Increasing evidence suggests that antibody-drug conjugates (ADCs) can enhance anti-tumor immunity and improve clinical outcome. Here, we elucidate the therapeutic efficacy and immune-mediated mechanisms of a novel HER2-targeting ADC bearing a potent anthracycline derivate as payload (T-PNU) in a human HER2-expressing syngeneic breast cancer model resistant to trastuzumab and ado-trastuzumab emtansine. Mechanistically, the anthracycline component of the novel ADC induced immunogenic cell death leading to exposure and secretion of danger-associated molecular signals. RNA sequencing derived immunogenomic signatures and TCRβ clonotype analysis of tumor-infiltrating lymphocytes revealed a prominent role of the adaptive immune system in the regulation of T-PNU mediated anti-cancer activity. Depletion of CD8 T cells severely reduced T-PNU efficacy, thus confirming the role of cytotoxic T cells as drivers of the T-PNU mediated anti-tumor immune response. Furthermore, T-PNU therapy promoted immunological memory formation in tumor-bearing animals protecting those from tumor rechallenge. Finally, the combination of T-PNU and checkpoint inhibition, such as α-PD1, significantly enhanced tumor eradication following the treatment. In summary, a novel PNU-armed, HER2-targeting ADC elicited long-lasting immune protection in a murine orthotopic breast cancer model resistant to other HER2-directed therapies. Our findings delineate the therapeutic potential of this novel ADC payload and support its clinical development for breast cancer patients and potentially other HER2 expressing malignancies.

Cancer Immunology Department of Biomedicine University of Basel Basel Switzerland

Celonic AG Eulerstrasse 55 4051 Basel Switzerland

Department of Biosystems Science and Engineering ETH Zürich Basel Switzerland

Department of Cancer Immunology and Immune Modulation Boehringer Ingelheim Pharma GmbH and Co KG Birkendorfer Str 65 88400 Biberach an der Riss Germany

Institute of Pathology University Hospital Basel Basel Switzerland

NBE Therapeutics Ltd Hochbergerstrasse 60C 4057 Basel Switzerland

NEXUS Personalized Health Technologies ETH Zürich Switzerland and Swiss Institute of Bioinformatics Zürich Switzerland

Sotio s a Jankovcova 1518 2 170 00 Prague 7 Czech Republic

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Detection of immunogenic cell death and its relevance for cancer therapy