This first-in-human study evaluated RO7122290, a bispecific fusion protein carrying a split trimeric 4-1BB (CD137) ligand and a fibroblast activation protein α (FAP) binding site that costimulates T cells for improved tumor cell killing in FAP-expressing tumors. Patients with advanced or metastatic solid tumors received escalating weekly intravenous doses of RO7122290 as a single agent ( n = 65) or in combination with a 1200-milligram fixed dose of the anti–programmed death-ligand 1 (anti–PD-L1) antibody atezolizumab given every 3 weeks ( n = 50), across a tested RO7122290 dose range of 5 to 2000 milligrams and 45 to 2000 milligrams, respectively. Three dose-limiting toxicities were reported, two at different RO7122290 single-agent doses (grade 3 febrile neutropenia and grade 3 cytokine release syndrome) and one for the combination (grade 3 pneumonitis). No maximum tolerated dose was identified. The pharmacokinetic profile of RO7122290 suggested nonlinearity in elimination. The observed changes in peripheral and tissue pharmacodynamic (PD) biomarkers were consistent with the postulated mechanism of action. Treatment-induced PD changes included an increase in proliferating and activated T cells in peripheral blood both in the single-agent and combination arms. Increased infiltration of intratumoral CD8 ⁺ and Ki67 ⁺ CD8 ⁺ T cells was observed for both treatment regimens, accompanied by the up-regulation of T cell activation genes and gene signatures. Eleven patients experienced a complete or partial response, six of whom were confirmed to be immune checkpoint inhibitor naive. These results support further evaluation of RO7122290 in combination with atezolizumab or other immune-oncology agents for the treatment of solid tumors.

• Angiogenesis is critical to the development and survival of solid tumors. • Vanucizumab inhibits both vascular endothelial growth factor and angiopoietin-2. • Vanucizumab exhibits anti-tumor, anti-angiogenic and anti-metastatic effects. • It markedly reduced blood-vessel markers in tumor/skin samples in cancer patients. • Skin biopsies are a valuable surrogate for studying angiogenesis-related mechanisms.

Antiangiogenic strategies have not shown striking antitumor activities in the majority of glioma patients so far. It is unclear which antiangiogenic combination regimen with standard therapy is most effective. Therefore, we compared anti-VEGF-A, anti-Ang2, and bispecific anti-Ang-2/VEGF-A antibody treatments, alone and in combination with radio- or temozolomide (TMZ) chemotherapy, in a malignant glioma model using multiparameter two-photon in vivo microscopy in mice. We demonstrate that anti-Ang-2/VEGF-A lead to the strongest vascular changes, including vascular normalization, both as monotherapy and when combined with chemotherapy. The latter was accompanied by the most effective chemotherapy-induced death of cancer cells and diminished tumor growth. This was most probably due to a better tumor distribution of the drug, decreased tumor cell motility, and decreased formation of resistance-associated tumor microtubes. Remarkably, all these parameters where reverted when radiotherapy was chosen as combination partner for anti-Ang-2/VEGF-A. In contrast, the best combination partner for radiotherapy was anti-VEGF-A. In conclusion, while TMZ chemotherapy benefits most from combination with anti-Ang-2/VEGF-A, radiotherapy does from anti-VEGF-A. The findings imply that uninformed combination regimens of antiangiogenic and cytotoxic therapies should be avoided.

The translatability of mouse models for the clinical development of cancer immunotherapies remains limited because of substantial differences between the murine and the human immune systems, as well as dissimilarities in the tumor biology. Non-human primates (NHPs) display good homologies towards the human immune system, based on the development of vaccines to several human pathogens. Tumor-bearing monkeys (TBMs) are NHPs that spontaneously developed cancer with progression patterns similar to humans, potentially bridging the experimental gap between early preclinical models and cancer patients treated with immunotherapeutic agents. TBMs are prevalently rhesus macaques (Macaca mulata) and the most common cancer types are of gastrointestinal, urogenital and endocrine origin. Rhesus macaques are genetically similar to humans and share many characteristics of aging. In both, humans and rhesus macaques, cancer incidence increases with age with the greatest incidence in those over 60 years of age and 20 years, respectively. TBMs were recently employed to assess the tumor targeting and the pharmacodynamics of the FAP-expressing tumor stroma-targeted immunocytokine FAP-IL2v (Evers et al, AACR 2014, Abstract 2592) and costimulatory agonist FAP-4-1BBL (Claus et al, Science translational Medicine, 2019). In the latter study, we could show targeting of FAP-4-1BBL to FAP-expressing tumor stroma and lymph nodes in a colorectal cancer-bearing rhesus monkey. These data were the basis to investigate tumor targeting of FAP-4-1BBL in an on-going clinical imaging study. In the present work, we validated further TBMs as translational models for cancer immunotherapy, by performing an imaging/biomarker study in animals exposed to a second FAP-targeted TNFRSF agonist. Two breast cancer-bearing rhesus monkeys (one triple-negative and one Luminal A) were first pre-immunized with a diphtheria/pertussis/tetanus (DTP) vaccine, and then exposed to a single injection of the TNFRSF agonist. In both animals, we could measure a strong systemic immune activation, induction of the TNFRSF agonist in T cells and also tumor regression. These observations validated the applied pre-immunization strategy to induce the TNFRSF protein expression, and confirmed the target to be pursued in the clinical setting. In conclusion, we consider TBMs as valuable translational animal models to generate proof-of-mechanism evidence in small “signal-seeking” preclinical studies. Tumor targeting, biodistribution, peripheral and intra-tumoral pharmacodynamic activity, pharmacokinetics, immunogenicity, intra-tumoral metabolic activity and tumor regression can all be assessed in TBMs. Because of the similar tumor stroma biology shared between humans and rhesus macaques, TBMs are particularly well suited to test FAP-targeting agents. We anticipate that testing cancer immunotherapy compounds in TBMs could be of high predictability for clinical behavior. Citation Format: Maurizio Ceppi, Michael Hettich, Volker Teichgräber, Wouter Driessen, Dietrich Tuerck, Estelle Marrer-Berger, Stefan Evers, Flavio Crameri, Bruno Gomes, Jürgen Bachl, Christian Klein, Christina Claus, Maria Amann, Oliver Krieter, Greg Dugan, David Caudell, Jason Grayson, Sai Kumar Solingapuram Kiran, Mark Cline. Tumor-bearing non-human primates: An unrivaled model for translational cancer immunology research [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6135.