bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2851: Reduced Tip60 expression in human gastric cancer
Authors: Chen Guangdi , Bai Rongpan , Zhao Zhongsheng , Zhou Jianwei , Weng Yu
Published: May 2015

The tumor suppressor Tip60 plays a major role in gene transcription, DNA damage response, apoptosis and cancer development, but its role in gastric cancer is unknown. In this study, we investigated the role of Tip60 in gastric cancer pathogenesis and assessed its prognostic value. Using tissue microarrays consisting of 567 cases of gastric cancers (78 for training set and 489 for validation set), we found that Tip60 expression was significantly reduced in advanced gastric cancer compared to early stages of gastric cancer (P < 0.001). Reduced Tip60 was correlated with gastric cancer lymph node metastasis (P < 0.001) although no significant correlation was found between Tip60 expression and gastric cancer distant metastasis. Kaplan-Meier survival curve analysis showed that reduced Tip60 expression was associated with a poorer five-year overall survival in gastric cancer patients from both training set (P = 0.010) and validation set (P < 0.001). Our data indicate that Tip60 may play an important role in gastric cancer metastasis and serve as a potential biomarker for gastric cancer patient outcome.

Note: This abstract was not presented at the meeting.

Citation Format: Guangdi Chen, Rongpan Bai, Zhongsheng Zhao, Jianwei Zhou, Yu Weng. Reduced Tip60 expression in human gastric 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 2851. doi:10.1158/1538-7445.AM2014-2851
Abstract 2854: Clinical significance of inhibin beta A expression in gastric cancer
Authors: Oshima Takashi , Imada Toshio , Yamamoto Naoto , Sato Tsutomu , Yukawa Norio , Shiozawa Manabu , Yoshikawa Takaki , Rino Yasushi , Kunisaki Chikara , Akaike Makoto , Masuda Munetaka
Published: May 2015

Objective: Inhibin beta A (INHBA), a glycoprotein belonging to the TGF-β superfamily, has recently been reported to participate in cancer-cell proliferation. We comprehensively studied the ratio of INHBA gene expression in gastric cancer tissue to that in adjacent normal mucosa by DNA microarray analysis and found that that INHBA gene expression was 12.8-fold higher in gastric cancer tissue. To extend our previous results, we investigated the clinical significance of INHBA gene expression in gastric cancer.

Methods: We studied 227 patients with gastric cancer followed up for at least 5 years postoperatively. The disease stage was I in 29 patients, II in 59, III in 88, and IV in 51. Of the 147 patients with stage II or III disease, 80 postoperatively received S-1 adjuvant chemotherapy, and 67 received surgery alone. We extracted mRNA from frozen specimens of gastric cancer and adjacent normal mucosa from these patients, calculated relative expression levels of the INHBA gene (quantitative PCR analysis), and studied the relations of INHBA expression to clinicopathological factors and treatment outcomes. We also examined the relation between S-1 adjuvant chemotherapy and INHBA gene expression in patients with Stage II or III gastric cancer, for which adjuvant chemotherapy is standard treatment.

Results: INHBA gene expression was significantly higher in gastric cancer than in adjacent normal mucosa (P<0.0001) and was significantly related to the clinicopathological factors of invasion depth, disease stage, and vascular invasion. High INHBA gene expression was significantly related to poor outcomes (P=0.0320). In stage II and III gastric cancer, survival rates did not differ according to INHBA gene expression in patients who underwent surgery alone. In contrast, survival was significantly poorer in gastric cancer with high INHBA gene expression in patients who received S-1 adjuvant chemotherapy (P=0.003).

Conclusions: INHBA gene expression was suggested to be a useful predictor of poor outcomes in patients with resected gastric cancer. In patients given S-1 adjuvant chemotherapy, INHBA gene expression may be a useful biomarker.

Citation Format: Takashi Oshima, Toshio Imada, Naoto Yamamoto, Tsutomu Sato, Norio Yukawa, Manabu Shiozawa, Takaki Yoshikawa, Yasushi Rino, Chikara Kunisaki, Makoto Akaike, Munetaka Masuda. Clinical significance of inhibin beta A expression in gastric 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 2854. doi:10.1158/1538-7445.AM2014-2854
Abstract 2856: Fibroblast growth factor receptor 1 gene amplification as an independent prognostic factor for recurrence in patients with resected squamous cell esophageal cancer
Authors: Kim Hyo Song , Lee Seung Eun , Bae Yoon Sung , Kim Joo Hang , Kim Hye Ryun , Kim Dae Joon , Kim Hyunki , Choi Yoon-La , Cho Byoung Chul
Published: May 2015

Purpose

To evaluate investigate the prognostic role of fibroblast growth factor receptor 1 (FGFR1) amplification in patients with surgically resected esophageal squamous cell carcinoma (ESCC).

Patients and methods

Gene copy number of FGFR1 was investigated in microarrayed tumors from 549 patients with ESCC who underwent curative esophagectomy. Gene copy number was evaluated by fluorescent in situ hybridization, and an FGFR1 amplified tumor (FGFR1 amp+) was prespecified if one of the following criteria is fulfilled: (I) FGFR1/centromer 8 (CEN8) ration is ≥ 2.0, (II) average gene copy nucleus is ≥ 6.0, and (III) percentage of tumor cells containing ≥ 15 gene copies or large cluster is ≥ 10%.

Results

FGFR1 amplification was detected in 8% (44 of 549) of ESCC. The average copy number per nucleus was 2.7 (range, 0-15.5) and the mean FGFR1/CEN8 ratio was 2.2 (range, 0-6). The 5-year disease free survival of the FGFR1 amp+ group was significantly shorter compared with that of the FGFR1amp- group (38.8% vs 63.7% P=0.012). In Cox proportional hazard model, patients with FGFR1 amp+ had a significantly greater risk of recurrence compare to amp- group after adjusting for sex, smoking status, pathologic stage, histology, and adjuvant chemotherapy (HR 1.74; 95% CI, 1.10-2.76, P=0.018). Of note, there was a trend toward worse overall survival for FGFR1 amp+ group compare to amp- group (5-year OS 32.0% vs 50.8%, P=0.11). FGFR1 amp+ group was significantly more likely to be smokers than FGFR1amp- group (97.7 % vs 2.3%, P=0.001). With increment of total cigarette smoking dosage, the incidence of FGFR1 amp+ significantly increased (P trend=0.005).

Conclusions

FGFR1 amplification is an independent prognostic factor for recurrence in surgically resected ESCC. FGFR1 amplification may have potential clinical application to the development of novel therapeutic strategies for ESCC

Citation Format: Hyo Song Kim, Seung Eun Lee, Yoon Sung Bae, Joo Hang Kim, Hye Ryun Kim, Dae Joon Kim, Hyunki Kim, Yoon-La Choi, Byoung Chul Cho. Fibroblast growth factor receptor 1 gene amplification as an independent prognostic factor for recurrence in patients with resected squamous cell esophageal 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 2856. doi:10.1158/1538-7445.AM2014-2856
Abstract 2864: RPS4X, a new prognostic and predictive biomarker of ovarian and breast cancer
Authors: Tsofack Serges P. , Meunier Liliane , Mes-Masson Anne-Marie , Lebel Michel
Published: May 2015

Epithelial ovarian cancer (EOC) is a disease responsible for more cancer deaths among women in the Western world than all other gynecologic malignancies and breast cancer is leading cause of cancer death among the women worldwide. A major problem with such cancers is chemoresistance; hence the need to identify predictive biomarkers. Interestingly, overexpression of YB-1 in ovarian and breast cancer cells induces cisplatin resistance. Platinum coumpounds like cisplatin is often used in the treatments of these cancers. Y box-binding protein 1 (YB-1) is a multifunctional protein that affects transcription, splicing and translation of mRNA. In this study, we used a tagged YB-1 construct to identify by mass spectrometry the proteins that interacted with YB-1 and required for cisplatin resistance. Using the combination of two bioinformatics databases (Oncomine public microarray and genomic hybridization), we focused on the YB-1 protein partners that are potentially involved in cancer progression. We used the siRNA technique to screen different partners of YB-1 that are directly involve in the cisplatin resistance of MCF7 and MDA-MB-231 cells lines. From this analysis, we found that the RPS4X protein, a new partner of YB-1, would be a good marker of cisplatin resistance in breast cancer patients. Interestingly, the YB-1/RPS4X complex was also found in ovarian cancer cells. Like in the breast cancer cell lines, the depletion of RPS4X protein induced the cisplatin resistance of ovarian cancer cells lines (SKOV3 and OVCAR3). Finally, we used a validate antibodies to assess by immunohistochemistry the protein levels of RPS4X and YB-1 in tumor tissue samples from 192 high-grade serous epithelial ovarian cancer patients. We found that RPS4X correlated significantly with ovarian cancer patient outcome.

Taken together, these results suggest that, the YB-1/RPS4X complex is a significant potential target to counteract cisplatin resistance in breast and ovarian cancers. Also, we have established that RPS4X is a new promising prognostic marker for patients with high-grade ovarian cancer. More importantly, RPS4X is shown to be predictive of cisplatin response. Additional immunohistochemistry studies on low-grade ovarian cancers and breast cancers are required to confirm the predictive value of RPS4X.

Citation Format: Serges P. Tsofack, Liliane Meunier, Anne-Marie Mes-Masson, Michel Lebel. RPS4X, a new prognostic and predictive biomarker of ovarian and 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 2864. doi:10.1158/1538-7445.AM2014-2864
Abstract 287: Characterizing the genetic basis of methylome diversity in histologically normal human lung tissue
Authors: Shi Jianxin , Marconett Crystal , Duan Jubao , Hyland Paula , Li Peng , Wang Zhaoming , Wheeler William , Campan Mihaela , Huang Jing , Zhou Weiyin , Triche Tim , Amundadottir Laufey , Hutchinson Amy , Chen Po-Han , Zhou Beiyun , Chung Brian , Bertazzi Pier Alberto , Bergen Andrew W. , Freedman Mathew , Lee Diane , Siegmund Kim , Warner Andrew Clay , Berman Ben , Pesatori Angela C. , Borok Zea , Consonni Dario , Chatterjee Nilanjan , Tucker Margaret , Caporaso Neil , Chanock Stephen J. , Laird-Offringa Ite A. , Landi Maria Teresa
Published: May 2015

The genetic regulation of the human epigenome is not fully appreciated. Here we describe the effects of genetic variants on the DNA methylome in human lung based on methylation-quantitative trait loci (meQTL) analyses. We report 34,304 cis- and 585 trans-meQTLs, a genetic-epigenetic interaction of surprising magnitude, including a regulatory hotspot. These findings are replicated in both breast and kidney tissues and show distinct patterns: cis-meQTLs mostly localize to CpG sites outside of genes, promoters, and CpG islands (CGIs) while trans-meQTLs are over-represented in promoter CGIs. meQTL SNPs are enriched in CTCF binding sites, DNaseI hypersensitivity regions and histone marks. Importantly, 4 of the 5 established lung cancer risk loci in European ancestry are cis-meQTLs and, in aggregate, cis-meQTLs are enriched for lung cancer risk in a genome-wide analysis of 11,587 subjects. Thus, inherited genetic variation may affect lung carcinogenesis by regulating the human methylome.

Citation Format: Jianxin Shi, Crystal Marconett, Jubao Duan, Paula Hyland, Peng Li, Zhaoming Wang, William Wheeler, Mihaela Campan, Jing Huang, Weiyin Zhou, Tim Triche, Laufey Amundadottir, Amy Hutchinson, Po-Han Chen, Beiyun Zhou, Brian Chung, Pier Alberto Bertazzi, Andrew W. Bergen, Mathew Freedman, Diane Lee, Kim Siegmund, Andrew Clay Warner, Ben Berman, Angela C. Pesatori, Zea Borok, Dario Consonni, Nilanjan Chatterjee, Margaret Tucker, Neil Caporaso, Stephen J. Chanock, Ite A. Laird-Offringa, Maria Teresa Landi. Characterizing the genetic basis of methylome diversity in histologically normal human lung tissue. [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 287. doi:10.1158/1538-7445.AM2014-287
Abstract 2870: ZGPAT gene expression in non-tumor hepatocellular carcinoma tissue is a likely biomarker for survival risk
Authors: Hishida Mitsuhiro , Nomoto Shuji , Inokawa Yoshikuni , Takano Nao , Kanda Mitsuro , Iwata Naoki , Tanaka Chie , Kobayashi Daisuke , Nishikawa Yoko , Yamada Suguru , Nakayama Goro , Fujii Tsutomu , Sugimoto Hiroyuki , Koike Masahiko , Fujiwara Michitaka , Kodera Yasuhiro
Published: May 2015

[Background & Aim]

Hepatocellular carcinoma (HCC) often recurs after surgery and multicentric occurrence is more common than intrahepatic metastasis. We therefore examined factors in non-tumor liver tissue in HCC.

[Material & Methods]

Control samples, termed super normal liver tissues (SN), were taken from 11 cases of metastatic liver cancer. One of corresponding samples of normal tissues (CN) was selected for comparison from a typical specimen with chronic hepatitis C and HCC. We performed expression profiling and methylation arrays on these samples, identified genes that differed between SN and CN. We analyzed the identified genes using quantitative real-time reverse-transcriptase polymerase chain reaction on SN from 11 cases of metastatic liver cancer and CN from 109 cases of HCC.

[Results]

We found the zinc finger, CCCH-type with G patch domain (ZGPAT) gene to have expression decreased 4.198-fold in CN, and higher methylation status in CN (0.718), than in SN (0.360). The average expression value of the ZGPAT gene (ZGPAT value ×103 / GAPDH) decreased in CN (9.94 ± 10.68) relative to SN (28.55 ± 22.05). Compared with other patients, those with strongly reduced ZGPAT expression (< 1.5) in their non-tumor liver samples had significantly shorter overall survival (P = 0.024), but not recurrence-free survival (P = 0.515).

[Discussion & Conclusion]

Our results indicate that expression of the ZGPAT gene in non-tumor liver tissue is a good biomarker for the risk of HCC development. ZGPAT gene is reportedly a transcription repressor that negatively regulates expression of epidermal growth factor receptor, which in turn, mediates cell proliferation, survival, migration. In treating HCC, detection of both prognostic factors from non-tumor liver tissue and malignant factors of the primary cancer lesion is important.

Citation Format: Mitsuhiro Hishida, Shuji Nomoto, Yoshikuni Inokawa, Nao Takano, Mitsuro Kanda, Naoki Iwata, Chie Tanaka, Daisuke Kobayashi, Yoko Nishikawa, Suguru Yamada, Goro Nakayama, Tsutomu Fujii, Hiroyuki Sugimoto, Masahiko Koike, Michitaka Fujiwara, Yasuhiro Kodera. ZGPAT gene expression in non-tumor hepatocellular carcinoma tissue is a likely biomarker for survival risk. [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 2870. doi:10.1158/1538-7445.AM2014-2870
Abstract 2875: ATAD2 overexpression indentifies aggressive endometrial carcinomas
Authors: Krakstad Camilla , Tangen Ingvild Løberg , Raeder Maria B. , Hoivik Erling , Kusonmano Kanthida , Zou June X. , Øyan Anne M. , Trovik Jone , Chen Hongwu , Kalland Karl Henning , Salvesen Helga B.
Published: May 2015

Background

Upregulation of the gene ATAD2 has been implicated in cancer development and progression in a number of tissues, and reported to be a member of prognostic gene-signatures in breast (van 't Veer et al, 2002; Wang et al, 2005) and endometrial cancers (Salvesen et al, 2009). We have investigated ATAD2 protein expression in relation to prognosis and gene expression alterations relevant for potential biological driver mechanisms and treatment targets in endometrial cancer.

Methods:

A total of 564 endometrial carcinoma primary lesions were prospectively collected and explored for ATAD2 protein expression in relation to survival and established markers for aggressive disease. Transcriptional alterations related to ATAD2 protein level were investigated by microarray analysis for 236 freshly frozen samples in parallel. Differentially expressed genes were identified using the significance analysis of microarray (SAM) method, and GSEA analysis was performed using the MSigDB version 4.0 C2 collection http://www.broadinstitute.org/gsea/msigdb/index.jsp).

Results:

High ATAD2 expression is significantly associated with the established clinical-pathological variables for aggressive endometrial cancer high FIGO stage, non-endometrioid subtype, high grade, aneuploidy, loss of hormone receptors and poor overall survival (all P-values≤0.001), also in the subset of ERα positive tumors. mRNA and protein expression for ATAD2 was highly correlated (p<0.001), suggesting that immunohistochemical staining may represent a more clinically applicable measure for mRNA levels for ATAD2 level in routinely collected formalin fixed paraffin embedded specimens. Gene expression alterations in samples with high ATAD2 expression revealed upregulation of several cancer-related genes (CDCs, E2Fs) and gene sets that previously has been linked to aggressive disease and potential for targeting new therapies.

Conclusion:

Our results support that immunohistochemical staining for ATAD2 may be a clinically applicable biomarker reflecting clinical phenotype and targetable alterations in endometrial carcinomas to be further explored in controlled clinical trials.

Salvesen HB et al (2009) Proc Natl Acad Sci U S A 106(12): 4834-9

van 't Veer LJ et al (2002) Nature 415(6871): 530-6

Wang Y et al (2005) Lancet 365(9460): 671-9

Citation Format: Camilla Krakstad, Ingvild Løberg Tangen, Maria B. Raeder, Erling Hoivik, Kanthida Kusonmano, June X. Zou, Anne M. Øyan, Jone Trovik, Hongwu Chen, Karl Henning Kalland, Helga B. Salvesen. ATAD2 overexpression indentifies aggressive endometrial carcinomas. [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 2875. doi:10.1158/1538-7445.AM2014-2875
Abstract 2877: Focal adhesion kinsase (FAK) protein overexpression and gene copy number gain correlate with better outcome in patients with surgically resected NSCLC tumors
Authors: Tang Ximing , Cai Yiran , Behrens Carmen , Lin Heather , Lee J. Jack , Wistuba Ignacio. I.
Published: May 2015

Purpose: Cancer cell migration mechanisms are important for invasion and metastasis development. Focal adhesion kinase (FAK) is a critical cancer-related signaling molecule that regulates cell motility and migration. We investigated the effect of tumor FAK protein expression and gene copy number gain in the prognosis of patients with surgically resected non-small cell lung cancer (NSCLC).

Material and Methods: We studied FAK protein expression by immunohistochemistry (IHC) in 216 NSCLC stages I-III archival tumors, including 151 adenocarcinomas (ACs) and 65 squamous cell carcinomas (SCCs). In 190 of these tumors (132 ACs and 58 SCCs), we examined FAK gene copy number by fluorescence in situ hybridization (FISH). FAK IHC expression was examined using the H-score, and a score >150 was defined as overexpression. We examined the correlation of FAK abnormalities with tumors' clinic-pathological and molecular features, and with patients' outcome, including recurrence free and overall survivals.

Results: FAK cytoplasmic protein expression score (P<0.0001) was significantly higher in AC (143 ± 71) compared to SCC (70 ± 42) histology, and in never smokers than smokers. FAK protein overexpression was more frequently (P=0.0005) detected in stage I (46%) than stage II (22%) and III (23%) tumors. In ACs, FAK protein overexpression was significantly higher (P=0.003) in tumors with EGFR mutation (11/13, 85%) compared with wild-type tumors (40/97, 41%). We found that ≥4 FAK gene copies in ≥20% of malignant cells were detected in 28.8% of ACs and 22.4% of SCCs. We did not find correlation between protein expression and gene copy number. After adjusting for stage and adjuvant therapy, NSCLC and AC patients with FAK expression higher than the median had better overall survival (NSCLC, HR 0.51 [CI 0.29-0.86], P=0.017; AC, HR 0.43 [CI 0.23-0.82], P=0.011) compared with patients without protein overexpression. Also, we found that, after adjusting for stage and adjuvant therapy, FAK gene copy number ≥4 in ≥20% of tumors cells correlated with better overall survival (HR 0.34 [CI 0.16-0.95], P=0.039) in patients with AC histology.

Conclusion: FAK protein overexpression and gene copy number gain were frequently detected in NSCLC tumors, and, interestingly, they correlated with better outcome in patients with surgically resected tumors.

Citation Format: Ximing Tang, Yiran Cai, Carmen Behrens, Heather Lin, J. Jack Lee, Ignacio. I. Wistuba. Focal adhesion kinsase (FAK) protein overexpression and gene copy number gain correlate with better outcome in patients with surgically resected NSCLC tumors. [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 2877. doi:10.1158/1538-7445.AM2014-2877
Abstract 2885: Therapeutic microparticle-based tumor lysate vaccination reduces spontaneous lung metastases in a murine breast cancer model
Authors: Gross Brett P. , Wongrapanich Amaraporn , Francis Meghan , Joshi Vijaya B. , Salem Aliasger K. , Norian Lyse A.
Published: May 2015

Although existing interventions are highly efficacious for the treatment of localized breast cancers, disseminated tumors remain incurable. Therapeutic induction of systemic anti-tumor immunity is a promising approach for treating metastatic disease. We have developed a novel heterologous prime/boost vaccination protocol that significantly reduces metastatic tumor burdens in a pre-clinical murine model of breast cancer.

In our studies, mice were orthotopically challenged with Luciferase-expressing 4T1 adenocarinoma cells. Six days later, mice received a priming therapy that consisted of parental 4T1 tumor lysates encapsulated in poly(lactic-co-glycolic) acid microparticles administered contralateral to the established tumor. On day eleven, mice received a boost therapy that consisted of 4T1 lysates and a cocktail of Toll-like receptor agonists. At the conclusion of the assay (day twenty-five), tumor burdens from excised lungs were quantified using bioluminescent imaging. Therapy-receiving mice showed a significant reduction in lung tumors (Student's T-test: p = 0.002, n = 37 control mice vs. 25 treated mice) with an average 46.66% decrease relative to no therapy control mice. Independent quantification of lung metastases was also performed using a standard 6-thioguanine assay. There was a strong correlation (R^2 = 0.8803) between bioluminescent intensity and 6-thioguanine colony number, demonstrating that the decrease in bioluminescence signal due to therapy is not merely explained by the in vivo loss of Luciferase expression by 4T1 cells.

We are currently characterizing the vaccination-induced differences in the localized immune environment of metastatic lung tumors. Effector T cell activation/exhaustion kinetics and the influx of immunosuppressive cell populations (myeloid-derived suppressor cells and regulatory T cells) are now being explored. These studies will help elucidate the immunological mechanism(s) responsible for the diminished metastatic tumor outgrowth observed with our therapy. In conclusion, our data demonstrate that therapeutic vaccination of tumor lysates is efficacious in controlling spontaneous lung metastases in an aggressive model of murine breast cancer.

Citation Format: Brett P. Gross, Amaraporn Wongrapanich, Meghan Francis, Vijaya B. Joshi, Aliasger K. Salem, Lyse A. Norian. Therapeutic microparticle-based tumor lysate vaccination reduces spontaneous lung metastases in a murine breast cancer model. [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 2885. doi:10.1158/1538-7445.AM2014-2885
Abstract 2890: Mechanistic evaluation of the anti-tumor activities of 2-fluorofucose alone and in combination with anti-idiotype vaccination
Authors: Okeley Nicole M. , Zeng Weiping , Levengood Matthew R. , McEarchern Julie A. , Jonas Mechthild , Duniho Steven , Senter Peter D. , Law Che L.
Published: May 2015

Fucose is a sugar that is widely incorporated in cellular glycoconjugates and has been implicated for roles in tumor progression and metastasis. An analog of fucose, 2-fluorofucose, has been shown to have activity in vitro and in vivo as an inhibitor of glycoconjugate fucosylation and has shown promising antitumor activity in cancer growth models as well as potentiation of the protective effect of a mouse B cell lymphoma idiotype vaccine against challenge by live A20 lymphoma cells (Okeley N.M. et al, PNAS 110, 5404-5409, 2013). The effects of 2-fluorofucose that mediate these activities was explored in vitro and in vivo. In vitro, we found that 2-fluorofucose is rapidly converted to GDP-2-fluorofucose, the active species that inhibits the de novo pathway for fucosylation of protein glycans. The molecule accumulates within cancer cells, and is retained with a half-life in the range of 20 hours. In vivo, evaluation of the effect of 2-fluorofucose treatment on the fucosylation and proliferation state of xenografts showed reduced LCA lectin binding by western blot and IHC, and reduced Ki-67 staining, suggesting a lower mitotic index in these tumors. Adoptive transfer experiments were undertaken in order to shed light onto potential mechanisms for the antitumor effects of 2-fluorofucose when used in a cancer vaccine setting. These studies were performed in mice that were immunized with an A20 idiotype vaccine in the presence or absence of 2-fluorofucose, prior to tumor challenge. The antitumor effects obtained when serum from 2-fluorofucose treated animals was injected into non-vaccinated mice suggested a role for defucosylated antibodies with enhanced ADCC activities. In addition, T cell depletion studies demonstrated that 2-fluorofucose stimulated a CD4+ T cell involvement in in the protection against tumor engraftment. These results suggest multifactorial roles for the activities of 2-fluorofucose as an anticancer agent and as a stimulator of vaccine activity.

Citation Format: Nicole M. Okeley, Weiping Zeng, Matthew R. Levengood, Julie A. McEarchern, Mechthild Jonas, Steven Duniho, Peter D. Senter, Che L. Law. Mechanistic evaluation of the anti-tumor activities of 2-fluorofucose alone and in combination with anti-idiotype vaccination. [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 2890. doi:10.1158/1538-7445.AM2014-2890
Abstract 2895: Small cell lung cancer cells express the late stage gBK tumor antigen: a possible immunotarget for the terminal disease
Authors: Hoa Neil T. , Ge Lisheng , Tajhya Rajeev B. , Beeton Christine , Cornforth Andrew N. , Abolhoda Amir , Lambrecht Nils , Dacosta-Iyer Maria , Ouyang Yi , Hickey Michelle J. , Erickson Kate L. , Kruse Carol A. , Jadus Martin R.
Published: May 2015

Big Potassium (BK) ion channels have several splice variants. One splice variant initially described within human glioma cells is called the glioma BK channel (gBK). This isoform consists of 33 amino acids inserted into the intracellular region of the BK ion channel. Using a gBK-specific antibody, we detected gBK within 3 human small cell lung cancer (SCLC) lines. Patch clamping revealed that functional membrane channels were found on the SCLC cells. Prolonged exposure to BK channel activators caused the SCLC cells to swell within 20 minutes and resulted in their cytotoxicity within 5 hours. Transfection of BK-negative HEK cells with gBK produced functional gBK channels. Quantitative RT-PCR analysis using primers specific for gBK, but not with a lung-specific marker, Sox11, confirmed that advanced, late-stage human SCLC tissues strongly expressed gBK mRNA. Normal human lung tissue and early, lower stage SCLC resected tissues very weakly expressed this transcript. Immunofluorescence using the gBK antibodies confirmed that SCLC cells taken at the time of autopsy intensely displayed this protein. gBK may represent a late-stage marker for SCLC. HLA-A*0201 restricted human CTL were generated in vitro by one-way mixed lymphocyte reactions or mixed lymphocyte dendritic cell reactions, the latter using gBK peptide pulsed dendritic cells. The exposure of SCLC cells to interferon-γ (IFN-γ) increased the expression of HLA; these treated cells were killed by the CTL better than non-IFN-γ treated cells even though the IFN-γ treated SCLC cells displayed diminished gBK protein expression. Prolonged incubation with recombinant IFN-γ slowed the in vitro growth and migration of the SCLC cells, suggesting it might inhibit tumor growth in vivo. Immunotherapy targeting gBK might possibly impede advancement to terminal state of SCLC.

Citation Format: Neil T. Hoa, Lisheng Ge, Rajeev B. Tajhya, Christine Beeton, Andrew N. Cornforth, Amir Abolhoda, Nils Lambrecht, Maria Dacosta-Iyer, Yi Ouyang, Michelle J. Hickey, Kate L. Erickson, Carol A. Kruse, Martin R. Jadus. Small cell lung cancer cells express the late stage gBK tumor antigen: a possible immunotarget for the terminal disease. [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 2895. doi:10.1158/1538-7445.AM2014-2895
Abstract 2901: Cell surface-associated anti-MUC1-derived signal peptide antibodies: implications for cancer diagnostics and therapy
Authors: Kovjazin Riva , Horn Galit , Smorodinsky Nechama I. , Shapira Michael Y. , Carmon Lior
Published: May 2015

The MUC1 tumor associated antigen is highly expressed on a range of tumors. Its broad distribution on primary tumors and metastases renders it an attractive target for immunotherapy. After synthesis MUC1 is cleaved, yielding a large soluble extracellular alpha subunit containing the tandem repeats array (TRA) domain specifically bound, via non-covalent interaction, to a smaller beta subunit containing the transmembrane and cytoplasmic domains. Thus far, inconclusive efficacy has been reported for anti-MUC1 antibodies directed against the soluble alpha subunit. Targeting the cell bound beta subunit, may bypass limitations posed by circulating TRA domains. MUC1's signal peptide (SP) domain promiscuously binds multiple MHC class II and Class I alleles, which upon vaccination, generated robust T-cell immunity against MUC1-positive tumors. This is a first demonstration of non-MHC associated, MUC1 specific, cell surfaces presence for MUC1 SP domain. Polyclonal and monoclonal antibodies generated against MUC1 SP domain specifically bind a large variety of MUC1-positive human solid and hematological tumor cell lines; MUC1-positive bone marrow derived plasma cells obtained from multiple myeloma (MM)-patients, but not MUC1 negative tumors cells, and normal naive primary blood and epithelial cells. Membranal MUC1 SP appears mainly as an independent entity but also co localized with the full MUC1 molecule. MUC1-SP specific binding in BM-derived plasma cells can assist in selecting patients to be treated with anti-MUC1 SP therapeutic vaccine, ImMucin. A therapeutic potential of the anti-MUC1 SP antibodies was suggested by their ability to support of complement-mediated lysis of MUC1-positive tumor cells but not MUC1 negative tumor cells and normal naive primary epithelial cells. Moreover in preliminary studies, anti-MUC1 SP antibodies delayed tumor growth of murine DA-3 cells transfected with human MUC1. These findings suggest a novel cell surface presence of MUC1 SP domain, a potential therapeutic benefit for anti-MUC1 SP antibodies in MUC1-positive tumors and a selection tool for MM patients to be treated with the anti-MUC1 SP vaccine, ImMucin.

Note: This abstract was not presented at the meeting.

Citation Format: Riva Kovjazin, Galit Horn, Nechama I. Smorodinsky, Michael Y. Shapira, Lior Carmon. Cell surface-associated anti-MUC1-derived signal peptide antibodies: implications for cancer diagnostics and 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 2901. doi:10.1158/1538-7445.AM2014-2901
Abstract 2903: IMAB362, a novel first-in-class monoclonal antibody for treatment of pancreatic cancer
Authors: Tuereci Oezlem , Woell Stefan , Jacobs Stefan , Mitnacht-Kraus Rita , Sahin Ugur
Published: May 2015

Pancreatic ductal adenocarcinoma (PDAC), the most frequent subtype (>80%) of pancreatic cancer (PC) is characterized by a generally lethal progress within a short period of time after primary diagnosis. Despite high efforts, the treatment options are very limited and mainly of palliative nature.

Therefore, we investigated whether IMAB362 might represent a potential treatment option in this patient population. IMAB362 is a highly potent and tumor-cell selective therapeutic antibody which is currently in clinical development in gastro-esophageal cancer (in phase II and IIb trials). IMAB362 is directed against the tight junction molecule CLDN18.2, a proliferation-promoting transmembrane protein, which is maintained during malignant transformation. Our previous studies demonstrated, that CLDN18.2 is expressed in >55% of patients with PDAC and frequently in metastases at considerable levels. To evaluate the anti-tumor activity of IMAB362 we performed in vitro and in vivo experiments on CLDN18.2 expressing PC cell lines.

IMAB362 exerts its anti-tumor function by antibody-dependant cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). ADCC was assessed in various cancer cell lines and peripheral blood mononuclear cells (PBMCs) at an E:T-ratio of 40:1 in the presence of IMAB362. The efficacy ranged from 67.1 ± 6.8 to 92.6 ± 5.5%. EC50 was determined between 23.4 ± 12.3 ng/ml and 5.7 ± 4.4 µg/ml. IMAB362's capacity to induce CDC was analyzed by incubation of various tumor cell lines with healthy human serum as source for complement. We observed a dose-dependent lysis with EC50 values ranging from 0.3 - 2.6 µg/ml.

In vivo studies in PC tumor bearing nude mice showed that administration of repeated doses of IMAB362 tends to result in inhibition of tumor growth and prolonged survival. Also, the anti-tumor effect of IMAB362 was strictly dependent on the expression of its target CLDN18.2. These results demonstrate that IMAB362 has antitumoral activity in vitro and in vivo against PC cell lines and PC xenografts. Most promising, the efficacy was due to IMAB362's single agent activity only. Combinations with established chemotherapies (e.g. 5-FU, gemcitabine) might deliver a strong rationale for clinical testing of the highly selective monoclonal antibody drug in patients with PDAC. Results from further in vivo testing of IMAB362 combinations with established chemotherapies will be presented at the meeting.

Citation Format: Oezlem Tuereci, Stefan Woell, Stefan Jacobs, Rita Mitnacht-Kraus, Ugur Sahin. IMAB362, a novel first-in-class monoclonal antibody for treatment of pancreatic 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 2903. doi:10.1158/1538-7445.AM2014-2903
Abstract 2924: MAPK pathway inhibition enhances the efficacy of an anti-endothelin B receptor drug conjugate by inducing target expression in melanoma
Authors: Asundi Jyoti , Polakis Paul , Lacap Jennifer A. , Nannini Michelle
Published: May 2015

Therapies targeting the MAP kinase pathway in melanoma have produced significant clinical responses; however, duration of response is limited by acquisition of drug resistance. Rationale drug combinations may improve outcomes in this setting. We assessed the therapeutic combination of an antibody drug conjugate (ADC) targeting the Endothelin B receptor (EDNRB) with small molecule inhibitors of the MAP kinase signaling pathway in melanoma. Cell lines and tumor models containing either mutant BRAF or NRAS, or wildtype for both, were exposed to small molecule inhibitors or BRAF and MEK. Expression of EDNRB was analyzed and the therapeutic impact of combining the anti-EDNRB ADC with the BRAF and MEK inhibitors was assessed. Increased expression of EDNRB in response to inhibition of BRAF and/or MEK was observed and augmented the antitumor activity of the ADC. Enhanced target expression and ADC anti-tumor activity were realized irrespective of the response of the tumor model to the BRAF or MEK inhibitors alone and could be achieved in melanoma with mutant NRAS, BRAF or neither mutation. Cells that acquired resistance to BRAF inhibition through long-term culture retained drug-induced elevated levels of EDNRB expression. Expression of EDNRB was not enhanced in normal human melanocytes by inhibition of BRAF and the combination of the ADC with MAPK inhibitors was well tolerated in mice. The anti-EDNRB ADC combines well with BRAF and MEK inhibitors and could have therapeutic utility in the majority of human melanoma cases.

Citation Format: Jyoti Asundi, Paul Polakis, Jennifer A. Lacap, Michelle Nannini. MAPK pathway inhibition enhances the efficacy of an anti-endothelin B receptor drug conjugate by inducing target expression in melanoma. [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 2924. doi:10.1158/1538-7445.AM2014-2924
Abstract 2927: Rapamycin decreases expression of thymidylate synthase and enhances the response to pemetrexed in preclinical studies and a Phase I/II clinical study of subjects with non-small cell lung cancer
Authors: Kawabata Shigeru , Chiang Chun-Te , Memmott Regan M. , Komiya Takefumi , Gills Joell J. , Dennis Phillip A.
Published: May 2015

Non-small-cell lung cancer (NSCLC) accounts for 80-85% of lung cancers and almost half of patients with newly diagnosed NSCLC have metastatic disease. Platinum-based chemotherapy regimens given as first-line treatment for advanced NSCLC patients with metastasis are standard but have modest response rates. Pemetrexed is well tolerated and inhibits several folate-dependent enzymes including thymidylate synthase, TS. Increased expression of TS confers resistance to pemetrexed in vitro and is a predictive factor for poor response to pemetrexed. Rapamycin is a mTOR inhibitor and suppresses protein synthesis. Here, we show that rapamycin decreases endogenous expression of TS in NSCLC cells. The combination of rapamycin and pemetrexed synergistically inhibits NSCLC cell proliferation and enhances the inhibition of 4E-binding protein 1 (4E-BP1) activation that is regulated by mTOR. Although pemetrexed as a single agent upregulated expression of TS, pretreatment with rapamycin suppressed pemetrexed-upregulated TS expression. In vivo, the combination of rapamycin and pemetrexed inhibited the growth of NSCLC xenografts, which correlated with decreased mTOR activity and inhibition of pemetrexed-induced upregulated expression of TS. Based on these preclinical data, we have conducted a Phase I/II clinical trial combining rapamycin and pemetrexed in subjects with relapsed NSCLC. This regimen was well tolerated, and resulted in a 22% partial response rate that is above what is expected for response to single agent pemetrexed (9%). In biomarker analyses, pretreatment with rapamycin decreased the expression of TS in peripheral blood mononuclear cells, which correlated with increased progression free survival. Collectively, these studies identify rapamycin and pemetrexed as an effective combination in NSCLC, and suggest that rapamycin enhances the efficacy of pemetrexed by suppressing TS expression.

Citation Format: Shigeru Kawabata, Chun-Te Chiang, Regan M. Memmott, Takefumi Komiya, Joell J. Gills, Phillip A. Dennis. Rapamycin decreases expression of thymidylate synthase and enhances the response to pemetrexed in preclinical studies and a Phase I/II clinical study of subjects with non-small cell 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 2927. doi:10.1158/1538-7445.AM2014-2927
Abstract 293: High-throughput methylation sequencing of targeted genes in breast cancer specimens using nanofluidic PCR prepared libraries
Authors: Wang Jun , Li Zibo , Wu Yepeng , Guo Xinwu , Li Shengyun , Xiao Zhi , Chen Feiyu , Deng Zhongping , Dai Lizhong , Yi Wenjun , Tang Lili
Published: May 2015

Breast cancer is a global public health issue as it is the most frequently diagnosed malignancy in women in the Western world with over a million new cases every year worldwide. Previous epigenetic analyses have identified aberrant DNA methylation signatures associated with breast cancer. Methylation alterations resulting in aberrant gene expression are key contributors to breast tumorigenesis. There are more than 100 candidate genes reported throughout the literatures as promoter hypermethylated in breast cancers (Pubmeth web resource). More recently, with the application of genome wide array-based methylation profile analysis and next generation sequencing, hundreds of new methylation candidate genes have been reported to be associated with breast cancer. It is crucial to develop an approach for targeted methylaton resequencing to validate these candidate genes in hundreds of patient samples and develop a breast cancer detection panel for early diagnostics.

We have developed a targeted gene bisulfite-sequencing approach using a nanofluidic platform, the Access Array™ system, which enables simultaneous amplification of 48 genetic regions (amplicon) from 48 samples in parallel. We have selected 48 breast cancer associated methylation candidate genes, each commonly reported by multiple literatures. Bisulfite-specific-sequencing (BSP) primers were designed for the promoter regions of these genes. In addition, we have applied a primer design strategy that incorporates sample-specific barcodes and Illumina (Miseq) sequencing adaptors into each amplicon, removing the need for additional library preparation before sequencing. We will present methylation sequencing data collected from amplicon libraries generated using the Access Array system. The 48 promoter regions have been amplified in paired tumor/normal tissues and plasma DNA from 48 breast cancer patients. Our results indicate the excellent utilization of this approach in the validation of methylation biomarkers and selection of detection panel for breast cancer.

Citation Format: Jun Wang, Zibo Li, Yepeng Wu, Xinwu Guo, Shengyun Li, Zhi Xiao, Feiyu Chen, Zhongping Deng, Lizhong Dai, Wenjun Yi, Lili Tang. High-throughput methylation sequencing of targeted genes in breast cancer specimens using nanofluidic PCR prepared libraries. [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 293. doi:10.1158/1538-7445.AM2014-293
Abstract 2935: RNAi screen identifies therapeutic targets in hepatocellular carcinoma
Authors: Huang Chun-Hao , Lujambio Amaia , Zuber Johannes , Kitzing Thomas , Tschaharganeh Darjus F. , Stanchina Elisa D. , Lowe Scott W.
Published: May 2015

Liver cancer is the third leading cause of cancer related mortality worldwide. Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is a very aggressive type of cancer that lacks effective treatment, in part, due to the undruggable nature of its main genetic drivers, such as oncogene MYC amplification or loss of tumor suppressor TP53. The introduction of the multikinase inhibitor Sorafenib represents the biggest therapeutic advance in the past decade, though its activity only increases the life expectancy of liver cancer patients from 8 to 11 months. To identify new therapeutic targets for HCC, we employ RNA interference (RNAi) screens as a non-biased means to identify genes required for tumor maintenance in genetically defined tumors. In the current study, we applied an optimized shRNA platform to perform a negative selection screen in genetically-defined murine hepatocellular carcinoma cell in vitro. Specifically, we have designed a library that contains 2,245 shRNAs targeting 442 “drugged” genes, including genes encoding proteins targeted by FDA-approved drugs, small molecules in clinical trials, or compounds under preclinical development. Countersceening and validation was performed in normal murine cells and human HCC cell lines and xenograft using shRNAs and corresponding small molecular drugs. Pharmacological or shRNA-mediated inhibition of certain targets led to robust antitumor effects, both in vitro and in vivo, and suggest new strategies to inhibit Myc function therapeutically. Our results highlight the utility of RNAi screening for studying cancer vulnerabilities that can be exploited for direct pharmacological intervention.

Citation Format: Chun-Hao Huang, Amaia Lujambio, Johannes Zuber, Thomas Kitzing, Darjus F. Tschaharganeh, Elisa De Stanchina, Scott W. Lowe. RNAi screen identifies therapeutic targets in hepatocellular carcinoma. [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 2935. doi:10.1158/1538-7445.AM2014-2935