Top

Published in:

01-06-2018 | Original Article

The Influence of Breast Tumour-Derived Factors and Wnt Antagonism on the Transformation of Adipose-Derived Mesenchymal Stem Cells into Tumour-Associated Fibroblasts

Authors: Malini Visweswaran, Kevin N. Keane, Frank Arfuso, Rodney J. Dilley, Philip Newsholme, Arun Dharmarajan

Published in: Cancer Microenvironment | Issue 1/2018

Abstract

Within the tumour stroma, a heterogeneous population of cell types reciprocally regulates cell proliferation, which considerably affects the progression of the disease. In this study, using tumour conditioned medium (TCM) derived from breast tumour cell lines – MCF7 and MDA MB 231, we have demonstrated the differentiation of adipose-derived mesenchymal stem cells (ADSCs) into tumour-associated fibroblasts (TAFs). Since the Wnt signalling pathway is a key signalling pathway driving breast tumour growth, the effect of the Wnt antagonist secreted frizzled-related protein 4 (sFRP4) was also examined. The response of ADSCs to TCM and sFRP4 treatments was determined by using cell viability assay to determine the changes in ADSC viability, immunofluorescence for mesenchymal markers, glucose uptake assay, and glycolysis stress test using the Seahorse Extracellular Flux analyser to determine the glycolytic activity of ADSCs. ADSCs have been shown to acquire a hyper-proliferative state, significantly increasing their number upon short-term and long-term exposure to TCM. Changes have also been observed in the expression of key mesenchymal markers as well as in the metabolic state of ADSCs. SFRP4 significantly inhibited the differentiation of ADSCs into TAFs by reducing cell growth as well as mesenchymal marker expression (cell line-dependent). However, sFRP4 did not induce further significant changes to the altered metabolic phenotype of ADSCs following TCM exposure. Altogether, this study suggests that the breast tumour milieu may transform ADSCs into a tumour-supportive phenotype, which can be altered by Wnt antagonism, but is independent of metabolic changes.

Junttila MR, de Sauvage FJ (2013) Influence of tumour micro-environment heterogeneity on therapeutic response. Nature 501(7467):346–354. https://doi.org/10.1038/nature12626 CrossRefPubMed

Jotzu C, Alt E, Welte G, Li J, Hennessy BT, Devarajan E, Krishnappa S, Pinilla S, Droll L, Song YH (2011) Adipose tissue derived stem cells differentiate into carcinoma-associated fibroblast-like cells under the influence of tumor derived factors. Cell Oncol 34(1):55–67CrossRef

Cho JA, Park H, Lim EH, Lee KW (2012) Exosomes from breast cancer cells can convert adipose tissue-derived mesenchymal stem cells into myofibroblast-like cells Int J Oncol:40

Cho JA, Park H, Lim EH, Kim KH, Choi JS, Lee JH (2011) Exosomes from ovarian cancer cells induce adipose tissue-derived mesenchymal stem cells to acquire the physical and functional characteristics of tumor-supporting myofibroblasts. Gynecol Oncol 123:379–386. https://doi.org/10.1016/j.ygyno.2011.08.005 CrossRefPubMed

Spaeth EL, Dembinski JL, Sasser AK, Watson K, Klopp A, Hall B, Andreeff M, Marini F (2009) Mesenchymal stem cell transition to tumor-associated fibroblasts contributes to fibrovascular network expansion and tumor progression. PLoS One 4(4):7CrossRef

Mishra P, Humeniuk R, Medina D, Alexe G, Mesirov J, Ganesan S, Glod J, Banerjee D (2008) Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells. Cancer Res 11:4331–4339CrossRef

Paunescu V, Bojin FM, Tatu CA, Gavriliuc OI, Rosca A, Gruia AT (2011) Tumour-associated fibroblasts and mesenchymal stem cells: more similarities than differences. J Cell Mol Med 15:635–646. https://doi.org/10.1111/j.1582-4934.2010.01044.x CrossRefPubMed

Matushansky I, Hernando E, Socci ND, Mills JE, Matos TA, Edgar MA, Singer S, Maki RG, Cordon-Cardo C (2007) Derivation of sarcomas from mesenchymal stem cells via inactivation of the Wnt pathway. J Clin Invest 117(11):3248–3257CrossRefPubMedPubMedCentral

Jian H, Shen X, Liu I, Semenov M, He X, Wang XF (2006) Smad3-dependent nuclear translocation of beta-catenin is required for TGF-beta1-induced proliferation of bone marrow-derived adult human mesenchymal stem cells. Genes Dev 20(6):666–674. https://doi.org/10.1101/gad.1388806 CrossRefPubMedPubMedCentral

10.

Sato M (2006) Upregulation of the Wnt/beta-catenin pathway induced by transforming growth factor-beta in hypertrophic scars and keloids. Acta Derm Venereol 86(4):300–307. https://doi.org/10.2340/00015555-0101 CrossRefPubMed

11.

DiRenzo DM, Chaudhary MA, Shi X, Franco SR, Zent J, Wang K, Guo L-W, Kent KC (2016) A crosstalk between TGF-β/Smad3 and Wnt/β-catenin pathways promotes vascular smooth muscle cell proliferation. Cell Signal 28(5):498–505. https://doi.org/10.1016/j.cellsig.2016.02.011 CrossRefPubMedPubMedCentral

12.

Zhou S (2011) TGF-β regulates β-catenin signaling and osteoblast differentiation in human mesenchymal stem cells. J Cell Biochem 112(6):1651–1660. https://doi.org/10.1002/jcb.23079 CrossRefPubMedPubMedCentral

13.

Akhmetshina A, Palumbo K, Dees C, Bergmann C, Venalis P, Zerr P, Horn A, Kireva T, Beyer C, Zwerina J, Schneider H, Sadowski A, Riener M-O, MacDougald OA, Distler O, Schett G, Distler JHW (2012) Activation of canonical Wnt signalling is required for TGF-β-mediated fibrosis. Nat Commun 3:735. https://doi.org/10.1038/ncomms1734 CrossRefPubMedPubMedCentral

14.

Saran U, Arfuso F, Zeps N, Dharmarajan A (2012) Secreted frizzled-related protein 4 expression is positively associated with responsiveness to cisplatin of ovarian cancer cell lines in vitro and with lower tumour grade in mucinous ovarian cancers. BMC Cell Biol 13(1):25CrossRefPubMedPubMedCentral

15.

Muley A, Majumder S, Kolluru GK, Parkinson S, Viola H, Hool L, Arfuso F, Ganss R, Dharmarajan A, Chatterjee S (2010) Secreted frizzled-related protein 4: an angiogenesis inhibitor. Am J Pathol 176(3):1505–1516CrossRefPubMedPubMedCentral

16.

Wolf V, Ke G, Dharmarajan AM, Bielke W, Artuso L, Saurer S, Friis R (1997) DDC-4, an apoptosis-associated gene, is a secreted frizzled relative. FEBS Letters 417 (3):385–389. doi:doi:https://doi.org/10.1016/S0014-5793(97)01324-0

17.

Warrier S, Balu SK, Kumar AP, Millward M, Dharmarajan A (2013) Wnt antagonist, secreted frizzled-related protein 4 (sFRP4), increases chemotherapeutic response of glioma stem-like cells. Oncol Res 21(2):93–102. https://doi.org/10.3727/096504013x13786659070154 CrossRefPubMed

18.

Perumal V, Pohl S, Keane KN, Arfuso F, Newsholme P, Fox S, Dharmarajan A (2016) Therapeutic approach to target mesothelioma cancer cells using the Wnt antagonist, secreted frizzled-related protein 4: metabolic state of cancer cells. Exp Cell Res 341(2):218–224. https://doi.org/10.1016/j.yexcr.2016.02.008 CrossRefPubMed

19.

Anderberg C, Pietras K (2009) On the origin of cancer-associated fibroblasts. Cell Cycle 8(10):1461–1462. https://doi.org/10.4161/cc.8.10.8560 CrossRefPubMed

20.

Hanley CJ, Noble F, Ward M, Bullock M, Drifka C, Mellone M, Manousopoulou A, Johnston HE, Hayden A, Thirdborough S, Liu Y, Smith DM, Mellows T, Kao WJ, Garbis SD, Mirnezami A, Underwood TJ, Eliceiri KW, Thomas GJ (2016) A subset of myofibroblastic cancer-associated fibroblasts regulate collagen fiber elongation, which is prognostic in multiple cancers. Oncotarget 7 (5):6159–6174. doi:https://doi.org/10.18632/oncotarget.6740

21.

Luo H, Tu G, Liu Z, Liu M (2015) Cancer-associated fibroblasts: a multifaceted driver of breast cancer progression. Cancer Lett 361(2):155–163. https://doi.org/10.1016/j.canlet.2015.02.018 CrossRefPubMed

22.

Subramaniam KS, Tham ST, Mohamed Z, Woo YL, Mat Adenan NA, Chung I (2013) Cancer-associated fibroblasts promote proliferation of endometrial cancer cells. PLoS One 8(7):e68923. https://doi.org/10.1371/journal.pone.0068923 CrossRefPubMedPubMedCentral

23.

Tyan SW, Kuo WH, Huang CK, Pan CC, Shew JY, Chang KJ, Lee EY, Lee WH (2011) Breast cancer cells induce cancer-associated fibroblasts to secrete hepatocyte growth factor to enhance breast tumorigenesis. PLoS One 6(1):e15313. https://doi.org/10.1371/journal.pone.0015313 CrossRefPubMedPubMedCentral

24.

Noma K, Smalley KS, Lioni M, Naomoto Y, Tanaka N, El-Deiry W, King AJ, Nakagawa H, Herlyn M (2008) The essential role of fibroblasts in esophageal squamous cell carcinoma-induced angiogenesis. Gastroenterology 134(7):1981–1993. https://doi.org/10.1053/j.gastro.2008.02.061 CrossRefPubMedPubMedCentral

25.

Kellermann MG, Sobral LM, da Silva SD, Zecchin KG, Graner E, Lopes MA, Kowalski LP, Coletta RD (2008) Mutual paracrine effects of oral squamous cell carcinoma cells and normal oral fibroblasts: induction of fibroblast to myofibroblast transdifferentiation and modulation of tumor cell proliferation. Oral Oncol 44(5):509–517. https://doi.org/10.1016/j.oraloncology.2007.07.001 CrossRefPubMed

26.

Jeon ES, Lee IH, Heo SC, Shin SH, Choi YJ, Park JH, Park DY, Kim JH (2010) Mesenchymal stem cells stimulate angiogenesis in a murine xenograft model of A549 human adenocarcinoma through an LPA1 receptor-dependent mechanism. Biochim Biophys Acta 1801(11):1205–1213. https://doi.org/10.1016/j.bbalip.2010.08.003 CrossRefPubMed

27.

Gabbiani G, Ryan GB, Majne G (1971) Presence of modified fibroblasts in granulation tissue and their possible role in wound contraction. Experientia 27(5):549–550CrossRefPubMed

28.

Katoh K, Kano Y, Masuda M, Onishi H, Fujiwara K (1998) Isolation and contraction of the stress Fiber. Mol Biol Cell 9(7):1919–1938CrossRefPubMedPubMedCentral

29.

Ronnov-Jessen L, Petersen OW (1993) Induction of alpha-smooth muscle actin by transforming growth factor-beta 1 in quiescent human breast gland fibroblasts. Implications for myofibroblast generation in breast neoplasia. Laboratory investigation; a journal of technical methods and pathology 68(6):696–707PubMed

30.

Jeon ES, Moon HJ, Lee MJ, Song HY, Kim YM, Cho M, Suh DS, Yoon MS, Chang CL, Jung JS, Kim JH (2008) Cancer-derived lysophosphatidic acid stimulates differentiation of human mesenchymal stem cells to myofibroblast-like cells. Stem Cells 26(3):789–797. https://doi.org/10.1634/stemcells.2007-0742 CrossRefPubMed

31.

Mazzocca A, Dituri F, Lupo L, Quaranta M, Antonaci S, Giannelli G (2011) Tumor-secreted lysophostatidic acid accelerates hepatocellular carcinoma progression by promoting differentiation of peritumoral fibroblasts in myofibroblasts. Hepatology 54(3):920–930. https://doi.org/10.1002/hep.24485 CrossRefPubMed

32.

Gottschling S, Granzow M, Kuner R, Jauch A, Herpel E, Xu EC (2013) Mesenchymal stem cells in non-small cell lung cancer–different from others? Insights from comparative molecular and functional analyses Lung Cancer 80:19–29. https://doi.org/10.1016/j.lungcan.2012.12.015 PubMedCrossRef

33.

Xu X, Zhang X, Wang S, Qian H, Zhu W, Cao H (2011) Isolation and comparison of mesenchymal stem-like cells from human gastric cancer and adjacent non-cancerous tissues. J Cancer Res Clin Oncol 137:495–504. https://doi.org/10.1007/s00432-010-0908-6 CrossRefPubMed

34.

Ding G, Shao J, Ding Q, Fang Z, Wu Z, Xu J (2012) Comparison of the characteristics of mesenchymal stem cells obtained from prostate tumors and from bone marrow cultured in conditioned medium Exp Ther Med:4

35.

Xu X, Zhang X, Wang S, Qian H, Zhu W, Cao H, Wang M, Chen Y, Xu W (2011) Isolation and comparison of mesenchymal stem-like cells from human gastric cancer and adjacent non-cancerous tissues. J Cancer Res Clin Oncol 137(3):495–504. https://doi.org/10.1007/s00432-010-0908-6 CrossRefPubMed

36.

Lacher MD, Siegenthaler A, Jager R, Yan X, Hett S, Xuan L, Saurer S, Lareu RR, Dharmarajan AM, Friis R (2003) Role of DDC-4//sFRP-4, a secreted frizzled-related protein, at the onset of apoptosis in mammary involution. Cell Death Differ 10(5):528–538CrossRefPubMed

37.

Drake JM, Friis RR, Dharmarajan AM (2003) The role of sFRP4, a secreted frizzled-related protein, in ovulation. Apoptosis 8(4):389–397CrossRefPubMed

38.

Hinz B, Celetta G, Tomasek JJ, Gabbiani G, Chaponnier C (2001) Alpha-smooth muscle actin expression upregulates fibroblast contractile activity. Mol Biol Cell 12(9):2730–2741CrossRefPubMedPubMedCentral

39.

Mar PK, Roy P, Yin HL, Cavanagh HD, Jester JV (2001) Stress Fiber Formation is Required for Matrix Reorganization in a Corneal Myofibroblast Cell Line. Experimental Eye Research 72 (4):455–466. doi:doi:https://doi.org/10.1006/exer.2000.0967

40.

Ford CE, Jary E, Ma SS, Nixdorf S, Heinzelmann-Schwarz VA, Ward RL (2013) The Wnt gatekeeper SFRP4 modulates EMT, cell migration and downstream Wnt signalling in serous ovarian cancer cells. PLoS One 8(1):e54362. https://doi.org/10.1371/journal.pone.0054362 CrossRefPubMedPubMedCentral

41.

Bhuvanalakshmi G, Arfuso F, Millward M, Dharmarajan A, Warrier S (2015) Secreted frizzled-related protein 4 inhibits glioma stem-like cells by reversing epithelial to mesenchymal transition, inducing apoptosis and decreasing cancer stem cell properties. PLoS One 10(6):e0127517. https://doi.org/10.1371/journal.pone.0127517 CrossRefPubMed

42.

Warburg O (1956) On the origin of cancer cells. Science 123(3191):309–314CrossRefPubMed

43.

Sotgia F, Martinez-Outschoorn UE, Pavlides S, Howell A, Pestell RG, Lisanti MP (2011) Understanding the Warburg effect and the prognostic value of stromal caveolin-1 as a marker of a lethal tumor microenvironment. Breast cancer research: BCR 13(4):213–213. https://doi.org/10.1186/bcr2892 CrossRefPubMedPubMedCentral

44.

Martinez-Outschoorn UE, Sotgia F, Lisanti MP (2012) Power surge: supporting cells "fuel" cancer cell mitochondria. Cell Metab 15(1):4–5. https://doi.org/10.1016/j.cmet.2011.12.011 CrossRefPubMed

45.

Guido C, Whitaker-Menezes D, Capparelli C, Balliet R, Lin Z, Pestell RG, Howell A, Aquila S, Ando S, Martinez-Outschoorn U, Sotgia F, Lisanti MP (2012) Metabolic reprogramming of cancer-associated fibroblasts by TGF-beta drives tumor growth: connecting TGF-beta signaling with "Warburg-like" cancer metabolism and L-lactate production. Cell Cycle 11(16):3019–3035. https://doi.org/10.4161/cc.21384 CrossRefPubMedPubMedCentral

46.

Zhang D, Wang Y, Shi Z, Liu J, Sun P, Hou X, Zhang J, Zhao S, Zhou BP, Mi J (2015) Metabolic reprogramming of cancer-associated fibroblasts by IDH3alpha downregulation. Cell Rep 10(8):1335–1348. https://doi.org/10.1016/j.celrep.2015.02.006 CrossRefPubMed

47.

G B AF, Millward M, Dharmarajan A, Warrier S (2015) Secreted frizzled-related protein 4 inhibits glioma stem-like cells by reversing epithelial to mesenchymal transition, inducing apoptosis and decreasing Cancer stem cell properties. PLoS One 10(6):e0127517. https://doi.org/10.1371/journal.pone.0127517 CrossRefPubMedCentral

Title: The Influence of Breast Tumour-Derived Factors and Wnt Antagonism on the Transformation of Adipose-Derived Mesenchymal Stem Cells into Tumour-Associated Fibroblasts
Authors: Malini Visweswaran
Kevin N. Keane
Frank Arfuso
Rodney J. Dilley
Philip Newsholme
Arun Dharmarajan
Publication date: 01-06-2018
Publisher: Springer Netherlands
Published in: Cancer Microenvironment / Issue 1/2018
Print ISSN: 1875-2292
Electronic ISSN: 1875-2284
DOI: https://doi.org/10.1007/s12307-018-0210-8

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2018

Extracellular ATP is Differentially Metabolized on Papillary Thyroid Carcinoma Cells Surface in Comparison to Normal Cells

Role of the Nervous System in Tumor Angiogenesis

Exosomes: Definition, Role in Tumor Development and Clinical Implications

Recovery of Immunoglobulin VJ Recombinations from Pancreatic Cancer Exome Files Strongly Correlates with Reduced Survival

Immunogenomics: A Negative Prostate Cancer Outcome Associated with TcR-γ/δ Recombinations

Positive Correlation between Matrix Metalloproteinases and Epithelial-to-Mesenchymal Transition and its Association with Clinical Outcome in Bladder Cancer Patients

Keynote webinar | Spotlight on antibody–drug conjugates in cancer