bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1302: Genetic analysis in a patient with nine primary malignant neoplasms: A rare case of Li-Fraumeni syndrome
Authors: Li Xiaoyuan , Kang Juan , Pan Qi , Yin Wenwei , Tong Shiwen , Zhao Dachun , Meng Changting , Bai Chunmei , Ren Hong , Ding Keyue 
Published: May 2015

Abstract
The identification of patients with hereditary cancer predisposition syndromes has important clinical implications for early diagnosis, prevention, and treatment. We applied whole exome sequencing to identify mutations underlying a patient diagnosed with Li-Fraumeni syndrome, who had a first-degree family antecedent of breast cancer at age 42 years. Nine primary metachronous malignant neoplasms were developed in this patient within a period of 38 years, including upper lip neuroblastoma (7 years) and rhabdomyosarcoma (7 years), breast cystosarcoma phyllodes carcinoma (28 years), bronchioalveolar cell carcinoma (29 years), breast infiltrating ductal carcinoma (39 years), and thyroid papillary carcinoma (39 years), gastric adenocarcinoma (41 years), maxillary (41 years) and sternal osteosarcoma (43 years), as well as acute myeloid leukemia (45 years). Peripheral blood lymphocyte DNA from the patient was obtained. Exome capture was performed using the SureSelect Human All Exon 51Mb System, and paired-end sequencing was performed using Illumina HiSeq90 platform. A bioinformatics pipeline was used to analyze sequencing reads and call single nucleotide variants (SNVs). We used a candidate strategy to analyze mutations in the case of only a single dominantly affected individual available. Whole-exome sequencing of peripheral blood lymphocyte DNA resulted in an output of 30 GB of raw sequence. After quality control, we obtained clean reads with an average depth of coverage of 105× and 99.4% exome regions were covered at least four times. Analysis of 451,409 SNVs revealed increased genetic risk for some cancers, including genes related to hereditary cancer disposition syndromes [e.g., TP53 (p.P72R and p.R273H), BRCA1, and BRCA2], and genes associated with sporadic cancers. Pathway analysis found that, of 68 genes in p53-signaling pathway, the number of mutated genes (n=14) was significantly higher than expected (P=0.02). Of novel mutations, five nonsense mutations in four genes and 242 missense mutations in 233 genes were discovered. Whole-exome sequencing can identify known and/or novel mutations in cancer susceptibility genes and establish mutation profile in a rare case of Li-Fraumeni syndrome.
Citation Format: Xiaoyuan Li, Juan Kang, Qi Pan, Wenwei Yin, Shiwen Tong, Dachun Zhao, Changting Meng, Chunmei Bai, Hong Ren, Keyue Ding. Genetic analysis in a patient with nine primary malignant neoplasms: A rare case of Li-Fraumeni syndrome. [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 1302. doi:10.1158/1538-7445.AM2014-1302
Abstract 1303: Genetic cancer incidence
Authors: Kim Min Kyu , Bae Soo Youn , Lee Jee Yeon 
Published: May 2015

Abstract
Objective: Hereditary Breast Ovarian Cancer (HBOC) is genetic malignancy associated with BRCA1/2 genes. Selecting confirmative tests for target population are not clearly defined and expensive. We undertook this study to find the detection rate of HBOC among Korean ovarian carcinoma patients based on family history and immunohistochemistry right after by surgeon.
Methods: This study examined 22 patients diagnosed with ovarian carcinomas by single surgeon and counselor. All patients were provided genetic counseling based on immunohistochemistry (IHC) of mismatch repair genes (BRCA1, BRCA2) and 1st, 2nd 3 rd relatives' family history of cancer. Additionally, direct full sequencing to verify germ line mutations was performed to all patients after consent.
Results: Median age was 54.Preop CA 125 was 1824(37.71∼16009). There were 3 refusals after counseling. Patients with family cancer history were 3/21(14%). The number of patients with negative IHC results for BRCA1/BRCA2 was 9 and 6.Both negative patients were 3/21(14%). Both normal staining patients were 4/21(19%).
There were 3/19 patients (16%) with germ line mutations (two BRCA1 and one BRCA2).Variation of unknown significance mutation patients were 5/19(26%).
Conclusion: Our data indicate approximately 17% of patients have a germ line mutation in this study group. Active genetic counseling by surgeon with IHC and family history can help detect HBOC among ovarian carcinoma patients.
Citation Format: Min Kyu Kim, Soo Youn Bae, Jee Yeon Lee. Genetic cancer incidence. [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 1303. doi:10.1158/1538-7445.AM2014-1303
Abstract 1312: Buformin, an anti-diabetic biguanide, inhibits proliferation, invasion and adhesion, and acts synergistically with paclitaxel in endometrial cancer cell lines
Authors: Kilgore Joshua , Jackson Amanda , Qiu Haifeng , Zhou Chunxiao , Gehrig Paola , Bae-Jump Victoria 
Published: May 2015

Abstract
Objectives: Epidemiologic and pre-clinical studies suggest that anti-diabetic biguanide drugs, such as metformin and buformin, may have anti-tumorigenic effects and behave as chemosensitizers. Thus, we evaluated the effects of buformin alone and in combination with paclitaxel on proliferation in human endometrial cancer (EC) cell lines as well as the impact of buformin on apoptosis and adhesion/invasion.
Methods: The ECC-1 and Ishikawa EC cell lines were treated with varying doses of buformin alone and in combination with paclitaxel. Cell growth was determined by MTT assay. Cell cycle progression was assessed by Cellometer. Apoptosis was evaluated by Annexin V-FITC assay. Invasion was demonstrated by transwell invasion assay. Adhesion was assessed by ELISA. Effects on AMPK and S6 expression were documented by Western blotting.
Results: Buformin inhibited proliferation in a dose-dependent manner in the EC cell lines (IC50 1000 μM for ECC-1, 10μM for Ishikawa, p=0.00001-0.0010). Treatment with buformin resulted in G1 arrest in the ECC-1 cell line and G2 arrest in the Ishikawa cell line. Buformin induced apoptosis in the Ishikawa cell line, but not in the ECC-1 cell line. Treatment with buformin resulted in inhibition of adhesion (p=0.42-0.0009) and invasion (p=0.00001-0.227) in both EC cell lines. Western blotting analysis demonstrated that buformin increased phosphorylation of AMPK and decreased phosphorylation of S6. Paclitaxel inhibited proliferation in a dose-dependent manner in both cell lines with IC50 values of 0.1-0.5 nM and 1-5 nM for Ishikawa and ECC-1 cells, respectively. To assess synergy of paclitaxel and buformin, the combination index (CI) was calculated by the method of Chou and Talalay. Simultaneous exposure of cells to various doses of paclitaxel in combination with buformin (0.1-1 μM) resulted in a significant synergistic anti-proliferative effect (CI &lt;1, range 0.009-0.85).
Conclusions: Buformin inhibited proliferation in EC cell lines, via AMPK activation and mTOR pathway inhibition. Inhibition of adhesion/invasion was also seen with buformin. Buformin acted synergistically with paclitaxel to inhibit cell proliferation. More work is needed to determine if buformin and other biguanides alone or in combination with cytotoxic agents will be beneficial in the treatment of women with EC.
Citation Format: Joshua Kilgore, Amanda Jackson, Haifeng Qiu, Chunxiao Zhou, Paola Gehrig, Victoria Bae-Jump. Buformin, an anti-diabetic biguanide, inhibits proliferation, invasion and adhesion, and acts synergistically with paclitaxel in endometrial 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 1312. doi:10.1158/1538-7445.AM2014-1312
Abstract 1315: CtIP is regulated by the APC/C-Cdh1 to mediate cell cycle-dependent control of DNA repair
Authors: Boer Harmen R. de , Lafranchi Lorenzo , Neugebauer Christine , Fehrmann Rudolf S.N. , Vries Elisabeth G.E. de , Sartori Alessandro A. , Vugt Marcel A.T.M. van 
Published: May 2015

Abstract
Human cells have evolved elaborate mechanisms for responding to DNA damage to maintain genome stability and prevent carcinogenesis. For instance, the cell cycle can be arrested at different stages to allow time for DNA repair. The APC/C-Cdh1 ubiquitin ligase regulates mitotic exit but is also implicated in the DNA damage-induced G2 arrest. However, it is unknown whether APC/C-Cdh1 also directly participates in DNA repair. Here we show that depletion of Cdh1 in untransformed RPE-1 cells or several cancer cell lines results in increased levels of genomic instability as well as enhanced sensitivity to DNA double-strand break (DSB)-inducing agents. Moreover, Cdh1 depletion results in aberrant homology-directed repair. Using an integrated SILAC proteomics and bioinformatics approach in RPE-1 cells, we identified CtIP as a novel APC/C-Cdh1 target. CtIP interacts with Cdh1 through a conserved KEN box, of which mutation prevents CtIP ubiquitination and impedes CtIP down-regulation both during G1 and after DNA damage in G2. Importantly, the inability of CtIP to interact with Cdh1 causes defective homology-directed repair. Combined, these results show that APC/C-Cdh1 safeguards genome integrity by controlling CtIP stability in a cell cycle- and DNA damage-dependent manner. Supported by European Research Councel grant -2011-293445; Dutch Cancer Society grant RUG-2011-5093; the Vontobel Stiftung and Swiss National Science Foundation grant 31003A_135507.
Citation Format: Harmen R. de Boer, Lorenzo Lafranchi, Christine Neugebauer, Rudolf S.N. Fehrmann, Elisabeth G.E. de Vries, Alessandro A. Sartori, Marcel A.T.M. van Vugt. CtIP is regulated by the APC/C-Cdh1 to mediate cell cycle-dependent control of DNA repair. [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 1315. doi:10.1158/1538-7445.AM2014-1315
Abstract 1319: Vulnerability and resistance in glioblastoma to G2-specific drugs
Authors: Dhruv Harshil D. , Nelson Andrew D. , Armstrong Brock , Boerner Julie L. , Sarkaria Jann N. , Tran Nhan L. , Berens Michael 
Published: May 2015

Abstract
Glioblastoma multiforme (GBM) is the most common and most lethal primary malignant brain tumor, affecting 25,000 patients per year. Approximately 87% of GBMs have abnormalities in the p53 pathway that affects G2 checkpoint. In the absence of p53, Wee1 kinase has been shown to regulate G2 checkpoint upon chemotherapeutic insult. Wee1 inhibits CDC2 function, thereby promoting G2 phase cell cycle arrest, thus preventing damaged cells from entering into premature mitosis without repairing the DNA. We hypothesize that in p53-deficient tumor cells, inhibitors of Wee1 would be effective to heighten DNA damage due to genetic and pharmacologic loss of both the G1 and G2 checkpoints, respectively. MK-1775 (Merck) is a potent and selective small molecule inhibitor of the Wee1 kinase that has demonstrated enhanced cytotoxic effect in combination with several chemotherapeutic agents in preclinical studies in vitro and in vivo. Here, we selected an array of short term cultures of glioma patient derived xenograft (PDX) models (GBM12, GBM43, GBM79, GBM84) and long term established glioma cell lines (U87, SF767, T98G) with aberrant and wild type G1 checkpoint pathways and investigated cytotoxic effects of MK-1775 as a single agent and in combination with Temozolomide (TMZ). Our results demonstrated differential sensitivity to MK-1775 and TMZ as single agents for different glioma cell lines in vitro. However, no correlation between MK-1775, TMZ sensitivity and p53 status of the cell lines was observed. Combination therapy with MK-1775 and TMZ showed a synergistic effect for one PDX model, GBM43, with combination index (CI) of 0.4. Combination therapy of all other glioma cell lines either showed an additive effect or antagonistic effect with CI of &gt;0.75 irrespective of their p53 status. Our results demonstrated that synergistic effects of MK-1775 in combination with chemotherapy (TMZ) are not solely dependent on p53 status of glioma cells and require a more comprehensivce understanding of the molecular mechanism that dictates GBM's dependence on Wee1 for escaping DNA damage therapy. Supported by the Ben & Catherine Ivy Foundation
Citation Format: Harshil D. Dhruv, Andrew D. Nelson, Brock Armstrong, Julie L. Boerner, Jann N. Sarkaria, Nhan L. Tran, Michael Berens. Vulnerability and resistance in glioblastoma to G2-specific drugs. [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 1319. doi:10.1158/1538-7445.AM2014-1319
Abstract 1327: Targeting the RB-pathway in sarcoma: Utility of CDK4/6 inhibitors
Authors: Francis Ashleigh M. , Carey Jason P. , Alexander Angela , Keyomarsi Khandan , Hunt Kelly K. 
Published: May 2015

Abstract
Background: Sarcomas are rare neoplasms of mesenchymal origin. Survival rates have not improved with standard chemotherapy; however, molecular insights have posed new drug targets. Aberrations in the RB-pathway, including loss of RB, CDK4 and cyclin D1 overexpression and p16 loss, have been identified in sarcomas. PD 03322991 (PD) is a second generation CDK4/6 inhibitor with the potential to reactivate the RB-pathway and suppress cell cycle progression. We evaluated the efficacy of PD in a panel of sarcoma cell lines.
Methods: Sarcoma cell lines (HT-1080, SK-LMS-1 and SaOS-2) were evaluated for RB-pathway alterations using western blot. Cells were treated with PD for 1, 2, 4, 6 and 8 days to determine IC50 values by MTT assay. Following the same treatment protocol at IC50 concentrations of PD, cells were harvested for cell cycle analysis, western blot and annexin cell death assay.
Results: HT-1080 and SK-LMS-1 were RB-positive with high expression of phospho-RB; SaOS-2 was RB-negative. CDK4/6 were uniformly expressed. HT-1080 and SK-LMS-1 had p16 loss while SaOS-2 overexpressed cyclin D1. PD was effective in all cell lines. Increasing treatment duration led to reduction of IC50 values. PD caused marked G1 arrest as early as day 1 in RB-positive HT-1080 (86.2% vs 74.2% [vehicle]) and SK-LMS-1 (91.4% vs 67.8% [vehicle]) but not RB-negative SaOS-2 (52.9-59.4% vs 57.0% [vehicle]). Annexin assay demonstrated minimal apoptosis with PD (HT-1080: 1.4-3.0% vs 41.2% [control], SK-LMS-1: 0.6-12.0% vs 23.4% [control], SaOS-2: 3.6-7.3% vs 78.7% [control]).
Conclusions: PD appears to have a cytostatic rather than cytotoxic effect in sarcoma. RB-negativity does not preclude PD efficacy but may require higher IC50 values. In RB-negative sarcomas, PD does not cause cell cycle arrest suggesting an alternate mechanism from reactivation of the RB-pathway is present. PD may serve as a potential therapy for sarcoma and should be studied in combination with other targeted therapies that can take advantage of its ability to cause G1 arrest.
Cell LinePD 991 IC50 Concentration (uM)Day 1Day 2Day 4Day 6Day 8HT-1080∼∼4.31.51.1SK-LMS-1∼∼∼2.41.2SaOS-2∼15.510.17.06.0
Citation Format: Ashleigh M. Francis, Jason P. Carey, Angela Alexander, Khandan Keyomarsi, Kelly K. Hunt. Targeting the RB-pathway in sarcoma: Utility of CDK4/6 inhibitors. [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 1327. doi:10.1158/1538-7445.AM2014-1327
Abstract 1330: Immunotherapy targeting folate receptor induces autophagy in ovarian cancer
Authors: Wen Yun-Fei , Graybill Whitney Spannuth ,   Department of Gyneocologic Oncology, Sood Anil 
Published: May 2015

Abstract
Objective Cancer cells are highly dependent on folate metabolism for replication of DNA, making these cells susceptible to drugs that inhibit folate metabolism. Herein, we investigated the functional mechanism underlying blockade of folate receptor (FR) in human ovarian cancer with MORAb-003 antibody, a humanized monoclonal antibody directly against folate receptor 1 (FRα).
Methods We first used FACS analysis and real-time PCR to compare the expression of FRα in several human ovarian cancer cells. We then examined the tumor growth in the orthotopic models of human ovarian cancer cells SKOV3 and A2780. Consequently, we performed in vitro genomic and proteomic profiling in these cells to identify the functional mechanism of MORAb-003.
Results Among 7 human ovarian cancer cell lines, SKOV3ip1 cells have the highest levels of FRα and A2780 cells have the lowest. In vivo, MORAb-003 monotherapy led to significantly decreased tumor growth in the SKOV3ip1 model, but not in the A2780 model. Combination of MORAb-003 and docetaxel resulted in significantly greater reduction in tumor growth in the SKOV3ip1 model, but not in the A2780 model. MORAb-003 reduced proliferation but had no significant effect on apoptosis. Reverse-phase array analysis in SKOV3 cells revealed MORAb-003 up-regulated autophagic factors including PEA-15, Beclin-1, and ATG3 and down-regulated mTOR. cDNA microarray further showed that MORAb-003 up-regulated the expression of BECN1, ATG8, ATG3 and LC3B. Levels of Beclin-1 and LC3B were substantially increased in SKOV3 tumors treated with MORAb-003 alone or in combination with docetaxel, but not in A2780 tumors. Using FACS analysis with acridine orange (AO) staining, we observed significantly more AO positive population in SKOV3 cells treated with MORAb-003 alone (16.9%) or MORAb-003+docetaxel (21.2%) than in cells treated with control IgG (5.24%). Such differences were not observed in A2780 cells. Finally, we measured conversion of LC3 from isoform I to isoform II in cells treated with MORAb-003, and observed the significantly increased LC3II in SKOV3 cells but not in A2780 cells.
Conclusion Taken together, our results indicate that MORAb-003 results in prominent anti-tumor activity through inducing sustained autophagy in ovarian cancer cells.
Citation Format: Yun-Fei Wen, Whitney Spannuth Graybill, Department of Gyneocologic Oncology, Anil Sood. Immunotherapy targeting folate receptor induces autophagy in ovarian 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 1330. doi:10.1158/1538-7445.AM2014-1330
Abstract 1332: Inhibition of G9a induces DUSP4-dependent autophagic cell death in head and neck squamous cell carcinoma
Authors: LI KAI-CHUN , HUA KUO-TAI , TAN CHING-TING , KUO MIN-LIANG 
Published: May 2015

Abstract
Emerging evidence indicates that alteration of epigenetic regulation might be a key event for cancer progression. Abnormal expression of histone methyltransferase G9a, which contributes to transcriptional repression of tumor suppressor genes, has been implicated in maintaining cancer cells proliferation. Recent study suggests that inhibition of G9a reduces cancer cell growth. However, its molecular process has not been fully explored. In this study, we show that G9a expression is positively correlated to poor prognosis in head and neck squamous cell carcinoma (HNSCC) patients. Knockdown of G9a or inhibition of its catalytic activity by bix-01294 in HNSCC cells reduced proliferation, colony formation and anchorage-independent growth without inducing necrosis or apoptosis. Instead, complementary approaches including immunoblot analysis, LC3-GFP-fluorescent microscopy and morphologic study by electron microscopy revealed inhibition of G9a inducing autophagy. Pre-treatment with autophagy inhibitor 3-methyladenine (3-MA) or chloroquine (CQ) antagonized the inhibition effect of bix-01294 on cells viability, demonstrating that autophagic cell death is the major consequences in G9a suppressed cells. Mechanistic investigation suggested that ERK phosphatase, DUSP4, may be a downstream target of G9a. Knockdown of G9a and then suppression of DUSP4 expression restored ERK phosphorylation and colony formation but inhibits autophagy induced by G9a silencing. This effect was further reversed by treatment with MAPK/ERK selective inhibitor U0126. Moreover, knockdown of G9a resulted in growth inhibition and autophagy in orthotopic tumor model in mice. Taken together, our results not only point out the mechanism of autophagic cell death induced by inhibition of G9a but also pave a way for new targeting therapy utility for HNSCC in clinical.
Citation Format: KAI-CHUN LI, KUO-TAI HUA, CHING-TING TAN, MIN-LIANG KUO. Inhibition of G9a induces DUSP4-dependent autophagic cell death in head and neck squamous cell 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 1332. doi:10.1158/1538-7445.AM2014-1332
Abstract 1333: Combining Afatinib and Cetuximab synergistically increases their cytotoxicity for EGFR T790M-harboring cells
Authors: Tanaka Nobuyuki , Watanuki Zenta , Fukuhara Tatsuro , Maemondo Makoto 
Published: May 2015

Abstract
Epidermal growth factor receptor (EGFR) is an important therapeutic target for non-small cell lung cancers (NSCLCs). Tyrosine kinase inhibitors (TKIs), such as gefitinib and erlotinib, are effective for patients with EGFR-activating mutations. However, these patients eventually develop resistance, usually through a secondary EGFR mutation, T790M: substitution of methionine for threonine at position 790 in the kinase domain. While second-generation irreversible TKIs have higher affinities for the double-mutant EGFRs, their therapeutic effect leaves room for improvement. Combining a second-generation TKI like afatinib with an anti-EGFR monoclonal antibody has been shown to improve clinical outcomes, although the mechanism is not fully understood. To investigate this mechanism, we used human erythroleukemia K562 cells, which are EGFR-negative, to establish cells with known EGFR mutations. Two activating EGFR mutations, Ex19del and L858R, were sensitive to first-generation TKIs. Acquisition of resistance to these TKIs was confirmed by adding the second EGFR mutation, T790M. Double-mutant EGFRs were moderately sensitive to afatinib, but were only minimally affected by the monoclonal anti-EGFR antibody cetuximab. Combining afatinib and cetuximab synergistically increased cytotoxity for K562 cells carrying double-mutant EGFRs. Apoptosis in these cells was preceded by induction of the pro-apoptotic protein BIM (Bcl2L11) and activation of Caspase-3 and PARP. Afatinib induced autophagy in these cells correlated with AKT and ERK1/2 signaling in the course of treatment. Accordingly, we found that cetuximab enhanced the afatinib-induced cytotoxicity through the autophagic cell death pathway. Our results indicate that two distinct mechanisms contribute to the synergistic effect exerted by afatinib and cetuximab against NSCLCs.
Citation Format: Nobuyuki Tanaka, Zenta Watanuki, Tatsuro Fukuhara, Makoto Maemondo. Combining Afatinib and Cetuximab synergistically increases their cytotoxicity for EGFR T790M-harboring cells. [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 1333. doi:10.1158/1538-7445.AM2014-1333