Spatial morphological and molecular differences within solid tumors may contribute to the failure of vascular disruptive agent treatments

BMC Cancer

Linh Nguyen¹  Theodora Fifis¹  Caterina Malcontenti-Wilson¹  Lie Sam Chan¹  Patricia Nunes Luiza Costa¹  Mehrdad Nikfarjam¹  Vijayaragavan Muralidharan¹  Christopher Christophi¹ 

[1] Department of Surgery, University of Melbourne, Austin Health, Heidelberg, Victoria, 3084, Australia

关键词: EMT;    Infiltrating cells;    Growth factor;    Hypoxia;    Tumor periphery;    OXi4503;    Vascular disruptive agent;   

Others  :  1134865
DOI  :  10.1186/1471-2407-12-522

 received in 2011-12-16, accepted in 2012-11-07,  发布年份 2012

【 摘 要 】

Background

Treatment of solid tumors with vascular disrupting agent OXi4503 results in over 90% tumor destruction. However, a thin rim of viable cells persists in the tumor periphery following treatment, contributing to subsequent recurrence. This study investigates inherent differences in the microenvironment of the tumor periphery that contribute to treatment resistance.

Methods

Using a murine colorectal liver metastases model, spatial morphological and molecular differences within the periphery and the center of the tumor that may account for differences in resistance to OXi4503 treatment were investigated. H&E staining and immunostaining were used to examine vessel maturity and stability, hypoxia and HIF1α levels, accumulation of immune cells, expression of proangiogenic factors/receptors (VEGF, TGF-β, b-FGF, and AT1R) and expression of EMT markers (ZEB1, vimentin, E-cadherin and β-catenin) in the periphery and center of established tumors. The effects of OXi4503 on tumor vessels and cell kinetics were also investigated.

Results

Significant differences were found between tumor periphery and central regions, including association of the periphery with mature vessels, higher accumulation of immune cells, increased growth factor expression, minimal levels of hypoxia and increased evidence of EMT. OXi4503 treatment resulted in collapse of vessels in the tumor center; however vasculature in the periphery remained patent. Similarly, tumor apoptosis and proliferation were differentially modulated between centre and periphery after treatment.

Conclusions

The molecular and morphological differences between tumor periphery and center may account for the observed differential resistance to OXi4503 treatment and could provide targets for drug development to totally eliminate metastases.

【 授权许可】

   
2012 Nguyen et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Folkman J: Anti-angiogenesis: new concept for therapy of solid tumors. Ann Surg 1972, 175:409-416.
• [2]Chaplin DJ, Hill SA: The development of combretastatin A4 phosphate as a vascular targeting agent. Int J Radiat Oncol Biol Phys 2002, 54(5):1491-1496.
• [3]Tozer GM, Kanthou C, Baguley BC: Disrupting tumour blood vessels. Nat Rev Cancer 2005, 5:423-435.
• [4]Siemann DW, Horsman MR: Vascular targeted therapies in oncology. Cell Tissue Res 2009, 335:241-248.
• [5]Rice L, Pampo C, Lepler S, Rojiani AM, Siemann DW: Support of a free radical mechanism for enhanced antitumor efficacy of the microtubule disruptor OXi4503. Microvasc Res 2011, 81:44-51.
• [6]Malcontenti-Wilson C, Chan L, Nikfarjam M, Muralidharan V, Christophi C: Vascular targeting agent Oxi4503 inhibits tumor growth in a colorectal liver metastases model. J Gastroenterol Hepatol 2008, 23:e96-e104.
• [7]Dalal S, Burchill SA: Preclinical evaluation of vascular-disrupting agents in Ewing’s sarcoma family of tumours. Eur J Cancer 2009, 45:713-722.
• [8]Ljuslinder I, Melin B, Henriksson ML, Oberg A, Palmqvist R: Increased epidermal growth factor receptor expression at the invasive margin is a negative prognostic factor in colorectal cancer. Int J Cancer 2011, 128:2031-2037.
• [9]Matsuda Y, Ishiwata T, Yamahatsu K, Kawahara K, Hagio M, Peng WX, Yamamoto T, Nakazawa N, Seya T, Ohaki Y, Naito Z: Overexpressed fibroblast growth factor receptor 2 in the invasive front of colorectal cancer: a potential therapeutic target in colorectal cancer. Cancer Lett 2011, 309:209-219.
• [10]Lin WX, Fifis T, Malcontenti-Wilson C, Nikfarjam M, Muralidharan V, Nguyen L, Christophi C: Induction of Th1Immune responses following laser ablation in a murine model of colorectal liver metastases. J Transl Med 2011, 9:83. BioMed Central Full Text
• [11]Allavena P, Sica A, Garlanda C, Mantovani A: The Yin-Yang of tumor-associated macrophages in neoplastic progression and immune surveillance. Immunol Rev 2008, 222:155-161.
• [12]DeNardo DG, Barreto JB, Andreu P, Vasquez L, Tawfik D, Kolhatkar N, Coussens LM: CD4(+) T cells regulate pulmonary metastasis of mammary carcinomas by enhancing protumor properties of macrophages. Cancer Cell 2009, 16:91-102.
• [13]Hagemann T, Robinson SC, Schulz M, Trumper L, Balkwill FR, Binder C: Enhanced invasiveness of breast cancer cell lines upon co-cultivation with macrophages is due to TNF-alpha dependent up-regulation of matrix metalloproteases. Carcinogenesis 2004, 25:1543-1549.
• [14]Jarnicki AG, Lysaght J, Todryk S, Mills KH: Suppression of antitumor immunity by IL-10 and TGF-beta-producing T cells infiltrating the growing tumor: influence of tumor environment on the induction of CD4+ and CD8+ regulatory T cells. J Immunol 2006, 177:896-904.
• [15]Kuruppu D, Christophi C, Bertram JF, O’Brien PE: Characterization of an animal model of hepatic metastasis. J Gastroenterol Hepatol 1996, 11:26-32.
• [16]Chan LS, Malcontenti-Wilson C, Muralidharan V, Christophi C: Effect of vascular targeting agent Oxi4503 on tumor cell kinetics in a mouse model of colorectal liver metastasis. Anticancer Res 2007, 27:2317-2323.
• [17]Lee TH, Bolontrade MF, Worth LL, Guan H, Ellis LM, Kleinerman ES: Production of VEGF165 by Ewing’s sarcoma cells induces vasculogenesis and the incorporation of CD34+ stem cells into the expanding tumor vasculature. Int J Cancer 2006, 119:839-846.
• [18]Hall AP: Review of the pericyte during angiogenesis and its role in cancer and diabetic retinopathy. Toxicol Pathol 2006, 34:763-775.
• [19]Rofstad EK, Maseide K: Radiobiological and immunohistochemical assessment of hypoxia in human melanoma xenografts: acute and chronic hypoxia in individual tumours. Int J Radiat Biol 1999, 75:1377-1393.
• [20]Brabletz T, Hlubek F, Spaderna S, Schmalhofer O, Hiendlmeyer E, Jung A, Kirchner T: Invasion and metastasis in colorectal cancer: epithelial-mesenchymal transition, mesenchymal-epithelial transition, stem cells and beta-catenin. Cells Tissues Organs 2005, 179:56-65.
• [21]Spaderna S, Schmalhofer O, Hlubek F, Berx G, Eger A, Merkel S, Jung A, Kirchner T, Brabletz T: A transient, EMT-linked loss of basement membranes indicates metastasis and poor survival in colorectal cancer. Gastroenterology 2006, 131:830-840.
• [22]Kaelin WG Jr: The von Hippel-Lindau protein, HIF hydroxylation, and oxygen sensing. Biochem Biophys Res Commun 2005, 338:627-638.
• [23]Chan LS, Malcontenti-Wilson C, Muralidharan V, Christophi C: Alterations in vascular architecture and permeability following OXi4503 treatment. Anticancer Drugs 2008, 19:17-22.
• [24]Brieger J, Schroeder P, Gosepath J, Mann WJ: VEGF-subtype specific protection of SCC and HUVECs from radiation induced cell death. Int J Mol Med 2005, 15:145-151.
• [25]Varey AH, Rennel ES, Qiu Y, Bevan HS, Perrin RM, Raffy S, Dixon AR, Paraskeva C, Zaccheo O, Hassan AB, Harper SJ, Bates DO: VEGF 165 b, an antiangiogenic VEGF-A isoform, binds and inhibits bevacizumab treatment in experimental colorectal carcinoma: balance of pro- and antiangiogenic VEGF-A isoforms has implications for therapy. Br J Cancer 2008, 98:1366-1379.
• [26]Cianchi F, Cuzzocrea S, Vinci MC, Messerini L, Comin CE, Navarra G, Perigli G, Centorrino T, Marzocco S, Lenzi E, Battisti N, Trallori G, Masini E: Heterogeneous expression of cyclooxygenase-2 and inducible nitric oxide synthase within colorectal tumors: correlation with tumor angiogenesis. Dig Liver Dis 2010, 42:20-27.
• [27]Koshikawa N, Mizushima H, Minegishi T, Eguchi F, Yotsumoto F, Nabeshima K, Miyamoto S, Mekada E, Seiki M: Proteolytic activation of heparin-binding EGF-like growth factor by membrane-type matrix metalloproteinase-1 in ovarian carcinoma cells. Cancer Sci 2011, 102:111-116.
• [28]Akishima-Fukasawa Y, Ishikawa Y, Akasaka Y, Uzuki M, Inomata N, Yokoo T, Ishii R, Shimokawa R, Mukai K, Kiguchi H, et al.: Histopathological predictors of regional lymph node metastasis at the invasive front in early colorectal cancer. Histopathology 2011, 59(3):470-481.
• [29]Scarpino S, Duranti E, Stoppacciaro A, Pilozzi E, Natoli G, Sciacchitano S, Luciani E, Ruco L: COX-2 is induced by HGF stimulation in Met-positive thyroid papillary carcinoma cells and is involved in tumour invasiveness. J Pathol 2009, 218:487-494.
• [30]Tan X, Egami H, Ishikawa S, Sugita H, Kamohara H, Nakagawa M, Nozawa F, Abe M, Ogawa M: Involvement of matrix metalloproteinase-7 in invasion-metastasis through induction of cell dissociation in pancreatic cancer. Int J Oncol 2005, 26:1283-1289.
• [31]Wente MN, Mayer C, Gaida MM, Michalski CW, Giese T, Bergmann F, Giese NA, Buchler MW, Friess H: CXCL14 expression and potential function in pancreatic cancer. Cancer Lett 2008, 259:209-217.
• [32]Schmalhofer O, Brabletz S, Brabletz T: E-cadherin, beta-catenin, and ZEB1 in malignant progression of cancer. Cancer Metastasis Rev 2009, 28:151-166.
• [33]Janda E, Lehmann K, Killisch I, Jechlinger M, Herzig M, Downward J, Beug H, Grunert S: Ras and TGF[beta] cooperatively regulate epithelial cell plasticity and metastasis: dissection of Ras signaling pathways. J Cell Biol 2002, 156:299-313.
• [34]Yang AD, Camp ER, Fan F, Shen L, Gray MJ, Liu W, Somcio R, Bauer TW, Wu Y, Hicklin DJ, Ellis LM: Vascular endothelial growth factor receptor-1 activation mediates epithelial to mesenchymal transition in human pancreatic carcinoma cells. Cancer Res 2006, 66:46-51.
• [35]Tozer GM, Prise VE, Wilson J, Locke RJ, Vojnovic B, Michael RL, Stratford MRL Dennis MF, Chaplin DJ: Combretastatin A-4 Phosphate as a Tumor Vascular-Targeting Agent: Early Effects in Tumors and Normal Tissues. Cancer Res 1999, 59:1626-1634.
• [36]Salmon BA, Siemann DW: Characterizing the tumor response to treatment with combretastatin A4 phosphate. Int J Radiat Oncol Biol Phys 2007, 68:211-217.
• [37]Wankhede M, Dedeugd C, Siemann DW, Sorg BS: In vivo functional differences in microvascular response of 4T1 and Caki-1 tumors after treatment with OXi4503. Oncol Rep 2010, 23:685-692.
• [38]Gaya A, Daley F, Taylor NJ, Tozer G, Qureshi U, Padhani A, Pedley RB, Begent R, Wellsted D, Stirling JJ, Rustin G: Relationship between human tumour angiogenic profile and combretastatin-induced vascular shutdown: an exploratory study. Br J Cancer 2008, 99:321-326.
• [39]Maddika S, Ande SR, Panigrahi S, Paranjothy T, Weglarczyk K, Zuse A, Eshraghi M, Manda KD, Wiechec E, Los M: Cell survival, cell death and cell cycle pathways are interconnected: implications for cancer therapy. Drug Resist Updat 2007, 10:13-29.
• [40]Nguyen DX, Massague J: Genetic determinants of cancer metastasis. Nat Rev Genet 2007, 8:341-352.
• [41]Gottesman MM, Fojo T, Bates SE: Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2002, 2:48-58.
• [42]Jain RK: Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 2005, 307:58-62.
• [43]Siemann DW, Chaplin DJ, Horsman MR: Vascular-targeting therapies for treatment of malignant disease. Cancer 2004, 100:2491-2499.
• [44]Jordan CT, Guzman ML, Noble M: Cancer stem cells. N Engl J Med 2006, 355:1253-1261.
• [45]Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L, Iovino F, Tripodo C, Russo A, Gulotta G, Medema JP, Stassi G: Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell 2007, 1:389-402.
• [46]Calabrese C, Poppleton H, Kocak M, Hogg TL, Fuller C, Hamner B, Oh EY, Gaber MW, Finklestein D, Allen M, Frank A, Bayazitov IT, Zakharenko SS, Gajjar A, Davidoff A, Gilbertson RJ: A perivascular niche for brain tumor stem cells. Cancer Cell 2007, 11:69-82.
• [47]Fodde R, Brabletz T: Wnt/beta-catenin signaling in cancer stemness and malignant behavior. Curr Opin Cell Biol 2007, 19:150-158.
• [48]Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, Bruns CJ, Heeschen C: Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 2007, 1:313-323.
• [49]Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, Campbell LL, Polyak K, Brisken C, Yang J, Weinberg RA: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 2008, 133:704-715.
• [50]Moody SE, Perez D, Pan TC, Sarkisian CJ, Portocarrero CP, Sterner CJ, Notorfrancesco KL, Cardiff RD, Chodosh LA: The transcriptional repressor Snail promotes mammary tumor recurrence. Cancer Cell 2005, 8:197-209.