thunder

Cancer remains an incurable disease for many patients with an unmet medical need for identifying and testing new diagnostics and therapeutics. Monoclonal antibodies, the major class of current biologicals, have revolutionized cancer therapy. With over 100 approved therapeutics, antibodies are currently the basis of nearly all immunotherapy approaches in oncology. However, their tetrameric structure and large molecular size limits tumor penetration and impedes the engineering of more effective drugs. Currently, there is a clear unmet need to further optimize antibody-based immunotherapy in oncology to turn incurable into curable diseases, and to overcome the current infrastructural limitations to provide high-level immunotherapeutics, such as CAR-T cells, to all patients at need. Nanobodies are next generation biologicals that promise to overcome limitations of conventional antibodies and to advance biologicals in clinical oncology to maintain response and overcome resistance. Nanobodies are robust single variable immunoglobulin domains derived from camelid heavychain antibodies. With their small size, nanobodies show good penetration into tumor tissues. Their high solubility facilitates bioengineering of innovative bispecific biologicals. The first nanobody-based drugs Caplacizumab and Cilta-cel (for thrombocytopenic purpura and multiple myeloma, respectively) provide proof of principle for the successful translation of nanobody technology into the clinic. Within THUNDER, we will combine the complementary expertise in nanobody research of two leading German medical centers in Hamburg and Bonn with the Comprehensive Cancer Centers of Excellence at these sites (UCCH, CIO). Together, we will establish a national innovation hub that provides a drug development pipeline to drive innovative nanobodybased cancer diagnostics and therapeutics into the clinic. Our track record in the field, with high quality publications, patent applications, and successful spinoff companies, underscores our ambition to drive innovative nanobody based anti-cancer drugs into the clinic. The interdisciplinary THUNDER team will provide momentum to the translational development of our lead candidates and to streamline our pipeline for the development of nanobody-based drugs in a truly synergistic manner at both sites. We will engineer nanobodies against eight validated targets (CD38, BCMA, CDCP1, EGFR, CD155, TIGIT, CD39, CD73) for targeting of different prototype tumors: we will target cell surface proteins expressed by i) hematological malignancies, ii) solid tumors, as well as iii) immune checkpoint proteins, and iv) purinergic checkpoint proteins. The interactions of lead nanobodies with their targets will be elucidated on the structural level by X-ray crystallography and electron microscopy. The developability of lead candidates will be validated with patient-derived cells, organoid cultures, and established mouse tumor models. Our novel llama-IgH-transgenic mice will facilitate the discovery of nanobodies against conserved epitopes of the selected targets antigens, thus allowing for generation of a completely new repertoire of nanobodies.

We will harness nanobody engineering technology to develop novel drug candidates based on nanobodies: cytotoxic and silent heavy-chain antibodies (hcAbs), biparatopic and bispecific nanobody dimers including bispecific killer or T cell engagers (Nb-BiKEs, Nb-BiTEs), hcAbs targeting different epitopes of one antigen or two different antigens, as well as bispecific chimeric antigen receptors (Nb-CARs) based on validated preclinical rationales. Bispecific strategies targeting two distinct tumor antigens with hcAbs or CARs aim to prevent immune escape of tumor cells by antigen loss, a central problem of current therapies. Moreover, nanobodies will be conjugated to small drugs to enrich highly toxic compounds selectively in tumor tissues, as well as to imaging tracers as theranostics for noninvasive imaging and therapy of cancer. Finally, we will specifically target tumor cells with adenoassociated viral vectors (AAV) that code for locally produced cytokines and/or nanobody-based checkpoint inhibitors in the tumor microenvironment. The complementary expertise of the THUNDER consortium in nanobody research and clinical oncology will be synergistically combined with existing early phase clinical trial units at UKE and UKB to provide a clear perspective for translation of nanobody innovations into clinical oncology.