novel therapeutic bispecific format based on synthetic orthogonal heterodimers enables T cell activity against Acute myeloid leukemia

Oncogene volume 42, pages 26–34 (2023)Cite this article

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Abstract

Many therapeutic bispecific T-cell engagers (BiTEs) are in clinical trials. A modular and efficient process to create BiTEs would accelerate their development and clinical applicability. In this study, we present the design, production, and functional activity of a novel bispecific format utilizing synthetic orthogonal heterodimers to form a multichain modular design. Further addition of an immunoglobulin hinge region allowed a stable covalent linkage between the heterodimers. As proof-of-concept, we utilized CD33 and CD3 binding scFvs to engage leukemia cells and T-cells respectively. We provide evidence that this novel bispecific T-cell engager (termed IgGlue-BiTE) could bind both CD3+ and CD33+ cells and facilitates robust T-cell mediated cytotoxicity on AML cells in vitro. In a mouse model of minimal residual disease, we showed that the novel IgGlue-BiTE greatly extended survival, and mice of this treatment group were free of leukemia in the bone marrow. These findings suggest that the IgGlue-BiTE allows for robust simultaneous engagement with both antigens of interest in a manner conducive to T cell cytotoxicity against AML. These results suggest a compelling modular system for bispecific antibodies, as the CD3- and CD33-binding domains can be readily swapped with domains binding to other cancer- or immune cell-specific antigens.

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Fig. 1: Design, predicted structure, in vitro expression and purification of IgGlue-BiTE.
Fig. 2: In vitro binding of IgGlue-BiTE with AML (HL-60) and T-cell (Jurkat).
Fig. 3: In vitro cytotoxicity of T-cells mediated by IgGlue-BiTE.
Fig. 4: IgGlue-BiTE mediated T-cell cytokine production.
Fig. 5: IgGlue-BiTE facilitates T cell clearance of AML from bone marrow in vivo and extends survival.
Fig. 6: No evidence of tumor in bone marrow aspirates.

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Acknowledgements

Research reported in this publication was performed in the CCTI Flow Cytometry Core, supported in part by the Office of the Director, National Institutes of Health under awards S10RR027050 (LSRII) and S10OD020056 (Influx sorter). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Mice imaging were performed using instrumentation maintained by the Cancer Center Small Animal Imaging Shared Resource with NIH grant #P30 CA013696 (National Cancer Institute).

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Author notes

  1. These authors contributed equally: Alan Burke, Florence Borot.

Authors and Affiliations

  1. Department of Medicine, Columbia University Irving Medical Center, Columbia University, New York, NY, 10032, USA

    Alan Burke, Florence Borot, Xing Du, Albert Mridul Grass, Abdullah Mahmood Ali & Siddhartha Mukherjee

  2. Department of Molecular Pharmacology and Therapeutics, Columbia University Irving Medical Center, Columbia University, New York, NY, 10032, USA

    Alan Burke

  3. Brahma Therapeutics Inc, Cambridge, MA, 02139, USA

    Michael Churchill, Jian Ding & Philip DeSouza

  4. Edward P. Evans Center for Myelodysplastic Syndromes, Columbia University Irving Medical Center, New York, NY, 10032, USA

    Abdullah Mahmood Ali & Siddhartha Mukherjee

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  1. Alan Burke
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Contributions

AB, FB, AMA, MC, and SM participated in the design of the experiments; AB, FB, MC, JD, and XD performed experiments. AB, FB, XD, JD, AMA, MC, and SM analyzed and interpreted data. AB, AMA, AMG, and SM wrote the manuscript. FB, XD, PD, MC, and JD edited the manuscript. SM conceived the idea. AMA and SM participated in the design of the study. All authors reviewed and checked the final version of the paper.

Corresponding authors

Correspondence to Abdullah Mahmood Ali or Siddhartha Mukherjee.

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Competing interests

Columbia University has granted an exclusive option to Brahma Therapeutics to license technology that is the subject of this study. AB, FB, AMA, and SM are co-inventors on pending patent applications filed by Columbia University and optioned to Brahma Therapeutics. SM has equity ownership and is on the Scientific Advisory Board of Brahma Therapeutics. MC, JD and PD is an employee of Brahma Therapeutics.

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Burke, A., Borot, F., Du, X. et al. A novel therapeutic bispecific format based on synthetic orthogonal heterodimers enables T cell activity against Acute myeloid leukemia. Oncogene 42, 26–34 (2023). https://doi.org/10.1038/s41388-022-02532-2

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