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Response to radiotherapy in pancreatic ductal adenocarcinoma is enhanced by inhibition of myeloid-derived suppressor cells using STAT3 anti-sense oligonucleotide

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Abstract

Pancreatic ductal adenocarcinoma (PDAC) has a heterogeneous tumor microenvironment (TME) comprised of myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages, neutrophils, regulatory T cells, and myofibroblasts. The precise mechanisms that regulate the composition of the TME and how they contribute to radiotherapy (RT) response remain poorly understood. In this study, we analyze changes in immune cell populations and circulating chemokines in patient samples and animal models of pancreatic cancer to characterize the immune response to radiotherapy. Further, we identify STAT3 as a key mediator of immunosuppression post-RT. We found granulocytic MDSCs (G-MDSCs) and neutrophils to be increased in response to RT in murine and human PDAC samples. We also found that RT-induced STAT3 phosphorylation correlated with increased MDSC infiltration and proliferation. Targeting STAT3 using an anti-sense oligonucleotide in combination with RT circumvented RT-induced MDSC infiltration, enhanced the proportion of effector T cells, and improved response to RT. In addition, STAT3 inhibition contributed to the remodeling of the PDAC extracellular matrix when combined with RT, resulting in decreased collagen deposition and fibrotic tissue formation. Collectively, our data provide evidence that targeting STAT3 in combination with RT can mitigate the pro-tumorigenic effects of RT and improve tumor response.

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Acknowledgements

We would like to acknowledge and thank Ionis Pharmaceuticals for providing the mouse surrogate STAT3 ASO. This work was funded in part by a Grant from AstraZeneca. This work was also supported by Cancer Center Support Grant (P30CA046934), R01-DE028282 (Karam), R01-DE028529 (Karam), Paul Sandoval Funds (Mueller), RSNA Resident Research Grant (Mueller), Cancer League of Colorado Grant (Mueller) and by the Wings of Hope Foundation (Karam, Goodman).

Funding

This work was supported by Cancer Center Support Grant (P30CA046934), R01-DE028282 (Karam), R01-DE028529 (Karam), Paul Sandoval Funds (Mueller), RSNA Resident Research Grant (Mueller), Cancer League of Colorado Grant (Mueller) and by the Wings of Hope Foundation (Karam, Goodman).

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Authors and Affiliations

  1. Department of Nuclear Medicine and Radiobiology, University of Sherbrooke, Sherbrooke, Canada

    Ayman J. Oweida & Sana D. Karam

  2. Bioscience, Oncology, IMED Biotech Unit, AstraZeneca, Boston, MA, USA

    Theresa Proia

  3. Department of Surgery, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Richard Schulick & Marco Del Chiaro

  4. Division of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Wells A. Messersmith

  5. Department of Anesthesiology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Eric Clambey

  6. Earle A. Chiles Research Institute, Providence Medical Center, Portland, OR, USA

    Michael J. Gough

  7. Department of Biochemistry, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Jason Williams & Kirk Hansen

  8. Department of Radiation Oncology, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Karyn Goodman

  9. Department of Immunology and Microbiology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Kimberly Jordan

  10. Department of Radiation Oncology, University of Colorado Anschutz Medical Campus, 1665 Aurora Court Suite 1032, Aurora, CO, 80045, USA

    Miles Piper, Dallin Milner, Benjamin Van Court, Shilpa Bhatia, Andy Phan, Thomas Bickett & Sana D. Karam

  11. Thomas Jefferson University, Bodine Center for Cancer Treatment, 1665 Aurora Court Suite 1032, Philadelphia, PA, USA

    Adam C. Mueller

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  1. Ayman J. Oweida
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Correspondence to Sana D. Karam.

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Ethical approval

All protocols for animal tumor models were approved by the IACUC of the University of Colorado Denver. Mice exhibiting signs of morbidity according to the guidelines set by the Institutional Animal Care and Use Committee (IACUC) were sacrificed immediately.

Informed consent

All human studies were performed after approval by the University of Colorado institutional review board (COMIRB16-1139) and written informed consent was obtained from all patients as dictated by the study protocol.

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262_2020_2701_MOESM1_ESM.pdf

Supplementary Figure 1 – (a) Identification and gating of immune populations on flow cytometry. For MDSC gating, the following criteria were used as previously described [33, 53]. In mouse samples, total MDSCs were defined as CD11b+Gr1+ cells. The two major subsets of MDSCs were differentiated by variable expression of the Gr1 marker (comprised of Ly6C and Ly6G). M-MDSCs were defined as CD11b+Ly6ChiLy6G– and G-MDSCs as CD11b+Ly6CloLy6G+. In human samples, only total MDSCs were analyzed and defined as CD11b+CD33+. (b) Effect of RT on the proportion of intratumoral T cells populations. (c) Analysis of pSTAT3 expression in various cell populations from FC1242 tumors. (d) Effect of RT on pSTAT3 expression in T cell populations. (PDF 295 kb)

262_2020_2701_MOESM2_ESM.pdf

Supplementary Figure 2 – Analysis of tumor growth in (a) FC1242 and (b) PK5L1940 tumor-bearing mice. (c) Analysis of tumor growth using STAT3 ASO alone. (PDF 227 kb)

262_2020_2701_MOESM3_ESM.pdf

Supplementary Figure 3 – (a) Relative M1/M2 ratios following STAT3 ASO + RT treatment. (b) T cell pSTAT3 expression levels following STAT3 ASO + RT treatment. (PDF 111 kb)

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Oweida, A.J., Mueller, A.C., Piper, M. et al. Response to radiotherapy in pancreatic ductal adenocarcinoma is enhanced by inhibition of myeloid-derived suppressor cells using STAT3 anti-sense oligonucleotide. Cancer Immunol Immunother 70, 989–1000 (2021). https://doi.org/10.1007/s00262-020-02701-w

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