ArticleMolecular therapy : the journal of the American Society of Gene Therapy2026
Efficient in vivo assembly of DNA encoded polyvalent BTEs for dual antigen targeting for broadening therapeutic impact in ovarian cancer.
Article in Molecular therapy : the journal of the American Society of Gene Therapy, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The clinical potential of bispecific T cell engagers (BTEs) is limited by their short serum half-life and the complexity and cost of recombinant protein manufacturing. Currently, BTEs rely on external production and repeat dosing, limiting scalability and patient access. Here, we present a synDNA platform that enables in vivo assembly and long-term secretion of bispecific antibodies directly from host muscle. The system includes complementary "knob" and "hole" Fc chains on separate plasmids that pair only when co-expressed, ensuring controlled heterodimer formation and reducing homodimer assembly. Using ovarian cancer (OC) as a model of antigen heterogeneity, we generated dKBTEs targeting follicle stimulating hormone receptor (dK_FSHRxCD3) and Her2 (dK_Her2xCD3). Artificial intelligence simulations supported successful knob-hole pairing and presented significant challenges for homodimer formation. Co-transfected Expi293F cells secreted fully assembled bispecifics that bound antigen and CD3, activated primary human T cells, and induced potent antigen-specific cytotoxicity. OC patient peripheral blood mononuclear cells responded strongly to dKBTEs in vitro. In mice, dKBTEs showed superior pharmacokinetics and tumor control over first-generation BTEs. Combination therapy with both dKBTEs further improved tumor control. synDNA delivery allows simultaneous in vivo production of two dKBTEs without compromising expression. These findings establish synDNA delivery as a programmable approach for sustained delivery of complex biologics.
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