Immune-phenotyping of pleomorphic dermal sarcomas suggests this entity as a potential candidate for immunotherapy
Pleomorphic dermal sarcomas (PDS) are sarcomas of the skin with local recurrences in up to 28% of cases, and distant metastases in up to 20%. Although recent evidence provides a strong rational to explore immunotherapeutics in solid tumors, nothing is known about the immune environment of PDS.
In the current study, a comprehensive immune-phenotyping of 14 PDS using RNA and protein expression analyses, as well as quantitative assessment of immune cells using an image-analysis tool was performed.
Three out of 14 PDS revealed high levels of CD8-positive tumor-infiltrating T-lymphocytes (TILs), also showing elevated levels of immune-related cytokines such as IL1A, IL2, as well as markers that were very recently linked to enhanced response of immunotherapy in malignant melanoma, including CD27, and CD40L. Using a multivariate analysis, we found a number of differentially expressed genes in the CD8-high group including: CD74, LYZ and HLA-B, while the remaining cases revealed enhanced levels of immune-suppressive cytokines including CXCL14. The “CD8-high” PDS showed strong MHC-I expression and revealed infiltration by PD-L1-, PD-1- and LAG-3-expressing immune cells. Tumor-associated macrophages (TAMs) predominantly consisted of CD68 + , CD163 + , and CD204 + M2 macrophages showing an accentuation at the tumor invasion front.
Together, we provide first explorative evidence about the immune-environment of PDS tumors that may guide future decisions whether individuals presenting with advanced PDS could qualify for immunotherapeutic options.
KeywordsImmune-phenotyping Major histocompatibility complex (MHC) NanoString Pleomorphic dermal sarcoma (PDS) RNA analysis Tumor-infiltrating lymphocytes (TILs)
Immune checkpoint inhibitors
Formalin-fixated and paraffin-embedded
Type 1 macrophages
Type 2 macrophages
Programmed cell death 1
Pleomorphic dermal sarcoma
T cell immunoreceptor with Ig and ITIM domains
We thank Wiebke Jeske and Susann Zupp for technical assistance performing the TMA, mRNA in-situ and immunohistochemical staining.
Sebastian Klein, Alexander Quaas and Doris Helbig designed the study, selected cases, conceived and carried out all experiments, analyzed and interpreted results, generated figures and tables, and performed literature research, and writing of the manuscript. Svenja Wagener-Ryczek and Maximilian Schoemmel were involved in performing the NanoString analyses. Cornelia Mauch and Reinhard Buettner were involved in designing the project. All the authors were involved in writing the paper and approved the final version of the manuscript.
This work was supported by the Deutsche Forschungsgemeinschaft through the SFB829 (Z4 to Doris Helbig and Cornelia Mauch) and the Else Kröner-Fresenius Stiftung (EKFS-2014-A06 and 2016_Kolleg.19 to Sebastian Klein).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The study protocol conformed to the ethical guidelines of the 1975 Declaration of Helsinki as reflected by the approval of the institution’s human research review committee (Ethics Committee of the Medical Faculty of University of Cologne: registration no. 15–307).
All patients gave written informed consent to the use of their tumors and their data for research.
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- 7.Borradori L, Sutton B, Shayesteh P, Daniels GA (2016) Rescue therapy with anti-programmed cell death protein 1 inhibitors (PD-1) of advanced cutaneous squamous cell carcinoma and basosquamous carcinoma: preliminary experience in 5 cases. Br J Dermatol 175(6):1382-1386 https://doi.org/10.1111/bjd.14642 CrossRefGoogle Scholar
- 12.Barberis I, Martini M, Iavarone F, Orsi A (2016) Available influenza vaccines: immunization strategies, history and new tools for fighting the disease. J Prev Med Hyg 57:E41–E46Google Scholar
- 13.Afanasiev OK, Yelistratova L, Miller N, Nagase K, Paulson K, Iyer JG, Ibrani D, Koelle DM, Nghiem P (2013) Merkel polyomavirus-specific T cells fluctuate with merkel cell carcinoma burden and express therapeutically targetable PD-1 and Tim-3 exhaustion markers. Clin Cancer Res 19:5351–5360. https://doi.org/10.1158/1078-0432.CCR-13-0035 CrossRefGoogle Scholar
- 22.Allred DC, Harvey JM, Berardo M, Clark GM (1998) Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol 11:155–168Google Scholar
- 26.Fehrenbacher L, Spira A, Ballinger M et al (2016) Atezolizumab versus docetaxel for patients with previously treated non-small-cell lung cancer (POPLAR): a multicentre, open-label, phase 2 randomised controlled trial. Lancet 387:1837–1846. https://doi.org/10.1016/S0140-6736(16)00587-0 CrossRefGoogle Scholar
- 27.Jensen TO, Schmidt H, Moller HJ, Hoyer M, Maniecki MB, Sjoegren P, Christensen IJ, Steiniche T (2009) Macrophage markers in serum and tumor have prognostic impact in American joint committee on cancer stage I/II melanoma. J Clin Oncol 27:3330–3337. https://doi.org/10.1200/JCO.2008.19.9919 CrossRefGoogle Scholar
- 34.Starnes T, Rasila KK, Robertson MJ, Brahmi Z, Dahl R, Christopherson K, Hromas R (2006) The chemokine CXCL14 (BRAK) stimulates activated NK cell migration: implications for the downregulation of CXCL14 in malignancy. Exp Hematol 34:1101–1105. https://doi.org/10.1016/j.exphem.2006.05.015 CrossRefGoogle Scholar
- 35.Kudjawu YC, Chatellier G, Decool E, de Maria F, Beltzer N, Gremy I, Meyer G, Eilstein D (2016) Timing in initiating lung cancer treatment after bronchoscopy in France: study from medico-administrative database. Lung Cancer 95:44–50. https://doi.org/10.1016/j.lungcan.2016.02.016 CrossRefGoogle Scholar
- 41.Roberts DJ, Franklin NA, Kingeter LM, Yagita H, Tutt AL, Glennie MJ, Bullock TN (2010) Control of established melanoma by CD27 stimulation is associated with enhanced effector function and persistence, and reduced PD-1 expression of tumor infiltrating CD8(+) T cells. J Immunother 33:769–779. https://doi.org/10.1097/CJI.0b013e3181ee238f CrossRefGoogle Scholar
- 42.Soong RS, Song L, Trieu J, Lee SY, He L, Tsai YC, Wu TC, Hung CF (2014) Direct T cell activation via CD40 ligand generates high avidity CD8+ T cells capable of breaking immunological tolerance for the control of tumors. PLoS One 9:e93162. https://doi.org/10.1371/journal.pone.0093162 CrossRefGoogle Scholar