Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages

JOURNAL OF HEPATOLOGY

卷:66

Article

Raggi, Chiara¹  Correnti, Margherita¹  Sica, Antonio^3,4  Andersen, Jesper B.⁵  Cardinale, Vincenzo⁶  Alvaro, Domenico⁶  Chiorino, Giovanna⁷  Forti, Elisa¹  Glaser, Shannon⁸  Alpini, Gianfranco⁸  Destro, Annarita⁹  Sozio, Francesca^10,11  Di Tommaso, Luca^9,12  Roncalli, Massimo^9,12  Banales, Jesus M.¹³  Coulouarn, Cedric¹⁴  Bujanda, Luis¹³  Torzilli, Guido¹⁵  Invernizzi, Pietro^1,2 

[1] Humanitas Clin & Res Ctr, Ctr Autoimmune Liver Dis, Rozzano, Italy

[2] Univ Milano Bicocca, Dept Med & Surg, Int Ctr Digest Hlth, Program Autoimmune Liver Dis, Via Cadore 48, I-20900 Monza, MB, Italy

[3] Humanitas Clin & Res Ctr, Lab Mol Immunol, Rozzano, Italy

[4] Univ Piemonte Orientale Amedeo Avogadro Novara, Dept Pharmaceut Sci, Novara, Italy

[5] Univ Copenhagen, Biotech Res & Innovat Ctr, Copenhagen, Denmark

[6] Sapienza Univ Rome, Dept Med Surg Sci & Biotechnol, Rome, Italy

[7] Fdn Edo & Elvo Tempia, Canc Genom Lab, Biella, Italy

[8] Texas A&M Hlth Sci Ctr, Scott & White Dept Med, Cent Texas Vet Hlth Care Syst, Scott & White Digest Dis Res Ctr,Res,Coll Med, Temple, TX USA

[9] Humanitas Res Hosp, Pathol Unit, Rozzano, Italy

[10] Humanitas Clin & Res Ctr, Leukocyte Migrat Lab, Rozzano, Italy

[11] Univ Brescia, Dept Mol & Translat Med, Brescia, Italy

[12] Univ Milan, Sch Med, Milan, Italy

[13] Univ Basque Country UPV EHU, Dept Liver & Gastrointestinal Dis, Ikerbasque, Biodonostia Res Inst Donostia Univ Hosp,CIBERehd, San Sebastian, Spain

[14] Hop Pontchaillou, INSERM, U991, F-35033 Rennes, France

[15] Humanitas Res Hosp, Dept Hepatobiliary & Gen Surg, Rozzano, Italy

关键词: Cholangiocarcinoma;    Cancer stem cells;    Tumor-associated macrophages;   

DOI  :  10.1016/j.jhep.2016.08.012

来源: Elsevier

【 摘 要 】

Background & Aims: A therapeutically challenging subset of cells, termed cancer stem cells (CSCs) are responsible for cholangiocarcinoma (CCA) clinical severity. Presence of tumor associated macrophages (TAMs) has prognostic significance in CCA and other malignancies. Thus, we hypothesized that CSCs may actively shape their tumor-supportive immune niche. Methods: CCA cells were cultured in 3D conditions to generate spheres. CCA sphere analysis of in vivo tumorigenicengraftment in immune-deficient mice and molecular characterization was performed. The in vitro and in vivo effect of CCA spheres on macrophage precursors was tested after culturing healthy donor cluster of differentiation (CD)14(+) with CCA-sphere conditioned medium. Results: CCA spheres engrafted in 100% of transplanted mice and revealed a significant 20.3-fold increase in tumor-initiating fraction (p = 0.0011) and a sustained tumorigenic potential through diverse xenograft-generations. Moreover, CCA spheres were highly enriched for CSC, liver cancer and embryonic stem cell markers both at gene and protein levels. Next, fluorescence activated cell sorting analysis showed that in the presence of CCA sphere conditioned medium, CD14(+) macrophages expressed key markers (CD68, CD115, human leukocyte antigen-D related, CD206) indicating that CCA sphere conditioned medium was a strong macrophage-activator. Gene expression profile of CCA sphere activated macrophages revealed unique molecular TAM-like features confirmed by high invasion capacity. Also, freshly isolated macrophages from CCA resections recapitulated a similar molecular phenotype of in vitro-educated macrophages. Consistent with invasive features, the largest CD163(+) set was found in the tumor front of human CCA specimens (n = 23) and correlated with a high level of serum cancer antigen 19.9 (n = 17). Among mediators released by CCA spheres, only interleukin (IL)13, IL34 and osteoactivin were detected and further confirmed in CCA patient sera (n = 12). Surprisingly, a significant association of IL13, IL34 and osteoactivin with sphere stem-like genes was provided by a CCA database (n = 104). In vitro combination of IL13, IL34, osteoactivin was responsible for macrophage-differentiation and invasion, as well as for in vivo tumor-promoting effect. Conclusion: CCA-CSCs molded a specific subset of stem-like associated macrophages thus providing a rationale for a synergistic therapeutic strategy for CCA-disease. Lay summary: Immune plasticity represents an important hallmark of tumor outcome. Since cancer stem cells are able to manipulate stromal cells to their needs, a better definition of the key dysregulated immune subtypes responsible for cooperating in supporting tumor initiation may facilitate the development of new therapeutic approaches. Considering that human cholangiocarcinoma represents a clinical emergency, it is essential to move to predictive models in order to understand the adaptive process of macrophage component (imprinting, polarization and maintenance) engaged by tumor stem-like compartment. (C) 2016 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

【 授权许可】

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