bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1820: Dynamics of HER-2 loss in mammary carcinoma of human HER-2 trangenic mice
Authors: Nanni Patrizia , Palladini Arianna , Landuzzi Lorena , Dall'Ora Massimiliano , Ianzano Marianna , Grosso Valentina , Ranieri Dario , Nicoletti Giordano , Laranga Roberta , Giovanni Carla D. , Iezzi Manuela , Lollini Pier-Luigi
Published: May 2015

Progression of HER-2+ breast cancer can result in the emergence of HER-2-negative tumor variants that activate alternative mitogenic pathways, either spontaneously or after therapy. We found that HER-2 loss occurs even in transgenic mouse models in which the oncogene is driven by viral promoters, thus mammary carcinoma of human HER-2 transgenic mice (huHER-2 mice) can be used to study not only the early phases of HER-2-driven mammary carcinogenesis, but also tumor progression beyond HER-2 addiction. Primary mammary carcinomas of huHER-2 mice express high levels of the oncogene, with a marked intratumoral heterogeneity. Cell lines grown from HER-2+ mammary carcinomas frequently undergo a progressive loss of expression. We have established a model system consisting of cell lines, clones and variants that exhibit one of three phenotypes: a) high and stable HER-2 expression in vitro and in vivo, b) high but labile HER-2 expression which is lost either during in vitro culture, after tumor growth in mice or after in vitro treatment with trastuzumab, and c) complete loss of HER-2 expression After HER-2 loss, most variants displayed a transition to an elongated, motile phenotype (epithelial to mesenchymal transition), an increased ability to generate mammospheres, a reduced expression of CD24 and an increased expression of CD44 (denoting mammary cancer stem cells). Tumorigenic and metastatic ability of HER-2-negative cells was increased in comparison to HER-2+ cells. As expected, HER-2 loss was accompanied by resistance to HER-2 targeted monoclonal antibodies and small molecule inhibitors. The study of therapeutic agents directed against downstream targets showed that HER-2-loss was accompanied by a loss of sensitvity to a Src inhibitor, whereas a PI3K inhibitor was highly effective regardless of HER-2 expression. Our results indicate that human HER-2 transgenic mice are a useful model to study the dynamics of HER-2 loss in advanced HER-2+ mammary carcinoma, and to analyze alternative therapeutic strategies.

Supported by grants from the Italian Association for Cancer Research (AIRC).

Citation Format: Patrizia Nanni, Arianna Palladini, Lorena Landuzzi, Massimiliano Dall'Ora, Marianna Ianzano, Valentina Grosso, Dario Ranieri, Giordano Nicoletti, Roberta Laranga, Carla De Giovanni, Manuela Iezzi, Pier-Luigi Lollini. Dynamics of HER-2 loss in mammary carcinoma of human HER-2 trangenic mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1820. doi:10.1158/1538-7445.AM2014-1820
Abstract 1823: Novel regulation of Jagged1 by ErbB2 in breast cancer: implications for anti-ErbB2 therapy
Authors: Osipo Clodia , Pandya Kinnari , Wyatt Debra
Published: May 2015

We have demonstrated that Notch1 is required for trastuzumab resistance in ErbB2 positive breast cancer. This indicates that ErbB2 suppresses Notch1 in breast cancer and therapeutic intervention targeting ErbB2 might have an unintended consequence which is aberrant up regulation of Notch1 which is a breast oncogene.

However, the mechanism of action by which ErbB2 restricts Notch1 activation is unknown. In this current study, we investigated the role of cis- and trans-activation of Notch signaling by Notch ligands which are developmentally conversed to tightly regulate Notch activation. To address this hypothesis, we performed co-culture studies using fibroblasts expressing no Notch ligands or over-expressing human Jagged1 or Deltalike1 and ErbB2 positive breast cancer cells. We performed flow cytometry to isolate breast cancer cells after co-culture and extracted RNA to measure expression of Notch gene targets as a measure of Notch activity. The results showed that trastuzumab, Lapatinib, or ErbB2 knockdown increased overall Notch activition. Similarly, Co-culture with Jagged1-expressing fibroblasts increased overall Notch activation. However, Knocked down of Jagged1 in the breast cancer cells had little effect on ligand-induced Notch activation relieving the possibility of cis-inhibition. In contrast, Jagged1 knocked down abrogated trastuzumab-induced Notch activation in the breast cancer cells. These results suggest that ErbB2 might restrict Notch activation by preventing Jagged1-mediated trans activation of Notch and not by promoting cis-inhibition. Confocal immunofluorescence showed that Jagged1 is localized with Notch1 when ErbB2 is hyperactive but is trafficked to the cell surface in response to trastuzumab. K44ADynamin abrogated Jagged1 expression on the cell surface as measured by IF and surface biotinylation studies. Furthermore, K44ADynamin expression abrogated trastuzumab-induced Notch1 activation. Importantly, we measured growth consequences of Jagged1-mediated Notch activation in response to trastuzumab and found that Jagged1 is necessary for survival of ErbB2 positive breast cancer cells and trastuzumab resistance as measured by cell cycle analysis and Annexin V staining. These results taken together indicate that ErbB2 restricts Notch by limiting Jagged1-mediated trans-activation.

Citation Format: Clodia Osipo, Kinnari Pandya, Debra Wyatt. Novel regulation of Jagged1 by ErbB2 in breast cancer: implications for anti-ErbB2 therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1823. doi:10.1158/1538-7445.AM2014-1823
Abstract 1825: Jab1/Csn5 a new target in the resistant mechanism to HER2-targeted therapies for breast cancer
Authors: Claret Francois X. , Vu Thuy , Shackleford Terry J. , Allensworth Jennifer , Zhang Qingxiu , Tian Ling , Zhang Ronghua
Published: May 2015

Her2 positive (Her2+) breast cancer (BC) accounts for 20% of all breast cancer subtypes and is associated with high risk of death. Trastuzumab (Ttzm), first FDA-approved targeted therapy for BC, represents a key milestone in the personalized treatment of Her2+ metastatic disease (MBC). However, 75% of Her2+ MBC are or become resistant to Ttzm and experience relapse or disease progression. This suggests that tumors acquire or possess an intrinsic mechanism of resistance that prevents from Her2 inhibition. Therefore, it is important to identify novel pathway to overcome the resistance. Recent studies have proposed various potential mechanisms leading to the resistance, including p27 rapid degradation. Jab1/Csn5 a novel candidate oncogene that we identified contributes to the progression of BC and is correlated with poor prognosis. Our study demonstrated that Jab1/Csn5 degrades p27 in MBC. These findings suggest that Jab1 overexpression contributes to Ttzm-resistance by facilitating p27-degradation. Jab1/Csn5 is overexpressed in 50% of primary and 90% of MBC while its expression is low or absent in normal adult breast tissues. We previously identified that high expression of Jab1 is associated with shorter progression-free survival in MBC patients. In this study, we showed that the knockdown of Jab1 sensitizes to Ttzm-treatment. Mechanistically, we found that Jab1/Csn5 overexpression is significantly correlated with the activation of Akt pathway in Her2+ MBC and xenograft models. Interestingly, activated Akt, due to PTEN-loss or PI3K activating mutation has been widely implicated in the potential mechanism to Ttzm-resistance. Therefore, our results suggest that targeting Jab1/Csn5 overcomes the resistance to Ttzm via interfering with PI3K/Akt pathway. Our study identifies Jab1 as a novel contributor to Ttzm-resistance and elucidates its potential mechanisms of actions. Jab1 could be used as a predictive marker of tumor response to Ttzm, which could assist clinicians in selecting the best treatment modalities for patients with MBC.

Note: This abstract was not presented at the meeting.

Citation Format: Francois X. Claret, Thuy Vu, Terry J. Shackleford, Jennifer Allensworth, Qingxiu Zhang, Ling Tian, Ronghua Zhang. Jab1/Csn5 a new target in the resistant mechanism to HER2-targeted therapies for breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1825. doi:10.1158/1538-7445.AM2014-1825
Abstract 1836: Erlotinib induces NF-kappa B dependence that promotes EGFR tyrosine kinase inhibitor resistance in lung adenocarcinoma
Authors: Blakely Collin M. , Pazarentzos Evangelos , Asthana Saurabh , Olivas Victor , Tan Irena , Fouts Timothy , Meshulam Jeffrey , Bivona Trever G.
Published: May 2015

The vast majority of patients with lung adenocarcinomas harboring activating mutation in EGFR respond to EGFR tyrosine inhibitors (TKI) (i.e. erlotinib). However, the magnitude of tumor regression is variable and responses are short-lived with a median duration of 9-12 months. We recently identified activation of the NF-κB (Nuclear Factor kappa-light-chain-enhancer of activated B cells) signaling pathway as a key mediator of de novo resistance to EGFR TKI therapy in cell lines and tumor xenograft models of lung adenocarcinoma. However, the role of NF-κB activation in mediating EGFR TKI acquired resistance in lung adenocarcinoma is not well defined. We have found that erlotinib treatment of EGFR TKI sensitive lung adenocarcinoma cell lines promotes the rapid phosphorylation and degradation of IκB (inhibitor of NF-κB) and subsequent activation of the NF-κB subunit RelA. RelA activation is accompanied by transcriptional activation of downstream NF-κB target genes, including IL6. Prolonged exposure of EGFR TKI sensitive lung adenocarcinoma cell lines to erlotinib leads to EGFR TKI acquired resistance that is accompanied by sustained NF-κB activation and IL6 expression. EGFR TKI acquired resistance can be overcome by treating cells with erlotinib in combination with PBS-1086 (rel∼MD, Inc.), a direct Rel inhibitor. Furthermore, concomitant treatment of EGFR TKI sensitive lung adenocarcinoma cells with erlotinib + PBS-1086 prevents the development of EGFR TKI acquired resistance. Together, these results demonstrate the molecular basis for the synthetic lethality of combined EGFR and NF-κB inhibition and provide mechanism-based rationale for polytherapies against both EGFR and NF-κB to enhance response in lung adenocarcinoma patients. Broadly, our findings provide novel insights into the biological and clinical consequences of the context-specific and dynamic functional interplay between EGFR and NF-κB signaling.

Citation Format: Collin M. Blakely, Evangelos Pazarentzos, Saurabh Asthana, Victor Olivas, Irena Tan, Timothy Fouts, Jeffrey Meshulam, Trever G. Bivona. Erlotinib induces NF-kappa B dependence that promotes EGFR tyrosine kinase inhibitor resistance in lung adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1836. doi:10.1158/1538-7445.AM2014-1836
Abstract 1837: A HSP90 inhibitor, AUY922, is effective to overcome the MET- and AXL-mediated resistance to EGFR-TKI in lung cancer
Authors: Rho Jin Kyung , Choi Yun Jung , Kime Seon Y. , So Gwang Sup , Choi Se Hoon , Choi Chang-Min , Lee Jae Cheol
Published: May 2015

Activation of bypass signals such as MET and AXL has been identified as one of possible resistant mechanisms to EGFR-TKI. Since various oncoproteins are dependent on HSP90 for its maturation and stability, we investigated the effect of AUY922, a newly developed HSP90 inhibitor of non-geldanamycin class, in lung cancer cells with MET- and AXL-mediated resistance. We established resistant cell lines with HCC827 harboring the deletion mutation on exon 19 of EGFR gene by long-term exposure to increasing concentrations of gefitnib and erlotinib (designated as HCC827/GR and HCC827/ER, respectively). The resistance of HCC827/GR was mediated by MET activation, whereas AXL activation caused resistance in HCC827/ER. AUY-922 treatment effectively suppressed the proliferation and induced cell death in both resistant cell lines. Accordingly, down-regulation of MET and AXL leading to decreased activation of Akt was noted. The inhibitory effect of AUY-922 on AXL and MET was also confirmed in AXL or MET-transfected LK-2 cells. In addition, AUY-922 led to reduced capability of invasion and migration of resistant cells. AUY-922 was effective to control the tumor growth in xenograft mouse models of HCC827/GR and HCC827/ER. Our study showed AUY-922 is one of promising therapeutic options for the MET- and AXL-mediated resistance to EGFR-TKI in lung cancer.

Citation Format: Jin Kyung Rho, Yun Jung Choi, Seon Ye Kime, Gwang Sup So, Se Hoon Choi, Chang-Min Choi, Jae Cheol Lee. A HSP90 inhibitor, AUY922, is effective to overcome the MET- and AXL-mediated resistance to EGFR-TKI in lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1837. doi:10.1158/1538-7445.AM2014-1837
Abstract 1840: Contribution of miR-205 in gefitinib-resistant lung cancer cell lines
Authors: Suzuki Toshihiro , Nagasawa Ikuko , Yamaoka Toshimitsu , Ohmori Tohru , Nishio Kazuto , Koyama Kiyotaka , Ogasawara Yuki
Published: May 2015

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), gefitinib is effective therapies for patients with advanced non-small cell lung cancer (NSCLC). Treatment with gefitinib frequently induces drug resistance in NSCLC. Two mechanisms of acquired resistance have been validated in patients. Secondly mutations in EGFR itself, EGFR T790M and amplification of the MET oncogene are observed in resistance case. However, it remains unclear how MET over expression contributes to TKI resistant NSCLC. Recently, micro RNA (miRNA) expression is noted and analyzed because the concern of these molecules in cancer pathogenesis and drug resistance has been elucidated. Circulating tumor cell (CTC) and miRNA are focused as sensitive and non-invasive biomarkers for cancer diagnosis. In this study, we aimed at the miRNA in the gefitinib-resistant cell lines, and evaluated the miRNAs being associated with resistance of gefitinib. We established gefitinib resistant cell lines, in which MET is amplified, PC-9/MET and PC-9/MET-1K by a stepwise escalation of gefitinib concentrations in vitro. Microarray and real time PCR analysis indicated that miR-205 is significantly elevated in both resistant cell lines, and ErbB3 was a significant target. We examine further to reveal that relation between miR-205 and EGFR family, such as ErbB2 and ErbB3. Our results suggest that miR-205 may function as a gefitinib resistance in NSCLC.

Citation Format: Toshihiro Suzuki, Ikuko Nagasawa, Toshimitsu Yamaoka, Tohru Ohmori, Kazuto Nishio, Kiyotaka Koyama, Yuki Ogasawara. Contribution of miR-205 in gefitinib-resistant lung cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1840. doi:10.1158/1538-7445.AM2014-1840
Abstract 1842: Synergistic inhibition of ALK inhibitor-resistant lung cancer cells by dual-targeting EGFR and ALK pathways
Authors: Kim Soyeon , Kim Tae Min , Ahn Yong-Oon , Keam Bhumsuk , Lee Se-Hoon , Heo Dae Seog , Kim Dong-Wan
Published: May 2015

Background: Despite the excellent initial responses of ALK-rearranged lung cancer to the ALK tyrosine kinase inhibitors (TKIs), most patients develop acquired resistance through several mechanisms, including ALK amplification, mutation or activation of alternative signaling pathways. In this study, we investigated the effect of dual-targeting EGFR and ALK pathways in NSCLC cells that are resistant to ALK inhibitors.

Methods: The cell proliferation, apoptosis and cell cycle analysis were measured by spectrophotometry and flow cytometry. The combination effect of EGFR TKI (afatinib or dacomitinib) and ALK TKI (crizotinib or LDK378) was evaluated using a combination index (CI). We analyzed EGFR ligand levels by ELISA and expression of ALK, EGFR and its downstream molecules were detected by Western blot assay after exposure to TKIs.

Results: NCI-H3122CR1 that was resistant to crizotinib and NCI-H3122 cell lines became resistant to LDK378 through the step-wise increase of the drug, resulting in H3122CR1LR1 and H3122LR1 cells, respectively. The resistant cell lines exhibited 10-14-fold higher IC50 values for crizotinib or LDK378 compared with parental cells. The genetic alterations of ALK or EGFR genes were not found in H3122CR1, H3122LR1, and H3122CR1LR1 cells. We analyzed EGFR signaling pathways in these cells to determine their roles in mediating ALK inhibitor resistance. We observed that H3122CR1, H3122LR1 and H3122CR1LR1 cells had higher phospho-EGFR than parental cells. In these cells, mRNA and protein levels of amphiregulin and EGF were up-regulated. Combined treatment with EGFR TKI and LDK378 synergistically inhibited cell growth (CI<1) and induced apoptosis in H3122LR1 and H3122CR1LR1 cells. Similar results were observed in H3122CR1 and SNU-2535 harboring ALK G1269A mutation cells, when exposed to afatinib and crizotinib. The combination of EGFR and ALK TKIs decreased the phosphorylation of several downstream molecules in H3122LR1 and H3122CR1LR1 cells, whereas EGFR or ALK TKI alone did not reduce the phosphorylation levels.

Conclusion: Our results suggest that dual inhibition of ALK and EGFR effectively abrogate ALK and its downstream signals and induce apoptosis in ALK TKIs-resistant, ALK-rearranged NSCLC cells. Therefore, dual-targeting of EGFR and ALK might be a promising strategy for ALK-rearranged NSCLC patients who showed acquired resistance to crizotinib and/or LDK378.

Citation Format: Soyeon Kim, Tae Min Kim, Yong-Oon Ahn, Bhumsuk Keam, Se-Hoon Lee, Dae Seog Heo, Dong-Wan Kim. Synergistic inhibition of ALK inhibitor-resistant lung cancer cells by dual-targeting EGFR and ALK pathways. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1842. doi:10.1158/1538-7445.AM2014-1842
Abstract 1845: Sensitivity of PC9 cells to erlotinib is affected by extracellular matrix
Authors: Onishko Halina M. , Zhao Jie , Jameson Katherine L. , Frick Peter L. , Tyson Darren R. , Quaranta Vito , Yankeelov Thomas E. , Rericha Erin C.
Published: May 2015

Tyrosine kinase inhibitors of epidermal growth factor receptor, such as erlotinib and gefinitib, have been effective in the initial treatment of non-small cell lung cancer. With time, however, initially responsive tumors almost invariably rebound and proliferate under therapy. Of general interest is the impact of tumor cell heterogeneity on rebound time. Previously, we isolated 89 single-cell derived discrete sublines (DS) from the commonly studied PC9 cell line by monitoring the distribution of cell fates (division, quiescence, and cell death) and the drug-induced proliferation (DIP) rate under erlotinib treatment. In this work we examine the role of the extracellular matrix (ECM) in determining proliferation behavior and the distribution of cell fates. Cells suspended in 3D matrices show increased sensitivity to erlotinb with the rate of cell death depending on the ECM composition. DS sublines that were highly resilient in 2D culture (i.e., positive DIP rate and <10% cell death under treatment) asymptote to about 40% cell death in Matrigel after ten days in 1 µM erlotinb. Both the rate of cell death and the total percentage of the cell population that dies increased in collagen I and in collagen I coated hydrogel. At asymptotic levels, the cell population contained both dividing and quiescent cells. In contrast, DS sublines with negative DIP rates in 2D culture reached 90-99% cell death in each ECM considered. We conclude that the pathways conferring resilience in these cells are sensitive to ECM conditions. Intriguingly, we find that resilient DS sublines tend to have diversity in cluster morphologies in the matrices, suggesting a possible correlation between ECM engagement and/or adhesion and resilience. Molecular studies should clarify mechanistic relationships between ECM and sensitivity to targeted therapy, and whether such relationship may extend to clinical tumors.

Citation Format: Halina M. Onishko, Jie Zhao, Katherine L. Jameson, Peter L. Frick, Darren R. Tyson, Vito Quaranta, Thomas E. Yankeelov, Erin C. Rericha. Sensitivity of PC9 cells to erlotinib is affected by extracellular matrix. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1845. doi:10.1158/1538-7445.AM2014-1845
Abstract 1851: Colorectal adenomatous polyps and mitochondrial DNA variants
Authors: Aikhionbare Felix O. , Mehrabi Sharifeh , Cohen Shakeria , Yao Xuebiao , Iyamu Osatohamwen
Published: May 2015

Patients with adenomatous polyps have a threefold higher risk of colon cancer over the general population, which increases to sixfold if the polyps are multiple and with lower survival among African American population. Up to now, only 6% of CRC can be ascribed to mutations in particular genes. Moreover, the optimal management of patients with colorectal adenomatous polyps depends on the accuracy of appropriate staging strategies because patients with similar colorectal adenocarcinoma architecture display heterogeneity in the course and outcome of the disease. In this study, using high resolution restriction endonucleases and PCR-based sequencing, we have potentially identified some distinctive mitochondrial DNA restriction sites and sequence polymorphisms that distinguished variants among colorectal adenomatous polyps and between pre-cancer and carcinoma in the regions of ND, CYTO b, ATPase and COX genes. The distribution of some of the observed mtDNA sequence variants includes:0% (0/161) tubular, 1.2% (12/161) tubulovillous, 54% (87/161) villous, and 94.4% (152/161) cancer . The restriction analysis, using HINC II enzyme, identified a number of band shifts indicating site gains or losses in differentiating between colorectal adenomatous polyps and cancer. Results are the first study to systemic survey mitochondrial genome alterations on the pathology and clinical significance of colonic adenocarcinoma with villous architecture. This abstract was supported by NIH-NIGMS # GM099663.

Citation Format: Felix O. Aikhionbare, Sharifeh Mehrabi, Shakeria Cohen, Xuebiao Yao, Osatohamwen Iyamu. Colorectal adenomatous polyps and mitochondrial DNA variants. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1851. doi:10.1158/1538-7445.AM2014-1851
Abstract 1854: P16 immunoreactivity in endocervical tubo-endometrial metaplasia (TEM) and correlation with high risk HPV (HRHPV) status by chromogenic in-situ hybridization (HPV-CISH) and hybrid capture 2 assay
Authors: Talukdar Shobhana , Juan Gomez-Gelvez , Buekers Thomas E , Zhang Ziying
Published: May 2015

Background

Several studies have shown that endocervical TEM can intensively over express p16 and involve more than 50% of the cells. It could be speculated that TEM may be a precursor lesion of cervical glandular intraepithelial neoplasm. The aim of this study is to evaluate endocervical TEM p16 overexpression pattern and further investigate its potential driven mechanisms by correlating with HRHPV status.

Method

Our search included cervical biopsy, LEEP, conization with TEM diagnosis from January 2005-May 2013. Only cases with concurrent Pap smear with HRHPV testing were selected. HRHPV detected in liquid based cytology is by Hybrid Capture 2 Assay. Surgical specimens containing concurrent squamous dysplasia were excluded due to potential confounding by HRHPV positivity in dysplastic squamous epithelium. P16 stain was performed and graded as negative, focal+ (<50% cytoplasmic and nuclear stain) and diffuse+ (>50% cytoplasmic and nuclear stain). HRHPV data was collected from Pap reports. HPV-CISH assays will be performed for all cased with positive HRHPV detected in liquid based cytology.

Result:

Our search generated 159 cases with TEM. Only 81 cases had concurrent Pap smear with HRHPV testing. 48 cases were excluded due to squamous dysplasia. A total of 33 patients with an age range of 20-59 were available for the study. Twenty nine out of 33 had negative HRHPV in liquid based cytology; in which, p16 stain results as: 8/29 negative, 14/29 focally +. 7/29 diffusely +. Four out of 33 had positive HRHPV detected in liquid based cytology; in which, p16 stain results as: 1/4 negative, 3/4 diffusely +. HPV-CISH assays were performed in all 4 cases and they were negative.

Conclusions

P16 overexpression in TEM can be intense and involve more than 50% of the cells but the pattern is heterogenous and positivity never involves 100% of the cells. Neither HPV-CISH nor Hybrid Capture 2 Assay detects HRHPV in TEM. P16 overexpression in TEM is most likely HRHPV-independent mechanisms.

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Note: This abstract was not presented at the meeting.

Citation Format: Shobhana Talukdar, Gomez-Gelvez Juan, Thomas E Buekers, Ziying Zhang. P16 immunoreactivity in endocervical tubo-endometrial metaplasia (TEM) and correlation with high risk HPV (HRHPV) status by chromogenic in-situ hybridization (HPV-CISH) and hybrid capture 2 assay. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1854. doi:10.1158/1538-7445.AM2014-1854
Abstract 1857: Prognostic biomarker for SNP (-617C-A) in ARE-like loci of the NRF2 gene in lung adenocarcinoma
Authors: Okano Yasuko , Ichikawa Yasushi , Miyagi Yohei , Kondo Keiichi , Natsumeda Yutaka
Published: May 2015

Purpose: We here report that SNP (c.-617C>A; rs6721961) in the ARE-like region of NRF2 gene is associated with Japanese non-smoking female patients with adenocarcinoma and their overall survival.

Experimental Design: We prepared genomic DNA samples from 387 Japanese patients with primary lung cancer and detected a SNP (-617C>A) in NRF2 gene by the rapid genetic testing method we developed in this study. We analyzed the association between the SNP in the NRF2 gene and overall survival.

Results:Patients harboring wild-type -617C/C, -617C/A, and -617A/A alleles numbered 216 (55.8%), 147 (38.0%), and 24 (6.2%), respectively. Multivariate logistic regression models revealed that SNP homozygote (-617A/A) was significantly related to gender. Its frequency was four-fold higher in female patients than in males (10.8% female vs 2.7% male) and was associated with female non-smokers with adenocarcinoma. Interestingly, lung cancer patients carrying the SNP homozygous alleles (-617A/A) and the 309T (WT) allele in the MDM2 gene exhibited remarkable survival over 1,700 days after surgical operation (log-rank p=0.021). Conclusion: SNP homozygous (-617 A/A) alleles in the NRF2 gene are associated with female non-smokers with adenocarcinoma and regarded as a prognostic biomarker for assessing overall survival of patients with lung adenocarcinoma.

Citation Format: Yasuko Okano, Yasushi Ichikawa, Yohei Miyagi, Keiichi Kondo, Yutaka Natsumeda. Prognostic biomarker for SNP (-617C-A) in ARE-like loci of the NRF2 gene in lung adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1857. doi:10.1158/1538-7445.AM2014-1857
Abstract 1861: S100P is a potential biomarker in distinguishing mucinous from non-mucinous pancreatic cysts and predicting invasive adenocarcinoma
Authors: Mi Wenyi , Diehl David L. , Shi Jianhi , Blansfield Joseph , Hunsinger Marie , Yang Wannian , Lin Fan
Published: May 2015

Introduction. Our previous publications demonstrated that S100P was a highly sensitive biomarker in identifying pancreatic mucinous neoplasms (PMNs) and invasive ductal adenocarcinomas (ADCs) on both cytological and surgical specimens. When working on pancreatic cystic lesions, there are two important undertakings: 1) distinguishing mucinous neoplasms from non-mucinous cysts and 2) predicting the presence of invasive adenocarcinomas. In this study, we investigated the potential utility of S100P for these purposes.

Design. Thirty pancreatic cystic fluid samples obtained by EUS-FNA biopsy were retrieved from Geisinger Biospecimen Bank. Diagnosis was based on definitive cytological diagnosis in all cases and additional surgical pathology confirmation in 11 cases. These 30 samples included 7 benign cysts in Group 1, 14 PMNs in Group 2, and 9 ADCs in Group 3. The S100P protein level of each sample was quantitatively measured by enzyme-linked immunoassay (ELISA) (CircuLex S100P ELISA kit, MBL International Corporation) according to the vendor's protocol.

Results. The results are summarized in Table 1. Marked elevation of S100P was seen in 6 of 9 ADCs (2129.81 + 1834.34 ng/ml), in contrast to the moderate increase in Group 2 (666.67 + 1439.91 ng/ml) and the minimal increase in Group 1 (22.27 + 18.62 ng/ml). Using 50 ng/ml as a cutoff value to differentiate a PMN/ADC (Group 2 and Group 3) from a benign cyst (Group 1), the sensitivity, specificity, positive predictive value and negative predictive value for identifying a PMN/ADC were 60.9%, 100%, 100%, and 43.8%, respectively.

Conclusion. These preliminary data suggest that S100P is a useful biomarker in identifying invasive adenocarcinomas and in distinguishing mucinous neoplasms from non-mucinous cysts. Caution should be taken since some ADCs may show only mildly increased S100P levels, similar to some PMNs and non-mucinous cysts. Additional study in a large series of cases is needed to substantiate the current findings.

Table 1.Summary of S100P protein level (ng/ml) in 30 pancreatic cysts by ELISACase #Benign CystsMucinous NeoplasmsAdenocarcinomas12.515.4812.6722.596.6413.2135.0410.8544.07428.0918.371849.20530.6219.802162.98643.2929.402831.67743.75120.483076.958137.954441.439158.684736.0810221.7711285.6012290.78133385.19144628.37

Citation Format: Wenyi Mi, David L. Diehl, Jianhi Shi, Joseph Blansfield, Marie Hunsinger, Wannian Yang, Fan Lin. S100P is a potential biomarker in distinguishing mucinous from non-mucinous pancreatic cysts and predicting invasive adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1861. doi:10.1158/1538-7445.AM2014-1861
Abstract 1869: Using Nanoparticle Tracking Analysis (NTA) to characterize cellular microvesicles
Authors: Capracotta Sonja , Carnell Pauline , Malloy Andrew , Hole Patrick , Carr Bob
Published: May 2015

Cellular microvesicles (100 nm-1µm) are shed into the circulation from many different cell types including; platelets, erythrocytes and endothelial cells. They have major potential as novel biomarkers as they have been shown to be implicated in a multitude of pathological conditions. However, developments in this area are constrained by limitations in the technology available for their measurement.

Nanoparticle Tracking Analysis (NTA) (NanoSight Ltd, Amesbury) offers the potential to both enumerate and speciate (through fluorescent markers) these microparticles in a rapid manner. In this method a laser beam passes through a suspension at a low angle. The particles scatter light which is collected onto a CCD (or sCMOS) camera by a microscope-type configuration. Particles between 10-2000 nm are tracked individually and their diffusion coefficient, and therefore size, calculated directly from their speed.

Therefore, this characterization method gives a direct measurement of the concentration and size distribution of the particles in the field of view. If the particles are fluorescently labelled, then the concentration and size distribution are those of the labelled particles only, allowing a proportion of labelled particles to be calculated. However, this requires a combination of aspects to be identified and optimised during the process. This talk will look at some of the challenges and considerations that need to be taken during the protocol development and measurement.

It will also review the application of the technique in this field, compare and contrast it to other applicable technologies such as various forms of electron microscopy (EM) and to flow cytometry.

Citation Format: Sonja Capracotta, Pauline Carnell, Andrew Malloy, Patrick Hole, Bob Carr. Using Nanoparticle Tracking Analysis (NTA) to characterize cellular microvesicles. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1869. doi:10.1158/1538-7445.AM2014-1869
Abstract 1879: Dual function HDAC and PI3K inhibitor, CUDC-907 affects cancer cells and the tumor microenvironment in hematological malignancies
Authors: Ma Anna W. , Atoyan Ruzanna , Younes Anas , Flinn Ian W. , Oki Yasuhiro , Copeland Amanda , Berdeja Jesus G. , Laliberte Robert , Viner Jaye , Samson Maria Elena S. , Dellarocca Steven , Yi Ling , Borek Mylissa , Zifcak Brian , Xu Guangxin , Wang Jing
Published: May 2015

Histone deacetylases (HDACs) and the phosphatidylinositol 3-kinase (PI3K)/AKT pathway are promising therapeutic targets in hematologic cancers and evidence of synergistic anti-cancer activity has recently emerged. CUDC-907, a small molecule drug candidate that is designed to target HDACs and PI3Ks in a single chemical entity, is currently in Phase 1 clinical testing for the treatment of patients with lymphoma or multiple myeloma. Preclinically, CUDC-907 has been shown to inhibit activation of PI3K/AKT, JAK/STAT and MAPK pathways in hematologic cancer cell lines such as Hodgkin's lymphoma, diffuse large B-cell lymphoma, and multiple myeloma. In this study, we report that in the setting of hematological malignancies, CUDC-907 targets not only the cancer cells but also the tumor microenvironment. Cytotoxicity against primary CLL cells was independent of the protective effects provided by stromal cells when primary CLL cells were co-cultured with nurse-like cells. CUDC-907 was shown in vitro to impair cytokine and chemokine production by immune cells in the tumor microenvironment and by Hodgkin's lymphoma cells. The ongoing first-in-human Phase 1 clinical study of CUDC-907 has yielded preliminary evidence of anti-cancer activity and impact on the tumor microenvironment as measured by cytokine and chemokine levels (e.g., thymus and activation regulated chemokine [TARC]). Ongoing analyses are probing the potential utility of selected cytokine and chemokine as predictive markers of CUDC-907 activity.

Citation Format: Anna W. Ma, Ruzanna Atoyan, Anas Younes, Ian W. Flinn, Yasuhiro Oki, Amanda Copeland, Jesus G. Berdeja, Robert Laliberte, Jaye Viner, Maria Elena S. Samson, Steven Dellarocca, Ling Yi, Mylissa Borek, Brian Zifcak, Guangxin Xu, Jing Wang. Dual function HDAC and PI3K inhibitor, CUDC-907 affects cancer cells and the tumor microenvironment in hematological malignancies. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1879. doi:10.1158/1538-7445.AM2014-1879
Abstract 1880: Pathologic response pattern of breast cancer after neoadjuvant chemotherapy: its correlation with molecular subtypes
Authors: Song In-Hye , Lee Hee Jin , Gong Gyungyub
Published: May 2015

Background: Several ways to assess pathologic response of breast cancer after neoadjuvant chemotherapy (NAC) are currently used, including Miller-Payne system and residual cancer burden. However, the relationship between pathologic response pattern and molecular subtype of breast cancer is unclear. Methods: We investigated the pathologic response pattern of breast cancer after NAC in each molecular subtype. Female breast cancer patients who received NAC and subsequent breast operation were retrospectively analyzed. The NAC regimen used for breast cancer was primarily anthracycline and taxane-based, depending on the axillary lymph node involvement. We evaluated the molecular subtypes by immunohistochemistry, pretreatment tumor sizes by MRI, absolute invasive tumor cellularities in pretreatment biopsy and operation specimens, pathologic complete response (pCR) rates, posttreatment pathologic tumor sizes, overall residual tumor cellularities, and grades for Miller-Payne system. Results: A total of 119 patients (age: 45.2 ± 9.5 yr) were enrolled. Sixty-six cases were hormone receptor positive (ER or PR positive), 18 cases were HER2 positive (ER and PR negative), and 35 cases were triple negative. There were no significant differences in pretreatment tumor size by MRI and absolute cellularity in biopsy between three groups. After NAC, hormone receptor positive tumors showed the lowest pCR rate (18.2 %) while triple negative tumors showed the highest pCR rate (45.7 %, p=0.013). In patients with residual carcinoma, hormone receptor positive tumors showed the largest ypT size (p=0.016), the least reduction of tumor size (p=0.001), and the least absolute invasive tumor cellularity (p=0.006). Overall residual tumor cellularity and Miller-Payne grade were not significantly different between three groups (p=0.4 and p=0.703, respectively). Conclusion: These results suggest that each molecular subtype of breast cancer shows characteristic pathologic response patterns following NAC.

Citation Format: In-Hye Song, Hee Jin Lee, Gyungyub Gong. Pathologic response pattern of breast cancer after neoadjuvant chemotherapy: its correlation with molecular subtypes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1880. doi:10.1158/1538-7445.AM2014-1880
Abstract 1887: Global gene expression analysis of tamoxifen-resistant breast cancer cell lines identified a gene signature predictive of clinical outcome of breast cancer patients treated with endocrine therapy
Authors: Roro Daniel Elias , Vever Henriette , Lykkesfeldt Anne , Ditzel Henrik
Published: May 2015

Objective: Altered gene expression associated with endocrine resistance in breast cancer cell lines may have potential prognostic/predictive values in patients with ER+ breast cancer receiving endocrine therapy. In this study, we investigated mRNA expression profiles associated with the tamoxifen-resistant phenotype using a panel of isogenic MCF-7-based breast cancer cell lines to elucidate the endocrine resistance mechanism and assess whether the pattern of gene expression predicted clinical outcome in breast cancer patients treated with endocrine therapy.

Method: Gene array was performed on a panel of clinically-relevant tamoxifen-resistant breast cancer cell lines (TamR) and their tamoxifen-sensitive parental cell line. Genes that showed significant alteration in expression (FDR 0.01) in TamRs vs parental cell lines were evaluated for their potential to stratify tamoxifen-treated breast cancer patients based on outcome using published gene expression data sets.

Results: Using a testing set consisting of microarray data from 163 ER+ primary breast cancer patients treated with tamoxifen we identified 20 genes among those that showed significant altered expression in TamR that could stratify patients according to clinical outcome. . The potential of this 20-gene signature to stratify patients was further confirmed in 2 independent data sets consisting of ER+ tamoxifen-treated breast cancer patients with a sensitivity and specificity 53% and 79% respectively in the first cohort (N=153), and sensitivity and specificity of 37% and 91% respectively in the second cohort (N=201). Evaluation of relapse-free survival of tamoxifen-treated patients classified by the signature as having bad or good outcomes showed significant differences in time-to-relapse, with median disease-free survival times of 5.1 vs 8.9 years in the first cohort (N=153, p=0.03) and 4.2 vs 7.7 years in the second (N=201, p=0.006).

Conclusions: The expression pattern of the 20-gene signature identified in this study appears to have a strong potential as a biomarker of clinical outcome of endocrine-treated breast cancer patients.

Citation Format: Daniel Elias Roro, Henriette Vever, Anne Lykkesfeldt, Henrik Ditzel. Global gene expression analysis of tamoxifen-resistant breast cancer cell lines identified a gene signature predictive of clinical outcome of breast cancer patients treated with endocrine therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1887. doi:10.1158/1538-7445.AM2014-1887
Abstract 1888: Target sequencing of papillary renal cell carcinoma, type 2, using custom-made kidney cancer panel
Authors: Kim Ji-Yeon , Lee Se-Hoon , Kim Jong-Il , Shin Jong-Yeon , Heo Dae seog
Published: May 2015

Purpose: Papillary renal cell carcinoma type2 (PRCC2) is 2nd most common kidney malignancy followed by clear cell renal cell carcinoma (CCRCC). PRCC2 has poor prognosis, treatment strategy is not established and the genetic alteration is poorly understood. We sequenced PRCC2 samples using custom-made kidney cancer panel to figure out genetic alterations of PRCC2.

Methods: We have reviewed publications about the genetics of kidney cancer including all subtypes and selected 55 candidate genes. This cancer panel consisted of 1022 regions by Agilent SureSelect Target Enrichment. We sequenced 12 PRCC2 tumor samples along with 6 paired normal tissue samples. The patients' age were ranged from 26 to 82 year-old, and 3 patients were female (25%). Genomic DNA was isolated from dissected tumor tissue samples (6 fresh, frozen and 6 formalin-fixed, paraffin-embedded) and normal tissue samples. Sequencing was performed by Illumina platform and aligned to H. sapiens, hg19, GRCh37. Total probe number was 4103 and size was 283.494 kbp. Presumed germline variants, which were present in matched normal tissue, were filtered out, and manual curation was performed for each area of variants.

Results: Fifty-four of 55 target genes had 100% coverage, in spite of 92.7% coverage of one target gene. We found two novel FH mutations (one single nucleotide variant and one 5bp indel) and one novel NFE2L2 mutation. Two out of 12 samples (17%) were altered in NRF2 pathway (1 NFE2L2 and 1 KEAP) which was already well-known as driver in PRCC2. Novel PBRM1 mutation in two samples and SETD2 mutation in one sample were identified. In addition, PTEN, TSC1, KDM5C and AKT1 mutation are observed in one case, respectively. No VHL mutation in PRCC2 was revealed.

Conclusions: We analyzed somatic mutation of PRCC2 with custom-made kidney cancer panel. We found several candidate driver mutations of PRCC2. Our result reveals that the genetics of PRCC2 is heterogeneous, therefore, the approach using kidney cancer panel could be used to characterize individual PRCC2.

The gene list of RCC panelABCB1BRAFERBB2MDM2NUDT11SDHBTP53ABL1CNTNAP4FHMDM4PBRM1SDHCTSC1AKR1B10CR1FLCNMETPCDHGA8SDHDTSC2AKT1CUL3HIF1AMTORPIK3CASTED2VEGFAALKDACH2KANK3MYCPOTECSIRT1VHLANKRD36EBPLKEAP1NBPF10PTENSPAM1WDR52ARID1AEGFRKRASNFE2L2RETSTAG3L2TSC1BAP1EPAS1KRT1NPNTROS1TERT

Citation Format: Ji-Yeon Kim, Se-Hoon Lee, Jong-Il Kim, Jong-Yeon Shin, Dae seog Heo. Target sequencing of papillary renal cell carcinoma, type 2, using custom-made kidney cancer panel. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1888. doi:10.1158/1538-7445.AM2014-1888
Abstract 1897: Application of a novel DNA methylation analysis method (MSE) for mucin expression in pancreatic juices for diagnosis of pancreatic neoplasms
Authors: Yonezawa Suguru , Yokoyama Seiya , Higashi Michiyo
Published: May 2015

Mucins play crucial roles in diagnostic and prognostic prediction and in carcinogenesis and tumor invasion. In our clinicopathological investigation for mucin expression in human pancreatic ductal adenocarcinomas (PDACs) and intraductal papillary mucinous neoplasms (IPMNs), we had reported that MUC1 (pan-epithelial membrane mucin) or MUC4 (tracheobronchial mambrane mucin) expression is related with the aggressive behavior and a poor outcome of the patients, whereas MUC2 (intestinal secretory mucin) expression tends to be related with the indolent behavior and a favorable outcome of the patients. We investigated the mechanism of mucin genes expression from the DNA methylation standpoint in human cancer cell lines using the mapping of methylated cytosines by the quantitative MassARRAY analysis, followed by the confirmation with MSP analysis and a novel DNA methylation analysis method ‘methylation specific electrophoresis (MSE, international patent open: WO 2011/132798)’, and compared the results with expression levels of mucin core protein and mRNA in the cancer cells. The results obtained showed that MUC1, MUC2 and MUC4 expression is regulated by DNA methylation at their proximal 5′ flanking regions in the cancer cells. The MSE method greatly decreases the amount of input DNA, although conventional analytical methods for DNA methylation require a large amount of DNA and have low sensitivity. The lower detection limit for distinguishing different methylation status is under 0.1% and the detectable minimum amount of DNA is 20 pg, which can be obtained from only a few cells. We applied the MSE method to human pancreatic juices from 45 patients with various pancreatic lesions. The analyses for DNA methylation status of MUC1, MUC2 and MUC4 combined with image data were useful for differential diagnosis of various human pancreatic neoplasms. The examination of 45 samples of human pancreatic juices by the MSE method combined with the image examination showed the following specificity and sensitivity by quadratic discrimination analysis: PDAC, 87% and 80%; gastric-type IPMN (safe type), 88% and 77%; intestinal-type IPMN (dangerous type), 100% and 88%; respectively. The MSE analysis of human pancreatic juices may give us very useful informations for the selection of the treatment methods for pancreatic neoplasms.

Citation Format: Suguru Yonezawa, Seiya Yokoyama, Michiyo Higashi. Application of a novel DNA methylation analysis method (MSE) for mucin expression in pancreatic juices for diagnosis of pancreatic neoplasms. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1897. doi:10.1158/1538-7445.AM2014-1897