Characterizing responsive and refractory orthotopic mouse models of hepatocellular carcinoma in cancer immunotherapy.

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and has a high mortality rate due to limited treatment options. Hence, the response of HCC to different cancer immunotherapies is being intensively investigated in clinical trials. Immune checkpoint blockers (ICB) show promising results, albeit for a minority of HCC patients. Mouse models are commonly used to evaluate new therapeutic agents or regimens. However, to make clinical translation more successful, better characterized preclinical models are required. We therefore extensively investigated two immune-competent orthotopic HCC mouse models, namely transplanted Hep-55.1c and transgenic iAST, with respect to morphological, immunological and genetic traits and evaluated both models' responsiveness to immunotherapies. Hep-55.1c tumors were characterized by rich fibrous stroma, high mutational load and pronounced immune cell infiltrates, all of which are features of immune-responsive tumors. These characteristics were less distinct in iAST tumors, though these were highly vascularized. Cell depletion revealed that CD8+ T cells from iAST mice do not affect tumor growth and are tumor tolerant. This corresponds to the failure of single and combined ICB targeting PD-1 and CTLA-4. In contrast, combining anti-PD-1 and anti-CTLA-4 showed significant antitumor efficacy in the Hep-55.1c mouse model. Collectively, our data comprehensively characterize two immune-competent HCC mouse models representing ICB responsive and refractory characteristics. Our characterization confirms these models to be suitable for preclinical investigation of novel cancer immunotherapy approaches that aim to either deepen preexisting immune responses or generate de novo immunity against the tumor.

New model for simulation of fracture repair in full-grown beagle dogs: model characterization and results from a long-term study with ibandronate.

INTRODUCTION: Given that bisphosphonates reduce bone turnover, it is important to establish that their long-term administration does not impair bone quality. This paper describes a new model for simulation of fracture repair to evaluate several aspects of bone quality following long-term administration (34 or 36 weeks) of ibandronate in full-grown beagle dogs. METHODS: The treatment schedule consisted of continuous daily subcutaneous administration of a pharmacologically active dose (1 microg/kg/day) and two cyclical intermittent regimens providing a similar total dose per animal at the end of the experiment. Seven or 8 weeks before study end, 10 holes were drilled in the left tibia and bone marrow ablation was performed in the ipsilateral femur. Serial measurements for blood biochemistry (osteocalcin and iso-alkaline phosphatase) and bone mineral density (BMD; whole body and L1-L7) by dual-energy X-ray absorptiometry (DEXA) were performed during the experiment. Bone quality was determined at the end of the experiment by assessing early and late stage defect healing and structural, cellular, and dynamic histomorphometry (femur, tibia, and lumbar vertebrae L3 and L4). RESULTS: Healing of the drill hole defects, which simulate the first stage of fracture healing, was neither qualitatively nor quantitatively influenced by ibandronate. The same was true for the activation of cortical remodeling that occurs in the later stage of fracture healing, which started in Week 4 after surgery and declined after Week 8 in all groups. Additionally, no difference was found between the various regimens and the controls with respect to DEXA analyses, trabecular bone volume, cancellous bone tissue area, cancellous bone perimeter, osteoclast count, serum osteocalcin, or bone-specific alkaline phosphatase. DISCUSSION: In conclusion, the presence of the first and second steps of fracture healing and the fact that the histological features closely resemble those of fracture repair validate the development and characterization of a new model for simulation of fracture repair. A long-term study with a therapeutically active dose of ibandronate shows that ibandronate does not impair BMD, bone structure, bone repair, coupling, and serum parameters for bone formation and turnover after long-term administration.