bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4364: S1P generated by SphK1 is important not only for primary tumor growth but also for tumor-induced hemangiogenesis and lymphangiogenesis
Authors: Nagahashi Masayuki , Ramachandran Subramaniam , Kim Eugene Y. , Allegood Jeremy C. , Rashid Omar M. , Yamada Akimitsu , Zhao Renping , Milstien Sheldon , Zhou Huiping , Spiegel Sarah , Takabe Kazuaki
Published: August 2012

INTRODUCTION: Sphingosine-1-phosphate (S1P), a pleiotropic bioactive lipid mediator, regulates many cellular processes important for breast cancer progression. The aim of this study is to investigate a role for S1P produced by sphingosine kinase 1 (SphK1) in breast cancer progression and tumor-induced hemangiogenesis and lymphangiogenesis. METHODS: An isozyme-specific inhibitor of SphK1 (SK1-I) was used. Tumor burden were quantified using an in vivo imaging system (IVIS 2000). S1P and SK1-I levels were measured by LC-ESI-MS/MS. Hemangiogenesis and lymphangiogenesis are determined by morphological analysis of microvessel density as well as by flow cytometric analysis of endothelial cells. RESULTS: A significant dose dependent effect of SK1-I on inhibition of 4T1-luc2 murine breast cancer cell growth was observed. We confirmed that SK1-I decreased the enzymatic activity of SphK1 and that downregulation of SphK1 with specific siRNA also suppressed growth of these cells. We found that S1P levels increased in the tumor and in the circulation after orthotopic implantation of 4T1-luc2 cells. Similarly, serum S1P levels were significantly elevated in stage IIIA breast cancer patients who have lymph node metastases, compared to age/ethnicity-matched healthy volunteers. SK1-I blocked serum S1P increases in in vivo model and reduced lymph node and lung metastases and overall tumor burden in vivo as well. Importantly, SK1-I also decreased hem- and lymphangiogenesis not only in the primary tumor, but also in lymph nodes, the host tumor microenvironment. Whereas supernatants from 4T1-luc2 cells significantly stimulated tube formation of HUVECs and HLECs, shRNA knockdown of SphK1 markedly reduced them, indicating that S1P produced by SphK1 in 4T1-luc2 breast cancer cells is an important contributor to hem- and lymphangiogenesis. CONCLUSIONS: S1P generated by SphK1 is important for tumor progression, tumor-induced hemangiogenesis and lymphangiogenesis, and lymph node metastais. Therefore targeting SphK1 and its product S1P would be a multi-pronged attack against breast cancer. This work was supported by Sumitomo Life Social Welfare Services Foundation grant to MN, NIH (K12HD055881) and Susan G. Komen for the Cure (KG090510) to KT, and NCI (R01CA61774) to SS.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4364. doi:1538-7445.AM2012-4364
Abstract 4392: Vaccination targeting IGF-IR sensitizes tumors to tamoxifen therapy in an anti-estrogen resistant mouse model
Authors: Cecil Denise L. , Park Kyong Hwa , Gad Ekram , Rastetter Lauren , Disis Mary L.
Published: August 2012

Tamoxifen is a standard treatment for estrogen receptor (ER)-positive breast cancer patients. However, acquired or de novo resistance to therapy is a major clinical problem. In hormone-resistant tumors, there is increased activation of insulin-like growth factor-1 receptor (IGF-IR) and subsequent downstream signaling molecules, such as AKT. We have determined that IGF-IR is immunogenic in breast cancer and is a potential target for active immunization. It has been demonstrated that natural immunogenic human epitopes can be predicted by high binding affinity across multiple class II alleles, thus, we used a combined scoring system from five algorithms for predicting class II binding to determine Th epitopes of IGF-IR. Of the 20 potentially immunogenic peptides identified, five peptides (p1166-1181, p1212-1227, p1301-1316, 1307-1322 and p1311-1326) in the C-terminal domain were determined to elicit a predominantly inflammatory Th1 response compared to an immunosuppressive Th2 response in human PBMC. Overexpression of AKT induces tamoxifen resistance, while inhibition restores tamoxifen sensitivity. The tumor suppressor, PTEN, actively inhibits AKT. Data we have generated has demonstrated that vaccination restored PTEN activity in tumor cells. We questioned if modulation of PTEN could render tumors in the anti-estrogen-resistant MMTV-neu mouse model sensitive to tamoxifen therapy. Vaccination demonstrated a robust Th1 response (p<0.001) and concomitant inhibition of tumor growth by 85% compared to adjuvant only control animals (p<0.001). Treatment with tamoxifen in vaccinated tumor-bearing mice resulted in a further 38% reduction in growth (p=0.02). Thus, active immunization targeting IGF-IR may induce both immunologic and biologic effects resulting in the sensitization of the tumor to tamoxifen therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4392. doi:1538-7445.AM2012-4392
Abstract 4397: A TLR4-interacting peptide (SPA4) reduces the lipopolysaccharide-stimulated NF-kappaB signaling, cytokines and migration of colon cancer cells
Authors: Madhusoodhanan Rakhesh , Ramani Vijay , Moriasi Cate , Anant Shrikant , Awasthi Shanjana
Published: August 2012

Persistent inflammation in patients with colitis can lead to cancer. However, molecular mechanisms responsible for inflammation-induced cancer are not understood. Activation of TLR4-NF-kappaB signaling is associated with inflammation. Thus, we hypothesized that inhibition of TLR4-NF-kappaB signaling will help reduce inflammatory response and inflammation-induced cancer progression. In this work, we studied the effects of a TLR4-interacting surfactant protein-A-derived (SPA4) peptide on lipopolysaccharide (LPS)-induced TLR4-NF-kappaB signaling and cancer progression in colon adenocarcinoma SW480 cells. The SW480 cells were challenged with Escherichia coli-derived LPS. Cells were then treated with varying amounts of SPA4 peptide. Any changes in the expression of TLR4, intracellular-signaling molecules (IKBalpha, p65, phosphorylated IKBalpha, phosphorylated p65, RelB, Cox-2) and cytokines (IL-1beta, IL-6), and activity of NF-kappaB were studied by immunocytochemistry, immunoblotting and NF-kappaB reporter assay, respectively. Simultaneously, cell cycle progression, viability and migratory pattern of SW480 cells were followed. We found that the binding of SPA4 peptide to TLR4 inhibits the LPS-induced expression of TLR4, phosphorylated p65, Cox-2, IL-1beta and IL-6, activity of NF-kappaB and migration of SW480 cells. Although there was no significant change in cell cycle progression, the SPA4 peptide significantly inhibited the viability of SW480 cells. In conclusion, our preliminary results suggest that the inhibition of TLR4-NF-kappaB signaling by SPA4 peptide may be useful in prevention and treatment of inflammation and inflammation-induced cancer in patients with colitis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4397. doi:1538-7445.AM2012-4397
Abstract 44: Enhancement endometrial cancer cells malignant transformation by mutual interaction with mesenchymal stem cells through TGF-beta and SDF-1 signaling pathways
Authors: Ding Dah-Ching , Chu Tang-Yuan
Published: August 2012

Designated to cyclic shedding, endometrial glandular stroma is rich for mesenchymal stem cells and may play crucial roles in cancer invasion. This study aimed to explore the interaction between endometrial cancer cells and endometrial mesenchymal stem cells (HEMSC). HEMSC were derived from human endometrium and were co-cultured with RL95-2 endometrial cancer cells. Transformation phenotypes including proliferation, migration, anchorage independent growth, colony formation and invasion were characterized. Cancer associated transforming growth factor (TGF)-beta1 and mesenchymal cell associated stromal cell-derived factor (SDF) and their down-stream signaling pathways were investigated. HEMSC had a similar surface marker profile as bone marrow mesenchymal stem cells, i.e. CD44(+), CD90(+) and CD34(-) and CD45(-) and readily be inducible to osteogenic and adipogenic differentiations. Upon coculture with HEMSC, RL95-2 cell secreted TGF-beta 1 which subsequently induced SDF1 secretion from HEMSC. The mutual interaction of these factors is associated with enhancement of cancer cell migration through the CXCR4 receptor and downstream transducer and activator of Stat3, with resulting over-expression of alpha-SMA and cytoskeleton re-organization. The study observed unprecedented interactions between cancer cells and mesenchymal stem cells with resulting enhancement of malignant transformation through mutual activation of cytokines and downstream signals. Mesenchymal stem cells either in adjacent to the lesion or home to the lesion can therefore be important activators of carcinogenesis and good therapeutic targets.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 44. doi:1538-7445.AM2012-44
Abstract 4403: Combined antagonism of the IL-4R and IKKα inhibits melanoma tumor growth in immunocompetent mice
Authors: Hawkins Oriana E. , Horton Linda , Fordyce Krystle , Ayers Gregory , Richmond Ann
Published: August 2012

With over 8500 deaths each year in the USA, malignant melanoma is the deadliest of all skin cancers. Constitutive activation of the nuclear factor kappa B (NF-κB) is common in melanoma, promoting tumor growth, survival and metastasis. In light of its role in tumorigenesis, targeting of the NF-κB pathway has been considered a possible therapeutic option for the treatment of melanoma. However, NF- αB activation is critical for the development, survival, migration and function of immune cells. We hypothesized that systemic inhibition of the activation of NF-κB might have a deleterious effect on the tumor immune response. Melanoma bearing C57BL/6 mice received 50 mg/kg of BMS-345541, a selective inhibitor of the inhibitor of αB kinase (IKKα), twice daily. Tumors, spleens and bone marrow were harvested at different time points and analyzed by flow cytometry for leukocyte composition and cytokine production. Intracellular cytokine staining revealed an early induction of Interleukin-4 (IL-4) within the myeloid and B cell components of the tumor, as a result of BMS-345541 treatment. The production of IL-4 by these cells suggests an early switch to a tumor promoting T helper 2 (Th2) and M2 driven immune response, which may counteract the anti-tumor effect of IKKα inhibition by generating a pro-tumorigenic microenvironment. These results, in part, explain the modest delay in tumor growth demonstrated. IL-4 receptor (IL-4R) expression on human melanoma cells has been previously documented; however the effects of IL-4 on these cells have generated contradictory reports. IL-4R signaling has been shown to either inhibit growth of tumor cells in vitro or to promote survival of tumor cells. In C57Bl/6 mice bearing a murine melanoma, BMS345541 (75 mg/kg bid) was given in combination with 100 µg of IL-4R blocking antibody on day 4 and day 12 of treatment. This combination significantly inhibited tumor growth compared to BMS345541 treatment alone. These data suggest an important role for IL-4 in promoting melanoma growth in vivo. Dual targeting of IL-4R and NF-κB signaling pathways neutralizes the tumor promoting immune component. Currently, IL-4R antagonists are in clinical trials for the treatment of asthma, which makes the translational prospects of this study extraordinarily viable. We are grateful for support from the NCI (CA116021-07, 5T32CA119925-03) and the Department of Veterans Affairs for these studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4403. doi:1538-7445.AM2012-4403
Abstract 4408: Targeting cancer stem cells as potential new therapy for pancreatic cancer
Authors: Cleary John P. , Raman Divya , Punzalan Rubio R. , Sharma Michael Vr , Sharma Cristian Vr , Sharma Sherven P. , Jani Jitesh , Sharma Bhuvnesh , Narayan Satya , Sharma Jay P.
Published: August 2012

Pancreatic cancer is the fourth leading cause of cancer mortality in the US, despite significant improvements in diagnostic imaging and operative modalities. The 5-year survival rate remains less than 6% because of microscopic or gross metastatic disease at time of diagnosis. Although the treatment of pancreatic cancer remains a huge challenge, it is entering a new era with the development of new strategies and trial designs. Because there is an increasing number of novel therapeutic agents and potential combinations available to test in patients with pancreatic cancer, the identification of robust prognostic and predictive markers and of new targets and relevant pathways is a top priority as well as the design of adequate trials incorporating molecular-driven hypothesis. Over the past decade, increasing evidence suggested that stem cells play a crucial role not only in the generation of complex multicellular organisms, but also in the development and progression of malignant diseases. Most tumors have been shown to contain a subset of distinct cancer cells that is responsible for tumor initiation and propagation. These cells are termed cancer stem cells or tumor-initiating cells and they are highly resistant to chemotherapeutic agents. Here, we examined the efficacy of combined treatments of CEP1101 and gemcitabine in human pancreatic cancer cells, and pancreatic CSCs (CD133, CD44, CXCR4, SSEA3/4, Oct4, ALDH, Telomerase & Nestin) from the same donors. CEP1101 inhibited the growth of CSCs, while gemcitabine suppressed the viability of non-CSCs. In vivo studies showed that CEP1101 combined with gemcitabine eliminate the engraftment of human pancreatic cancer and CSCs, more effectively than the individual agents. These data demonstrate that administration of CEP1101, which targets CSCs, may constitute a potential therapeutic strategy for improving the efficacy of gemcitabine to eradicate pancreatic cancer. This study shows potential molecular therapeutic targets to eradicate the tumor - and metastasis-initiating cells and their progenies for the evaluation of new effective combination therapies against locally advanced and metastatic pancreatic cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4408. doi:1538-7445.AM2012-4408
Abstract 4411: Farletuzumab (mAb003), a neutralizing monoclonal antibody to folate receptor alpha, enhances paclitaxel efficacy in an ovarian cancer model
Authors: Maddage Christopher , Rybinski Katherine , Zhou Yuhong , Grasso Luigi , Nicolaides Nicholas C. , Lin JianMin
Published: August 2012

Farletuzumab, MORAb-003 is a humanized monoclonal antibody (mAb) that recognizes the folate receptor alpha (FRα) which is over-expressed in various malignancies including ovarian and lung cancers. This presents a unique approach to an unmet medical need in ovarian cancer therapy by targeting this receptor. Farletuzumab is currently in clinical trials in combination with paclitaxel for metastatic ovarian and lung cancer. In this study we evaluated potential benefit that farletuzumab offers when added to a traditional chemotherapy in a pre-clinical murine model. In a human ovarian cancer model, female athymic nude mice were implanted intraperitoneally with IGROV cells which express FRα. One week post tumor establishment, these mice were treated with Paclitaxel, MORAb-003, or the combination of the two, respectively. A group of the tumor-bearing mice received PBS as control. Tumors found in peritoneal cavity were harvested and weighed, then recorded as tumor burden. In animals treated with MORAb-003 at 25mg/kg twice weekly for 4 weeks, tumor growth was reduced near 30%. With treatment of paclitaxel at 0.5 mg/kg weekly for 4 weeks, the tumor burden in these mice was reduced ∼65%. An enhancement of therapeutic effect on tumor burden was clearly observed when MORAb-003 was added to the chemo treatment and resulted in a growth inhibition of greater than 80%. Our results demonstrate the ability of MORAb003, as a mono-agent, in inhibiting the growth of ovarian cancer in a murine model. When added to the chemotherapeutic regimen, it enhances the therapeutic efficacy of traditional chemotherapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4411. doi:1538-7445.AM2012-4411
Abstract 4413: Rosiglitazone prevents graft-versus-host disease (GVHD)
Authors: Song Eun-Kee , Yhim Ho-Young , Yim Jun-Mo , Yim Joo-Yun , Song Min-Young , Lee Na-Ri , Kwak Jae-Yong , Yim Chang-Yeol , Sohn Myung-Hee
Published: August 2012

GVHD is a host tissue injury mediated by donor-host immune reactions, and a major complication in allogeneic hematopoietic stem cell transplantation. Peroxisome proliferator-activated receptor-α (PPARδ) plays an important role in regulating immune responses. We investigated the role of rosiglitazone, a synthetic PPARδ agonist, in a murine parent-to-F1 GVHD model. During mixed lymphocyte reactions (MLR), rosiglitazone inhibited the proliferative response and cytotoxicity of responder cells, and induced more apoptosis in CD4+, CD8+, and B220+ cells than in NK1.1+, Mac-1+, CD4/CD25+ and CD3/NK1.1+ cells. The in vivo administration of rosiglitazone demonstrated histologic improvements of GVHD in the liver, skin, spleen and intestine. Rosiglitazone treatment inhibited GVHD-induced increases in serum levels of TNFα, IFNα, IL-6, and IL-12, and the decreases in TGFβ and IL-10. Splenic leukocyte immunophenotyping demonstrated that while the proportions of both donor and host CD4/CD25+ and CD3/NK1.1+ cells were increased by the treatment with rosiglitazone, both donor and host CD8+ cells were decreased. Rosiglitazone treatment inhibited GVHD-induced decreases in the expression of phosphate and tensin homologue deleted on chromosome ten (PTEN), and the increases in the expression of p-Akt and nuclear NF-κB in the spleen. These results indicate that rosiglitazone and activation of PPARδ may be useful to protect the host from GVHD.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4413. doi:1538-7445.AM2012-4413
Abstract 4476: Molecular typing of breast cancer cases among a Hispanic population and its association with body mass index
Authors: Diaz-Santana Mary V. , Ramos Gilberto , Nazario Cruz M.
Published: August 2012

Background: Breast cancer is a heterogeneous disease which can be divided into distinct molecular subtypes based on patterns of gene expressions and tumor marker staining. The prevalence of breast cancer molecular subtypes (BCMS), as well as specific risk factors by subtypes varies by ethnicity, highlighting the relevance of research focused in specific populations. Although the BCMS have been studied, there is scarce evidence of the association between breast cancer risk factors and molecular subtypes and the prevalence of each subtype among Hispanics populations. In Puerto Rico, breast cancer is the most frequently diagnosed malignancy and the leading cause of cancer death among females. However, little is known about the breast cancer subtypes distribution and its etiologic profile among Puerto Rican women. Objectives: We aimed to determine the prevalence of BCMS and assess whether obesity differs according to these subtypes among a female population of breast cancer cases in Puerto Rico. Methods: A case-case analysis was conducted using all cases of invasive breast carcinoma consulted to the UPR-School of Medicine Immunoperoxidase Laboratory, from 2008-2010, for which hormonal receptors and HER2 oncoprotein analysis were performed. To determine molecular subtypes we characterized 172 tumor specimens based on immunohistochemical (IHC) surrogates for ER, PR and HER-2+ status. The molecular subtypes were defined as: luminal (ER+ and/or PR+), non-luminal (ER-, PR-). Body Mass Index (BMI) was analyzed using the following categories: normal weight (BMI <25.0 kg/m2), overweight (BMI 25.0-29.9 kg/m2), and obese (BMI α30.0 kg/m2). Logistic regression was used to estimate the associations between BMI and breast cancer molecular subtypes. Results: The luminal subtype was the most prevalent in our sample (75.0%) compared with the non-luminal (25.0%) subtype. Association with Body Mass Index, evaluated using unadjusted logistic regression model, differed by tumor subtypes. Using non-luminal subtype as the reference group we observed that overweight women were 1.87 (95% CI: 0.73, 4.77) times more likely to be diagnosed with the luminal disease compared to women with normal weight. However, obese women were 4% (OR: 0.96, 95% CI: 0.40, 2.30) less likely to be diagnosed with the luminal subtype compared to women with normal weight. Conclusion: Even though this is a small pilot study, it is very suggestive of an important association between body mass index and luminal breast cancer subtype. Future studies that further assess the association between BMI and BCMS will add to the biologic understanding of this disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4476. doi:1538-7445.AM2012-4476
Abstract 4480: Associations between phytanic acid and alpha-methylacyl-CoA racemase (AMACR) expression in the normal human prostate
Authors: Kataria Yachana , Gann Peter , Enk Erika , Moser Ann , Deaton Ryan , Lu Yi , Nonn Larisa , Sharifi Roohollah , Deane Leslie , Ananthanarayanan Vijayalakshmi , Wright-Geise Margaret
Published: August 2012

Alpha-methylacyl-CoA racemase (AMACR) is an enzyme that is overexpressed in prostate cancer, with lower levels present in normal prostatic tissues. AMACR is involved in the metabolism of phytanic acid - a branched chain saturated fatty acid that has been linked with elevated risks of prostate cancer in several epidemiologic studies. Although considerable research has been done on AMACR as a clinical biomarker in prostate cancer, no study has evaluated the possible link between phytanic acid and AMACR gene expression in the normal human prostate, and thus the possibility that AMACR plays a causal role in prostate carcinogenesis. Fasting blood and histologcally benign prostate tissue samples were obtained from 31 men undergoing radical prostatectomy for the treatment of localized disease. Concentrations of phytanic acid in serum and tissues were determined by isotope dilution gas chromatography-mass spectrometry. RNA was extracted from epithelial cells isolated from fresh-frozen prostate tissues by laser capture microdissection, and AMACR mRNA levels were quantified using real time polymerase chain reaction. Tissue phytanic acid concentrations were not significantly correlated with serum levels nor with prostatic AMACR mRNA levels. Notably, AMACR was significantly inversely correlated with body mass index (Spearman correlation coefficient (r)= –0.43, p=0.02) and directly correlated with Gleason score (r=0.35, p=0.05). Stratification by body mass index revealed an unexpected inverse correlation between wAMACR gene expression levels and phytanic acid concentrations in the prostate (r= –0.95, p= 0.001) in obese men. In conclusion, blood and tissue levels of phytanic acid were not correlated, which suggests that blood measures should not be utilized as a surrogate measure of this fatty acid in the target organ. Contrary to expectation, tissue phytanic acid levels were significantly inversely associated with AMACR gene expression levels in the prostate, but only among obese men.These findings require confirmation in other, larger studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4480. doi:1538-7445.AM2012-4480
Abstract 4484: A novel objective, image-based phenotypic assay reveals higher α-radiation-induced γ-H2AX level in peripheral blood lymphocytes as a risk predictor for lung cancer
Authors: He Yonggang , Gong Yilei , Gu Jian , Lin Jie , Chang David W. , Lippman Scott M. , Roth Jack , Wu Xifeng
Published: August 2012

Intermediate phenotypic biomarker measures the summary results of genetic variations and may enhance predictive power in cancer risk assessment. In this study, we developed a novel objective, image-based phenotypic assay to measure α-radiation induced DNA double strand breaks (DSBs) in peripheral blood lymphocytes (PBLs) and used it as a potential biomarker for lung cancer risk prediction. γ-H2AX formation in the chromatin flanking DNA DSBs is one of the most sensitive DNA damage markers. In an ongoing lung cancer case-control study, the baseline and α-radiation-induced γ-H2AX immunofluorescence intensities in the PBLs from 403 untreated lung cancer patients and 307 controls subjects were assessed, and the ratio of the induced γ-H2AX level to the baseline γ-H2AX level was used as a variable to evaluate the sensitivity to mutagen challenge. The mean γ-H2AX ratio was significantly higher in cases than controls (case vs. control: 1.46±0.14 vs. 1.41±0.12, p < 0.001). A significantly decreasing trend in γ-H2AX ratio with age was observed for both cases (p for trend =0.003) and controls (p for trend =0.011). Dichotomized at the median level of the γ-H2AX ratio in controls, individuals with high ratio exhibited a significantly increased risk of lung cancer (OR=2.33, 95%CI: 1.67-3.23). There was also a dose-response relationship with lung cancer risk in tertile and quartile analysis (p for trend < 0.001 for both). Analysis of the joint effects of risk factors revealed an apparently elevated risk of lung cancer for subjects with a higher γ-H2AX ratio who were ever smokers, compared with subjects with a low ratio who were never smokers (OR=12.68, 95%CI: 6.84-23.51, p < 0.001). This is the first study applying relative quantification of the γ-H2AX level in PBL to assess the risk for developing lung cancer. Susceptibility to DNA DSBs might help to identify individuals at high risk for this cancer. α-radiation-induced DNA damage as determined with the new γ-H2AX assay could be a novel phenotypic risk factor of lung cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4484. doi:1538-7445.AM2012-4484
Abstract 4494: Preoperative serum levels of matrix metalloproteinase-2 (MMP-2) and survival of breast cancer among Korean women
Authors: Song Nan , Sung Hyuna , Jeon Sujee , Lee Yunhee , Choi Ji-Yeob , Park Sue K. , Lee Kyoung-Mu , Yoo Keun-Young , Noh Dong-Young , Ahn Se-Hyun , Lee Sang-Ah , Kang Daehee
Published: August 2012

Objective: We evaluated whether preoperative serum levels of matrix metalloproteinase-2 (MMP-2) work as a prognostic biomarker in breast cancer prognosis. Methods: Three hundred and three women with histologically confirmed breast cancer were recruited. The follow-up time for all patients was 4.24 years. The MMP-2 levels were quantitatively measured by enzyme-linked immunosorbent assay (ELISA) using the preoperative serum. The relationship of MMP-2 to survival was investigated using Cox's proportional hazard model adjusted for the TNM stage and estrogen receptor (ER) status. Results: In the multivariate analysis, disease-free survival (DFS) was worse among patients with the third tertile of MMP-2 compared to the first tertile of MMP-2 (hazard ratio (HR)=1.80, 95% confidence interval (CI)=1.04-3.11, P=0.04). Furthermore, when the patients were stratified by histological grade and nuclear grade, the worse DFS was predicted by high levels of MMP-2 (HR=2.90 and 95% CI=1.42-5.92 in histological grade III vs. I-II and HR=2.61 and 95% CI=1.26-5.39 in nuclear grade III vs. I-II). In ER negative patients, high levels of MMP-2 also tended to have a worse prognosis (HR=2.75 and 95% CI=1.32-5.73). Conclusions: Our results suggest that the preoperative serum levels of MMP-2 were associated with the survival of breast cancer as a potential prognostic biomarker.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4494. doi:1538-7445.AM2012-4494
Abstract 4495: Clinical implications of family history of prostate cancer in an active surveillance cohort
Authors: Goh Chee L. , Saunders Edward , Leongamornlert Daniel , Tymrakiewicz Malgorzata , Thomas Karen , Selvadurai Elizabeth , Woode-Amissah Ruth , Dadaev Tokhir , Mahmud Nadiya , Castro Elena , Olmos David , Guy Michelle , Govindasami Koveela , O'Brien Lynne , Hall Amanda , Wilkinson Rosemary , Sawyer Emma , Olama Ali Amin Al , Easton Douglas , Kote-Jarai Zsofia , Parker Christopher , Eeles Rosalind
Published: August 2012

Background Prostate cancer (PCa) is one of the most heritable cancers and a positive family history (FH) of PCa increases risk to 1st degree relatives by over 2 fold. The search for causative genetic variants has led to genome-wide association studies, which have so far identified over 45 susceptibility loci associated with PCa. However, the clinical utility of FH status or PCa risk SNP profiles to predict disease progression or aggression is yet undefined. We explore their potential prognostic role in patients undergoing Active Surveillance (AS). Methods Those enrolled in the Royal Marsden Hospital AS study were eligible. FH was obtained using questionnaires. For those with missing/incomplete data, hospital records were used. Patients with positive FH were further stratified according to the involvement of 1st and/or 2nd degree relatives. For risk score analyses, DNA was genotyped using the Sequenom MassARRAY iPLEX platform and Taqman assays. The cumulative risk scores for each patient were calculated by summing risk alleles for each loci using the weighted effect as estimated in previous studies. FH status and the SNP risk scores were analysed separately against defined adverse outcomes in AS, including adverse histology on repeat biopsy and time to treatment, to determine their prognostic value. Results 439 patients were eligible for FH analyses. 31% of those have since undergone treatment and 13% have histological upgrade on repeat biopsies. On univariate analysis, there was no significant relationship between FH of PCa in any degree of relation, and biopsy upgrade or time to treatment. For risk score analyses, 391 patients’ DNA were studied. There was also no relationship between the calculated genetic risk scores and biopsy upgrade or time to treatment (p=0.57 and p=0.83 respectively). When analysing differences between the higher and lower genetic risk groups (defined as the top 25% and lowest 25% of the genetic risk distribution), there was again no relationship (p=0.85 and p=0.91 respectively). Conclusion FH status or currently known PCa SNP profile risk scores have not been shown to be prognostic factors and should not be used to influence decisions in AS. This is the first study analysing the implications of PCa risk SNPs in AS.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4495. doi:1538-7445.AM2012-4495
Abstract 4504: Retrospective study of clinicopathologic factors associated with ALK rearrangement and survival outcome in Chinese patients with NSCLC
Authors: Zhang Xu-chao , Blanckmeister Carolyn A. , Yang Jinji , Tian Hong-Xia , Chen Jie , Xuan Jian-wei , Xie Zhi , Su Jian , Luo Xin-lan , Yan Hong-hong , Liu Yan-hui , Wu Yi-Long
Published: August 2012

Background: ALK rearrangement and EGFR mutation are significant molecular subtypes of non-small cell lung cancer (NSCLC) that are targets for different tyrosine kinase inhibitors (TKIs). We tested a large cohort of NSCLC patients for ALK, EGFR and KRAS abnormalities and correlated clinical factors with the ALK fusion gene and overall survival (OS). Methods: ALK status was detected by RACE sequencing; EGFR and KRAS status were also detected by direct DNA sequencing. OS was estimated by the Kaplan-Meier method. Results: 294 consecutive, unselected, banked NSCLC cases were tested; 27 (9.2%), 79 (29.2%) and 26 (9.6%) patients were ALK-, EGFR- and KRAS-positive, respectively. ALK rearrangement and EGFR mutation were largely exclusive (r=−0.15, P=0.007) although two cases were both ALK- and EGFR-positive. In non-parametric tests, ALK-positivity was associated with female gender (χ2=5.189; P=0.023), non/light-smoking (χ2=6.067; P=0.014) and adenocarcinoma (χ2=4.301; P=0.038). The ALK fusion partner was EML4 in all 27 cases; EML4-ALK fusion variants V1, V2 and V3 (V3a/V3b/V3c) made up the majority (24/27) of EML4-ALK fusions. There were no significant OS differences between ALK-positive, EGFR-mutant, KRAS-mutant and triple-negative groups, between ALK-positive and -negative patients, or when ALK-positive cases were matched 1:2 or 1:1 with (non-TKI-treated) controls balancing for disease stage, gender, histology and EGFR/KRAS status. In ALK-positive patients, bivariate analysis found that non- or light-smoking status (P=0.016) and prior surgery (P=0.024) were associated with increased OS and a multivariate Cox regression model found that disease stage and surgical treatment were prognostic for OS. In a multivariate logistic regression model, female gender (HR=2.793; P=0.041), wild type EGFR (HR=15.147; P=0.001), non/light-smoking (HR=3.572; P=0.020) and adenocarcinoma (HR=5.713; P=0.024) were significantly associated with ALK-rearrangement. Conclusion: Overall, the ALK fusion gene occurred in 9.2% of NSCLC patients, and was associated with non/light-smoking history, adenocarcinoma histology and female gender. ALK fusion variants were mostly V1-3, which may inform ALK screening by RT-PCR. ALK rearrangement was not a favorable prognostic factor in NSCLC, although smoking history may influence OS in ALK-positive patients. A TKI (crizotinib) now exists for ALK-positive NSCLC and may improve OS in these patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4504. doi:1538-7445.AM2012-4504
Abstract 4520: Elevated serum antibodies against insulin-like growth factor-binding protein-2 allow detecting early stage cancers: Evidences from glioma and colorectal carcinoma studies
Authors: Song Sonya W. , Li Yuchen , Zhang Jing , Zhang Bin , Yang Wei , You Gan , Xu Kaiyu , Wu Jing , Luo Chenghua , Jiang Tao
Published: August 2012

Background: Tumor-specific immunity of insulin-like growth factor binding protein-2 (IGFBP-2) has been reported in several cancers. We aimed to assess the role of serum IGFBP-2 antibodies (IGFBP-2-Abs) in early cancer detection. Patients and Methods: Glioma and colorectal carcinoma (CRC) were used as models. Serum IGFBP-2 and IGFBP-2-Abs were measured in 260 tumor patients (145 gliomas, 45 colorectal polyps, and 70 CRCs) and 141 controls. Receiver operating characteristic curves were applied. Results: Serum IGFBP-2-Ab levels were significantly elevated in tumors (mean: 82.1 ng/ml, median: 17 ng/ml, range: 0-1,387 ng/ml) compared with controls (10.6 ng/ml, 0 ng/ml, 0-212 ng/ml) (P < 0.0001), and higher in early than advanced cancers opposite of serum IGFBP-2 levels. IGFBP-2-Abs effectively discriminated between controls and grade II and III gliomas [area under the curve (AUC): 0.821-0.864; 95% confidence interval (CI) = 0.762-0.936; P < 0.0001], and CRC I-II (AUC: 0.668; 95% CI = 0.566-0.770; P = 0.002) as well as indicative of advanced polyps at high risk of CRC (AUC: 0.72; 95% CI = 0.630-0.811; P < 0.0001). The sensitivity and specificity for diagnosing grade II-III gliomas reached 66.2% - 83.8% and 81%. Combined serum IGFBP-2 and IGFBP-2-Abs augmented the discriminative power of all stage tumors (AUC: 0.823), gliomas (AUC: 0.800), and CRCs (AUC = 0.917). Conclusions: Our results first demonstrate IGFBP-2-Abs for early cancer detection, and in combination of serum IGFBP-2 for improved cancer diagnosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4520. doi:1538-7445.AM2012-4520
Abstract 4529: Prognostic implications of TSPO in human colorectal cancer
Authors: McKinley Eliot T. , Ayers Gregory D. , Smith Ralph A. , Zhao Ping , Washington Mary K. , Manning Henry C.
Published: August 2012

Translocator protein (TSPO), previously referred to as peripheral benzodiazepine receptor, is an 18kDa outer mitochondrial membrane protein involved in a number of cellular processes including steroid synthesis, cholesterol metabolism, proliferation, and apoptosis. Elevated TSPO expression has been linked with disease progression, diminished survival, and aggressiveness in a number of cancers. Studies conducted in our laboratory have demonstrated that positron emission tomography (PET) imaging with TSPO targeted probes can be used to non-invasively quantify TSPO levels in tumors. These imaging studies suggest that TSPO ligand PET may be a valuable clinical cancer imaging biomarker. In the current studies, TSPO expression in human colon cancer and paired liver metastases was evaluated using tissue microarrays. These arrays were constructed using 99 primary colon cancers and 95 matched primary colon cancers and liver metastases from Vanderbilt patients with known clinical outcomes. Among these, we found that nearly 2/3 of the colon cancers were positive for TSPO expression (64%). Similar to previous studies in other organ sites, elevated TSPO expression was significantly associated with higher tumor grade in the Vanderbilt cohort of CRC patients. Among patients with grade 2 or grade 3 tumors, an individual with elevated TSPO expression was 3 times more likely to have a grade 3 tumor than a grade 2 tumor. In comparing matched primary colon cancers and associated liver metastases, we found a strong concordance between TSPO levels and localization across matched specimens. No correlation was observed between TSPO expression and Ki67 or pEGFR levels suggesting the potentially unique information provided by the measurement of TSPO levels. In conclusion, clinically relevant prognostic information can be obtained by the measurement of TSPO expression in colorectal cancer. Thus, TSPO targeted PET may be a promising-non invasive biomarker in this setting.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4529. doi:1538-7445.AM2012-4529
Abstract 4574: FGFR1 amplification in high grade, estrogene receptor positive and metastasized breast cancers
Authors: Pfaltz Katrin , Schneider Sandra , Güth Uwe , Kilic Ergin , Eppenberger-Castori Serenella , Tapia Coya F.
Published: August 2012

Background: The FGFR1-gene (fibroblast growth factor receptor 1) is located at chromosome 8p12 encoding for a tyrosine kinase. FGFR1 is involved in cell cycle, differentiation, survival, apoptosis, and angiogenesis. Recently, it was shown that breast cancer cell lines harboring a FGFR1 amplification were more sensitive to treatment with an FGFR inhibitor. To estimate if FGFR inhibitors can be a therapeutic option in human breast cancer we investigated the prevalence and the histological subtypes of FGFR1 amplification in a large cohort of breast cancers. Methods: Tissue micro-arrays with 907 breast cancers were hybridized using the commercially available fluorescent in-situ hybridization probe (FGFR1/CEN8; ZytoVision®). The results were interpreted as following: a normal gene status was considered as a ratio (FGFR1/CEN8): 0.8-1.9, an amplification was defined as a ratio β2.0, a polysomy was defined as more than >4 FGFR1 and CEP8 signals. Results: FGFR1 amplification was observed in 8.9% (n=81), a normal gene status was found in 80.7% (n=732), and a polysomy was detected in 10.2% (n=93) tumors. FGFR1 amplified breast cancers showed the following features: expression of estrogen receptor 85% (67/78), ductal 72.8% (59/81), high grade (G3) 45.5% (35/77), her2 over-expression (score2+/3+) 16% (12/75), and 18% (13/72) recurrence. Comparing T (T1, T2, T3) and N (N0 and >N0)-categories between FGFR1 amplified and non-amplified tumors the following results were seen: T1: 25% (19/77) vs. 34% (245/719), T2: 56% (43/77) vs. 48% (345/719), T3: 9% (7/77) vs. 6% (43/719), N0: 44.5% (28/63) vs 53% (338/641), >N0: 55.5% (35/63) vs 47% (303/641). We could observe a slight better long term overall survival in patients treated with anti-hormonal therapy and FGFR1 amplified breast cancers (p=0.085). Conclusions: FGFR1 amplification is prevalent (8.9%) in breast cancer, especially among the most frequent subtypes such as estrogen receptor positive breast cancers (10%) and ductal type (8%). Further, FGFR1 amplified breast cancer showed often unfavorable/aggressive features such as a high tumor grade (45.5%), larger tumor diameter, and metastasis (55.5%). These patients are a clinically relevant group since they need aggressive adjuvant treatment regimen. The detection of FGFR1 amplification could help in the identification of some patients already at higher risk which might benefit from a new therapeutic option with FGFR1 inhibitors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4574. doi:1538-7445.AM2012-4574
Abstract 4598: Allelic loss of NDST4, a putative tumor suppressor gene, is associated with tumor progression of colorectal cancer
Authors: Tzeng Sheng-Tai , Tsai Ming-Hong , Lee Jing-Xing , Yang Ya-Chien
Published: August 2012

Colorectal cancer (CRC) is one of the most common malignancies in the world. Many molecular abnormalities have been reported in CRC, of which accumulated alterations of tumor suppressor genes (TSGs) and proto-oncogenes are required for the disease development. Genomic deletion is reflected in a genetic mechanism called loss of heterozygosity (LOH). Allelic deletion at tumor suppressor loci is common in all human cancers, and contributes to TSG silencing. By allelotyping with 22 polymorphic microsatellite markers spanning from chromosome 4pter to 4qter, we first defined the most frequent LOH region at 4q27, which occurred allelic imbalance in 32.1% (34/106) of colorectal carcinomas. After fine deletion mapping in these carcinomas, we further identified a minimal region harboring one gene with known functions, named NDST4. By quantitative RT-PCR, NDST4 expression could hardly be detected in 10 of 11 CRC cell lines, and was downregulated in 57.7% (30/52) of CRC tumors. Importantly, NDST4 expression was significantly decreased in tumors compared with matched normal mucosa or adenomas (P<0.01). In addition, immunohistochemistry staining of CRC tissue array also demonstrated decreased protein levels in Dukes’ stages C and D tumors. We further directly determined the genetic loss of NDST4 in colorectal tumors by use of 3 microsatellite markers located upstream of and inside NDST4 gene. Sixty (29.9%) of 201 carcinomas exhibited LOH at one or more microsatellite loci. Allelic loss of NDST4 was increased in patients with late stage of tumor extension (T3 and T4), lymph node involvement and distant metastasis (P=0.011, 0.053 and 0.058, respectively). Notably, LOH at MS5979 locus, a newly identified microsatellite marker within NDST4 gene, was significantly associated with disease progression of Dukes’ stages (P<0.01). Taken together, NDST4 could be a CRC-associated TSG and its genetic loss might be involved in tumor progression. In addition, the LOH assay established in the study could be a potential diagnostic and prognostic test in CRC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4598. doi:1538-7445.AM2012-4598