Vasculature analysis of patient derived tumor xenografts using species-specific PCR assays: evidence of tumor endothelial cells and atypical VEGFA-VEGFR1/2 signalings

BMC Cancer

Ivan Bieche⁶  Sophie Vacher³  David Vallerand¹  Sophie Richon⁸  Rana Hatem³  Ludmilla De Plater⁵  Ahmed Dahmani⁵  Fariba Némati⁵  Eric Angevin⁷  Elisabetta Marangoni⁵  Sergio Roman-Roman⁵  Didier Decaudin⁴  Virginie Dangles-Marie² 

[1] Roche SAS, 30, cours de l'Ile Seguin, 92650 Boulogne-Billancourt, Cedex, France

[2] Université Paris Descartes, Sorbonne Paris Cité, 4 avenue de l’Observatoire, Paris, France

[3] Laboratoire d’Oncogénétique, 35 rue Dailly, Institut Curie - Hôpital Rene Huguenin, St Cloud, France

[4] Département d’Oncologie Médicale, Institut Curie, 26 rue d’Ulm, Paris, France

[5] Département de Recherche Translationnelle, Laboratoire d’Investigation Préclinique, Paris, France

[6] INSERM UMR745, Sorbonne Paris Cité, 4 avenue de l’Observatoire, Paris, France

[7] Institut de Cancérologie Gustave Roussy, 39 rue Camille Desmoulins, Villejuif, France

[8] CNRS, UMR 144, Centre de Recherche, Institut Curie, 26 rue d’Ulm, Paris, France

关键词: VEGFA-VEGFR1/2 signalings;    Endothelial markers;    Species-specific PCR assays;    Patient-derived xenografts;    Tumor vasculature;   

Others  :  858978
DOI  :  10.1186/1471-2407-14-178

 received in 2013-09-29, accepted in 2014-01-27,  发布年份 2014

【 摘 要 】

Background

Tumor endothelial transdifferentiation and VEGFR1/2 expression by cancer cells have been reported in glioblastoma but remain poorly documented for many other cancer types.

Methods

To characterize vasculature of patient-derived tumor xenografts (PDXs), largely used in preclinical anti-angiogenic assays, we designed here species-specific real-time quantitative RT-PCR assays. Human and mouse PECAM1/CD31, ENG/CD105, FLT1/VEGFR1, KDR/VEGFR2 and VEGFA transcripts were analyzed in a large series of 150 PDXs established from 8 different tumor types (53 colorectal, 14 ovarian, 39 breast and 15 renal cell cancers, 6 small cell and 5 non small cell lung carcinomas, 13 cutaneous melanomas and 5 glioblastomas) and in two bevacizumab-treated non small cell lung carcinomas xenografts.

Results

As expected, mouse cell proportion in PDXs -evaluated by quantifying expression of the housekeeping gene TBP- correlated with all mouse endothelial markers and human VEGFA RNA levels. More interestingly, we observed human PECAM1/CD31 and ENG/CD105 expression in all tumor types, with higher rate in glioblastoma and renal cancer xenografts. Human VEGFR expression profile varied widely depending on tumor types with particularly high levels of human FLT1/VEGFR1 transcripts in colon cancers and non small cell lung carcinomas, and upper levels of human KDR/VEGFR2 transcripts in non small cell lung carcinomas. Bevacizumab treatment induced significant low expression of mouse Pecam1/Cd31, Eng/Cd105, Flt1/Vegfr1 and Kdr/Vefr2 while the human PECAM1/CD31 and VEGFA were upregulated.

Conclusions

Taken together, our results strongly suggest existence of human tumor endothelial cells in all tumor types tested and of both stromal and tumoral autocrine VEGFA-VEGFR1/2 signalings. These findings should be considered when evaluating molecular mechanisms of preclinical response and resistance to tumor anti-angiogenic strategies.

【 授权许可】

   
2014 Bieche et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Ellis LM, Hicklin DJ: VEGF-targeted therapy: mechanisms of anti-tumour activity. Nat Rev Cancer 2008, 8:579-591.
• [2]Hicklin DJ, Ellis LM: Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol 2005, 23:1011-1027.
• [3]Olsson AK, Dimberg A, Kreuger J, Claesson-Welsh L: VEGF receptor signalling - in control of vascular function. Nat Rev Mol Cell Biol 2006, 7:359-371.
• [4]Ping YF, Bian XW: Consice review: Contribution of cancer stem cells to neovascularization. Stem Cells 2011, 29:888-894.
• [5]Knizetova P, Ehrmann J, Hlobilkova A, Vancova I, Kalita O, Kolar Z, Bartek J: Autocrine regulation of glioblastoma cell cycle progression, viability and radioresistance through the VEGF-VEGFR2 (KDR) interplay. Cell Cycle 2008, 7:2553-2561.
• [6]Lichtenberger BM, Tan PK, Niederleithner H, Ferrara N, Petzelbauer P, Sibilia M: Autocrine VEGF signaling synergizes with EGFR in tumor cells to promote epithelial cancer development. Cell 2010, 140:268-279.
• [7]Hamerlik P, Lathia JD, Rasmussen R, Wu Q, Bartkova J, Lee M, Moudry P, Bartek J Jr, Fischer W, Lukas J, Rich JN, Bartek J: Autocrine VEGF-VEGFR2-Neuropilin-1 signaling promotes glioma stem-like cell viability and tumor growth. J Exp Med 2012, 209:507-520.
• [8]Tentler JJ, Tan AC, Weekes CD, Jimeno A, Leong S, Pitts TM, Arcaroli JJ, Messersmith WA, Eckhardt SG: Patient-derived tumour xenografts as models for oncology drug development. Nat Rev Clin Oncol 2012, 9:338-350.
• [9]Bieche I, Onody P, Laurendeau I, Olivi M, Vidaud D, Lidereau R, Vidaud M: Real-time reverse transcription-PCR assay for future management of ERBB2-based clinical applications. Clin Chem 1999, 45:1148-1156.
• [10]Baird SM, Beattie GM, Lannom RA, Lipsick JS, Jensen FC, Kaplan NO: Induction of lymphoma in antigenically stimulated athymic mice. Cancer Res 1982, 42:198-206.
• [11]Eberhard A, Kahlert S, Goede V, Hemmerlein B, Plate KH, Augustin HG: Heterogeneity of angiogenesis and blood vessel maturation in human tumors: implications for antiangiogenic tumor therapies. Cancer Res 2000, 60:1388-1393.
• [12]Fukumura D, Xavier R, Sugiura T, Chen Y, Park EC, Lu N, Selig M, Nielsen G, Taksir T, Jain RK, Seed B: Tumor induction of VEGF promoter activity in stromal cells. Cell 1998, 94:715-725.
• [13]Liang WC, Wu X, Peale FV, Lee CV, Meng YG, Gutierrez J, Fu L, Malik AK, Gerber HP, Ferrara N, Fuh G: Cross-species vascular endothelial growth factor (VEGF)-blocking antibodies completely inhibit the growth of human tumor xenografts and measure the contribution of stromal VEGF. J Biol Chem 2006, 281:951-961.
• [14]Hendrix MJ, Seftor EA, Hess AR, Seftor RE: Vasculogenic mimicry and tumour-cell plasticity: lessons from melanoma. Nat Rev Cancer 2003, 3:411-421.
• [15]Alvero AB, Fu HH, Holmberg J, Visintin I, Mor L, Marquina CC, Oidtman J, Silasi DA, Mor G: Stem-like ovarian cancer cells can serve as tumor vascular progenitors. Stem Cells 2009, 27:2405-2413.
• [16]Bussolati B, Bruno S, Grange C, Ferrando U, Camussi G: Identification of a tumor-initiating stem cell population in human renal carcinomas. Faseb J 2008, 22:3696-3705.
• [17]Bussolati B, Grange C, Sapino A, Camussi G: Endothelial cell differentiation of human breast tumour stem/progenitor cells. J Cell Mol Med 2009, 13:309-319.
• [18]Hardee ME, Zagzag D: Mechanisms of Glioma-Associated Neovascularization. Am J Pathol 2012, 181:1126-1141.
• [19]Ricci-Vitiani L, Pallini R, Biffoni M, Todaro M, Invernici G, Cenci T, Maira G, Parati EA, Stassi G, Larocca LM, De Maria R: Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells. Nature 2010, 468:824-828.
• [20]Soda Y, Marumoto T, Friedmann-Morvinski D, Soda M, Liu F, Michiue H, Pastorino S, Yang M, Hoffman RM, Kesari S, Verma IM: Transdifferentiation of glioblastoma cells into vascular endothelial cells. Proc Natl Acad Sci U S A 2011, 108:4274-4280.
• [21]Wang R, Chadalavada K, Wilshire J, Kowalik U, Hovinga KE, Geber A, Fligelman B, Leversha M, Brennan C, Tabar V: Glioblastoma stem-like cells give rise to tumour endothelium. Nature 2010, 468:829-833.
• [22]Ghanekar A, Ahmed S, Chen K, Adeyi O: Endothelial cells do not arise from tumor-initiating cells in human hepatocellular carcinoma. BMC Cancer 2013, 13:485. BioMed Central Full Text
• [23]Julien S, Merino-Trigo A, Lacroix L, Pocard M, Goere D, Mariani P, Landron S, Bigot L, Nemati F, Dartigues P, Weiswald LB, Lantuas D, Morgand L, Pham E, Gonin P, Dangles-Marie V, Job B, Dessen P, Bruno A, Pierre A, De Thé H, Soliman H, Nunes M, Lardier G, Calvet L, Demers B, Prevost G, Vrignaud P, Roman-Roman S, Duchamp O, et al.: Characterization of a large panel of patient-derived tumor xenografts representing the clinical heterogeneity of human colorectal cancer. Clin Cancer Res 2012, 18:5314-5328.
• [24]Reyal F, Guyader C, Decraene C, Lucchesi C, Auger N, Assayag F, De Plater L, Gentien D, Poupon MF, Cottu P, De Cremoux P, Gestraud P, Vincent-Salomon A, Fontaine JJ, Roman-Roman S, Delattre O, Decaudin D, Marangoni E: Molecular profiling of patient-derived breast cancer xenografts. Breast Cancer Res 2012, 14:R11. BioMed Central Full Text
• [25]Fonsatti E, Altomonte M, Nicotra MR, Natali PG, Maio M: Endoglin (CD105): a powerful therapeutic target on tumor-associated angiogenetic blood vessels. Oncogene 2003, 22:6557-6563.
• [26]Angevin E, Glukhova L, Pavon C, Chassevent A, Terrier-Lacombe MJ, Goguel AF, Bougaran J, Ardouin P, Court BH, Perrin JL, Vallancien G, Triebel F, Escudier B: Human renal cell carcinoma xenografts in SCID mice: tumorigenicity correlates with a poor clinical prognosis. Lab Invest 1999, 79:879-888.
• [27]Cottu P, Marangoni E, Assayag F, de Cremoux P, Vincent-Salomon A, Guyader C, de Plater L, Elbaz C, Karboul N, Fontaine JJ, Chateau-Joubert S, Boudou-Rouquette P, Alran S, Dangles-Marie V, Gentien D, Poupon MF, Decaudin D: Modeling of response to endocrine therapy in a panel of human luminal breast cancer xenografts. Breast Cancer Res Treat 2012, 133:595-606.
• [28]de Plater L, Lauge A, Guyader C, Poupon MF, Assayag F, de Cremoux P, Vincent-Salomon A, Stoppa-Lyonnet D, Sigal-Zafrani B, Fontaine JJ, Brough R, Lord CJ, Ashworth A, Cottu P, Decaudin D, Marangoni E: Establishment and characterisation of a new breast cancer xenograft obtained from a woman carrying a germline BRCA2 mutation. Br J Cancer 2010, 103:1192-1200.
• [29]Leuraud P, Taillandier L, Medioni J, Aguirre-Cruz L, Criniere E, Marie Y, Kujas M, Golmard JL, Duprez A, Delattre JY, Sanson M, Poupon MF: Distinct responses of xenografted gliomas to different alkylating agents are related to histology and genetic alterations. Cancer Res 2004, 64:4648-4653.
• [30]Nemati F, Bras-Goncalves R, Fontaine JJ, de Pinieux G, De Cremoux P, Chapelier A, Daniel C, Laurent-Puig P, Livartowski A, Judde JG, Bordier V, Poupon MF, Decaudin D: Preclinical assessment of cisplatin-based therapy versus docetaxel-based therapy on a panel of human non-small-cell lung cancer xenografts. Anticancer Drugs 2009, 20:932-940.
• [31]Nemati F, Daniel C, Arvelo F, Legrier ME, Froget B, Livartowski A, Assayag F, Bourgeois Y, Poupon MF, Decaudin D: Clinical relevance of human cancer xenografts as a tool for preclinical assessment: example of in-vivo evaluation of topotecan-based chemotherapy in a panel of human small-cell lung cancer xenografts. Anticancer Drugs 2010, 21:25-32.
• [32]Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ, Double JA, Everitt J, Farningham DA, Glennie MJ, Kelland LR, Robinson V, Stratford IJ, Tozer GM, Watson S, Wedge SR, Eccles SA: Guidelines for the welfare and use of animals in cancer research. Br J Cancer 2010, 102:1555-1577.
• [33]Bieche I, Parfait B, Le Doussal V, Olivi M, Rio MC, Lidereau R, Vidaud M: Identification of CGA as a novel estrogen receptor-responsive gene in breast cancer: an outstanding candidate marker to predict the response to endocrine therapy. Cancer Res 2001, 61:1652-1658.