Blood and tissue biomarker analysis in dogs with osteosarcoma treated with palliative radiation and intra-tumoral autologous natural killer cell transfer

Sean J. Judge, Mio Yanagisawa, Ian R. Sturgill, Sarah B. Bateni, Alicia A. Gingrich, Jennifer A. Foltz, Dean A. Lee, Jaime F. Modiano, Arta M. Monjazeb, William T.N. Culp, Robert B. Rebhun, William J. Murphy, Michael S. Kent, Robert J. Canter

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1 Scopus citations

Abstract

We have previously reported radiation-induced sensitization of canine osteosarcoma (OSA) to natural killer (NK) therapy, including results from a first-in-dog clinical trial. Here, we report correlative analyses of blood and tissue specimens for signals of immune activation in trial subjects. Among 10 dogs treated with palliative radiotherapy (RT) and intra-tumoral adoptive NK transfer, we performed ELISA on serum cytokines, flow cytometry for immune phenotype of PBMCs, and PCR on tumor tissue for immune-related gene expression. We then queried The Cancer Genome Atlas (TCGA) to evaluate the association of cytotoxic/ immune-related gene expression with human sarcoma survival. Updated survival analysis revealed five 6-month survivors, including one dog who lived 17.9 months. Using feeder line co-culture for NK expansion, we observed maximal activation of dog NK cells on day 17–19 post isolation with near 100% expression of granzyme B and NKp46 and high cytotoxic function in the injected NK product. Among dogs on trial, we observed a trend for higher baseline serum IL-6 to predict worse lung metastasis-free and overall survival (P = 0.08). PCR analysis revealed low absolute gene expression of CD3, CD8, and NKG2D in untreated OSA. Among treated dogs, there was marked heterogeneity in the expression of immune-related genes pre- and post-treatment, but increases in CD3 and CD8 gene expression were higher among dogs that lived > 6 months compared to those who did not. Analysis of the TCGA confirmed significant differences in survival among human sarcoma patients with high and low expression of genes associated with greater immune activation and cytotoxicity (CD3e, CD8a, IFN-γ, perforin, and CD122/IL-2 receptor beta). Updated results from a first-in-dog clinical trial of palliative RT and autologous NK cell immunotherapy for OSA illustrate the translational relevance of companion dogs for novel cancer therapies. Similar to human studies, analyses of immune markers from canine serum, PBMCs, and tumor tissue are feasible and provide insight into potential biomarkers of response and resistance.

Original languageEnglish (US)
Article numbere0224775
JournalPloS one
Volume15
Issue number2
DOIs
StatePublished - Feb 1 2020

Bibliographical note

Funding Information:
This work was supported in part by National Institute for Health/National Cancer Institute grant R01 CA189209 (WJM) and U01 CA224166?01 (RJC, RBR). Additional funding was provided by the Society for Surgical Oncology Foundation (RJC), the Sarcoma Foundation of America (RJC), and the University of California Coordinating Committee for Cancer Control CRR-13-201,404 (RJC).

Publisher Copyright:
© 2020 Judge et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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