Peter J. Hamer

Elevated plasma tissue inhibitor of metalloproteinase-1 level predicts decreased response and survival in metastatic breast cancer.

Tissue inhibitors of metalloproteinase (TIMPs) have at least 2 different functions. They inhibit the catalytic activity of matrix metalloproteinases, and they act as growth factors.

Serum TIMP-1 and Response to the Aromatase Inhibitor Letrozole Versus Tamoxifen in Metastatic Breast Cancer

PURPOSE To determine the effect of elevated serum TIMP-1 on the response of patients with metastatic breast cancer to an aromatase inhibitor versus tamoxifen. PATIENTS AND METHODS Five hundred twenty-two patients estrogen receptor-positive metastatic breast cancer were randomly assigned to receive first-line hormone therapy with letrozole or tamoxifen. Serum tissue inhibitor of metalloproteinases-1 (TIMP-1) levels were measured using an enzyme-linked immunosorbent assay. RESULTS Pretreatment serum TIMP-1 was elevated in 120 (23%) of 522 patients. Patients with elevated serum TIMP-1 had a significantly reduced objective response rate (19.2% v 30.6%; odds ratio, 0.54; P = .01), duration of response (median, 15.5 v 26.2 months; P = .001), time to treatment progression (TTP; median, 4.5 v 9.2 months; HR, 1.78; P = .0001), time to treatment failure (median, 3.5 v 9.0 months; HR, 1.77; P = .0001), and overall survival (median, 20.3 v 35.8 months; HR, 1.77; P = .0001) compared with patients with normal pretreatment TIMP-1 levels. Letrozole was superior to tamoxifen in both the normal serum TIMP-1 group (median TTP, 11.8 v 8.6 months; P = .003) and in the elevated serum TIMP-1 group (median, 6.1 v 3.2 months; P = .03) In multivariate analysis, elevated serum TIMP-1 remained an independent predictor of both shorter TTP (HR, 1.46; P = .002) and survival (HR, 1.44; P = .002), as did serum HER-2. Combined analysis of both serum TIMP-1 and HER-2/neu conferred additional ability to predict significantly different clinical outcomes compared to using either biomarker alone. CONCLUSION Patients with elevated pretreatment serum TIMP-1 had a significantly reduced response and survival. Serum TIMP-1 was an independent predictive and prognostic factor. Blockade of TIMP-1 and HER-2/neu activity may be beneficial in a subset of patients with breast cancer.

Serum epidermal growth factor receptor/HER-2 predicts poor survival in patients with metastatic breast cancer.

Epidermal growth factor receptor (EGFR, HER‐1, and erbB1) is overexpressed in primary breast cancer and had been identified as a poor prognostic factor.

Elevated plasma tissue inhibitor of metalloproteinase‐1 levels predict decreased survival in castration‐resistant prostate cancer patients

Tissue inhibitor of metalloproteinase‐1 (TIMP‐1) has paradoxical multifunctional roles in tumorigenesis: inhibition of the catalytic activity of matrix metalloproteinases and apoptosis as well as promotion of angiogenesis and tumor growth. Elevated TIMP‐1 levels have been associated with a poorer prognosis in multiple cancers.

High levels of serum TIMP-1 correlate with advanced disease and predict for poor survival in patients with multiple myeloma treated with novel agents

Tissue inhibitor of metalloproteinase-1 (TIMP-1) was evaluated in the pre-treatment serum of 55 newly diagnosed patients with symptomatic myeloma. TIMP-1 was elevated in 47% of patients and correlated with lytic bone disease and increased bone resorption. Importantly, TIMP-1 correlated with ISS stage (p=0.005) and was an independent prognostic covariate for survival [HR: 1.003 (1-1.006), p=0.004] in these patients who were all treated with novel agents (bortezomib and/or IMiDs) during their disease course. Our study provides evidence that pre-treatment serum TIMP-1 is associated with advanced myeloma and suggests the further evaluation of this molecule to better determine its prognostic potential in MM.

Abstract 826: Expression levels of circulating soluble-EphA2 receptor in cancer patients

EphA2 is a receptor tyrosine kinase that critically controls many aspects of cell behavior. It is expressed in a manner that is consistent with the regulation of cell growth, migration and invasion. Analysis by IHC has shown that elevated expression of EphA2 exhibits predictive value in lung cancer survival and recurrence. Importantly, current data suggests that targeting EphA2 could be beneficial and is currently being tested in an ovarian cancer setting. We developed a prototype ELISA for circulating EphA2 receptor in order to investigate expression levels of the soluble form of EphA2 in certain cancer types. We have developed a prototype ELISA which measures circulating human EphA2 receptor. 96 well Nunc plates were coated with an affinity-purified goat polyclonal antibody specific for the extracellular domain (ECD) of human EphA2. Human serum and plasma samples were diluted 5-fold in a sample diluent with blocking reagents. The standard was a mammalian cell-derived form of the ECD for the human EphA2 receptor. Standards and samples were incubated on the plates for 2 Hours, 37C. The detector antibody was a biotinylated affinity-purified goat polyclonal antibody and was added to the plates for 1 Hour, 37C. Plates were washed and a streptavidin-HRP conjugate was added to the plates for 30 minutes, 22C. A final wash was performed and a TMB substrate was added to the plates for color development. The reaction was stopped with sulfuric acid and plates were read at an absorbance of 450nm. Data generated to date using this prototype ELISA indicates that over-expression of soluble-EphA2 occurs in many cancers versus normal samples tested. The 95% CI of total normal plasma and serum was 672_1306 and 894_1429pg/mL, respectively. Capture Inhibition studies revealed specificity of the assay in human samples (100% inhibition). Parallelism studies revealed acceptable performance in serially diluted samples. Significant over-expression of soluble-EphA2 was observed in benign lung cancers versus normals (100%, 10/10) while staged lung cancers exhibited significantly lower levels of circulating EphA2 (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 826.

Abstract 2276: Serum endoglin and uPA change predicts PFS and overall survival in first line trastuzumab-treated breast cancer

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Background: About half of HER2-positive breast cancer patients will respond to first-line trastuzumab-containing therapy, but of these most will progress within a year with acquired resistance. Since trastuzumab treatment is now used in the HER2-positive adjuvant breast cancer setting, trastuzumab resistance will continue to be a recurring clinical problem, and better predictive and prognostic biomarkers are urgently needed. Methods: Serum TIMP-1, uPA, carbonic anhydrase 9 (CA9), and endoglin (co-receptor for TGF- β) were measured using ELISA in 60 metastatic breast cancer patients before starting first-line trastuzumab-containing therapy, and at the closest 1 month post-trastuzumab blood draw. The TIMP-1, uPA and CA9 ELISAs were from Oncogene Science/Siemens Healthcare Diagnostics, and the endoglin ELISA was from R&D Systems. Progression-free (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox modeling. Results: When serum biomarker change (pretreatment to 1 month post-trastuzumab treatment) was analyzed on a continuous basis, change in serum TIMP-1 and CA 9 were not significant for PFS or OS. However the change in serum endoglin (HR = 1.017, p = 0.013) and serum uPA (HR 1.018, p = 0.006) were significant predictive factors for PFS to first-line trastuzumab treatment. The change in serum endoglin (HR = 1.021, p = 0.004) and serum uPA (HR 1.016, p = 0.03) were also significant prognostic factors for OS: declining serum endoglin or uPA predicted for increased PFS and OS. Changes in serum endoglin and uPA were positively correlated (r = 0.7, p <0.0001). Due to the significant correlation between the change in serum endoglin and uPA levels, these 2 serum biomarkers were analyzed separately along with the change in serum TIMP-1 and CA9 in multivariate analysis. In these analyses, only the change in serum endoglin or uPA was significant. Conclusions: The change in serum endoglin and uPA level from pretreatment to 1 month post-trastuzumab treatment predicted PFS and overall survival in metastatic breast cancer patients treated with first-line trastuzumab-containing therapy. Endoglin (CD105) is a co-receptor for TGF-β, is expressed by human vascular endothelial cells, and plays a major role in angiogenesis. These angiogenesis-related serum biomarkers deserve further study in larger trials of HER2-targeted breast cancer treatment. Supported by a grant from Komen for the Cure. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2276. doi:10.1158/1538-7445.AM2011-2276

Use of serum uPA change to predict PFS and overall survival in first-line trastuzumab-treated breast cancer.

1050 Background: About half of patients with HER2-positive breast cancer will respond to first-line trastuzumab-containing therapy, but of these most will progress within a year with acquired resistance. Since trastuzumab treatment is now used in the HER2-positive adjuvant breast cancer setting, trastuzumab resistance will continue to be a recurring clinical problem, and better predictive and prognostic biomarkers are urgently needed. Methods: Serum TIMP-1, uPA, and carbonic anhydrase 9 (CA9) were measured using ELISA in 60 patients with metastatic breast cancer before starting first-line trastuzumab-containing therapy, and at the closest 1 month post-trastuzumab blood draw. The TIMP-1, uPA, and CA9 ELISAs were from Oncogene Science/Siemens Healthcare Diagnostics, Cambridge, MA. Progression-free survival (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox modeling. Results: When serum biomarker change (pretreatment to 1 month post-trastuzumab treatment) was analyzed on a co...

Abstract 2254: A panel of circulating biomarkers in advanced pancreatic cancer

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Background: Pancreatic cancer is one of the deadliest forms of cancer as its five year survival rate is only 4%. There is currently a dearth of circulating biomarkers that have been investigated for pancreatic cancer. We analyzed plasma and serum banks for a variety of circulating biomarkers with potential significance in this cancer or biomarkers known to be targets for new therapeutics in development. Increased PDGFR-B has been seen in pancreatic tumors as compared to normal tissue. Increased TIMP-1 or uPA has been associated with an unfavorable prognosis in many cancers including those of the colon, bladder and breast. Mutations in the K-ras gene occur in a high percentage of pancreatic tumors, therefore we analyzed a subset of samples for circulating Ras p21, as well as for VEGF-165, VEGF-R2 and EphA2. Methods: 52 EDTA Plasma samples from two pancreatic patient cohorts were analyzed for TIMP-1 and PDGFR-β levels. In addition, 25 pancreatic cancer patient sera were analyzed for TIMP-1, uPA and PDGFR-β. A subset of 20 EDTA plasma samples from one of the cohorts of pancreatic cancer patients were analyzed for circulating Ras p21, VEGF-165, VEGF-R2 and EphA2. The majority of this subset of patients (70%) had stage 3 pancreatic cancer while the remainder had stage 2 or stage 4 disease. ELISA assays used were from Oncogene Science/Siemens Healthcare Diagnostics (Cambridge, MA). Normal ranges and cutoff values were established for each analyte using in-house normal samples. Statistical methods included one-way ANOVA, using Kruskal-Wallis test and Dunns multiple comparison test. Results: Data from this study showed that several putative biomarkers were elevated in a large percentage of the advanced pancreatic patients. Of the samples from the plasma cohort, 90% of them were elevated for TIMP-1. Similarly, in the serum cohort, 70% of the samples were elevated for TIMP-1. Of the plasma subset, 45% of the patients had increased EphA2 levels and 60% were increased for VEGF-165 while 77% of the patients had elevated Ras p21 levels. Conclusions: A high percentage of pancreatic cancer patients have elevated levels of several circulating biomarkers such as Ras p21, VEGF-165, EphA2 and TIMP-1. It is known that a high percentage of pancreatic cancer patients have mutated K-ras genes and the introduction of an assay for circulating ras p21 may assist in deciphering what role Ras p21 plays in pancreatic cancer. In addition, VEGF-165 is the target of many current and future anti-angiogenesis drugs. A panel of markers such as those analyzed in this study of pancreatic cancer has the potential for use as a selection tool for targeted therapies such as EphA2 targeted therapies under development. In addition, biomarker panels, which may include traditional tumor markers, may have utility in monitoring patient therapy. 1 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2254.

Abstract 5081: Dysregulation of soluble EGFr, TIMP-1, EphA2 and CAIX levels in advanced ovarian cancer

Background: In recent years, survival rates for patients with ovarian cancer have been improving, however, management of patients with advanced ovarian cancer continues to be a difficult clinical problem. Due to tumor heterogeneity, patients often receive expensive therapies with dangerous side-effects to which their tumors do not respond. Soluble biomarkers, particularly when assayed in panels, have the potential to assist clinicians in selecting and monitoring appropriate therapies. Increases in soluble TIMP-1 and CAIX and decreases in soluble EGFR levels have been reported in many cancers, and along with EphA2, are targets for novel cancer therapeutics. To address the utility of using these in a panel for ovarian cancer, we determined serum and plasma levels of all four biomarkers. Methods: Plasma and sera from stage I -IV ovarian cancer patients were analyzed using the TIMP-1, EGFR, CAIX and EphA2 ELISA assays from Oncogene Science. Levels were then compared to those found in healthy, normal females. Statistical methods used included Kruskal-Wallis test and Dunn9s multiple comparison test. Results: Soluble levels of all four biomarkers were dysregulated in the serum and/or plasma of ovarian cancer patients as compared with levels found in normal healthy, females. For our analysis of serum results, stages I and II were combined and stages III and IV were combined due to the low number of stages I and IV samples in this cohort. Serum levels of TIMP-1 and CAIX were significantly elevated in stages I/II (p = Conclusions: EGFR, TIMP-1, CAIX and EphA2 levels in ovarian cancer patients are significantly different from the levels found in normal, healthy females. These markers are dysregulated in early as well as later stages and changes involve both decreases and increases in levels. The fact that these biomarkers behave differently from one another may reflect different biological pathways which could contribute to ovarian tumor heterogeneity. Using these markers as a panel may assist clinicians in selecting the most appropriate therapy for their patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5081.