The Blockade of Tumoral IL1β-Mediated Signaling in Normal Colonic Fibroblasts Sensitizes Tumor Cells to Chemotherapy and Prevents Inflammatory CAF Activation
Abstract
:1. Introduction
2. Results
2.1. IL1β mRNA Expression in Colorectal Cell Lines, Fibroblasts and Cocultures
2.2. IL1β Receptor Expression in Colorectal Cell Lines and NCFs
2.3. Differential Impact of IL1β on the Proliferation of Tumor Cells and Fibroblasts
2.4. Exploring the Effect of IL1β and Its Targets on Fibroblast and Tumor Cell Migration and Recruitment
2.5. Chemoresistance Induced by IL1β-Responsive Soluble Factors
3. Discussion
4. Methods
4.1. Culture of Primary Fibroblasts and Preparation of Conditioned Medium (CM)
4.2. Colorectal Cancer Cell Lines
4.3. Cocultures of NCFs with Colorectal Cancer Cell Lines (CCCL)
4.4. RNA Isolation and Quantitative Real-Time PCR (qRT-PCR)
4.5. Cell Proliferation and Viability
4.6. Wound-Healing Assay and Directional Migration
4.7. Silencing IL1β by Lentiviral shRNA
4.8. Cytokines and Chemicals
4.9. Western Blot Analysis
4.10. Cytokine Arrays and ELISA
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
- Bissell, M.J.; Hines, W.C. Why don’t we get morecancer? A proposed role of the microenvironment in restraining cancer progression. Nat. Med. 2011, 17, 320. [Google Scholar] [CrossRef] [Green Version]
- Hanahan, D.; Coussens, L.M. Accessories to the crime: Functions of cells recruited to the tumor microenvironment. Cancer Cell 2012, 21, 309. [Google Scholar] [CrossRef] [Green Version]
- Hanahan, D.; Weinberg, R.A. Hallmarks of cancer: The next generation. Cell 2011, 144, 646. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kerbel, R.S. Tumor angiogenesis. N. Engl. J. Med. 2008, 358, 2039. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kalluri, R.; Zeisberg, M. Fibroblasts in cancer. Nat. Rev. Cancer 2006, 6, 392. [Google Scholar] [CrossRef] [PubMed]
- Yamaguchi, H.; Sakai, R. Direct Interaction between Carcinoma Cells and Cancer Associated Fibroblasts for the Regulation of Cancer Invasion. Cancers 2015, 7, 2054. [Google Scholar] [CrossRef]
- Li, J.; Chen, L.; Qin, Z. Multifaceted tumor stromal fibroblasts. Cancer Microenviron. 2012, 5, 187. [Google Scholar] [CrossRef] [Green Version]
- Webber, J.; Steadman, R.; Mason, M.D.; Tabi, Z.; Clayton, A. Cancer exosomes trigger fibroblast to myofibroblast differentiation. Cancer Res. 2010, 70, 9621. [Google Scholar] [CrossRef] [Green Version]
- Roulis, M.; Nikolaou, C.; Kotsaki, E.; Kaffe, E.; Karagianni, N.; Koliaraki, V.; Salpea, K.; Ragoussis, J.; Aidinis, V.; Martini, E.; et al. Intestinal myofibroblast-specific Tpl2-Cox-2-PGE2 pathway links innate sensing to epithelial homeostasis. Proc. Natl. Acad. Sci. USA 2014, 111, E4658. [Google Scholar] [CrossRef] [Green Version]
- Tjomsland, V.; Spångeus, A.; Välilä, J.; Sandström, P.; Borch, K.; Druid, H.; Falkmer, S.; Falkmer, U.; Messmer, D.; Larsson, M. Interleukin 1α sustains the expression of inflammatory factors in human pancreatic cancer microenvironment by targeting cancer-associated fibroblasts. Neoplasia 2011, 13, 664. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kogan-Sakin, I.; Cohen, M.; Paland, N.; Madar, S.; Solomon, H.; Molchadsky, A.; Brosh, R.; Buganim, Y.; Goldfinger, N.; Klocker, H.; et al. Prostate stromal cells produce CXCL-1, CXCL-2, CXCL-3 and IL-8 in response to epithelia-secreted IL-1. Carcinogenesis 2009, 30, 698. [Google Scholar] [CrossRef] [Green Version]
- Dudás, J.; Fullár, A.; Bitsche, M.; Schartinger, V.; Kovalszky, I.; Sprinzl, G.M.; Riechelmann, H. Tumor-produced, active interleukin-1β regulates gene expression in carcinoma-associated fibroblasts. Exp. Cell Res. 2011, 317, 2222. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Berdiel-Acer, M.; Sanz-Pamplona, R.; Calon, A.; Cuadras, D.; Berenguer, A.; Sanjuan, X.; Paules, M.J.; Salazar, R.; Moreno, V.; Batlle, E.; et al. Differences between CAFs and their paired NCF from adjacent colonic mucosa reveal functional heterogeneity of CAFs, providing prognostic information. Mol. Oncol. 2014, 8, 1290. [Google Scholar] [CrossRef] [PubMed]
- Bauer, M.; Su, G.; Casper, C.; He, R.; Rehrauer, W.; Friedl, A. Heterogeneity of gene expression in stromal fibroblasts of human breast carcinomas and normal breast. Oncogene 2010, 29, 1732. [Google Scholar] [CrossRef] [Green Version]
- Wu, T.; Hong, Y.; Jia, L.; Wu, J.; Xia, J.; Wang, J.; Hu, Q.; Cheng, B. Modulation of IL-1β reprogrammes the tumor microenvironment to interrupt oral carcinogenesis. Sci. Rep. 2016, 6, 20208. [Google Scholar] [CrossRef] [Green Version]
- Voronov, E.; Shouval, D.S.; Krelin, Y.; Cagnano, E.; Benharroch, D.; Iwakura, Y.; Dinarello, C.A.; Apte, R.N. IL-1 is required for tumor invasiveness and angiogenesis. Proc. Natl. Acad. Sci. USA 2003, 100, 2645. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xu, D.; Matsuo, Y.; Ma, J.; Koide, S.; Ochi, N.; Yasuda, A.; Funahashi, H.; Okada, Y.; Takeyama, H. Cancer cell-derived IL-1α promotes HGF secretion by stromal cells and enhances metastatic potential in pancreatic cancer cells. J. Surg. Oncol. 2010, 102, 469. [Google Scholar] [CrossRef]
- Mendoza-Rodríguez, M.; Arévalo Romero, H.; Fuentes-Pananá, E.M.; Ayala-Sumuano, J.T.; Meza, I. IL-1β induces up-regulation of BIRC3, a gene involved in chemoresistance to doxorubicin in breast cancer cells. Cancer Lett. 2017, 390, 39. [Google Scholar] [CrossRef]
- Zhang, D.; Li, L.; Jiang, H.; Li, Q.; Wang-Gillam, A.; Yu, J.; Head, R.; Liu, J.; Ruzinova, M.B.; Lim, K.H.; et al. Tumor-Stroma IL1β-IRAK4 Feedforward Circuitry Drives Tumor Fibrosis, Chemoresistance, and Poor Prognosis in Pancreatic Cancer. Cancer Res. 2018, 78, 1700 LP. [Google Scholar] [CrossRef] [Green Version]
- Neuzillet, C.; Tijeras-Raballand, A.; Ragulan, C.; Cros, J.; Patil, Y.; Martinet, M.; Erkan, M.; Kleeff, J.; Wilson, J.; Apte, M.; et al. Inter- and intra-tumoral heterogeneity in cancer-associated fibroblasts of human pancreatic ductal adenocarcinoma. J. Pathol. 2019, 248, 51–65. [Google Scholar] [CrossRef] [Green Version]
- Hsia, L.; Ashley, N.; Ouaret, D.; Wang, L.M.; Wilding, J.; Bodmer, W.F. Myofibroblasts are distinguished from activated skin fibroblasts by the expression of AOC3 and other associated markers. Proc. Natl. Acad. Sci. USA 2016, 113, E2162. [Google Scholar] [CrossRef] [Green Version]
- Lewis, M.P.; Lygoe, K.A.; Nystrom, M.L.; Anderson, W.P.; Speight, P.M.; Marshall, J.F.; Thomas, G.J. Tumor-derived TGF-beta1 modulates myofibroblast differentiation and promotes HGF/SF-dependent invasion of squamous carcinoma cells. Br. Cancer J. 2004, 90, 822–832. [Google Scholar] [CrossRef] [Green Version]
- Scotton, C.J.; Chambers, R.C. Molecular targets in pulmonary fibrosis: The myofibroblast in focus. Chest 2007, 132, 1311. [Google Scholar] [CrossRef]
- Fuschiotti, P. Role of IL-13 in systemic sclerosis. Cytokine 2011, 56, 544. [Google Scholar] [CrossRef] [PubMed]
- Kendall, R.T.; Feghali-Bostwick, C.A. Fibroblasts in fibrosis: Novel roles and mediators. Front. Pharmacol. 2014, 5, 123. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Öhlund, D.; Elyada, E.; Tuveson, D. Fibroblast heterogeneity in the cancer wound. J. Exp. Med. 2014, 211, 1503. [Google Scholar] [CrossRef]
- Costa, A.; Kieffer, Y.; Scholer-Dahirel, A.; Pelon, F.; Bourachot, B.; Cardon, M.; Sirven, P.; Magagna, I.; Fuhrmann, L.; Bernard, C.; et al. Heterogeneity and Immunosuppressive Environment in Human Breast Cancer. Cancer Cell 2018, 33, 463. [Google Scholar] [CrossRef] [Green Version]
- Lauri, D.; Bertomeu, M.C.; Orr, F.W.; Bastida, E.; Sauder, D.; Buchanan, M.R. Interleukin-1 increases tumor cell adhesion to endothelial cells through an RGD dependent mechanism: In vitro and in vivo studies. Clin. Exp. Metastasis 1990, 8, 27. [Google Scholar] [CrossRef] [PubMed]
- Giavazzi, R.; Garofalo, A.; Bani, M.R.; Abbate, M.; Ghezzi, P.; Boraschi, D.; Mantovani, A.; Dejana, E. Interleukin 1-induced augmentation of experimental metastases from a human melanoma in nude mice. Cancer Res. 1990, 50, 4771. [Google Scholar]
- Erez, N.; Truitt, M.; Olson, P.; Hanahan, D. Cancer-Associated Fibroblasts Are Activated in Incipient Neoplasia to Orchestrate Tumor-Promoting Inflammation in an NF-kappaB-Dependent Manner. Cancer Cell 2010, 17, 125. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Arlt, A.; Vorndamm, J.; Müerköster, S.; Yu, H.; Schmidt, W.E.; Fölsch, U.R.; Schäfer, H. Autocrine production of interleukin 1beta confers constitutive nuclear factor kappaB activity and chemoresistance in pancreatic carcinoma cell lines. Cancer Res. 2002, 62, 910. [Google Scholar] [PubMed]
- Young, H.; Rowling, E.J.; Bugatti, M.; Giurisato, E.; Luheshi, N.; Arozarena, I.; Acosta, J.C.; Kamarashev, J.; Frederick, D.; Cooper, T.; et al. An adaptative signaling network in melanoma inflammatory niches confers tolerance to MAPK signaling inhibition. J. Exp. Med. 2017, 214, 1691–1710. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Su, S.; Chen, J.; Yao, H.; Liu, J.; Yu, S.; Lao, L.; Wang, M.; Luo, M.; Xing, Y.; Chen, F.; et al. CD10+GPR77+ Cancer-Associated Fibroblasts Promote Cancer Formation and Chemoresistance by Sustaining Cancer Stemness. Cell 2018, 172, 841. [Google Scholar] [CrossRef] [PubMed]
- Öhlund, D.; Handly-Santana, A.; Biffi, G.; Elyada, E.; Almeida, A.S.; Ponz-Sarvise, M.; Corbo, V.; Oni, T.E.; Hearn, S.A.; Lee, E.J.; et al. Distinct populations of inflammatory fibroblasts and myofibroblasts in pancreatic cancer. J. Exp. Med. 2017, 214, 579. [Google Scholar] [CrossRef] [PubMed]
- Gonçalves-Ribeiro, S.; Díaz-Maroto, N.G.; Berdiel-Acer, M.; Soriano, A.; Guardiola, J.; Martínez-Villacampa, M.; Salazar, R.; Capellà, G.; Villanueva, A.; Martínez-Balibrea, E. Carcinoma-associated fibroblasts affect sensitivity to oxaliplatin and 5FU in colorectal cancer cells. Oncotarget 2016, 7, 59766. [Google Scholar] [CrossRef] [Green Version]
- Rider, P.; Carmi, Y.; Cohen, I. Biologics for Targeting Inflammatory Cytokines, Clinical Uses, and Limitations. Int. Cell J. Biol. 2016, 2016, 9259646. [Google Scholar] [CrossRef] [Green Version]
- Grassi, A.; Perilli, L.; Albertoni, L.; Tessarollo, S.; Mescoli, C.; Urso, E.D.L.; Fassan, M.; Rugge, M.; Zanovello, P. A coordinate deregulation of microRNAs expressed in mucosa adjacent to tumor predicts relapse after resection in localized colon cancer. Mol. Cancer 2018, 17, 17. [Google Scholar] [CrossRef] [Green Version]
- Sanz-Pamplona, R.; Berenguer, A.; Cordero, D.; Molleví, D.G.; Crous-Bou, M.; Sole, X.; Paré-Brunet, L.; Guino, E.; Salazar, R.; de Santos, C.; et al. Aberrant gene expression in mucosa adjacent to tumor reveals a molecular crosstalk in colon cancer. Mol. Cancer 2014, 13, 46. [Google Scholar] [CrossRef] [Green Version]
- Elyada, E.; Bolisetty, M.; Laise, P.; Flynn, W.; Courtois, E.; Burkhart, R.; Teinor, J.; Belleau, P.; Biffi, G.; Lucito, M.; et al. Cross-species single-cell analysis of pancreatic ductal adenocarcinoma reveals antigen-presenting cancer-associated fibroblasts. Cancer Discov. 2019, 9, 1102–1123. [Google Scholar] [CrossRef] [Green Version]
- Dominguez, C.; Müller, S.; Keerthivasan, S.; Koeppen, H.; Hung, J.; Gierke, S.; Breart, B.; Foreman, O.; Bainbridge, T.; Castiglioni, A.; et al. Single-cell RNA sequencing reveals stromal evolution into LRRC15+ myofibroblasts as a determinant of patient response to cancer immunotherapy. Cancer Discov. 2020, 10, 232–253. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Guillen, N.; Sanz-Pamplona, R.; Berdiel-Hacer, M.; Cimas, F.J.; García, E.; Gonçalves-Ribeiro, S.; Albert, N.; García-Vicién, G.; Capella, G.; Moreno, V.; et al. Noncanonical TGFβ pathway relieves the blockade of IL1β/TGFβ-mediated crosstalk between tumor and stroma: TGFBR1 and TAK1 inhibition in colorectal cancer. Clin. Cancer. Res. 2019, 25, 4466–4479. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Biffi, G.; TOni, E.; Spielman, B.; Hao, Y.; Elyada, E.; Park, Y.; Preall, J.; Tuveson, D.A. Il1-induced JAK/STAT signaling is antagonized by TGFb to shape CAF heterogeneity in pancreatic ductal adenocarcinoma. Cancer Discov. 2019, 9, 282–301. [Google Scholar] [CrossRef] [Green Version]
- Hurwitz, H.; Van Cutsem, E.; Bendell, J.; Hidalgo, M.; Li, C.; Garrido Salvo, M.; Macarulla, T.; Sahai, V.; Sama, A.; Greeno, E.; et al. Ruxolitinib+ capecitabine in advanced/metastatic pancreatic cancer after disease progression/intolerance to first-line therapy: JANUS 1 and 2 randomized phase III studies. Investig. New Drugs 2018, 36, 683–695. [Google Scholar] [CrossRef] [PubMed]
- Hurwitz, H.I.; Uppal, N.; Wagner, S.A.; Bendell, J.C.; Beck, J.T.; Wade III, S.M.; Nemunaitis, J.J.; Stella, P.J.; Pipas, J.M.; Wainberg, Z.A.; et al. Ranzomized, double-blind, phase II study of Ruxolitinib or placebo in combination with Capecitabine in patients with metastatic pancreatic cancer from whom therapy with Gemcitabine has failed. Clin. J. Oncol. 2015, 33, 4039. [Google Scholar] [CrossRef] [PubMed]
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Díaz-Maroto, N.G.; Garcia-Vicién, G.; Polcaro, G.; Bañuls, M.; Albert, N.; Villanueva, A.; Molleví, D.G. The Blockade of Tumoral IL1β-Mediated Signaling in Normal Colonic Fibroblasts Sensitizes Tumor Cells to Chemotherapy and Prevents Inflammatory CAF Activation. Int. J. Mol. Sci. 2021, 22, 4960. https://doi.org/10.3390/ijms22094960
Díaz-Maroto NG, Garcia-Vicién G, Polcaro G, Bañuls M, Albert N, Villanueva A, Molleví DG. The Blockade of Tumoral IL1β-Mediated Signaling in Normal Colonic Fibroblasts Sensitizes Tumor Cells to Chemotherapy and Prevents Inflammatory CAF Activation. International Journal of Molecular Sciences. 2021; 22(9):4960. https://doi.org/10.3390/ijms22094960
Chicago/Turabian StyleDíaz-Maroto, Natalia Guillén, Gemma Garcia-Vicién, Giovanna Polcaro, María Bañuls, Nerea Albert, Alberto Villanueva, and David G. Molleví. 2021. "The Blockade of Tumoral IL1β-Mediated Signaling in Normal Colonic Fibroblasts Sensitizes Tumor Cells to Chemotherapy and Prevents Inflammatory CAF Activation" International Journal of Molecular Sciences 22, no. 9: 4960. https://doi.org/10.3390/ijms22094960