Abstract A012: Humanized patient-derived xenografts (PDXs) recapitulate clinical responses in microsatellite stable (MSS) and unstable (MSI-H) colorectal cancer (CRC)
Authors: Capasso Anna , Lang Julie , Pitts Todd M. , Lieu Christopher H. , Kopetz Scott , Davis Sarah L. , Jordan Kimberly , Bagby Stacey M. , Messersmith Wells A. , Pelanda Roberta , Eckhardt S. Gail 
Published: August 2018

Introduction: Recent data have demonstrated that patients with MSI-H metastatic CRC are more likely to respond to a PD-1 inhibitor as a single agent compared to patients with MSS CRC. Effective immunotherapy approaches for MSS CRC remain a critical unmet need. There is great interest in investigating immune checkpoint inhibition in combination with novel agents, but preclinical studies have been hampered by current models. To gain a better understanding of immune responses and facilitate preclinical evaluation of combination strategies with immunotherapy, we developed a “hematopoietic” humanized mouse model with the intent of leveraging this model for the development of rational combinations in MSS CRC patients. Methods: BRGS (in house breeding) newborn pups were transplanted with CD34+ cells purified from (HLA unmatched) umbilical cord blood at 48 hours. In order to better understand the immune context of these novel animal models, we selected an MSI-high CRC (MDA-C0999-203) and a MSS (CRC172) humanized PDX model. At 16 weeks, MDA-C0999-203 (MSI-H) and CRC172 (MSS) were implanted on both flanks of humanized mice and nonhumanized controls. When the average tumor size reached ~150-300 mm3, both groups of mice were randomized into vehicle and nivolumab (30mg/kg IP twice weekly) stratified by % chimerism (human CD45+ &gt;45%). At the end of the treatment, a portion of the tumor was immediately fixed in formalin for immunohistochemistry and pharmacodynamic analyses with seven-color multispectral imaging using the Perkin Elmer Vectra- 3 instrument, and compared to a surgical specimen from an untreated patient with an MSI-H tumor. Results: In the MSI-H model we observed tumor growth inhibition in the nivolumab-treated, humanized mice with respect to the humanized vehicle-treated control and the nonhumanized nivolumab-treated group (TGII 3.5% vs 76%). In a separate experiment, an MSS CRC humanized PDX model (CRC172) was treated with nivolumab as described above. Although initial control of tumor growth was observed, this was followed by rapid tumor progression (TGII 138.4%). In addition to flow cytometry, an MSI-H CRC tumor directly from a patient was stained for a variety of immunologic markers and analyzed by Vectra. The tumors contained both CD4- and CD8-positive cells, indicating T-cell infiltration. These data are consistent with what was observed by flow cytometry. Similar results were observed in an MSI-H CRC tumor grown as a xenograft in humanized mice both with and without nivolumab. However, in the MSS CRC PDX model, very few CD4 and CD8 cells were observed in the tumor or in the surrounding stroma. Conclusions: We have successfully established in vivo MSI-H and MSS humanized CRC PDXs. All mice were highly chimeric and the MSI-H model demonstrated high TILs, and responding tumors exhibited IFNγ production, high CD8%, higher effector memory % (HLADR+, CD45RO+), and decreased PD-L1 expression by flow cytometry. Interestingly, we also observed greater numbers of T cells in the lymph nodes of the MSI-H PDX. Moreover, immune infiltrates were observed in the MSI-H PDX compared to the MSS by immunohistochemistry. These data suggest that humanized PDX models may be useful in the development of rational combinations of immunotherapy.

Citation Format: Anna Capasso, Julie Lang, Todd M. Pitts, Christopher H. Lieu, Scott Kopetz, Sarah L. Davis, Kimberly Jordan, Stacey M. Bagby, Wells A. Messersmith, Roberta Pelanda, S. Gail Eckhardt. Humanized patient-derived xenografts (PDXs) recapitulate clinical responses in microsatellite stable (MSS) and unstable (MSI-H) colorectal cancer (CRC) [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr A012.