bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3437: P276, a cyclin-dependent kinase inhibitor with antitumor activity against high-risk human cervical cancer.
Authors: Shangary Sanjeev Kumar , Deshpande Gandhali , Sonawane Vinay , Venkatasubbaiah Venkatesha , Jalota-Badhwar Archana , Joshi Asavari , Mayekar Manoj , Shirsath Nitesh , Pandey Prashant , Agarwal Veena R.
Published: August 2014

Cervical cancer is the third most common cancer worldwide in women. While preventive vaccines have been approved for cervical cancer, treatment options are limited to chemotherapy drugs. An unmet need, thus, exists for the treatment of cervical cancer. High-risk human papilloma viruses (HPVs), type 16 and 18, account for 70% of the cervical cancer. Viral oncoproteins E6 and E7 bind and inhibit the p53 and Rb tumor suppressors, respectively. One attractive strategy for the treatment of cervical cancer is to attenuate the expression of E6 and E7 viral oncoproteins by a pharmacological inhibitor of cyclin-dependent kinases (CDKs). P276 is a CDKs inhibitor with potent activity against CDK4, CDK1 and CDK9 (Joshi et al, Mol. Cancer Ther. 2007; 6:918-25). The therapeutic potential and mechanism of action of P276 in cervical cancer was investigated using high-risk HPV type 16 (SiHa and Caski) and type 18 (HeLa) human cervical cancer cell line. P276 inhibited cell growth with sub-micromolar IC50 values (0.5-0.8 μM), induced cell cycle arrest and apoptosis in cervical cancer lines. E6 negatively regulates p53 protein stability in HPV-infected cervical cancer. P276 downregulated mRNA and protein expression of E6 in SiHa cells with concomitant upregulation of p53 protein and its downstream target p21, indicating activation of the p53 pathway. It also downregulated E7 mRNA expression in SiHa and HeLa cell lines. Further, it is known that hyperphosphorylation of Rb relieves its interaction with E2F, leading to Rb inactivation and E2F activation. Treatment with P276 dephosphorylated Rb at Ser780, indicating Rb activation. P276 demonstrated significant (p<0.01) in vivo efficacy in HPV type 16 SiHa human cervical cancer xenograft model when administered by i.p. route, Treatment with 25 mg/kg of P276 for 21 days induced 59% of tumor growth inhibition without body weight loss in this model. Conclusion: P276, a CDKs inhibitor, is efficacious against high-risk cervical cancer.

Citation Format: Sanjeev Kumar Shangary, Gandhali Deshpande, Vinay Sonawane, Venkatesha Venkatasubbaiah, Archana Jalota-Badhwar, Asavari Joshi, Manoj Mayekar, Nitesh Shirsath, Prashant Pandey, Veena R. Agarwal. P276, a cyclin-dependent kinase inhibitor with antitumor activity against high-risk human cervical cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3437. doi:10.1158/1538-7445.AM2013-3437
Abstract 3439: Suppression of pancreatic cancer cell growth by NISC-6 through activation of Par-4 and death receptor 5.
Authors: Karelia Deepkamal , Pandey Manoj K. , Irby Rosalyn , Amin Shantu , Sharma Arun K.
Published: August 2014

Pancreatic cancer is one of the deadliest cancers, with a median survival of 6 months and a 5 year survival rate of 3-5%. The poor prognosis of pancreatic cancer is attributable to its tendency for late presentation, aggressive local invasion, early metastases, and poor response to chemotherapy. Currently used drugs result in marginal survival advantage and are associated with multiple adverse events and drug resistance. Thus, there is a need for novel therapeutic agents and strategies involving less toxic agents that can sensitize pancreatic cancer cells to chemotherapy. Through extensive structure-activity relationship studies, we have recently developed an Akt pathway inhibitor, NISC-6, designed by incorporating an isoselenocyanate (-N=C=Se) functionality into naphthalimide structure, rendering it lethal to cancer cells at a dose that is non-toxic to normal cells. Our hypothesis is that the Akt inhibitor, NISC-6, in conjunction with Prostate apoptosis response protein 4 (Par-4), will induce apoptosis in pancreatic tumor cells through the TRAIL pathway. This is based on the observation that Par-4 causes apoptosis in cancer, but not in normal, cells, often in response to an apoptotic agent and is often inactivated in cancer cells through interaction with Akt1. In pancreatic cancer, Par-4 expression is often reduced through deletion of a chromosome segment containing 12q21, the location of the PAR-4 gene. Activation of Par-4 through inhibition of Akt by NISC-6 thus holds promise as a possible strategy for pancreatic cancer treatment. Our studies have shown NISC-6 to be effective in reducing cell proliferation in three different pancreatic cancer cell lines e.g. MiaPaCa-2, Panc-1, and BxPC-3. The treatments of NISC-6 for 48 h reduced the cell viabilities of pancreatic cancer cells with an IC50 of between 1-2.5 μM. In addition, treatment of MiaPaCa-2 cells with NISC-6 for 24 h showed significant activation of Par-4 and death receptor 5 (DR5). This activation was associated with the down-regulation of anti-apoptotic protein Bcl-xL and Survivin. Furthermore, NISC-6 potentiated the apoptotic effects of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/APO-2L). Taken together, these results suggest that NISC-6 may represent a novel therapeutic agent to sensitize pancreatic cancer cells to TRAIL and chemotherapeutic drugs. Detailed results of these investigations will be presented.

Citation Format: Deepkamal Karelia, Manoj K. Pandey, Rosalyn Irby, Shantu Amin, Arun K. Sharma. Suppression of pancreatic cancer cell growth by NISC-6 through activation of Par-4 and death receptor 5. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3439. doi:10.1158/1538-7445.AM2013-3439
Abstract 3455: Elafin is a candidate biomarker that underlies the similarities between basal-like breast cancer and ovarian cancer.
Authors: Labidi-Galy Intidhar , Clauss Adam , Ng Vivian , Duraisamy Sekhar , Ganesan Shridar , Drapkin Ronny
Published: August 2014

We recently reported that Elafin, a serine proteinase inhibitor encoded on chromosome 20q12-13.2, is overexpressed and secreted by High grade serous carcinoma of the ovary (HGS-OvCa) and is associated with poor overall survival and chemotherapy resistance. Here we show that Elafin confers a proliferative advantage to HGS-OvCa cells through activation of the MAP kinase pathway. Mutations in the anti-proteinase domain of Elafin did not interfere with its ability to stimulate cell growth. This mitogenic effect can be neutralized by RNA interference, specific antibodies targeting Elafin and small molecule inhibitor against MEK kinase. We also observed that IC50 to cisplatin and paclitaxel are significantly higher in OvCa cell lines (n=11) highly expressing Elafin. A phosphoproteome array among 20 cell lines derived from patient's ascites samples showed that expression of Elafin significantly correlates with increased expression of RAD50 and STAT5A, two proteins involved in platinum resistance.

HGS-OvCa shared genomic features with basal-like breast cancer (BLBC) including frequent TP53 mutations, genomic instability, and poor prognosis. Thus, we looked at the expression of Elafin in BC. A meta-analysis of gene-expression profiling from 7 publicly available BC datasets including more than 1,300 tumors showed that Elafin is significantly highly expressed in BLBC compared to luminal and HER-2 subtypes and this observation has been confirmed by IHC on BC tumors (n=25) and on BC cell lines. Additionally, we observed that RNA interference against Elafin reduces proliferation of BLBC cell lines.

Citation Format: Intidhar Labidi-Galy, Adam Clauss, Vivian Ng, Sekhar Duraisamy, Shridar Ganesan, Ronny Drapkin. Elafin is a candidate biomarker that underlies the similarities between basal-like breast cancer and ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3455. doi:10.1158/1538-7445.AM2013-3455
Abstract 3460: Tumor budding is a predictive feature for lymph node metastasis in invasive ductal breast cancer.
Authors: Trippel Mafalda , Pfaltz Katrin , Salhia Bodour , Grogg André , Zlobec Inti , Tapia Coya F.
Published: August 2014

Background: Tumor budding is a phenomenon associated with epithelial mesenchymal transition and correlates with tumor invasiveness and metastasis in colon cancer. In breast cancer this histomorphological feature has not yet been determined. In this study we investigated the prognostic value of peritumoral budding (PTB) and intratumoral budding (ITB) in invasive ductal breast cancer and its correlation with lymph node status. Method: Two pathologists counted tumor buds within the tumor (ITB) and at the periphery (PTB) of 90 primary invasive ductal breast cancers. ITB and PTB were scored on whole tissue sections stained with a PanCK antibody. Budding was defined as isolated single cancer cells or microscopic cell clusters composed of ≥1 or up to 5 cells. A tumor was designated to have high-grade budding if it had an average of >10 buds. PTB and ITB were then correlated with pathological features of breast cancer. Results: Forty-eight percent (n=43) and 50% (n=45) were considered high-grade for either ITB or PTB, respectively. High-grade PTB was significantly (p=0.0044) associated with lymph node metastasis and ITB was significantly (p=0.0018) associated with hormone receptor positive tumors. ITB and PTB were frequent in Her2 negative (50% and 53%) tumors and breast cancer with higher T-category >T3 (57%). Neither PTB nor ITB showed an association with tumor grade or proliferation index as measured by Mib-1 staining. Conclusion: For the first time, we report that PTB is a strong (p=0.0044) predictive tumor feature of lymph node metastasis in invasive ductal breast cancer. The association of PTB with metastatic disease may be due to an increased occurrence of lymph vessels found at the tumor periphery. Additional studies are ongoing to determine if tumor budding can select for more aggressive subtypes of invasive breast cancer.

Citation Format: Mafalda Trippel, Katrin Pfaltz, Bodour Salhia, André Grogg, Inti Zlobec, Coya F. Tapia. Tumor budding is a predictive feature for lymph node metastasis in invasive ductal breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3460. doi:10.1158/1538-7445.AM2013-3460
Abstract 3466: Reduced expression of CYLD is an independent prognostic factor in breast cancer and confers treatment resistance and migratory activity in vitro.
Authors: Hayashi Mitsuhiro , Shinriki Satoru , Jono Hirofumi , Nakamura Takuya , Guo Jianying , Yamamoto Yutaka , Iwase Hirotaka , Ando Yukio
Published: August 2014

Background: CYLD was originally identified as a tumor suppressor that is mutated in familial cylindromatosis. CYLD has deubiquitinase activity and can negatively regulate NF-κB pathways. Recent study has revealed that CYLD plays important roles in tumor progression as well as immune response and osteoclastogenesis. CYLD loss of function has been associated with several tumors. Despite the broad relevance of CYLD to various tumors, its clinical and biological significances in breast cancer are still largely unknown.

Materials and Methods: We assessed CYLD expression in 171 primary breast cancer tissues by immunohistochemistry and examined the correlations with CYLD expression and clinicopathological factors as well as patients’ prognosis. In addition, we investigated effects of CYLD knockdown by using siRNA on malignant state in breast cancer cell lines.

Results: Our immunohistochemical analysis revealed a significant correlation between loss of CYLD expression and poor patients’ disease free survival (log-rank, p = 0.004), particularly in luminal and triple negative breast cancer patients. Multivariate analysis revealed that loss of CYLD expression was an independent poor prognosis factor (Hazard ratio 2.95, 95% CI 1.20 - 7.98, p = 0.02) in addition to hormone receptor expression status. Loss of CYLD expression correlated with high nuclear grade but not hormone and HER2 status.

Consistent with our clinical data, repression of CYLD increased CD44 high/ CD24 low cancer stem-like cell population with acquisition of resistance to apoptosis induced by conventional therapy in MDA-MB-231 cells. Moreover, CYLD knockdown promoted migration through downregulation of Maspin which is a potent suppressor of breast cancer metastasis.

Conclusions: CYLD expression was an independent prognostic factor in primary breast cancer. Reduced CYLD function may promote breast cancer progression and treatment resistance.

Citation Format: Mitsuhiro Hayashi, Satoru Shinriki, Hirofumi Jono, Takuya Nakamura, Jianying Guo, Yutaka Yamamoto, Hirotaka Iwase, Yukio Ando. Reduced expression of CYLD is an independent prognostic factor in breast cancer and confers treatment resistance and migratory activity in vitro. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3466. doi:10.1158/1538-7445.AM2013-3466
Abstract 3469: The expression and prognostic significance of neutrophil gelatinase-asssociated lipocalin (NGAL) in primary breast cancer.
Authors: Kim Lee-Su , Kim Hae Sung
Published: August 2014

Introduction:

Neutrophil gelatinase-associated lipocalin (NGAL; also known as lipocalin 2), a member of the lipocalin family that transports small lipophilic ligands, has gained recent attention as both a potential biomarker and a modulator of human cancers including breast cancer. In this study, we examined the expression of

NAGL in breast carcinoma and evaluated the prognostic value of its expression.

Methods:

Tissue microarray (TMA) blocks were made with formalin-fixed paraffin-embedded tissues from 207 breast cancer patients and the immunohistochemical staining was done using an anti-NGAL antibody. The correlation between NGAL expression and the clinicopathologic parameters such as age, tumor size, lymph node status, Ki-67 labeling index, histologic grade, hormone receptors and HER2/neu expression as well as clinical outcome was evaluated.

Results:

NGAL was expressed in 60 breast carcinomas (21.9%). A higher NGAL expression was noted in the younger (p=0.018) and premenopausal women (p=0.036). NGAL expression correlated with a high Ki-67 labeling index (p=0.001) and poor histologic grade (p=0.002). No significant association between NGAL expression and hormone receptor status and HER2/neu overexpression. With a median follow-up of

63 months, NGAL expression was associated with decreased disease-free survival, especially for patients with estrogen receptor negative group (p=0.013) and lymph node positive group (p=0.005), but not associated with overall survival.

Conclusion:

NGAL is expressed in approximately one-fifth of the breast carcinomas and its expression is correlated with several poor prognostic markers and decreased disease-free survival. This results show that NGAL expression may be a predictor of poor prognosis in primary human breast cancer, especially with more aggressive clinical features

Citation Format: Lee-Su Kim, Hae Sung Kim. The expression and prognostic significance of neutrophil gelatinase-asssociated lipocalin (NGAL) in primary breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3469. doi:10.1158/1538-7445.AM2013-3469
Abstract 3473: Intra-person and inter-person biomarker variation for modeling longitudinal values of ovarian cancer early detection biomarkers.
Authors: Raamanathan Archana , Lu Zhen , Bast Robert C. , Skates Steven J.
Published: August 2014

In 2012, an estimated 22,280 women will be diagnosed with ovarian cancer in the United States and 15,500 women will die from this disease. Though localized ovarian cancers have a survival rate of 92%, only 15-20% of ovarian cancers are detected at this stage. Developing an effective strategy for early detection could reduce mortality. While individual values of CA125 are neither sufficiently sensitive nor specific for early detection, monitoring CA125 over time shows greater specificity. The risk of ovarian cancer algorithm, ROCA, analyzes the trend of annual values for CA125, utilizing each woman's own baseline to refer a small fraction (<1-2%) of women to transvaginal sonography. Using this strategy, no more than three operations are required for each case of ovarian cancer detected. Use of multiple biomarkers might increase sensitivity for preclinical disease. Based on a multiplex assay of 96 biomarkers, a four biomarker panel was identified of CA125, HE4, MMP-7 and CA 72-4 with greater sensitivity than CA125 alone. To estimate the utility of these biomarkers for longitudinal screening, we assessed five annual longitudinal control serum samples from 200 women who did not develop ovarian cancer from the MDACC SPORE normal risk ovarian cancer screening study (NROSS). Utilizing WinBUGS, the inter-person and intra-person CVs were evaluated. Upon log transformation for CA125, HE4 and MMP-7 and log(log()+1) transformation for CA72-4, both the inter-person and intra-person CVs follow a log-normal distribution. The median inter-person CVs for CA125, HE4, MMP-7 and CA 72-4 were 49%, 20%, 35% and 84% respectively and the median intra-person CVs were 15%, 25%, 21% and 21% respectively. The ratios of intra-person CVs to inter-person CVs derived for CA125, HE4, MMP-7 and log(CA 72-4) were 0.31, 1.25, 0.6 and 0.31. We anticipate that the lower the ratio, the higher the contribution of each individual marker to early detection during longitudinal measurements. The intra-person CV's should permit development of a multi-marker algorithm. Assessment of longitudinal samples derived from 50 cases of preclinical ovarian cancer and 10 controls per case (UKCTOCS trial) are underway in order to establish such a longitudinal multi-marker algorithm for assessment of ovarian cancer risk.

Citation Format: Archana Raamanathan, Zhen Lu, Robert C. Bast, Steven J. Skates. Intra-person and inter-person biomarker variation for modeling longitudinal values of ovarian cancer early detection biomarkers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3473. doi:10.1158/1538-7445.AM2013-3473
Abstract 3474: GDF15 as a serum marker of cisplatin response in ovarian cancer cells.
Authors: Izaguirre Daisy I. , Kwan Suet Yan , Zu Zhifei , Tsang-Lee Yvonne T. , Wong Kwong-Kwok
Published: August 2014

Ovarian cancer treatment consists of surgery followed by a chemotherapy combination entailing a platinum agent (cisplatin/carboplatin) and a taxane (paclitaxel). While 70-80% of patients initially respond to the standard treatment, the majority will relapse, and of these patients, 70-90% die because of drug resistance. Therefore, identifying markers that can predict therapeutic response and/or therapeutic targets is of great interest. To identify genes involved in chemoresistance, a microarray study of an ovarian cancer cell line (A2780) treated with varying doses of cisplatin and at varying time points was conducted. This study showed GDF15 had the highest fold-induction by cisplatin treatment. GDF15 induction by cisplatin was further validated by RT-PCR and Western blot analysis in the A2780 and 12 other cell lines with varying sensitivity/resistance to cisplatin. To further test whether GDF15 plays a role in cisplatin resistance, we knocked down its expression by esiRNA. GDF15 knockdown revealed an increase in the cisplatin IC50, suggesting that GDF15 sensitizes ovarian cancer cells to cisplatin. To determine whether GDF15 was also induced in vivo we injected mice intraperitoneally with either A2780 (cisplatin sensitive) or RMG1 (cisplatin resistant) cells. Once the tumors developed, the mice were treated with either 2.5 mg/kg or 5 mg/kg cisplatin and both the tumors and sera collected 24 and 48 hours after treatment. ELISA results on the serum showed an increase in GDF15 in mice after cisplatin treatment. Although there was no correlation between the two cisplatin doses used and GDF15 there was a correlation between GDF15 and tumor weight. Thus, our results suggest that GDF15 is induced by cisplatin both in vitro and in vivo and has a potential role as a serum biomarker to predict chemotherapeutic response.

Citation Format: Daisy I. Izaguirre, Suet Yan Kwan, Zhifei Zu, Yvonne T. Tsang-Lee, Kwong-Kwok Wong. GDF15 as a serum marker of cisplatin response in ovarian cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3474. doi:10.1158/1538-7445.AM2013-3474
Abstract 348: LKB1 deficiency confers lung adenocarcinoma phenotypic plasticity with squamous transdifferentiation potential.
Authors: Han Xiangkun , Li Fuming , Fang Zhaoyuan , Gao Yijun , Fang Rong , Li Fei , Sun Yihua , Xia Jufeng , Yao Shun , Li Li , Chen Haiquan , Hou Yingyong , Liu Xinyuan , Ji Hongbin
Published: August 2014

The existence and related mechanisms of phenotypic plasticity and dynamic lineage conversion of lung cancers, as implicated by clinical observation of mixed adenocarcinoma (ADC) and squamous cell carcinoma (SCC) pathologies in adenosquamous cell carcinoma (Ad-SCC), remain as fundamental yet unsolved questions. Here we find that about 13% human lung Ad-SCC harbor genetic alterations of tumor suppressor LKB1. Through lineage tracing experiments and pathological analyses in mouse models of human lung cancer, we further show that LKB1 deficiency confers lung adenocarcinoma phenotypic plasticity to progressively transdifferentiate into squamous cell carcinoma (SCC), via pathologically mixed Ad-SCC as intermediate. Down-regulation of lysyl oxidase (LOX) following reduced hypoxia level in LKB1-deficient lung adenocarcinoma decreases collagen disposition and triggers extracellular matrix remodeling and eventually up-regulates p63 expression, the SCC lineage survival oncogene. Moreover, pharmacological inhibition of Lox enzymatic activity promotes the transdifferentiation, whereas ectopic Lox expression conversely increases collagen deposition and significantly inhibits this process. ADC and SCC show differential response to pharmacological Lox inhibition. Notably, human lung Ad-SCC with LKB1 genetic alterations show pathology-specific patterns of LOX expression and collagen deposition in adenomatous and squamous portions as seen from animal models. These data uncover an essential role and related molecular mechanism of LKB1 deficiency in determining lung adenocarcinoma phenotypic plasticity.

Citation Format: Xiangkun Han, Fuming Li, Zhaoyuan Fang, Yijun Gao, Rong Fang, Fei Li, Yihua Sun, Jufeng Xia, Shun Yao, Li Li, Haiquan Chen, Yingyong Hou, Xinyuan Liu, Hongbin Ji. LKB1 deficiency confers lung adenocarcinoma phenotypic plasticity with squamous transdifferentiation potential. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 348. doi:10.1158/1538-7445.AM2013-348
Abstract 3481: Development of personalized paclitaxel therapy (IG-001) for ovarian cancer.
Authors: Hwang Larn , Hsiao Chao , Motamed Kouros , Trieu Vuong
Published: August 2014

Introduction: IG-001, like nab-paclitaxel, is a polymer bound nanoparticle paclitaxel. It is being developed for difficult to perfuse hypoxic tumors such as pancreatic cancer and ovarian cancer. IG-001 has shown promising activity against ovarian cancer in phase II clinical trials. In the planned phase III trial of IG-001, serum biomarkers as well as blood paclitaxel will be monitored to as prognostic indicators of response. The goal of this study is to developed point-of-care testing devices for PD biomarkers including CA125, B-type natriuretic peptide (BNP), Follicle Stimulating Hormone (FSH) and blood paclitaxel.

Methods: Serum samples collected at time of diagnosis of ovarian cancer were tested using rapid and quantitative point-of-care (POC) devices for blood biomarkers (CA125, FSH, BNP, and paclitaxel) and the data were evaluated using JMP9 statistical analysis software.

Results: 41 ovarian cancer patients were analyzed for CA125 prior to surgery of which 3 (7%), 11 (27%), and 27 (66%) were clear cell carcinoma, cystadenocarcinoma and adenocarcinoma, respectively. CA125 was not diagnostic of ovarian cancer. However, incidence of elevated (BNP > 25 pg/ml) was higher for patients (14 of 19, 74%) vs. normal controls (3 of 10, 30%, p = 0.02, Chi-square). FSH level was higher (median=151.6 mU/ml) vs. normal controls (median of 13.4 mU/ml, p = 0.01, Wilcoxon). Paclitaxel POC device was developed to monitored paclitaxel level down to 20ng/mL for paclitaxel.

Conclusions: Rapid and quantitative point-of-care tests for PD biomarker testing and paclitaxel PK therapeutic drug monitoring have been developed for use in conjunction with IG-001 development against ovarian cancer. IG-001 is ideal for paclitaxel therapeutic drug monitoring (TDM) due to its dose proportional PK. Paclitaxel TDM should allow for more effective dosing of the patients and thereby improving effectiveness of IG-001. These POCTs are also patient-centric, inviting better compliance and patient participation in personalizing his/her treatment. Plasma or serum samples obtained via repetitive venipuncture represent the accepted gold standard for monitoring the pharmacodynamics (PD) and pharmacokinetics (PK) of anticancer agents. Unfortunately for research subjects, the burden of frequent venipuncture is high; and, for researchers, venipuncture samples require immediate processing and storage facilities with freezers. The work presented here resolved the operational difficulty associated with personalized therapy in general and IG-001 in specific.

Citation Format: Larn Hwang, Chao Hsiao, Kouros Motamed, Vuong Trieu. Development of personalized paclitaxel therapy (IG-001) for ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3481. doi:10.1158/1538-7445.AM2013-3481
Abstract 3482: Directly Detect High Risk HPV Oncogenes E6/E7 mRNAs from Pap Smear Samples without RNA Purification and RT-PCR.
Authors: Zhang Aiguo , Zhang Lulu , Chuang Rachel , Zhu Xining , Diaz Rachel , Chen Lily
Published: August 2014

Cervical cancer is one of the most common cancers in women, with about 450,000 new cases diagnosed each year. Nearly all the cervical cancers are from Papillomaviruses (HPV) infection. HPV are DNA-based viruses that infect the skin and mucous membranes of humans and many other animals. Most cancers of the vulva and vagina are induced by oncogenic HPV types. In precancerous lesions, most HPV genomes persist in an episomal state whereas, in many high-grade lesions and carcinomas, genomes are found integrated into the host chromosome. Two viral genes, E6 and E7, are invariably expressed in HPV-positive cancer cells. Their gene products are known to inactivate the major tumour suppressors, p53 and retinoblastoma protein (pRB), respectively. In addition, E6 oncoprotein is also capable to up regulate the expression of inhibitors of apoptosis, and E6 and E7 cooperate to effectively immortalise primary epithelial cells. It has been demonstrated that HPV E6/E7 expression level plays a key role in the progression of invasive carcinoma of the uterine cervix via the deregulation of cellular genes controlling tumour cell proliferation. HPV E6/E7 oncogenes have been proved to be good biological markers for prognosis assessment and specific therapy of the disease. We have developed a sandwich nucleic acid hybridization assay using branched DNA to amplify the signals. The assay can simultaneously detect E6/E7 mRNAs of all 14 high risk HPV subtypes directly from Pap smear samples without RNA purification and RT-PCR. All HPV mRNA targets are captured through cooperative hybridization of multiple probes and probe set design determines the specificity of each HPV subtype of E6/E7 mRNA. Probe set oligonucleotides bind a contiguous region of the target E6/E7 mRNAs and selectively capture target RNAs to a solid surface during hybridization. Signal amplification is performed via sequential hybridization of Pre-Amplifier, Amplifier and label probe. The assay is highly specific and sensitive, which can detect as low as 50 transcript molecules.

Citation Format: Aiguo Zhang, Lulu Zhang, Rachel Chuang, Xining Zhu, Rachel Diaz, Lily Chen. Directly Detect High Risk HPV Oncogenes E6/E7 mRNAs from Pap Smear Samples without RNA Purification and RT-PCR. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3482. doi:10.1158/1538-7445.AM2013-3482
Abstract 3483: Modeling circulating tumor cells in the peripheral blood and CSF of breast cancer patients.
Authors: Allen Joshua E. , Patel Akshal S. , Dicker David T. , Sheehan Jonas M. , Glantz Michael , El-Deiry Wafik S.
Published: August 2014

The enumeration of circulating tumor cells (CTCs) using the CellSearch system (Veridex LLC) is a valuable prognostic clinical tool in several epithelial malignancies. Surveillance methods for metastatic disease involving the central nervous system (CNS) lack adequate sensitivity, specificity, and reproducibility in the clinic. We previously reported a method to enumerate CTCs in the cerebrospinal fluid (CSF) of breast cancer patients with metastases involving the CNS. The enumeration of CTCs in CSF by this method is highly sensitive, accurate, reproducible, and correlates with disease burden by several measures. Here, we concomitantly monitor CTCs in the peripheral blood and CSF of five patients with neoplastic meningitis receiving intrathecal (IT) chemotherapy. In each patient, CTC counts have been more sensitive than conventional cytology, and have predicted the course of symptoms and overall survival more precisely than conventional cytology. We observed a striking inverse relationship between CTC counts in the peripheral blood and CSF. This observation generated the hypothesis that CTCs migrate from the blood to the CSF and the converse as sanctuary sites during compartmentalized therapy to result in systemic disease recurrence. To test this hypothesis we created mouse models of these clinical phenomena including injection of triple-negative breast cancer into the peripheral blood or intracranially. Injection of luciferase-infected MDA-MB-231 or MDA-MB-468 human triple-negative breast cancer cells into the brain of athymic nude mice resulted in primary tumors at the site of injection. The MDA-MB-231, unlike the MDA-MB-468, generated detectable tumor cell dissemination into the CSF of inoculated mice that correlated with a significantly declined survival. CSF collected from the cisterna magna of inoculated mice corroborated these observations. Current studies are examining the impact of compartmentalized therapy on the migration of these tumor cells as well as profiling unique genetic drivers of the subset of tumors cells that disseminate into the CSF. The intercompartmental cycling of tumor cells may represent an important mechanism for disease persistence and recurrence, and suggests that concurrent systemic and CNS-directed therapy may be warranted.

Citation Format: Joshua E. Allen, Akshal S. Patel, David T. Dicker, Jonas M. Sheehan, Michael Glantz, Wafik S. El-Deiry. Modeling circulating tumor cells in the peripheral blood and CSF of breast cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3483. doi:10.1158/1538-7445.AM2013-3483
Abstract 3489: A novel monoclonal antibody to native HER2 epitope detects HER2-positive CTCs, HER2 in blood, and HER2 in tissue: realistic approach for monitoring breast carcinoma.
Authors: Alper Özge , Akoa Achille , Alper Özgül M. , Chen Cui P.
Published: August 2014

Background: Amplification of the HER2 gene and/or overexpression of its protein product have been reported in 25% to 30% of breast carcinomas and have been associated with poorer outcomes. However, there are still highly controversial aspects of HER2 testing. In the present study, we generated a novel monoclonal antibody against human native epitope of HER2 and investigated its clinical potential in managing breast carcinoma.

Methods and findings: We generated a novel monoclonal antibody (mAb), AB-HER2 using hybridoma technology and native folding of the protein as an immunogen. This mAb is of IgG1 (kappa) type. It specifically recognized both native and denatured human HER2 protein and displayed high affinity as assessed by Circulating Tumor Cells (CTCs), ELISA, immunohistochemistry assay, western blot and flow cytometry applications. We have developed a CTCs assay to detect specific HER2-positive tumor cells circulating in blood, while we did not observe any background staining in blood cells. Using AB-HER2 mAb in ELISA, we showed soluble HER2 level is significantly increased in breast cancer patients’ plasma which directly correlated with HER2 expression pattern in breast cancer tissues. Tissue expression of HER2 was determined by using immunohistochemical techniques. Most of normal tissues showed negative staining, while different levels of membrane staining were observed in cancer sections. To evaluate the immunoreactivity of HER2 associated with tumor histological types, the staining intensity in membranes were scored according to the guidelines of ASCO/CAP. We also have compared AB-HER2 mAB with commercial HER2 antibodies in all immunoassays.

Conclusion: The novel AB-HER2 mAb showed improved accuracy in all immunoassays and clinical outcome. Circulating HER2-positive tumor cells, HER2 protein levels in plasma and in breast tissues significantly increased in patients with breast carcinoma compared to normal healthy controls. Our data suggest the potential clinical use of AB-HER2 mAB as a powerful immunoreagent in measuring real-time HER2 levels in managing breast cancer.

Citation Format: Özge Alper, Achille Akoa, Özgül M. Alper, Cui P. Chen. A novel monoclonal antibody to native HER2 epitope detects HER2-positive CTCs, HER2 in blood, and HER2 in tissue: realistic approach for monitoring breast carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3489. doi:10.1158/1538-7445.AM2013-3489
Abstract 3500: Analysis of the prevalence and prognostic significance of 92 endpoints using high throughput multiplexed qPCR In archival tissue from multiple NSCLC trials.
Authors: Do An D. , Sumiyoshi Teiko , Wang Yulei , Schleifman Erica , Xiao Yuanyuan , Klughammer Barbara , Wong Cheryl V. , Abbas Ilma , Fu Ling , Amler Lukas C. , Raja Rajiv , Shames David S.
Published: August 2014

Over one third of all cancer related deaths worldwide are caused by non-small cell lung cancer (NSCLC). Until recently, tumor histology was rarely a factor in treatment decisions. However, this paradigm is shifting with the recent emergence of molecularly targeted therapies and next-generation chemotherapies. To better define the molecular sub-types of NSCLC that associate with active therapies and to evaluate their prognostic significance in historical clinical trials, we developed a microfluidics-based multiplexed PCR platform that enables analyses of 96 gene transcripts in 96 samples using material from as little as a single biopsy slide. The multiplexed assay panel consists of Taqman assays for markers of histology, chemo-resistance, prognosis, oncogenic pathway activity, epithelial vs. mesenchymal biology, as well as NSCLC drug targets.

We report here on the development and validation of this 96-gene transcript-profiling panel. Using titrations of several lung biopsy formalin-fixed paraffin embedded (FFPE) tissues, we show that as little as 10 ng of FFPE RNA is feasible and yields robust gene expression data across all assays of the profiling panel.

Using this technology, we tested archival lung biopsy samples from multiple Phase II and III studies to assess the prevalence and potential prognostic significance of 92 endpoints in parallel. We identified several markers that were strongly associated with histology as defined by TTF1 or p63 expression. In addition, several of the these markers were also associated with longer survival regardless of therapy, and will be discussed. These data suggest that retrospective analyses of clinical trial samples using Fluidigm-based expression analysis can be used to better inform future trial design in NSCLC.

Citation Format: An D. Do, Teiko Sumiyoshi, Yulei Wang, Erica Schleifman, Yuanyuan Xiao, Barbara Klughammer, Cheryl V. Wong, Ilma Abbas, Ling Fu, Lukas C. Amler, Rajiv Raja, David S. Shames. Analysis of the prevalence and prognostic significance of 92 endpoints using high throughput multiplexed qPCR In archival tissue from multiple NSCLC trials. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3500. doi:10.1158/1538-7445.AM2013-3500
Abstract 3505: Development of robust copy number assays for tumor FFPE tissues.
Authors: Huw Ling-Yuh , Patel Rajesh , O'Brien Carol , Fu Ling , Raja Rajiv , Amler Lukas , Hampton Garret , Lackner Mark
Published: August 2014

Somatic copy number variations (CNVs) are frequently observed in cancer, likely a result high genomic and structural instability characteristic of cancer. CNVs have been shown to be important drivers for many oncogenic pathways and are associated with drug sensitivity and resistance. Therefore, these genetic alterations are valuable candidate biomarkers for patient stratification and predictors for therapeutic responses to targeted therapies. A fundamental challenge for copy number determination in tumor FFPE DNA is that the material is often degraded and modified by fixation, processing and storage. Here we present a PCR-based copy number assay utilizing target specific pre-amplification and a microfluidics detection system. 45 genes were selected based on literature and their potential relevance to therapeutic targets. This high-throughput platform showed excellent specificity and dynamic range from 1 to 100ng input genomic DNA. A panel of matched frozen and FFPE DNAs from cell lines were used for cross -validation with copy number identified by array CGH. A medium-center method is employed to calculate copy number and assess the extent of degradation in sample DNAs. To evaluate clinical utility of the technology, copy number alteration from a collection of matched primary and metastatic breast tumors as well as correlation between copy number changes and gene expression will be reported. We will also report results from a survey of other tumor types, including gastric cancer, mesothelioma, and HER2+ breast cancer.

Citation Format: Ling-Yuh Huw, Rajesh Patel, Carol O'Brien, Ling Fu, Rajiv Raja, Lukas Amler, Garret Hampton, Mark Lackner. Development of robust copy number assays for tumor FFPE tissues. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3505. doi:10.1158/1538-7445.AM2013-3505
Abstract 3510: Evaluation of PI3K Inhibitor chemosensitivity in human tumor explants using the Mosaic Blue assay and relationship to biomarkers by immunohistochemistry.
Authors: Dauffenbach Lisa , Cash Patricia A. , Sia Gela C. , Girees Sherif K. , Lim Ryan S. , Zheng Jianping , Olsen Eric P. , Kerfoot Christopher A.
Published: August 2014

The AKT protein family plays an important role in cellular survival by inhibiting apoptotic processes and is one of the major downstream targets of the phosphatidylinositol 3-kinase (PI3K) pathway. Mammalian target of rapamycin (mTOR) is a key kinase acting downstream of the activation of PI3K. Several chemotherapeutic agents target this pathway and are in clinical trials to treat patients with solid tumors. The use of chemosensitivity testing using human cancer explants is very useful in identifying biomarkers that relate to the intrinsic sensitivity of the tumor to a chemotherapetuic agent. The Mosaic Blue assay is a soft agar, ex vivo tumor response assay that utilized non-expanded patient cancer cells.

Human tumors with matched fresh tissue and formalin-fixed, paraffin embedded (FFPE) tissue were used for this study. Fresh tissues were disaggregated into single cells and exposed to various inhibitors of the PI3K pathway. Samples were classified as resistant or sensitive. The corresponding FFPE tissue block was stained by immunohistochemistry for several phosphorylated proteins, which included AKT, PRAS40, 4EBP1, S6 kinase, S6 ribosomal protein, and PTEN in accordance with validated protocols. Additional fresh cells were also treated with inhibitors, harvested, and phophorylated proteins were evaluated in the pre and post-dose tumor cells. Results were compared the chemosensitivity results and 2 relationships were identified. First, biomarker status of the archival tissue was evaluated in order to understand the intrinsic chemosensitivity. Second, biomarker status upon treatment was evaluated to understand modulation of proteins due to therapy.

In summary, use of soft agar ex vivo tumor response assays such as the Mosaic Blue assay is a useful tool to predict relative sensitivity across tumor types and allow investigation of potential biomarkers.

Citation Format: Lisa Dauffenbach, Patricia A. Cash, Gela C. Sia, Sherif K. Girees, Ryan S. Lim, Jianping Zheng, Eric P. Olsen, Christopher A. Kerfoot. Evaluation of PI3K Inhibitor chemosensitivity in human tumor explants using the Mosaic Blue assay and relationship to biomarkers by immunohistochemistry. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3510. doi:10.1158/1538-7445.AM2013-3510
Abstract 3514: A dose-escalation study of anastrozole and everolimus in patients with advanced gynecologic and breast malignancies: tolerance, biological activity, and molecular alterations in the PI3K/AKT/mTOR pathway.
Authors: Wheler Jennifer J. , Moulder Stacey L. , Naing Aung , Janku Filip , Piha-Paul Sarina A. , Falchook Gerald S. , Zinner Ralph , Tsimberidou Apostolia M. , Fu Siqing , Hong David S. , Atkins Johnique T. , Cronin Maureen , Yelensky Roman , Stephens Phil
Published: August 2014

Background

Combining aromatase inhibitors with PI3K/AKT/mTOR inhibitors is synergistic and may overcome hormone resistance.

Patients and Methods

We evaluated anastrozole and everolimus in patients with metastatic estrogen receptor (ER) and/or progesterone receptor (PR)-positive breast and gynecologic tumors. Endpoints were safety, antitumor activity and molecular correlates.

Results

Fifty-five patients were enrolled. Full doses of anastrozole (1 mg PO daily) and everolimus (10 mg PO daily) were well tolerated. Fifty patients were evaluable for response with a median age of 57 (range: 24-82) and a median number of prior therapies of 3 (range: 0-13). Twelve of 50 patients (24%) achieved stable disease (SD) ≥ 6 months/partial response (PR)/complete response (CR), including 9 of 32 patients (28%) with breast cancer, 2 of 9 patients (22%) with ovarian cancer and 1 of 6 patients (17%) with uterine cancer. Six of 8 patients (75%) with SD ≥ 6 months/PR/CR with molecular testing demonstrated at least one alteration in the PI3K/AKT/mTOR pathway including mutations in PIK3CA (n=3) and AKT1 (n=1) or PTEN loss (n=3). All 3 patients who had next generation sequencing demonstrated additional alterations: amplifications in CCNE1, IRS2, MCL1, CCND1, FGFR1 and MYC and a rearrangement in PRKDC. Six of 22 patients (27%) with molecular alterations in the PI3K/AKT/mTOR pathway achieved SD ≥ 6 months/PR/CR.

Conclusion

Combination anastrozole and everolimus is well tolerated at full approved doses and active in heavily pretreated patients with ER and/or PR-positive breast, ovarian and endometrial cancers. Patients with SD ≥ 6 months/PR/CR frequently had alterations in the PI3K/AKT/mTOR pathway.

Citation Format: Jennifer J. Wheler, Stacey L. Moulder, Aung Naing, Filip Janku, Sarina A. Piha-Paul, Gerald S. Falchook, Ralph Zinner, Apostolia M. Tsimberidou, Siqing Fu, David S. Hong, Johnique T. Atkins, Maureen Cronin, Roman Yelensky, Phil Stephens. A dose-escalation study of anastrozole and everolimus in patients with advanced gynecologic and breast malignancies: tolerance, biological activity, and molecular alterations in the PI3K/AKT/mTOR pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3514. doi:10.1158/1538-7445.AM2013-3514