bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3612: Detection and quantification of tumor-derived microparticles in a murine orthotopic tumor progression model
Authors: Korman Abraham , Green Lisa , White Sandy , Burch John , Clutinger Catherine , Li Li , Neubauer Blake L. , Landschulz Katherine T.
Published: August 2012

Approximately 85% of newly identified pancreatic cancer patients are estimated to die within one year of diagnosis. This poor prognosis is attributable in part to inadequate early detection methods, with the presence of pancreatic cancer often becoming evident only upon metastasis. Development of non-invasive early detection technologies that permit earlier initiation of treatment might increase the likelihood of patient survival. Our research centered on the task of developing a method for the detection of tumor-derived microparticles (MPs) using a murine orthotopic pancreatic tumor model. MPs are 100nm to 1.5µm membrane blebs released from cells via budding upon cell activation or during apoptosis which then circulate in the bloodstream. Cell-derived MPs retain characteristics, including cell surface antigens, of their parent cells. These antigens serve as cellular “fingerprints” of MP origin. In this way, the presence of cell-specific (e.g. tumor-specific) MPs signals the presence of a tumor, and therefore, serves as a biomarker of disease. The Mia-Paca-2 human pancreatic cell line expresses the cell surface tumor marker MUC-1 and this model has been widely used as a murine xenograft model to evaluate experimental therapeutics. In a novel orthotopic model of tumor growth, 5x106 Mia-Paca-2 cells were injected directly into the pancreas of athymic nude mice, while control mice were sham-injected. Treated and control mice groups of mice were sacrificed every two weeks for 8 weeks, at which points, each pancreas was weighed, tumor weight calculated, and blood drawn. Blood-derived MUC-1 expressing MPs were enriched from a subset of Mia-Paca-2-injected mice representing treatment and compared by flow cytometry with MPs enriched from control mice. Our findings suggested a correlation between tumor size and the concentration of detected MPs in early time-points following injection of tumor cells. This relationship changed with advanced tumor progression, where a significant drop in detectable MPs was observed. These preliminary findings support the contention that measurement of circulating tumor-derived MPs may be used for the early detection of pancreatic cancer. While sensitivity remains to be confirmed and further characterization is needed, this potentially translational method is non-invasive and compatible with multiple, longitudinal measurements. Further work will be needed to elucidate the relationship between MPs generation and tumor growth, and the relationship between MPs and metastatic tumor progression.

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 3612. doi:1538-7445.AM2012-3612
Abstract 3616: Validation and fitness testing of a quantitative immunoassay for HIF1α in biopsy specimens
Authors: Park Sook Ryun , Kinders Robert J. , Khin Sonny , Hollingshead Melinda , Parchment Ralph E. , Tomaszewski Joseph E. , Doroshow James H.
Published: August 2012

HIF1α is an important marker of hypoxia in human tumors that is altered in a number of cancers, but a reproducible method to determine changes in HIF1α protein expression in human tumor biopsy specimens has not been available. While HIF1α is being used as a pharmacodynamic marker in clinical trials in the NCI (CTEP# 8610, 8856, 8880), an important limitation in measuring the change in HIF1α induced by cancer therapeutic agents is the lability of the protein, in the presence of oxygen, upon collection of the sample. We have devised a method of specimen collection, handling, and extraction that preserves and stabilizes HIF1α levels in tumor biopsies. Degassed collection buffer yielded a significant increase of recovered HIF1α levels compared to the standard method of flash-freezing. The addition of 2-hydroxyglutarate, an inhibitor of prolyl hydroxylase domain enzyme that is involved in the degradation pathway of HIF1α under normoxic conditions, to the collection buffer showed a trend for improved yield of the HIF1α recovery. Sonication and homogenization with ceramic beads were superior to homogenization with metallic beads or tissue grinding. Employing this specimen-handling method allowed validation of a two-site immunoassay for HIF1α quantitation in solid-tissue extracts generated from tumor biopsies. Intra-assay variability was less than 10%, and inter-assay variability was less than 20%. Accuracy, assessed by spike recovery, was 99 +/− 7%. HIF1α readings declined linearly with decreasing sample load over a range of 1 to 10 µg protein per well. Fitness for purpose was demonstrated by quantifying a reduction in HIF1α protein levels following topotecan or indenoisoquinoline NSC 743400 treatment in an A375 xenograft model, followed by assessment of other xenograft models. HIF1α was also demonstrated to be upregulated under low oxygen tension culture conditions in DU145 human prostate cancer cells. Funded by NCI Contract No. HHSN261200800001E.

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 3616. doi:1538-7445.AM2012-3616
Abstract 3618: Rapid and quantitative lateral flow point-of-care Therapeutic Drug Monitoring (TDM) assays for LH and FSH
Authors: Hwang Larn , Hsiao Chao , Motamed Kouros , Trieu Vuong
Published: August 2012

Introduction: Hormone manipulation therapies, such as long-acting gonadotropin-releasing hormone (GnRH) agonists, are used in the treatment of estrogen-dependent forms of breast cancer, as well as prostate cancer, because down-regulation of Luteinizing Hormone (LH) and Follicle Stimulating Hormone (FSH) leads to a decline in hormonal support to the ovary and testis. Effectiveness of hormone manipulation therapies can be monitored by quantitation of LH and/or FSH. Methods & Results: Traditionally, these hormones are measured by ELISA assays that require blood to be drawn at either a doctor's office or a diagnostic laboratory. Here, we describe the development of rapid point-of-care tests for detection of these hormones that are quantitative and applicable to plasma, serum, as well as urine. These assays exhibit dynamic ranges of 17-170 mIU/ml for LH and 0.01-10 IU/ml for FSH. They exhibit linear correlation between nominal and actual values, with correlation coefficients greater than 0.95. They comply with FDA guidance for bioanalytical assays. And they utilize cassettes that are read using an optical reader with radio-frequency identification (RFID) capability, and the data can be printed out or stored on the reader for uploading onto the hospital database. The reader utilizes confocal optics with a low distance-to-target ratio. The reflectometric measurement is converted to activity units, using an established calibration curve embedded in the RFID. The cassettes are made to be stable for up to 72 hours and can be shipped to a central lab or doctor's office for quantitation if the patient does not have access to the reader. Conclusion: Rapid and quantitative point-of-care tests (POCTs) for LH and FSH have been developed for field deployment directly at home, and the cassettes can be read directly by the patient or can be shipped to the central lab/doctor's office for reading. As TDM, these tests should allow for more effective dosing of the patients and thereby improving effectiveness of hormone manipulation therapy. The tests are also patient-centric, inviting better compliance and patient participation in personalizing his/her treatment. The simplicity of the assays would allow for their deployment in underdeveloped regions lacking access to central laboratories with specialized and expensive equipments. Clinical studies are ongoing to evaluate the usefulness of these assays.

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 3618. doi:1538-7445.AM2012-3618
Abstract 3622: Predicting response to EC145 therapy using the folate receptor-specific radiodiagnostic imaging agent, 99mTc-EC20
Authors: Leamon Christopher P. , Reddy Joseph A. , Bloomfield Alicia , Dorton Ryan , Nelson Melissa , Klein Patrick J. , Vlahov Iontcho R. , Vetzel Marilynn
Published: August 2012

EC145 is a potent folate-targeted Vinca alkaloid conjugate that is currently being evaluated in a global phase 3 trial for the treatment of platinum-resistant ovarian cancer. By design, response to EC145 should be dependent on folate receptor (FR) expression. Therefore, characterizing the presence and functionality of the FR in real time is critical for the selection of patients for whom FR-targeted therapy may be beneficial. A promising method that's currently being co-developed with EC145 is to image patients prior to therapy using 99mTc-EC20, which is a small molecular weight folate-targeted companion imaging agent. Similar to EC145, 99mTc-EC20 binds to the FR with high affinity (Kd ∼ 3 nM) in a manner that is both saturable and independent of the cell type. 99mTc-EC20's binding specificity has been confirmed using validated preclinical models where it was found that i) binding of 99mTc-EC20 to cells or accumulation within solid tumors is dependent on FR expression (but independent of isoform), and ii) excess folic acid can competitively block 99mTc-EC20's cell/tissue binding. Furthermore, EC145 was also found to displace pre-bound 99mTc-EC20 from FR-positive tumors and kidneys in mice, strongly suggesting that EC145 and 99mTc-EC20 distribute similarly in vivo. Taken together, these findings indicate that the FR mediates tissue uptake of 99mTc-EC20. Since 99mTc-EC20 provides a non-invasive, real-time assessment of functionally active as well as anatomically-accessible FRs, this radiodiagnostic agent may predict what tissues will accumulate and potentially respond to folate-targeted drugs, such as EC145.

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 3622. doi:1538-7445.AM2012-3622
Abstract 3623: Clinical significance of serum tumor markers in patients with esophageal cancer
Authors: Hagiwara Nobutoshi , Miyashita Masao , Nomura Tsutomu , Makino Hiroshi , Matsutani Takeshi , Yamashita Naoyuki , Akagi Ichiro , Shinozuka Eriko , Kato Shunji , Fujita Itsuro , Kanazawa Yoshiichi , Sakurazawa Nobuyuki , Onodera Hiroyuki , Uchida Eiji
Published: August 2012

Background: Esophageal cancer is still a disease with a high degree of fatality despite resent advances in surgical techniques and various therapies. Several serum tumor markers have been applied to diagnose and to follow-up in patients with esophageal cancer in clinical practice in Japan. The aim of this study was to investigate the diagnostic and prognostic significance of various serum tumor markers in patients with esophageal cancer. Methods: Eighty one patients with esophageal cancer were recruited for this study. Serum level of tumor markers, squamous cell carcinoma antigen(SCC-Ag), p53 antibody(p53-Ab) carcinoembryonic antigen(CEA), and CYFRA 21-1, were measured in subjects before treatment. The results of these tumor markers were compared with the clinicopathological findings of the subjects. Results: The positive rates of SCC-Ag, p53-Ab, CEA and CYFRA 21-1 were 44%, 34%, 17%, and 16% patients with esophageal cancer, respectively. SCC-Ag and p53-Ab showed positive with a relatively high frequency in cases of early stage esophageal cancer or superficial invasion, while CEA and CYFRA 21-1 could rarely be detected in such cases. Positive for SCC-Ag was significantly higher in patients with tumor invading into or through the musclaris propria than the patients within the submucosal layer. Substantially, the positive rate of SCC-Ag was associated with an advanced stage of disease. Values of p53-Ab in serum virtually demonstrated gradual decrease after treatment, not rapid depletion. All negative cases for p53-Ab before treatment showed negative at any point of treatment or follow-up on a consistent basis. CYFRA 21-1 showed positive in cases with distant metastasis. Conclusion: Each tumor marker for esophageal cancer presented different features in a clinicopathological analysis. The use of several combination of serum tumor markers seems to be most important for the detection and follow-up for of esophageal cancer at present.

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 3623. doi:1538-7445.AM2012-3623
Abstract 3626: Quantitative analysis of microRNAs by in situ hybridization reveals prognostic value of miRNAs in melanoma and breast cancer
Authors: Hanna Jason A. , Wimberly Hallie , Kumar Salil , Agarwal Seema , Rimm David L.
Published: August 2012

Introduction: Most miRNA measurement methods require total RNA extraction which destroys spatial information and makes standardization a challenge. In situ hybridization (ISH) however, allows the direct assessment of miRNA expression levels in tissue and importantly allows for the evaluation of expression in malignant cells as well as stromal cells. We have developed and validated a novel method for the quantitative analysis of miRNA expression by ISH on formalin fixed paraffin embedded tissue microarrays (TMAs) using miR-21, miR-92a, miR-34a, miR-221, miR-205, and Let-7a. Methods: In order to quantitatively measure miRNA expression by ISH, DIG labeled LNA modified miRNA probes were employed and multiplexed with DAPI and cytokeratin or S100/GP100 immunofluorescence to apply the AQUA technology. This co-localization based, quantitative approach allows for the measurement of miRNA expression in subcellular compartments. The assay allows standardization of miRNA measurement and potential for rapid large cohort assessment. We also assessed protein targets of each miRNA by AQUA to investigate the hypothesized inverse relationship between miRNAs and target proteins. Results: In hundreds of tissue samples a broad dynamic range and a variable expression pattern specific to each miRNA was observed. Specificity of the assay was validated with anti-miR and miRNA mimic transfected cell lines, miR-21 knock out mouse tissue, and blocking oligo experiments. The assays were performed on near serial sections and were found to have high reproducibility with R*2>0.8. miR-221 and miR-205 were found to be prognostic in breast cancer and melanoma respectively (p<0.05). Finally, the inverse relationship between miRNAs and putative targets were not seen in the large population cohorts. Conclusions: This specific and reproducible method for the quantitative analysis of miRNA expression provides proof of concept for the use of miRNAs as tissue biomarkers using quantitative ISH. Future studies will further determine the prognostic value of miR-221 and miR-205, and the relationship between miRNAs and target protein expression.

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 3626. doi:1538-7445.AM2012-3626
Abstract 3630: Non-enrichment based method for analysis of androgen receptor expression in circulating tumor cells (CTCs) in patients with metastatic castrate resistant prostate cancer
Authors: Gross Mitchell E. , Lazar Daniel C. , Cho Edward H. , Luttgen Madelyn , Uson Maria L. , Kuhn Peter , Torrey Melissa L.
Published: August 2012

Background: We have established a fluid phase biopsy approach that identifies CTCs which preserves cytologic features in high-definition (HD) for diagnostic pathology without using immune or surface receptor-based enrichment. HD-CTCs identified with this approach can be used for enumeration and molecular characterization. Methods: Blood was collected from metastatic prostate cancer patients and normal donors in Cyto-Chex® tubes (Streck, Omaha, NE) as part of IRB approved protocols at each site. Following erythrocyte lysis, 3 million nucleated cells were deposited on a glass slide. Samples were incubated with a pan-cytokeratin (CK), CD45, and androgen receptor (AR) antibodies and counter-stained with DAPI. LNCaP cells were spiked into normal blood. Images were obtained with a fluorescent scanning microscope and analyzed with a computer algorithm. Candidate HD-CTCs were subsequently verified by expert readers. Slides were re-imaged for quantitative analysis using at a fixed exposure and gain. Results: A total of 227 CTCs from ten patients were compared to 20 LNCaP cells. The median (range) HD-CTCs in this cohort was: 9 (1-62) cells/ml. The mean ± standard deviation measurements in HD-CTCs were observed: CK intensity 60.4±154; total cell area 89.0 ± 53.8 μm2; nuclear area 61.1 ± 36.0 μm2. LNCaP cells spiked into normal blood gave the following values: CK intensity 1166+/−306; total cell area 143 ± 48.1 μm2; nuclear area 63.1 ± 18.6 μm2. CTCs were additionally classified as either AR positive (AR+) or AR negative (AR-). 37 of the 227 (16.3%) HD-CTCs were AR+. The average CK intensity was significantly higher in AR+ versus AR- cells at 174.23 and 39.86, respectively (p<0.001). The AR expression intensity in AR+ HD-CTCs and LNCaP cells was comparable at 979.4 and 902.2, respectively (p=0.824). Conclusions: We find a positive association between AR and CK expression on a per cell basis. Further, we find AR is expressed at comparable levels in CTCs from patients and human prostate cancer cells in culture. The HD-CTC based approach may be used for enumeration and molecular interrogation of CTCs in patients with prostate 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 3630. doi:1538-7445.AM2012-3630
Abstract 3633: Prognostic value of chromogranin A and alkaline phosphatase in patients with advanced neuroendocrine tumor
Authors: Ter-Minassian Monica , Chan Jennifer A. , Hooshmand Susanne M. , Brais Lauren K. , Daskalova Anastassia , Heafield Rachel , Qian Zhiron , Lin Xihong , Christiani David C. , Kulke Matthew H.
Published: August 2012

BACKGROUND: Serum chromogranin A (CgA) or alkaline phosphatase (ALP) are often assessed in patients with advanced neuroendocrine tumor (NET) patients but their prognostic significance continues to be debated. We evaluated whether elevated CgA or ALP levels were associated with shorter survival in a large, prospectively collected cohort of NET patients. METHODS: We identified patients with metastatic NET enrolled in an institutional database between 2003-10. We used the single measurement of serum CgA or ALP levels collected with shortest time after metastatic diagnosis and classified them as binary variables (elevated/normal) based on clinical laboratory specification of elevated value. Elevated biomarker levels were correlated with overall survival using the log-rank test. Hazard ratios for each biomarker were calculated using multivariate Cox regression, adjusting for baseline characteristics of age, gender, tumor histology, and tumor subtype. Each biomarker was modeled as a time-varying covariate to account for variability in time from initial metastatic diagnosis to time of biomarker test. RESULTS: We identified 526 patients who presented at our institution with metastatic NET. Of these, 349 had available CgA values and 350 had available ALP values. There were 146 and 153 deaths among those tested for CgA and ALP, respectively. The median survival time from time of metastatic diagnosis (yrs, (95%CI)) for patients with elevated CgA was 5.98 (5.04, 7.3), as compared to 13.27 (9.51, -) for those at normal CgA limits (p<0.0001). The median survival time from time of metastatic diagnosis (yrs, (95%CI)) for patients with elevated ALP was 4.51 (3.27, 5.93), as compared to 8.87 (7.07, 9.64) for those at normal ALP levels (p=0.0001). Using the adjusted time-varying model, either elevated CgA (aHR 3.97 (2.73, 5.75) p=3.15E-13) or elevated ALP (aHR 2.4 (1.7, 3.4) p=7.96E-07) were adverse prognostic factors for patients with advanced NET. Elevated CgA was significantly associated with shorter survival in all NET subgroups (pancreatic NET, carcinoid, other NET). Elevated ALP was also a predictor of shorter survival in advanced NET patients, with the exception of those with advanced pancreatic NET. CONCLUSION: Elevated serum CgA or elevated ALP is an independent adverse prognostic factor in patients with advanced NET.

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 3633. doi:1538-7445.AM2012-3633
Abstract 3641: The predictive and prognostic roles of DNA damage response genes in epithelial ovarian cancer
Authors: Alkema Nicolette G. , Everts Marieke , van der Zee Ate G.J. , Hoor Klaske A. ten , Hollema Harry , de Jong Steven , van Vugt Marcel A.T.M. , Wisman Bea G.B.A.
Published: August 2012

Background: Despite platinum-based chemotherapy, 5-year survival of advanced stage ovarian cancer patients is only 15-30%. Exposure to platinum-based chemotherapeutics induces double strand DNA breaks (DSBs) and subsequently leads to activation of the DNA damage response (DDR). Therefore inhibition of components of the DDR may lead to better response to therapy. Aim of this study was to investigate in a large series of ovarian cancer patients the predictive and prognostic role of the activation status of four DDR-proteins- ATM, a key controller in the DDR, and three ATM substrates, including Chk2, 53BP1 and γ-H2AX, of which the latter is a direct readout for DNA damage. Methods: Expression of phospho-ATM, Chk2, phospho-Chk2, 53BP1, phospho-53BP1 and γ-H2AX was immunohistochemically assessed in 309 patients with chemo-naive ovarian cancer. Expression levels were related to clinicopathological characteristics and survival. Response to platinum-based chemotherapy was analyzed by defining two populations with either an optimal therapy response or a very poor therapy response. Both groups consisted of patients with advanced stage ovarian cancer, >2 cm residual disease after primary surgery and were all treated with platinum-based chemotherapy. The responder group had a well-defined PFS of more than 18 months while the non-responder group had a PFS of less than 6 months. Results: In the two well-defined groups with the largest contrast regarding response to treatment, positive γ-H2AX expression (OR=0.193; p=0.036) and positive Chk2 expression (OR=0.149; p=0.011) were related to a good response to platinum-based chemotherapy. In advanced stage patients, phospho-53BP1 expression was independently related to a worse disease-specific survival (HR=1.905; p= 0.044). Conclusion: High DDR protein expression in advanced stage ovarian cancers suggests a high degree of genomic instability, while the negative prognostic impact of phospho-53BP1 may be due to enhanced repair capacity of chemotherapy-induced DSBs on the long term.

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 3641. doi:1538-7445.AM2012-3641
Abstract 3644: The PSP94αprostasin pathway and early detection of ovarian cancer
Authors: Guo Yi , Yan Bingxue , Mueller Michael D. , Yu Jing Jie
Published: August 2012

Objective: Our long-term goal is to better understand the mechanism of tumor development in human epithelial ovarian cancer. The short-term goal is to identify a biomarker for early detection of this malignancy. Methods: qPCR was performed for PSP94 expression using SYBR® Green reagents kit in 3 pairs of tumor and normal samples of ovarian cancer patients and 4 ovarian cancer cell lines on ABI 7500 Fast Real-Time PCR System. Overexpression of PSP94 or prostasin by cDNA transfection was performed in ovarian cancer cell lines. The expression levels of PSP94 and prostasin after cDNA transfection were determined by western blot analysis. Results: In gene expression analysis by qPCR, we found that PSP94 expresses in tumor tissues of ovarian cancer patients and cancer cell lines but not in normal ovarian specimens. Our investigation of the relationship between PSP94 and prostasin by cDNA transfection assay demonstrated that expression of prostasin was increased in the PSP94 overexpressed cells. In contrast, PSP94 expression did not change in the prostasin-forced-expression cells. This suggests that the two genes most likely share part of a regulation network and that PSP94 appears to be an upstream regulator of prostasin. Conclusions: We conclude that PSP94 expression occurs only in tumor samples and PSP94 and prostasin share a signaling pathway. We hypothesize that alteration of PSP94-prostasin pathway may be an early event in the development of epithelial ovarian cancer; PSP94 appears to be an upstream regulator that modulates prostasin expression. Thus, PSP94 seems to play an important role in tumorigenesis and may hold great promise as a biomarker for early detection of human epithelial ovarian 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 3644. doi:1538-7445.AM2012-3644
Abstract 3647: Integrin β5 as a new target in ovarian cancer treatment
Authors: Ohyagi Chifumi , Sawada Kenjiro , Kinose Yasuto , Tomita Yasuhiko , Morishige Ken-ichirou , Kamiura Shoji , Kimura Tadashi
Published: August 2012

[Background] Ovarian cancer is a highly metastatic characterized by widespread peritoneal dissemination andascites, and is the leading cause of death from gynecological malignancies. Previously, we identified integrin β5 as a key molecule for peritoneal dissemination. In this study, we aimed to determine the predictivevalue of integrin β5 in Japanese ovarian cancer patients.[Methods] Ovarian carcinoma samples were collected from 148 patients who were treated at Osaka Medical Center for Cancer and Cardiovascular Disease (Osaka, Japan) between 2000 and 2005 and were subject to the tissue microarray (TMA) slides. Satisfactory spots and clinical data were obtained from 114patients. The slides were stained with an antibody against human integrin β5 and each sample was scored based on the percentage of positive cells (0, α10%;1, 10-25%; 2, 25-50%; 3, Δ50%) and the intensity of the staining (0, none; 1, very weak; 2, weak; 3, strong). Overexpression was defined if the total scoreof density and intensity is Δ5. “Low” expression was defined if the total score of density and intensity scores was α4. Survivalestimates were computed using the Kaplan-Meier method, and comparisons between groups were analyzed using the log-rank test.[Results] Of 114 patients, 33(29.8%) had integrin β5 overexpression, including 6 of 30 serous (20.0%), 1 of 12 mucinous (8.3%), 1of 13 endometrioid (7.7%), 13 of 17 clear cell (75.6%), 7 of 26 low potential malignant (26.9%) and 5 of 18 others (27.8%) ovarian carcinomas. Among patients with advanced stages (stage lll/lV), those who had integrin β5 overexpression showed significantly worse overall survival than those who had low expression (11.2 months vs 40.0months, P=0.05).[Conclusion] Integrin β5 overexpression was seen in most of clear cell carcinoma, which is extremely resistantto the current chemotherapies. Besides, in advanced ovarian cancer patients, integrin β5 was an independent prognostic maker. These datasuggested that targeting this integrin could be a promising treatment option, at least, for a subset of ovarian cancer such as clear cell carcinoma and needs to be explored for future clinical application.

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 3647. doi:1538-7445.AM2012-3647
Abstract 3653: Cancerous inhibitor of PP2A (CIP2A) expression for bladder cancer diagnostics
Authors: Huang Lisa P. , Adelson Martin , Mordechai Eli , Trama Jason
Published: August 2012

Bladder cancer is one of the most common cancers in the United States. In 2011, the National Cancer Institute estimates approximately 70,000 cases of bladder cancer will be diagnosed; of those, more than 15,000 are expected to die. According to the American Cancer Society, the five-year survival rate for patients diagnosed with bladder cancer is 98% (at stage 0), 88% (at stage I), 63% (at stage II), 46% (at stage III), and 15% (at stage IV). These statistics highlight the fact that early detection of bladder cancer is critical for improving prognosis and long-term survival. Numerous markers have been tested for bladder cancer detection and surveillance. These markers include complement factor H (BTA-Stat/TRAK), nuclear matrix proteins (NMP22), mucin-like antigens, hyaluronic acid, hyaluronidase, survivin, soluble Fas, telomerase and detection of chromosomal aneuploidy and deletion using fluorescence in situ hybridization (UroVysion®). However, none of them are accepted as a routine tool for bladder cancer diagnostics and surveillance. There exists a continuing need for an assay that detects the presence of bladder cancer in humans. It would be advantageous to develop an assay with a protein that is associated with the development of bladder cancer. We sought to identify cancerous inhibitor of PP2A (CIP2A) as a novel bladder cancer biomarker. In this study, Western blot and ELISA were used to access the level of CIP2A protein and other known bladder cancer protein biomarkers (i.e., survivin) in bladder cancer cell lines, bladder cancer patient tissues, and bladder cancer patient urines. Our studies indicate CIP2A protein was abundantly expressed in bladder cancer cell lines and it was not expressed in non-tumor epithelial cell lines. CIP2A protein was specifically expressed in bladder tumor tissue at different cancer stages, and not in adjacent non-tumor bladder tissue. Our data showed that CIP2A protein detection in bladder cancer tissues had a sensitivity of 75% and specificity of 100%. Our study also suggests there was no detectable level of CIP2A protein in bladder cancer patient urine, in contrast to that of other bladder cancer biomarkers such as survivin. In conclusion, our studies suggest that CIP2A protein is specifically expressed in human bladder cancer tissue and not in adjacent tissue. It will be important to extend this finding and to develop non-invasive assays for early detection or for surveillance of bladder cancer in humans.

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 3653. doi:1538-7445.AM2012-3653
Abstract 366: Enhanced tumor detection of fluoroscence laparoscopy improves laparoscopic resection of primary pancreatic cancer in mouse models
Authors: Metildi Cristina A. , Kaushal Sharmeela , Snyder Cynthia S. , Luiken George A. , Hoffman Robert M. , Bouvet Michael
Published: August 2012

The aim of this study was to evaluate the utility of fluorescence laparoscopy for resection of primary pancreatic cancer. Orthotopic mouse models of human pancreatic cancer were established with the brightly-fluorescent BxPC-3 RFP human pancreatic cancer cell line. Two weeks after implantation, mice were randomized to undergo either bright light laparoscopic resection (BLR) or fluorescence-guided laparoscopic resection (FLR). Fluorescence laparoscopy (FL) was performed with a Stryker L9000 LED light source 24 hours after tail vein injection of CEA antibodies conjugated to Alexa 488. Bright light laparoscopy (BL) was performed with a Stryker X8000 xenon light source. Pre- and postoperative images were taken with the Olympus OV-100 Small Animal Imaging System to assess completeness of resection. Postoperatively, whole body images were obtained to assess for recurrence and follow tumor progression. All resected tumors were collected for histologic review. Fluorescence laparoscopy with a 495-nm emission filter and an LED light source enabled real-time identification and localization of the brightly-fluorescent BxPC-3 RFP tumor additionally labeled with anti-CEA-Alexa 488. The combination of red and green fluorophores optimized the fluorescence signal of the tumor thereby enabling accurate distinction of tumor margins from normal surrounding tissue without compromising background illumination. Furthermore, adequate visualization of surrounding structures enabled surgical navigation for resection of primary pancreatic tumor. Tumor was more readily detected and resected under fluorescence guidance than under standard bright light. In addition, sub-millimeter lesions within the pancreas that were undetected by bright light laparoscopy were easily identified and resected by fluorescence laparoscopy. This study demonstrates the capabilities of laparoscopic fluorescence-guided surgery of primary pancreatic cancer. The optimal combination of fluorophores with the use of an LED light source enhanced the fluorescence tumor-to-background ratio while permitting enough light leakage for background illumination for surgical navigation.

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 366. doi:1538-7445.AM2012-366
Abstract 3662: FGFR2 gene amplification and overexpression frequently occur in squamous cell carcinomas of the lung
Authors: Kikuchi Ryutaro , Huang Cheng-long , Sonobe Makoto , Kobayashi Masashi , Imamura Naoto , Horiuchi Marika , Kitamura Jiro , Ishikawa Masashi , Date Hiroshi
Published: August 2012

Introduction: To date, only a few genetic abnormalities have been reported to be involved in tumorigenesis of squamous cell carcinomas (SqCCs) of the lung. Recently, fibroblast growth factor receptor (FGFR) 1 gene amplification was identified as a potential target for treatment in approximately 20% of SqCCs. In order to identify the clinical significance of FGFR family (FGFR1, FGFR2, FGFR3, and FGFR4) in SqCC, we performed a clinical study on the FGFR family in patients with non-small cell lung cancers (NSCLCs). Methods: Consecutive 138 NSCLC patients were studied. Gene expressions of FGFR family were evaluated by quantitative real-time RT-PCR. Intratumoral protein expressions of FGFR1, FGFR2, and Ki-67 were evaluated by immunohistochemistry. Gene amplifications of FGFR1 and FGFR2 were investigated by FISH. In addition, mutations of EGFR (exon 19-22) and KRAS (exon 1) were also investigated using either PCR-SSCP or PCR-RFLP, confirmed by direct sequencing. Results: Among 138 NSCLCs, 91 tumors were adenocarcinoma, 37 tumors were SqCCs, and 10 tumors were other types. Regarding FGFR gene expressions in NSCLCs, The FGFR2 gene expression was significantly higher in NSCLCs with both wild-type EGFR and wild-type KRAS than in NSCLCs with either mutant EGFR or mutant KRAS (P = 0.0012). FGFR2 gene expression was significantly higher in SqCCs than in adenocarcinomas (P < 0.0001). Furthermore, the FGFR2 gene expression was the highest among the gene expressions of FGFR family in SqCC. In contrast, no significant difference was observed in FGFR1 gene expression in relation to tumor histology. Regarding FGFR protein expressions in NSCLCs, the intratumoral FGFR2 protein expression significantly correlated with the FGFR2 gene expression (P < 0.0001, r = 0.5492), and the intratumoral FGFR2 protein expression was also significantly higher in SqCCs than in adenocarcinomas (P = 0.0008). Furthermore, the FGFR2 protein expression significantly correlated with Ki-67 proliferation index (P < 0.0001, r = 0.4677). In FISH analysis, FGFR1 gene amplification was detected in 34.3% (12/35) of SqCCs, and FGFR2 gene amplification was detected in 22.9% (8/35) of SqCCs. Furthermore, the Ki-67 proliferation index was significantly higher in SqCCs with FGFR2 gene amplification than in SqCCs without FGFR2 gene amplification (P = 0.0069). Conclusions: FGFR2 gene amplification and overexpression frequently occur and correlate with the tumor proliferation in SqCCs of the lung. FGFR2 could be a potential molecular target for the treatment of SqCCs of the lung as well as FGFR1.

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 3662. doi:1538-7445.AM2012-3662
Abstract 3667: Inactivating BRAF mutations confer dasatinib sensitivity in lung cancer
Authors: Sen Banibrata , Peng Shaohua , Tang Ximing , Erickson Heidi , Galindo Hector , Mazumdar Tuhina , Stewart David J. , Wistuba Ignacio , Johnson Faye M.
Published: August 2012

NSCLC is a lethal disease for which personalized therapies that target specific genetic aberrations have been markedly effective in subsets of patients. An important approach for discovering effective cancer therapeutic targets is to characterize responsive tumors. We conducted a phase II trial of the tyrosine kinase inhibitor dasatinib in stage IV NSCLC. The dramatic response of one patient, who remains cancer free four years later, led us to examine the molecular characteristics of his tumor. We performed a comprehensive analysis of this NSCLC patient's tumor including mutational analysis of 40 genes, array comparative genomic hybridization, and immunohistochemistry. We discovered a novel, inactivating BRAF mutation (Y472CBRAF) in the patient's tumor; no inactivating BRAF mutations were found in the non-responding patients. Cells transfected with Y472CBRAF exhibited CRAF, MEK, and ERK activation, which were identical to signaling changes that occur with previously known inactivating BRAF mutants. Dasatinib induced senescence in NSCLC cells with endogenous inactivating BRAF mutations. Transfection of cells with inactivating BRAF mutations led to increased dasatinib sensitivity; conversely, cells transfected with an activating BRAF mutation were more resistant. Likewise, BRAF inhibition in NSCLC cells expressing wild-type BRAF enhanced dasatinib sensitivity. Dasatinib sensitivity may depend upon CRAF since dasatinib led to decreased CRAF activity and only NSCLC cells with inactivating BRAF mutations were sensitive to CRAF inhibition. We hypothesize that patient's BRAF mutation was likely responsible for his marked response to dasatinib and suggests that tumors bearing inactive BRAF mutations will be exquisitely sensitive to dasatinib.

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 3667. doi:1538-7445.AM2012-3667
Abstract 3669: Autocrine interleukin/STAT3 signaling in breast implant-associated T cell anaplastic large cell lymphoma cell lines suggests a mechanism for tumor progression and effective therapy
Authors: Church Connor H. , Lechner Melissa G. , Megiel Carolina , Bass Rikki S. , Russell Sarah M. , Epstein Alan L.
Published: August 2012

Recently, over 80 cases of anaplastic large-cell kinase (ALK)-negative, T-cell, anaplastic, non-Hodgkin lymphomas (T-ALCL) have been identified worldwide in patients with textured saline and silicone breast implants. These breast implant-associated T-ALCL cases are striking for their homogeneous clinical presentation and pathology that is distinct from established classes of ALCL. In order to understand the nature of this newly emerged clinical entity, we have established three new cell lines, designated T cell Breast Lymphoma (TLBR) -1, -2, and -3, from patient primary tumor biopsy specimens. Characterization of these pre-clinical models confirmed fidelity to the original tumor biopsy specimens and highlighted unique features that may aid the development of effective treatments for these cancers. Gene expression analysis demonstrated significant up-regulation of survivin and down-regulation of pro-apoptotic genes (BID, BAK, BBC3) by all TLBR cell lines relative to healthy donor T cells. This preliminary finding prompted examination of upstream regulators of cell survival in T cells, namely the interleukin signaling associated with the JAK/STAT pathway. TLBR cell lines were found to secrete high levels of IL-6 and IL-10, and were strongly positive for the cognate receptors. The TLBR cell lines also showed elevated levels of STAT3 and pSTAT3 by immunoblotting techniques, along with a simultaneous reduction in expression of the pSTAT3 phosphatase, SHP-1. These results implicate an unregulated autocrine STAT3 pathway present in the TLBR cell lines that strongly contributes to their tumorigenesis. Assays of various inhibitors of this pathway support this hypothesis since targeting of STAT3 with the tyrosine kinase inhibitor, Sunitinib, led to dramatic in vitro cell death in all TLBR cell lines. More specific inhibition of STAT3 with S3I-201 had comparable results with Sunitinib. Furthermore, IL-6 neutralization by a monoclonal antibody or SHP-1 restoration by 5-azacytidine treatment also demonstrated moderate efficacy in inhibiting TLBR cell line growth in vitro. These discoveries strongly suggest the use of these reagents in the clinical treatment of Breast Implant Associated T-ALCL. The TLBR cell lines closely resemble the primary breast implant-associated lymphomas from which they were derived, and as such provide valuable preclinical models to study the unique biology and provide clinicians with improved knowledge for treatment and 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 3669. doi:1538-7445.AM2012-3669
Abstract 3671: A distinct subset of atypical Spitz tumors is characterized by BRAF mutation and loss of BAP1 expression
Authors: Wiesner Thomas , Murali Rajmohan , Fried Isabella , Cerroni Lorenzo , Kutzner Heinz , Bastian Boris C.
Published: August 2012

We recently reported that germline mutations in BAP1 cause a familial tumor syndrome characterized by high penetrance for melanocytic tumors with distinctive clinical and histologic features. Melanocytic neoplasms in affected individuals harbored BRAF mutations, showed loss of BAP1 expression, and histologically resembled so-called “atypical Spitz tumors” (ASTs). ASTs are an ill-defined and probably heterogenous group of melanocytic tumors that display histologic features seen in both Spitz nevi and melanomas. Their biologic behavior cannot be reliably predicted. Based on the histologic similarities of the familial tumors and ASTs, we hypothesized that a subset of ASTs might harbor genetic alterations seen in the familial tumors. To address this hypothesis, we analyzed 32 sporadic ASTs for BRAF mutations, and for BAP1 expression. To determine the utility of immunohistochemistry to assess BAP1 status, we analyzed samples from 42 patients with the above syndrome as well as 46 melanocytic nevi and found that loss of BAP1 expression was restricted to patients with the syndrome. Nine (28%) sporadic ASTs showed loss of BAP1 expression, of which of 8 (89%) had concomitant BRAF mutations. Only one of the BAP1-positive ASTs (4%) had a BRAF mutation (p<0.0001). BRAF mutated, BAP1-negative tumors were primarily located in the dermis, and were composed entirely or predominantly of epithelioid melanocytes with abundant amphophilic cytoplasm and well-defined cytoplasmic borders. Nuclei were commonly vesicular, and exhibited substantial pleomorphism and conspicuous nucleoli. The combination of BRAF mutation and loss of nuclear BAP1 expression thus characterizes a subset of ASTs with distinctive histologic features. The typical morphology of these tumors and BAP1 immunohistochemistry provide pathologic clues that will enable accurate identification of this subset. Future studies are necessary to determine whether this subset has a predictable clinical behavior.

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 3671. doi:1538-7445.AM2012-3671
Abstract 3674: Next generation cancer gene sequencing for clinically actionable mutations
Authors: Wen Yu-Ye , Douglas Ganka , Hu Xiafeng , Millham Michele , Cifuentes Francisco , Peters Garrick , Carmack Condie Edwin , Li Marilyn
Published: August 2012

Cancer gene mutations are important diagnostic, prognostic, and predictive biologic markers. Mutation detection in cancer tissues has been a real challenge for researchers as a cancer is often associated with multiple mutations and new mutations may occur as the disease progresses and during treatments. Essentially, all cancer tissues are a mosaic mixture of normal and abnormal cell clones. Therefore, a new technology that can target many mutations simultaneously with high sensitivity is exceedingly desirable. Targeted next generation sequencing permits deep sequencing of hundreds of mutations concurrently. We have used a novel next generation sequencing technology that merges multiplex PCR with ion semiconductor sequencing (AmpliSeq, Life Technology) to sequence a panel of clinically actionable cancer gene mutations on 12 tumor samples including 6 archived FFPE samples, 2 bone marrow samples, and 4 cell lines with known mutations, and a normal blood sample as a control. Each sample was run three times to assay for inter-run variability. The panel contains 739 mutations including clinically actionable mutations such as EGFR mutations in exons 18-21, BRAF V600E, KRAS mutations at codons 12, 13, and 61, and many more. Using this technology we successfully identified all known mutations previously detected using Pyposequencing or Sanger sequencing technologies. The most common somatic mutations were also confirmed by Competitive Allele-Specific TaqMan® PCR (castPCR, Life Technology). To determine the sensitivity of the test, we studied mutation positive FFPE samples with serial dilutions. Our results showed that the test can detect mutations at frequencies as low as 5% with 99% confidence. Analysis of mutation calls in different runs showed little inter-run variation. Other benefits of this novel technology are its fast turn around time, which can be as short as two days, and its low cost. Our experience demonstrates that this technology holds great potential clinical utility including diagnostic and therapeutic applications.

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 3674. doi:1538-7445.AM2012-3674