Article | Published:

Heterogeneous cancer-associated fibroblast population potentiates neuroendocrine differentiation and castrate resistance in a CD105-dependent manner

Oncogenevolume 38pages716730 (2019) | Download Citation

Abstract

Heterogeneous prostatic carcinoma-associated fibroblasts (CAF) contribute to tumor progression and resistance to androgen signaling deprivation therapy (ADT). CAF subjected to extended passaging, compared to low passage CAF, were found to lose tumor expansion potential and heterogeneity. Cell surface endoglin (CD105), known to be expressed on proliferative endothelia and mesenchymal stem cells, was diminished in high passage CAF. RNA-sequencing revealed SFRP1 to be distinctly expressed by tumor-inductive CAF, which was further demonstrated to occur in a CD105-dependent manner. Moreover, ADT resulted in further expansion of the CD105+ fibroblastic population and downstream SFRP1 in 3-dimensional cultures and patient-derived xenograft tissues. In patients, CD105+ fibroblasts were found to circumscribe epithelia with neuroendocrine differentiation. CAF-derived SFRP1, driven by CD105 signaling, was necessary and sufficient to induce prostate cancer neuroendocrine differentiation in a paracrine manner. A partially humanized CD105 neutralizing antibody, TRC105, inhibited fibroblastic SFRP1 expression and epithelial neuroendocrine differentiation. In a novel synthetic lethality paradigm, we found that simultaneously targeting the epithelia and its microenvironment with ADT and TRC105, respectively, reduced castrate-resistant tumor progression, in a model where either ADT or TRC105 alone had little effect.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Bhowmick NA, Chytil A, Plieth D, Gorska AE, Dumont N, Shappell S, et al. TGF-beta signaling in fibroblasts modulates the oncogenic potential of adjacent epithelia. Science. 2004;303:848–51.

  2. 2.

    Ayala G, Tuxhorn JA, Wheeler TM, Frolov A, Scardino PT, Ohori M, et al. Reactive stroma as a predictor of biochemical-free recurrence in prostate cancer. Clin Cancer Res. 2003;9:4792–801.

  3. 3.

    Hayward SW, Wang Y, Cao M, Hom YK, Zhang B, Grossfeld GD, et al. Malignant transformation in a nontumorigenic human prostatic epithelial cell line. Cancer Res. 2001;61:8135–42.

  4. 4.

    Olumi AF, Grossfeld GD, Hayward SW, Carroll PR, Tlsty TD, Cunha GR. Carcinoma-associated fibroblasts direct tumor progression of initiated human prostatic epithelium. Cancer Res. 1999;59:5002–11.

  5. 5.

    Gleave M, Hsieh JT, Gao CA, von Eschenbach AC, Chung LW. Acceleration of human prostate cancer growth in vivo by factors produced by prostate and bone fibroblasts. Cancer Res. 1991;51:3753–61.

  6. 6.

    Placencio VR, Sharif-Afshar AR, Li X, Huang H, Uwamariya C, Neilson EG, et al. Stromal transforming growth factor-beta signaling mediates prostatic response to androgen ablation by paracrine Wnt activity. Cancer Res. 2008;68:4709–18.

  7. 7.

    Qi J, Tripathi M, Mishra R, Sahgal N, Fazli L, Ettinger S, et al. The E3 ubiquitin ligase Siah2 contributes to castration-resistant prostate cancer by regulation of androgen receptor transcriptional activity. Cancer Cell. 2013;23:332–46.

  8. 8.

    De Sousa EMF, Vermeulen L, Fessler E, Medema JP. Cancer heterogeneity—a multifaceted view. EMBO Rep. 2013;14:686–95.

  9. 9.

    Kiskowski MA, Jackson RS 2nd, Banerjee J, Li X, Kang M, et al. Role for stromal heterogeneity in prostate tumorigenesis. Cancer Res. 2011;71:3459–70.

  10. 10.

    Bhowmick NA, Ghiassi M, Aakre M, Brown K, Singh V, Moses HL. TGF-beta-induced RhoA and p160ROCK activation is involved in the inhibition of Cdc25A with resultant cell-cycle arrest. Proc Natl Acad Sci USA. 2003;100:15548–53.

  11. 11.

    Brennen WN, Chen S, Denmeade SR, Isaacs JT. Quantification of mesenchymal stem cells (MSCs) at sites of human prostate cancer. Oncotarget. 2013;4:106–17.

  12. 12.

    Fuyuhiro Y, Yashiro M, Noda S, Kashiwagi S, Matsuoka J, Doi Y, et al. Upregulation of cancer-associated myofibroblasts by TGF-beta from scirrhous gastric carcinoma cells. Br J Cancer. 2011;105:996–1001.

  13. 13.

    Fonsatti E, Altomonte M, Nicotra MR, Natali PG, Maio M. Endoglin (CD105): a powerful therapeutic target on tumor-associated angiogenetic blood vessels. Oncogene. 2003;22:6557–63.

  14. 14.

    Beer TM, Armstrong AJ, Rathkopf DE, Loriot Y, Sternberg CN, Higano CS, et al. Enzalutamide in metastatic prostate cancer before chemotherapy. N Engl J Med. 2014;371:424–33.

  15. 15.

    Beltran H, Tagawa ST, Park K, MacDonald T, Milowsky MI, Mosquera JM. et al. Challenges in recognizing treatment-related neuroendocrine prostate cancer. J Clin Oncol. 2012;30:e386–89.

  16. 16.

    Aggarwal RR, Feng FY, Small EJ. Emerging categories of disease in advanced prostate cancer and their therapeutic implications. Oncology. 2017;31:467–74.

  17. 17.

    Dayyani F, Varkaris A, Araujo JC, Song JH, Chatterji T, Trudel GC, et al. Increased serum insulin-like growth factor-1 levels are associated with prolonged response to dasatinib-based regimens in metastatic prostate cancer. Prostate. 2013;73:979–85.

  18. 18.

    Banerjee J, Mishra R, Li X, Jackson RS 2nd, Sharma A, et al. A reciprocal role of prostate cancer on stromal DNA damage. Oncogene. 2014;33:4924–31.

  19. 19.

    True LD, Zhang H, Ye M, Huang CY, Nelson PS, von Haller PD, et al. CD90/THY1 is overexpressed in prostate cancer-associated fibroblasts and could serve as a cancer biomarker. Mod Pathol. 2010;23:1346–56.

  20. 20.

    Zhao H, Peehl DM. Tumor-promoting phenotype of CD90hi prostate cancer-associated fibroblasts. Prostate. 2009;69:991–1000.

  21. 21.

    R2: Genomics Analysis and Visualization Platform (http://r2.amc.nl).

  22. 22.

    Erho N, Crisan A, Vergara IA, Mitra AP, Ghadessi M, Buerki C, et al. Discovery and validation of a prostate cancer genomic classifier that predicts early metastasis following radical prostatectomy. PLoS ONE. 2013;8:e66855.

  23. 23.

    Beltran H, Rickman DS, Park K, Chae SS, Sboner A, MacDonald TY, et al. Molecular characterization of neuroendocrine prostate cancer and identification of new drug targets. Cancer Discov. 2011;1:487–95.

  24. 24.

    Li X, Placencio V, Iturregui JM, Uwamariya C, Sharif-Afshar AR, Koyama T, et al. Prostate tumor progression is mediated by a paracrine TGF-beta/Wnt3a signaling axis. Oncogene. 2008;27:7118–30.

  25. 25.

    Nolan-Stevaux O, Zhong W, Culp S, Shaffer K, Hoover J, Wickramasinghe D, et al. Endoglin requirement for BMP9 signaling in endothelial cells reveals new mechanism of action for selective anti-endoglin antibodies. PLoS ONE. 2012;7:e50920.

  26. 26.

    Zhu Y, Xu Y, Helseth DL Jr., Gulukota K, Yang S, Pesce LL, et al. Zodiac: a comprehensive depiction of genetic interactions in cancer by integrating TCGA data. J Natl Cancer Inst. 2015;107:djv129.

  27. 27.

    Kanayama M, Hayano T, Koebis M, Maeda T, Tabe Y, Horie S, et al. Hyperactive mTOR induces neuroendocrine differentiation in prostate cancer cell with concurrent up-regulation of IRF1. Prostate. 2017;77:1489–98.

  28. 28.

    Sun F, Zhang ZW, Tan EM, Lim ZLR, Li Y, Wang XC, et al. Icaritin suppresses development of neuroendocrine differentiation of prostate cancer through inhibition of IL-6/STAT3 and Aurora kinase A pathways in TRAMP mice. Carcinogenesis. 2016;37:701–11.

  29. 29.

    Romero D, O’Neill C, Terzic A, Contois L, Young K, Conley BA, et al. Endoglin regulates cancer-stromal cell interactions in prostate tumors. Cancer Res. 2011;71:3482–93.

  30. 30.

    Ghajar CM, Peinado H, Mori H, Matei IR, Evason KJ, Brazier H, et al. The perivascular niche regulates breast tumour dormancy. Nat Cell Biol. 2013;15:807–17.

  31. 31.

    Bussard KM, Mutkus L, Stumpf K, Gomez-Manzano C, Marini FC. Tumor-associated stromal cells as key contributors to the tumor microenvironment. Breast Cancer Res. 2016;18:84.

  32. 32.

    Wadosky KM, Koochekpour S. Molecular mechanisms underlying resistance to androgen deprivation therapy in prostate cancer. Oncotarget. 2016;7:64447–70.

  33. 33.

    Mu P, Zhang Z, Benelli M, Karthaus WR, Hoover E, Chen CC, et al. SOX2 promotes lineage plasticity and antiandrogen resistance in TP53- and RB1-deficient prostate cancer. Science. 2017;355:84–88.

  34. 34.

    Joesting MS, Cheever TR, Volzing KG, Yamaguchi TP, Wolf V, Naf D, et al. Secreted frizzled related protein 1 is a paracrine modulator of epithelial branching morphogenesis, proliferation, and secretory gene expression in the prostate. Dev Biol. 2008;317:161–73.

  35. 35.

    Zheng L, Sun D, Fan W, Zhang Z, Li Q, Jiang T. Diagnostic value of SFRP1 as a favorable predictive and prognostic biomarker in patients with prostate cancer. PLoS ONE. 2015;10:e0118276.

  36. 36.

    Esteve P, Sandonis A, Ibanez C, Shimono A, Guerrero I, Bovolenta P. Secreted frizzled-related proteins are required for Wnt/beta-catenin signalling activation in the vertebrate optic cup. Development. 2011;138:4179–84.

  37. 37.

    Siamakpour-Reihani S, Caster J, Bandhu Nepal D, Courtwright A, Hilliard E, Usary J, et al. The role of calcineurin/NFAT in SFRP2 induced angiogenesis—a rationale for breast cancer treatment with the calcineurin inhibitor tacrolimus. PLoS ONE. 2011;6:e20412.

  38. 38.

    Sun Y, Zhu D, Chen F, Qian M, Wei H, Chen W, et al. SFRP2 augments WNT16B signaling to promote therapeutic resistance in the damaged tumor microenvironment. Oncogene. 2016;35:4321–34.

  39. 39.

    Bovolenta P, Esteve P, Ruiz JM, Cisneros E, Lopez-Rios J. Beyond Wnt inhibition: new functions of secreted Frizzled-related proteins in development and disease. J Cell Sci. 2008;121:737–46.

  40. 40.

    Kawano Y, Diez S, Uysal-Onganer P, Darrington RS, Waxman J, Kypta RM. Secreted Frizzled-related protein-1 is a negative regulator of androgen receptor activity in prostate cancer. Br J Cancer. 2009;100:1165–74.

  41. 41.

    Joesting MS, Perrin S, Elenbaas B, Fawell SE, Rubin JS, Franco OE, et al. Identification of SFRP1 as a candidate mediator of stromal-to-epithelial signaling in prostate cancer. Cancer Res. 2005;65:10423–30.

  42. 42.

    Miyata Y, Mitsunari K, Asai A, Takehara K, Mochizuki Y, Sakai H. Pathological significance and prognostic role of microvessel density, evaluated using CD31, CD34, and CD105 in prostate cancer patients after radical prostatectomy with neoadjuvant therapy. Prostate. 2015;75:84–91.

  43. 43.

    Degraff DJ, Aguiar AA, Sikes RA. Disease evidence for IGFBP-2 as a key player in prostate cancer progression and development of osteosclerotic lesions. Am J Transl Res. 2009;1:115–30.

  44. 44.

    Laskaratos FM, Rombouts K, Caplin M, Toumpanakis C, Thirlwell C, Mandair D. Neuroendocrine tumors and fibrosis: an unsolved mystery? Cancer. 2017;123:4770–90.

  45. 45.

    Zikusoka MN, Kidd M, Eick G, Latich I, Modlin IM. The molecular genetics of gastroenteropancreatic neuroendocrine tumors. Cancer. 2005;104:2292–309.

  46. 46.

    Igarashi A, Fukagai T, Morita M, Hayashi K, Koshikiya A, Ogawa Y, et al. Initial experience of the enzalutamide treatment for castration-resistant prostate cancer. Nihon Hinyokika Gakkai Zasshi. 2016;107:155–61.

  47. 47.

    Karzai FH, Apolo AB, Cao L, Madan RA, Adelberg DE, Parnes H, et al. A phase I study of TRC105 anti-endoglin (CD105) antibody in metastatic castration-resistant prostate cancer. BJU Int. 2015;116:546–55.

  48. 48.

    Franco OE, Jiang M, Strand DW, Peacock J, Fernandez S, Jackson RS 2nd, et al. Altered TGF-beta signaling in a subpopulation of human stromal cells promotes prostatic carcinogenesis. Cancer Res. 2011;71:1272–81.

  49. 49.

    Stark HJ, Baur M, Breitkreutz D, Mirancea N, Fusenig NE. Organotypic keratinocyte cocultures in defined medium with regular epidermal morphogenesis and differentiation. J Invest Dermatol. 1999;112:681–91.

  50. 50.

    Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO et al.Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal. 2013; 6:pl1.

  51. 51.

    Cerami E, Gao J, Dogrusoz U, Gross BE, Sumer SO, Aksoy BA. et al.The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data.Cancer Discov. 2012;2:401–04.

Download references

Acknowledgements

We thank Charles Theurer at Tracon Pharmaceuticals, Inc. for the TRC105 and M1043 neutralizing antibodies. The RNA-sequencing and histology slide scanning was performed at the Cedars-Sinai Genomics Core and Microscopy Core, respectively. This work was supported by grants from the National Cancer Institute (CA108646) and Veterans Affairs (BX001040) to NAB.

Author contributions

Conceptualization, MK, VRP-H, and NAB; Methodology and Project Administration, MK and VRP-H; Investigation, MK, VRP-H, AM, SH, KR-R, MT, RM, SB, BS, PA, FD, and BA; Formal analysis, DH, JZ; Writing, MK, VRP-H, and NAB; supervision and funding, NAB.

Author information

Author notes

  1. These authors contributed equally: Manabu Kato, Veronica R. Placencio-Hickok

Affiliations

  1. Nephro-Urologic Surgery and Andrology, Mie University Hospital, Tsu, Mie, Japan

    • Manabu Kato
  2. Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, 90048, USA

    • Veronica R. Placencio-Hickok
    • , Anisha Madhav
    • , Subhash Haldar
    • , Manisha Tripathi
    • , Sandrine Billet
    • , Rajeev Mishra
    • , Bethany Smith
    • , Krizia Rohena-Rivera
    • , Priyanka Agarwal
    • , Frank Duong
    • , Bryan Angara
    • , David Hickok
    • , Zhenqiu Liu
    •  & Neil A. Bhowmick

Authors

  1. Search for Manabu Kato in:

  2. Search for Veronica R. Placencio-Hickok in:

  3. Search for Anisha Madhav in:

  4. Search for Subhash Haldar in:

  5. Search for Manisha Tripathi in:

  6. Search for Sandrine Billet in:

  7. Search for Rajeev Mishra in:

  8. Search for Bethany Smith in:

  9. Search for Krizia Rohena-Rivera in:

  10. Search for Priyanka Agarwal in:

  11. Search for Frank Duong in:

  12. Search for Bryan Angara in:

  13. Search for David Hickok in:

  14. Search for Zhenqiu Liu in:

  15. Search for Neil A. Bhowmick in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Neil A. Bhowmick.

Electronic supplementary material

About this article

Publication history

Received

Revised

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/s41388-018-0461-3