Tumor Tissue Factor Research Articles

Abstract P6-11-07: A Phase IV Trial of Caelyx as First Line Chemotherapy (CT) in Women 65 Years or Older with Locally Advanced or Metastatic Breast Cancer: In Search for Treatment Predictive Factors

Abstract Background: Anthracyclines belong to the most effective therapies for patients (pts) with breast cancer. Toxicity limits its use in elderly women where, beside nausea, vomiting, hair loss and the risk for infections, cardiotoxicity is of special concern. Caelyx is pegylated liposomal doxorubicin, a formulation diminishing the risk for the mentioned side effects but with a risk for skin toxicity known as Palmar-Plantar Erythrodysesthesia (PPE). As Caelyx is generally well-tolerated, we investigated its value as first-line CT in women 65 years or older. We also collected a variety of tumor and patient markers in search of predictors of outcome and toxicity. Methods: Women age ≥65 years, with inoperable locally advanced or metastatic breast cancer with performance status (PS) WHO 0-2, measurable disease according to RECIST or WHO criteria for bone metastasis or non-measurable disease, Left Ventricular Ejection Fraction (LVEF) ≥50% and normal organ function, received 40 mg/m2 Caelyx every 28 days. Pts underwent tumor evaluation including LVEF every 3 months. Primary endpoint was time to treatment failure (TTF) defined as progression of disease or unacceptable toxicity. Secondary endpoints were safety, response rate (RR), time to progression (TTP), overall survival (OS), and analysis of treatment predictive factors in tumor tissue. Also the ABCB1 (=Multi Drug Resistance gene 1) single nucleotide polymorphism (SNP) was investigated. Results: 25 women were included in the study. Median age was 72 years (range 65-81). The median TTF was 5.5 months and the mean number of cycles received was 7.4 (range 1-21). 22 pts were followed by RECIST or a combination of RECIST and WHO. Of these one achieved a partial response (PR) and 11 stable disease (SD). The median TTP was 5.7 months and the median OS was 20.6 months. The majority of adverse events (AE's) were mild to moderate and transient, the main toxicities being nausea 60% (grade I/II/III 24/32/4), fatigue 56% (40/12/4) and PPE 52% (24/20/8). Clinically significant changes in LVEF were noted in three pts. Severe AE's occurred in 8 pts. And the dose was reduced due to toxicity in 4 pts. Analysis of the ABCB1 SNPs revealed that pts with homozygous mutation for SNP G2677T had significantly shorter time to treatment failure and time to progression as compared to the rest of the pts. Those with the 3435T/T genotype were more susceptible to moderate to severe PPE (5 out of 8 with SNP 3435T/T as compared to 2 out of 16 non-3435T/T pts). Conclusion: Breast cancer in elderly patients in need of CT can be challenging because of the risk of toxicity, reduced quality of life and also the lack of tumor response. To be able to select the right patient for a certain therapy would be essential. We treated 25 elderly women with advanced breast cancer with Caelyx with acceptable toxicity and efficacy. We also found that SNPs in ABCB1 correlated with both TTP and the risk of PPE, thereby indicating that the ABCB1 genotype could be a predictive factor for treatment with Caelyx in elderly women. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-11-07.

Increased Tissue Factor Expression Is Associated with Reduced Survival in Non–Small Cell Lung Cancer and with Mutations of TP53 and PTEN

Tissue factor (TF), the main initiator of blood coagulation, is also a signaling protein that regulates cancer progression. TF synthesis was recently shown to be affected by tumor suppressor genes (TSGs) in tumor cell lines. We therefore studied TF gene (F3) expression and the status of genes coding for tumor protein p53 (TP53), phosphatase and tensin homolog (PTEN), and serine/threonine kinase 11 (STK11) in non-small cell lung cancer (NSCLC). Heparanase (HPSE) gene expression was also measured because this endo-beta-D-glucuronidase was recently shown to enhance TF gene expression. TF and heparanase mRNA expression was measured by real-time PCR in 53 NSCLC tumors. Exons 5-8 of TP53 were sequenced from genomic DNA. Mutations of PTEN and STK11 were screened by multiplex ligation-dependent probe amplification. TF mRNA levels were significantly higher in T(3)-T(4) tumors (P = 0.04) and in stages III-IV of NSCLC (P = 0.03). Mutations of TP53, STK11, and PTEN were identified in 20 (37.7%), 21 (39%), and 20 (37.7%) of tumors, respectively. TF expression was higher in mutated TP53 (TP53(Mut)) (P = 0.02) and PTEN(Mut) (P = 0.03) samples. Moreover, TF mRNA increased from 2700 copies (no mutation) to 11 6415 when 3 TSG were mutated. Heparanase gene expression did not differ according to TF gene (F3) expression or TSG mutation. The median survival time was shorter in patients with tumor TF mRNA levels above median values (relative risk 2.2; P = 0.03, multivariate analysis) and when TP53 was mutated (relative risk 1.8; P = 0.02). These results provide clear evidence that combined oncogene events affecting TSG dramatically increase TF gene expression in lung tumors. Moreover, this study suggests that TF gene expression could be used as a prognostic marker in NSCLC.

Alterations of P53 and PTEN Genes Are Associated with Increased TF Expression and Reduced Survival in Non Small Cell Lung Cancer

Introduction: Tissue Factor (TF) is the main initiator of blood coagulation and also a signalling protein that regulates cancerous cell migration, angiogenesis and metastasis. TF is frequently synthesized by many tumor cells and TF gene expression can be modulated by tumor suppressor genes (TSG) such as P53 and PTEN. Moreover, heparanase is an endo-b-D-glucuronidase also implicated in tumor development and was recently described to enhance TF expression and activity. Finally, another TSG, LKB1 that regulates the mTOR pathway with P53 and PTEN, has also been recently shown to modulate cancer differentiation and metastasis.Aim of the study: To look for a relationship between TF and heparanase genes expression in NSCLC and to investigate whether these genes can be regulated by P53, PTEN and LKB1.Material and Methods: TF and heparanase mRNA levels were measured by real-time PCR in lung tumors surgically removed in 53 Caucasian patients with NSCLC. Direct sequencing of exons 5–8 of P53 was performed on genomic DNA. Mutations of PTEN and LKB1 were screened by multiplex ligation dependant probe amplification using specific SALSA-MLPA kits (MRC-Holland, Amsterdam, Holland). Results were analyzed and compared to clinicopathological features using the Mann Whitney U-test and Fisher's exact test. Moreover, survival analysis was evaluated using the Kaplan Meier method and differences assessed by the Log-rank test.Results: TF mRNA levels were variable from one sample to another (median: 11540 copies/108 copies of 18S RNA; range: 10–310 000) but significantly higher in T3–T4 tumors (19150 vs 8530 copies in T1–T2 tumors; p=0.04) and in stages III–IV of NSCLC (17710 vs 9780 copies in stages 1–2; p=0.03) as previously reported (Regina et al, J Thorac Oncol, 2008). Mutations of P53, LKB1 and PTEN were identified in 37.7 % (20/53), 49 % (26/53) and in 37.7 % (20/53) of tumors, respectively. TF mRNA levels were higher in samples mutated for P53 (median in P53Mut: 15655 vs 6225 copies/108 copies of 18S RNA in P53WT tissues; p=0.02), and also in cancerous tissues in whom PTEN mutations had been evidenced (median in PTENMut: 17370 vs 8530 copies in PTENWT; p=0.03). On the other hand, TF gene expression was unaffected in tumors when LKB1 mutations were detected alone but it dramatically increased if combined genetic alterations were present, with a number of mRNA copies varying from 3500 when no mutation was detected to 11600, 15800, and 78 000 copies when 1, 2 and 3 TSG were mutated (Spearman correlation p=0.01). Heparanase mRNA levels measured in lung tumors were also variable (median: 2922 copie/108 copies of 18S RNA; range: 80–39750), without any correlation with TF gene expression, TSG mutations and clinicopathological features. In contrast, the median survival time was shorter in patients with tumor TF mRNA levels above median value (26 vs 66 months; Hazard ratio: 1.96; CI95%:1.01–3.8; p=0.04) and when P53 was mutated (33 vs 43 months; Hazard ratio: 1.5; CI95%:1.02–2.5; p=0.05).Conclusion: This study provides evidence that combined oncogene events affecting TSG, such as P53, PTEN and LKB1, dramatically affect TF gene expression in NSCLC, this process contributing to cancer progression. TSG directly stimulate the mTOR pathway that could also be activated by TF/VIIa/XA complexes, favoring tumor cell migration.

Sharing Tissue Factor

Existence of a close relationship between increased clotting and malignancy has been clearly recognized for over a century. Many tumor cell types express tissue factor (TF), a procoagulant protein, on their cell surfaces. Tissue factor is a transmembrane cellular receptor for coagulation factor VII/VIIa. Binding of factor VIIa to TF triggers the activation of coagulation cascade leading ultimately to the generation of thrombin which in turn stimulates platelet activation and cleaves fibrinogen. Thus, there is no surprise that the activation of coagulation by tumor cell TF or by membrane fragments shed from tumor cells carrying TF contributes to cancer-associated thrombotic disorders.1,2 The close association between cancer and TF has also raised the possibility that TF may contribute to the pathogenesis of cancer beyond thrombosis. In fact, there are a number of reports in the literature showing that TF expressed by tumor cells plays an important role in both primary tumor growth and metastasis (see reviews3–5). The prometastatic effect of TF involves both activation of the coagulation pathway (eg, mechanisms linked to thrombin generated by TF) and effects mediated through the direct signaling activity of TF, probably via the cytoplasmic domain of TF.6 Recent studies have shown that direct TF-FVIIa protease-induced cell signaling is a major contributor to tumor growth in breast cancer.7 However, there are a couple of convincing reports in the literature that provide evidence that is contrary to the general belief that tumor-derived TF is crucial for primary tumor growth or metastasis. Toomey et al,8 using embryonic stem (ES) cells that do not express TF, showed that tumor-derived TF was not required for either tumor growth or metastasis. In a more recent study, Palumbo et al9 showed that tumor TF was neither required for primary tumor growth nor necessary for the initial …

Microparticle-associated tissue factor activity: a link between cancer and thrombosis?

Cancer, in particular mucinous adenocarcinoma, is associated with venous thromboembolism (VTE). Tissue factor (TF), initiator of coagulation, plays a central role in the paradigm that clotting and tumor growth form a vicious circle, in which hypercoagulability facilitates the aggressive biology of cancer and vice versa. Expression of TF in tumors is associated with poor differentiation and poor prognosis. We investigated the association between clinically manifest VTE and procoagulant properties of circulating microparticles (MP) isolated from blood of unselected pancreatic and breast adenocarcinoma patients' consecutive subjects, who presented with ultrasound or CT-scan confirmed VTE, and healthy subjects. Patients with disseminated breast and pancreatic cancer had significantly increased levels of MP-associated TF activity compared with healthy controls, subjects with idiopathic acute VTE and non-metastatic cancer patients. Patients with both high MP-associated TF-activity and MP-associated epithelial mucin (MUC1) had a lower survival rate at 3-9 months follow-up than those with low TF-activity and no MUC1 expression: the likelihood of survival was 0.42 (95% CI: 0.19- 0.94) for an individual with these two predictor variables present, after adjustment for other factors (age cohort, type of cancer, VTE) in the Cox proportional hazards model. Our results suggest an important role for MP-associated TF and MUC1 in the pathogenesis of thrombosis in disseminated mucinous adenocarcinoma patients. Future studies should reveal the mechanism underlying the observed associations.

Celecoxib Up-Regulates the Expression of the ζ Chain of T Cell Receptor Complex in Tumor-Infiltrating Lymphocytes in Human Cervical Cancer

We evaluated the effects of celecoxib treatment on tumor-infiltrating lymphocyte (TIL) subsets [CD3(+), CD4(+),CD8(+), CD25(+), and T cell receptor (TCR)-zeta-expressing cells] and tryptase-positive mast cells in cervical tumors. Circulating levels of cytokines [interleukin (IL)-1beta, IL-10, tumor necrosis factor-alpha, IL-6, and IL-12] and angiogenesis-modulating factors (vascular endothelial growth factor and endostatin) have also been analyzed. Cervical tumor biopsies and blood samples were obtained at the time of diagnosis and after 10 days of celecoxib treatment (400 mg b.i.d., at 8:00 a.m. and 8:00 p.m.) in 27 cases. Immunohistochemistry and ELISA assays were used to assess the expression of biological factors in tumor tissue and circulating levels of cytokines and angiogenic molecules. We showed a statistically significant increase in the percentage of TIL expressing the TCR-zeta chain after celecoxib treatment: indeed, in cases exposed to celecoxib, the percentage of TCR-zeta(+) cells ranged from 5.0 to 50.0 (median, 22.5) with respect to baseline expression (range, 3.0-50.0; median, 10.0; P = 0.0016). There was no significant treatment-related difference in the percentage of CD3(+), CD4(+), CD8(+), and CD25(+) TIL as well as in tryptase-positive cells. IL-12 levels were significantly reduced in posttreatment samples with respect to baseline levels (P = 0.002). We also found a reduction in the circulating levels of vascular endothelial growth factor, and a statistically significant increase of serum endostatin levels (P = 0.035). We reported the first evidence in humans that celecoxib restores zeta expression by TIL in primary cervical tumors, suggesting that a positive modulation of immune function may serve as an additional mechanism supporting the antitumor effect of this class of drugs.

Association of Interleukin-8 and Plasminogen Activator System in the Progression of Colorectal Cancer

The aim of this study was to investigate the relationship of Interleukin-8 (IL-8) with vascular endothelial growth factor (VEGF) and plasminogen activator system (PA system) in the progression of colorectal cancer (CRC). In eighty-seven patients with CRC, the levels of IL-8, and VEGF as representative angiogenic factors and urokinase-type plasminogen activator (uPA), uPA receptor (uPAR), plasminogen activator inhibitor-1 (PAI-1), and PAI-2 as representative invasive factors were quantitatively assayed in tumor and adjacent normal tissues. The levels of IL-8, VEGF, and PA system factors in tumor tissues were all significantly higher than those in normal tissues. The IL-8 level was significantly associated with tumor size, depth of infiltration, Dukes stage, and liver metastasis, and also significantly correlated with the levels of VEGF, uPAR, uPA, and PAI-1. The VEGF level was significantly associated with tumor size, vascular involvement. The levels of uPAR and PAI-1 were significantly associated with tumor size and depth of infiltration, and the uPAR level was associated with liver metastasis. The VEGF level was significantly correlated with the levels of uPAR and PAI-1. These results reveal that IL-8, VEGF, and PA system factors are contributed to tumor growth, invasion, and metastasis in CRC. Univariate analysis revealed that high levels of IL-8, VEGF, and uPAR were significantly associated with a shorter overall survival time; however, multivariate analysis identified only liver metastasis as an independent prognostic factor. In conclusion, IL-8 is responsible to tumor progression and liver metastasis of CRC, and the activation of PAS induced by IL-8 as well as VEGF may play an important role in the progression of CRC.

Tissue factor and fibrin in tumor angiogenesis.

The hypercoagulability exhibited by most cancer patients leads to serious complications such as venous thromboembolism and contributes to the pathogenesis of tumor growth and metastasis by promoting angiogenesis. The key player in this vicious cycle is tissue factor (TF), the initiator of blood coagulation. Although TF normally safeguards vascular integrity by inducing hemostasis upon injury, abnormal expression of TF in different tumors and related vascular endothelial cells contributes to unnecessary clot formation in cancer patients. Clotting-dependent induction of tumor angiogenesis is primarily mediated by TF-induced generation of thrombin and subsequent deposition of cross-linked fibrin. A cross-linked fibrin network provides a provisional proangiogenic matrix that facilitates blood vessel infiltration. Clotting-independent mechanisms of TF-induced tumor angiogenesis have also been described, mediated primarily by the cytoplasmic tail of the TF receptor. TF activation could contribute to the venous thromboembolism that has been reported as a complication of the use of novel antiangiogenic agents in combination with chemotherapy. Anticoagulants, such as low-molecular-weight heparin, may act to prevent these complications both by interfering with TF-mediated activation of clotting and by directly down-regulating angiogenesis. Thus, TF may prove to be a novel target for cancer therapy.

The role of the hemostatic system in tumor growth, metastasis, and angiogenesis: tissue factor is a bifunctional molecule capable of inducing both fibrin deposition and angiogenesis in cancer.

Cancer patients are prone to venous thromboembolism (VTE), and this hypercoagulability favors tumor growth and metastasis. After a brief review of the clinical aspects of VTE and cancer, we discuss the pathogenesis of hypercoagulability with an emphasis on the role of tissue factor (TF). The discovery that, in addition to tumor cells, TF is expressed by tumor-associated macrophages and tumor-associated endothelial cells led to studies of the role of TF in the regulation of tumor angiogenesis. In human lung cancer, melanoma, and breast cancer, TF and vascular endothelial growth factor (VEGF) co-localize in tumor cells; a close correlation exists between TF and VEGF synthesis (P = .001) in tumor cell lines and with angiogenesis in vivo in a severe, combined immunodeficient mouse model. Transfection of a TF/VEGF low-producing human tumor cell line with full length TF complementary DNA (cDNA) results in conversion to a high producer of TF and VEGF; transfection of a deletion-mutant TF cDNA lacking cytoplasmic serine residues restores full TF procoagulant activity but not VEGF synthesis to the cells. These results suggest that the cytoplasmic tail of TF is necessary for tumor cell VEGF synthesis. Targeting of TF in tumors and tumor-associated blood vessels is discussed as a strategy for drug delivery and rational anti-cancer and anti-angiogenesis drug design.

Tissue factor expression in breast cancer tissues: its correlation with prognosis and plasma concentration.

Tissue factor (TF), an initiator of the extrinsic coagulation cascade, is expressed in a wide range of cancer cells and plays important roles in cancer progression and metastasis. Recently, the intracellular function of TF has been revealed to be involved in cancer invasion, independent of the blood coagulation pathway. To evaluate the clinical significance of TF expression, we performed an enzyme-linked immunosorbent assay (ELISA) in the plasma of 67 breast cancer patients and immunohistochemistry in 213 breast cancer tissues. In the ELISA study, we showed an up-regulation of plasma TF concentration in breast cancer patients compared with normal controls. Immunohistochemistry demonstrated that TF was expressed in tumour cells and stromal cells and tumour TF expression closely correlated with stromal TF expression (P = 0.0005). The concentration of plasma TF was associated with tissue TF expression in both tumour and stroma. The multivariate analysis demonstrated that tumour TF expression was an independent prognostic indicator for overall survival (P = 0.0452). Our data show that plasma TF concentration reflects tissue TF expression and tumour TF expression can provide some predictive value for prognosis and distant metastasis, which indicates the importance of TF function in tumour progression. © 2000 Cancer Research Campaign

Larger and more invasive colorectal carcinoma contains larger amounts of plasminogen activator inhibitor type 1 and its relative ratio over urokinase receptor correlates well with tumor size

Considering recent findings that both urokinase plasminogen activator receptor (uPAR) and plasminogen activator inhibitors (PAIs) are involved in tumor growth through an urokinase-type plasminogen activator (uPA) activity-independent mechanism, the relation between the presence of these factors in tumor tissue and the clinicopathologic variables in colorectal carcinoma was reevaluated. In 100 colorectal carcinoma patients, antigen levels of u-PA, uPAR, and PAI-1 and PAI-2 were assayed in both tumor tissues and their normal counterparts. Plasma levels of soluble uPAR also were determined. All uPAR, uPA, PAI-1, and PAI-2 antigen levels in tumor tissue were significantly higher than those in normal tissue. Levels of both uPAR and PAI-1 were significantly higher (3.09 +/- 1.37 and 6.63 +/- 7.49, respectively) in large tumors (>/=50 mm in greatest dimension) than those in smaller tumors (< 50 mm) (2.50 +/- 1.07 and 2.72 +/- 2.70, respectively) (P < 0.05). Significant positive correlation coefficients (r) were obtained between tumor size and the calculated ratios of PAI-1/uPAR (r = 0.490; P < 0.0001) and PAI-1/uPA (r = 0. 469; P < 0.0001). In addition to liver metastases (P = 0.004) and lymph node involvement (P = 0.04), high levels of uPAR (P = 0.05) also were found to be of independent prognostic value by multivariate analysis. Higher expression of uPAR was related to poor prognosis of patients with colorectal carcinoma and excess amounts of PAI-1 over uPAR or uPAR-bound uPA appeared to play an important role in tumor progression.