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DNA-encoded bispecific T cell engagers and antibodies present long-term antitumor activity
Alfredo Perales-Puchalt, … , Kar Muthumani, David B. Weiner
Alfredo Perales-Puchalt, … , Kar Muthumani, David B. Weiner
Published April 18, 2019
Citation Information: JCI Insight. 2019;4(8):e126086. https://doi.org/10.1172/jci.insight.126086.
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Research Article Immunology Oncology

DNA-encoded bispecific T cell engagers and antibodies present long-term antitumor activity

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Abstract

Specific antibody therapy, including mAbs and bispecific T cell engagers (BiTEs), are important new tools for cancer immunotherapy. However, these approaches are slow to develop and may be limited in their production, thus restricting the patients who can access these treatments. BiTEs exhibit a particularly short half-life and difficult production. The development of an approach allowing simplified development, delivery, and in vivo production would be an important advance. Here we describe the development of a designed synthetic DNA plasmid, which we optimized to permit high expression of an anti-HER2 antibody (HER2dMAb) and delivered it into animals through adaptive electroporation. HER2dMAb was efficiently expressed in vitro and in vivo, reaching levels of 50 μg/ml in mouse sera. Mechanistically, HER2dMAb blocked HER2 signaling and induced antibody-dependent cytotoxicity. HER2dMAb delayed tumor progression for HER2-expressing ovarian and breast cancer models. We next used the HER2dMAb single-chain variable fragment portion to engineer a DNA-encoded BiTE (DBiTE). This HER2DBiTE was expressed in vivo for approximately 4 months after a single administration. The HER2DBiTE was highly cytolytic and delayed cancer progression in mice. These studies illustrate an approach to generate DBiTEs in vivo, which represent promising immunotherapies for HER2+ tumors, including ovarian and potentially other cancers.

Authors

Alfredo Perales-Puchalt, Elizabeth K. Duperret, Xue Yang, Patricia Hernandez, Krzysztof Wojtak, Xizhou Zhu, Seang-Hwan Jung, Edgar Tello-Ruiz, Megan C. Wise, Luis J. Montaner, Kar Muthumani, David B. Weiner

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Figure 1

Design, expression, and binding of HER2dMAb.

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Design, expression, and binding of HER2dMAb.
(A) Schematic of DNA constr...
(A) Schematic of DNA construct encoding HER2dMAb. (B) Western blot analysis of HER2dMAb or FSHR constructs expressed in 293T cells (2 independent experiments). Numbers indicate molecular weight (kDa). (C) Western blot of human IgG from mouse sera electroporated with HER2dMAb or pVAX control 14 days after DNA injection and electroporation (n = 5 mice per group). (D) Expression levels of human IgG quantified by ELISA from sera of nude mice electroporated with HER2dMAb (n = 5 mice per group, 2 independent experiments). (E) Binding ELISA of sera from mice expressing HER2dMAb or pVax after coating the plate with HER2 protein (n = 5 mice per group, 2 independent experiments). (F) Flow cytometry plot showing binding of HER2dMAb to mouse breast cancer cell lines with and without HER2 expression (representative of triplicates).

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