bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 370: Expansion of CTCs from early stage lung cancer patients using a microfluidic co-culture model
Authors: Zhang Zhuo , Shiratsuchi Hiroe , Lin Jules , Chen Guoan , Reddy Rishindra M. , Azizi Ebrahim , Fouladdel Shamileh , Chang Andrew C. , Lin Lin , Jiang Hui , Waghray Meghna , Simeone Diane M. , Wicha Max S. , Beer David G. , Luker Gary , Ramnath Nithya , Nagrath Sunitha
Published: August 2015

Establishing the role of circulating tumor cells (CTCs) in tumor progression and metastasis depends both on enumeration and on obtaining sufficient numbers of cells for downstream assays. The numbers of CTCs are few in early stages of cancer, limiting detailed molecular characterization. One approach to overcome this limitation is to expand CTCs, i.e. increase their numbers as a result of cell division. Herein, we have developed a novel in-situ capture and culture methodology for ex-vivo expansion of CTCs using a three dimensional co-culture model, simulating a tumor microenvironment to support tumor development. We have successfully identified CTCs isolated from 14 of 19 early stage lung cancer patients. Expanded lung CTCs expressed tumor specific markers and formed spheroids in culture. CTCs derived xenograft demonstrated tumorigenic capability of these cells. Furthermore, CTCs carried mutations of the TP53 gene identical to those observed in the matched primary tumors. Next-generation sequencing further revealed additional matched mutations between primary tumor and CTCs of cancer-related genes. This strategy sets the stage to further characterize the biology of CTCs derived from patients with early lung cancers, thereby leading to a better understanding of these putative drivers of metastasis.

Citation Format: Zhuo Zhang, Hiroe Shiratsuchi, Jules Lin, Guoan Chen, Rishindra M. Reddy, Ebrahim Azizi, Shamileh Fouladdel, Andrew C. Chang, Lin Lin, Hui Jiang, Meghna Waghray, Diane M. Simeone, Max S. Wicha, David G. Beer, Gary Luker, Nithya Ramnath, Sunitha Nagrath. Expansion of CTCs from early stage lung cancer patients using a microfluidic co-culture model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 370. doi:10.1158/1538-7445.AM2015-370
Abstract 3715: Survival of patients with mucinous ovarian carcinoma and ovarian metastases: A population-based cancer registry study
Authors: Simons Michiel , Ezendam Nicole , Bulten Johan , Nagtegaal Iris , Massuger Leon
Published: August 2015

Objectives: Patients with mucinous ovarian carcinoma (MOC) generally have a favorable prognosis, although in advanced stage, prognosis is significantly worse compared to patients with serous ovarian carcinomas (SOC). This might be due to the difficulties in the distinction of MOC from metastatic tumors. In the current study we investigate prognosis of MOC compared to other types of ovarian cancer and to synchronous metastases to the ovary (sMO).

Materials and methods: Age, laterality, FIGO stage, tumor grade, treatment and survival were extracted from the Eindhoven Cancer registry for all patients diagnosed with ovarian carcinomas or sMO between 1990 and 2012. 5-year survival analysis and Cox proportional hazard analysis were conducted.

Results: 3,556 patients with primary ovarian carcinoma (of which 474 mucinous) and 289 with sMO were identified. In advanced stage, 5-year survival of patients with MOC was comparable to survival of patients with sMO (11% vs. 11%, p = 0.32) and decreased compared to patients with SOC (26%, p<0.01). For MOC there was no clinically significant effect on 5-year survival of either debulking (12% vs. 8%, p<0.01) or chemotherapy (12% vs. 10%, p = 0.02) as compared to patients with SOC (28 vs. 13% after chemotherapy, p<0.01 and 29% vs. 5% after debulking, p<0.01). Multivariate regression analysis for age, tumor grade and subtype including sMO was carried out, showing a significantly better overall survival for SOC compared to MOC (HR 0.51, p<0.01), whereas there was no difference for sMO compared with MOC (HR 1.16, p = 0.19).

Conclusion: Patients with advanced stage MOC have a worse prognosis than advanced stage SOC. Survival is almost identical to that of patients with sMO, which might indicate that (a large part of) advanced stage MOC are in fact metastases. This may partly explain the limited effects of chemotherapy and debulking in patients with MOC. Methods to differentiate between primary MOC and metastatic disease are needed to provide optimal treatment and insight in prognosis.

Citation Format: Michiel Simons, Nicole Ezendam, Johan Bulten, Iris Nagtegaal, Leon Massuger. Survival of patients with mucinous ovarian carcinoma and ovarian metastases: A population-based cancer registry study. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3715. doi:10.1158/1538-7445.AM2015-3715
Abstract 3716: The origin of metastatic ovarian tumors: A national population-based analysis
Authors: Simons Michiel , Bruls Jolien , Overbeek Lucy , Bulten Johan , Massuger Leon , Nagtegaal Iris
Published: August 2015

Objective: A significant part of ovarian malignancies are metastatic tumors with a wide variety in site of origin. Differentiating between a primary and metastatic malignancy of the ovaries can be difficult, especially in case of mucinous carcinomas, and misdiagnosis might have considerable impact on both treatment and prognosis. To further examine the origin of metastatic tumors in the ovary, we performed a large scale, nationwide search for metastatic ovarian malignancies in the Dutch Pathology Registry (PALGA).

Materials and methods: All pathology reports concerning metastatic ovarian carcinomas and associated primary tumors in the Netherlands from 2000-2010 were collected. Age, year of diagnosis, tumor type, location of the primary tumor and laterality of the ovarian tumor were extracted from the database. Tumors without a proven primary tumor were excluded. The Chi-square test was used to compare multiple nominal variables.

Results: 2,312 patients were identified. The most common primary sites were colon (33.2%), endometrium (17.1%), breast (14.3%), appendix (7.3%) and stomach (4.5%). Bilateral tumors were the most frequent mode of presentation (46.3%) followed by right (26.7%) and left side metastases (19.8%). In 7.2% of cases, laterality was unknown. Mucinous carcinomas were the most common histological type (46.7%), followed by signet-ring cell carcinomas (7.9%) and endometrioid carcinomas (7.7%). Of colorectal tumors, only 40.2% metastasized bilaterally, compared to 64% of breast, 63% of gastric and 59% of appendix tumors. Left sided ovarian tumors originated more often from the left colon than the right colon (69.4%, p<0.0001). Right sided ovarian tumors did not originate more often from the right side of the colon than from the left side. Appendiceal primary tumors did give rise to more right sided ovarian metastases (71.6%, p = 0.016).

Conclusion: The most common histological tumor type in metastatic ovarian carcinomas is mucinous carcinoma. The colon is the most frequent primary site of metastatic malignancies to the ovaries, followed by endometrium, breast, appendix and stomach. Breast, stomach and appendix tumors metastasized mostly bilateral, whereas colorectal tumors metastasized mostly unilateral and ipsilateral. The mechanisms underlying these preferred sites for metastasis or laterality are still unclear.

Citation Format: Michiel Simons, Jolien Bruls, Lucy Overbeek, Johan Bulten, Leon Massuger, Iris Nagtegaal. The origin of metastatic ovarian tumors: A national population-based analysis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3716. doi:10.1158/1538-7445.AM2015-3716
Abstract 3730: Feasibility of office hysteroscopy for evaluation of women with postmenopausal bleeding and association with improved pathological diagnosis
Authors: Shaw Jacquelyn M. , Mehta Sukrant , Chudnoff Scott , Levie Mark , Einstein Mark H. , Goldberg Gary L. , Nevadunsky Nicole S.
Published: August 2015

Objectives: Endometrial sampling is critical in the evaluation of women for cancers of the uterine corpus. Hysteroscopy in the operating room is associated with monetary and time expense to the patient and health care system. The purpose of this study was to compare histopathological diagnoses of endometrial biopsies obtained by office hysteroscopy (OH) versus endometrial pipelle (EP).

Methods: After IRB approval, records of all women with ICD-9 diagnosis (627.1) for postmenopausal bleeding who had OH attempted in the last 5 years at a single medical institution were evaluated. Demographic data collected included age, BMI, medical comorbidities, and histopathologic diagnosis. “Inadequate tissue” for pathologic evaluation was defined as stated in the pathology report or absence of endometrial tissue on biopsy specimen. Statistical analysis was performed using chi-square and Fisher's exact testing with SPSS software.

Results: 85 OHs were attempted and 50 had prior attempted EPs. 98% of OHs and 94% of EPs were successfully performed. However, adequate pathology was obtained in 93% of OHs verses 38% of EPs (p<0.01). Median age was 61 years (range 48-85) and median BMI was 32 kg/m2 (range 19-71). 5% had a BMI<25. The prevalence of diabetes mellitus, hypertension, hyperlipidemia, and cardiovascular disease were 32%, 74%, 44%, and 21% respectively. There was a significant difference in pathologic diagnoses between OH and EP (p <0.01). Six patients were diagnosed with endometrial hyperplasia, and 3 with endometrial cancers by OH that were not identified by EP.

Conclusions: OH is a feasible procedure that may enhance clinicians’ ability to diagnose benign, pre-malignant and malignant causes of post-menopausal bleeding. Further research is needed to understand the association of cost and quality of life with OH versus hysteroscopy in the operating room.

Table 1.Pathology findings from OH vs. EPPathologyOffice Hysteroscopy (OH)%Endometrial Pipelle Biopsy (EP)%n = 79n = 19Endometrial Cancer34%00%Endometrial Hyperplasia68%00%Benign Endometrium1924%1053%Atrophic Endometrium45%421%Benign Endometrial Polyp4759%526%

Citation Format: Jacquelyn M. Shaw, Sukrant Mehta, Scott Chudnoff, Mark Levie, Mark H. Einstein, Gary L. Goldberg, Nicole S. Nevadunsky. Feasibility of office hysteroscopy for evaluation of women with postmenopausal bleeding and association with improved pathological diagnosis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3730. doi:10.1158/1538-7445.AM2015-3730
Abstract 3731: The course of fatigue and its correlates in colorectal cancer survivors: A prospective cohort study of the PROFILES registry
Authors: Husson Olga , Mols Floortje , van de Poll-Franse Lonneke , Thong Melissa
Published: August 2015

Objective: Cancer survivors who remain fatigued during long-term follow-up are at risk for worse health outcomes. To achieve personalized management of cancer-related fatigue, insight into its correlates among long-term survivors is needed to identify survivors at risk of remaining fatigued and also the correlates for intervention.

Methods: Colorectal cancer (CRC) survivors diagnosed between 2000-2009, as registered in the population-based Eindhoven Cancer Registry, completed three annual surveys that included the Fatigue Assessment Scale, the EORTC Quality of Life (EORTC-QLQ-C30) questionnaire, Hospital Anxiety and Depression Scale, and physical activity levels. Linear mixed-model were used to assess the course of fatigue and identify its correlates.

Results: 1734 (66%) CRC survivors completed at least 2 surveys. Fatigue levels were relatively stable over time. Being female, young (≤65 years old), single, low education, chemotherapy treatment, or having ≥1 comorbid condition was associated with higher fatigue scores. Years since diagnosis, radiotherapy treatment and disease stage were not related to fatigue scores. Significant between- and within-subjects effects were found for well-being factors (social, emotional, and cognitive function, and global quality of life), symptoms (anxiety, depression, pain, and insomnia) and functional status (physical and role function, and physical activity levels) in relation to fatigue. Differences in fatigue could be largely attributed to behavior/well-being (59%) and functional status (37%), and to some extent to sociodemographic (4%) and clinical (8%) factors.

Conclusion: This study showed that behavior/well-being and functional status explained more variance in fatigue levels among CRC survivors than sociodemographic and clinical factors.

Citation Format: Olga Husson, Floortje Mols, Lonneke van de Poll-Franse, Melissa Thong. The course of fatigue and its correlates in colorectal cancer survivors: A prospective cohort study of the PROFILES registry. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3731. doi:10.1158/1538-7445.AM2015-3731
Abstract 3744: Single-cell analysis of adaptive resistance and fractional responses of melanoma cells to RAF/MEK inhibition
Authors: Fallahi-Sichani Mohammad , Moerke Nathan J. , Niepel Mario , Zhang Tnghu , Gray Nathanael S. , Sorger Peter K.
Published: August 2015

Treatment of BRAFV600E melanomas with drugs, such as vemurafenib, that inhibit RAF/MEK signaling is effective in the short term, but remission is not durable. Acquired drug resistance is thought to involve short-term adaptive responses that compensate for RAF/MEK inhibition via up-regulation of other pro-growth mechanisms. Thus, understanding and ultimately preventing adaptive responses is a key to durable therapy. Systematic data comparing BRAFV600E tumor cells is generally lacking and it is not known whether adaptation is fundamentally similar across cell types or among individual cells within a cell population.

We apply a systematic approach to studying the responses of human melanoma cell lines to five drugs, RAF and MEK inhibitors, with the overall goal of (i) characterizing variability in adaptation with time, dose, cell type and across individual cells, (ii) discovering new or poorly characterized adaptive mechanisms, and (iii) demonstrating the effectiveness of a high-throughput approach involving multiplex measurement, single-cell analysis and computational modeling. The data involves time-course measurement of total level and activity of signaling proteins and cell state markers using array-based methods and single-cell immunofluorescence assays as well as measurement of apoptosis and cell viability under the same conditions. Statistical modeling using partial least squares regression (PLSR) revealed which of the changes in the ∼200,000 point dataset were phenotypically consequential.

We found that responses to RAF inhibitors are remarkably diverse and involve multiple pathways that can be up or down-regulated over time, with significant variability across cell types and individual cells. We identified a role for JNK/c-Jun signaling in altering the cell-cycle distribution of melanoma cells, causing apoptosis-resistant cells to accumulate and drug maximal effect (Emax) to fall; co-drugging with RAF and JNK inhibitors or JUN knockdown reverse this effect. The primary effect of JNK inhibitors is to minimize the cell-to-cell variability in pS6 suppression, promoting the induction of apoptosis.

Our study shows that a systems-level approach (combining high density time-dependent measurements, quantitative modeling and single-cell analysis) may provide a general framework for evaluating new drugs with adaptive and paradoxical response, and identifying potentially useful combination therapies.

Citation Format: Mohammad Fallahi-Sichani, Nathan J. Moerke, Mario Niepel, Tnghu Zhang, Nathanael S. Gray, Peter K. Sorger. Single-cell analysis of adaptive resistance and fractional responses of melanoma cells to RAF/MEK inhibition. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3744. doi:10.1158/1538-7445.AM2015-3744
Abstract 3748: Transcriptome landscape of brain tumor progression at single-cell resolution
Authors: Kim Kyu-Tae , Lee Jin-Ku , Park Dong-Hyun , Nam Do-Hyun , Lee Hae-Ock , Park Woong-Yang
Published: August 2015

BACKGROUND: Tumor progression within a brain is the process of acquiring malignant genetic mutations from glial progenitor cells, dodging their original developmental fate and deteriorating functional connectivity of CNS. In order to devise effective targeted therapeutic strategies, understanding distinct molecular and phenotypic evolutionary features of cancer clones is eminent.

METHODS: To examine molecular signatures associated with malignant glioma progression, we analyzed transcriptome dynamics of 311 single-cells derived from 3 patients showing distinct disease patterns. Combined with single-cell level signatures, clonal architecture of tumors was reconstructed based on Bayesian-model clustering with integrated information of SSNVs and SCNAs identified from bulk population-level using aCGH and WES.

RESULTS: We found evidence of branched evolutionary patterns within and across two histopathological separated tumor regions. Both tumors have high clonal probable somatic mutations, and evolutionary inference showed clonal lineage between two tumor regions was divided at early time during tumor progression. For all patients, the gene expression signatures within a tumor were heterogeneous but converged towards mesenchymal phenotype in the aggressive region. In the first case, the left tumor was self-limiting and showed mixed proneural and classical gene expression signatures whereas the right side tumor has expanded to the corpus callosum with EGFR overexpression and mesenchymal gene expression. In a case of a monocentric tumor with PDGFRA amplification, cancer cells disseminated from the core mass with massive immune cell infiltration underwent transcriptome switching from proneural to mesenchymal gene expression, with a concomitant FGFR1 copy number gain and mutation. Lastly, in a secondary glioblastoma with IDH1 mutation, GLIOLAN positive glioblastoma was surrounded by grade III oligodendroglioma. The two tumor areas showed intermingled heterogeneous gene expression signatures, with upregulation of astrocytic/mesenchymal genes in the GLIOLAN positive tumor cells.

CONCLUSIONS: Our results show distinct genetic patterns of clonal expansion across patients undergoing malignant glioma progression. We also provide the transcriptome-wide view of gene expression signatures directed by genomic alterations. Overall our study reveals heterogeneity of individual tumor but elucidates common genomic features of glioblastoma progression.

Citation Format: Kyu-Tae Kim, Jin-Ku Lee, Dong-Hyun Park, Do-Hyun Nam, Hae-Ock Lee, Woong-Yang Park. Transcriptome landscape of brain tumor progression at single-cell resolution. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3748. doi:10.1158/1538-7445.AM2015-3748
Abstract 3758: A combined machine learning and bioinformatic analysis approach identifies biological pathways that predict clinical stage and survival outcome in neuroblastoma patients
Authors: Carlisle Alex , Caceres Ivan , Mehta Sonali , Schindler Jay , Sharma Jonathan
Published: August 2015

Neuroblastoma is an aggressive pediatric cancer that displays tremendous clinical and biological heterogeneity. Approximately 40% of patients present with metastatic disease and have poor outcome; new biomarkers that improve risk classification and prognostication are desperately needed. Applying a machine learning-based approach to analyzing transcriptome-wide signals in multiple populations of neuroblastoma patients spanning clinical stage we identified a combined set of 14 clinical and molecular variables that predicted survival outcome and tumor stage with over 87% accuracy. A systems biology-based analysis of the molecular variables from our predictive set identified 5 major networks with hubs defining 3 major ontological processes: apoptosis, nucleotide biosynthesis, and RNA metabolism.

Citation Format: Alex Carlisle, Ivan Caceres, Sonali Mehta, Jay Schindler, Jonathan Sharma. A combined machine learning and bioinformatic analysis approach identifies biological pathways that predict clinical stage and survival outcome in neuroblastoma patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3758. doi:10.1158/1538-7445.AM2015-3758
Abstract 3768: Identification of new alternative lengthening of telomeres (ALT)-positive cancer cell lines using the C-circle assay
Authors: Malykh Andrei G. , Doyle Jonathan R. , Tian Fang , Lee Joyce HY , Henson Jeremy D. , Reddel Roger R.
Published: August 2015

Alternative lengthening of telomeres (ALT) is a telomere maintenance mechanism used by approximately 10% of human cancers, including poor prognosis childhood and adult cancers for which development of better treatments is a priority. Currently there are only about ten published ALT+ cancer cell lines available for research, only four of which are available from international repositories. The development of the C-Circle Assay has made it possible to accurately and conveniently test a large number of cell lines for ALT activity. We have tested a panel of 297 cancer cell lines from the American Type Culture Collection (ATCC) with the C-Circle Assay for ALT and identified 34 new ALT+ cell lines. Importantly, we have identified for the first time ALT+ cancer cell lines derived from breast carcinoma, hepatocellular carcinoma, melanoma and leukemia. The overall proportion of ALT+ cancer cell lines was 11.4% (n = 297), which included 19.5% of leukemia (n = 46), 19.0% of melanoma (n = 21), 12.7% of non-small cell lung carcinoma (n = 53) and 3.3% of breast carcinoma (n = 37) cell lines. These results identify additional cancer cell line resources for research into the ALT mechanism, including cell lines from cancer types where ALT has not previously been studied in detail.

Citation Format: Andrei G. Malykh, Jonathan R. Doyle, Fang Tian, Joyce HY Lee, Jeremy D. Henson, Roger R. Reddel. Identification of new alternative lengthening of telomeres (ALT)-positive cancer cell lines using the C-circle assay. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3768. doi:10.1158/1538-7445.AM2015-3768
Abstract 3784: Regulation of chromatin condensation by mitotic checkpoint protein MAD2
Authors: Shandilya Jayasha , Roberts Stefan GE
Published: August 2015

The process of cell division is stringently regulated by spindle and mitotic checkpoint components mainly MAD2, MAD1, CDC20, BUBR1, BUB3 and MAD3. These checkpoint proteins detect unattached or improperly attached kinetochore-microtubule binding sites and generate an anaphase-wait signal by inhibiting the anaphase-promoting complex (APC/C). Activation of checkpoint signaling arrests the cells at metaphase until the attachment errors are rectified and all the chromosomes are properly bi-oriented at the metaphase plate. Together with these checkpoint proteins, the expression of checkpoint-regulatory factors such as p31 (comet) and recently identified, WT1 can hypo- or hyper-activate the signaling pathway, respectively and regulate the timing of mitotic exit. Recently we found that WT1 can interact with MAD2 and regulate the mitotic checkpoint function. Furthermore our studies show that MAD2 and WT1 can also influence the chromatin condensation process required for mitotic progression. We analyzed the effect of MAD2 and WT1 perturbations on the epigenetic mark (phosphorylation of histone H3) associated with condensed chromatin state. Our results show that MAD2 regulates the recruitment of chromatin modifying enzyme Aurora kinase B during mitosis. We further show that MAD2 is an important factor required for synchronization of key events beyond the checkpoint signaling sites to ensure the fidelity of chromosome segregation.

Citation Format: Jayasha Shandilya, Stefan GE Roberts. Regulation of chromatin condensation by mitotic checkpoint protein MAD2. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3784. doi:10.1158/1538-7445.AM2015-3784
Abstract 3808: Targeting cholesterol for increased chemotherapy efficacy
Authors: Yamaguchi Ryuji
Published: August 2015

The PI3K-AKT pro-survival signal present in most cancer cells is the major obstacle to effective chemotherapies. However, most chemotherapeutics targeting the PI3K-AKT pathway have undesirable side effects. β-cyclodextrin (βCD) and its derivatives were found to diminish PI3K-AKT activities in cancer cells with limited side effects. Treatments of cultured cells with βCD depleted cholesterol from the plasma membrane abolishing their coated pits. Even though growth factors such as IGF1 and insulin activated their receptors, their signals were not transduced to downstream signals such as PI3K-AKT pro-survival pathway. The inhibitory effect lasted as long as βCD was present in the medium. Injected into a mouse, βCD attenuated IGF1-induced blood glucose drop for several hours.

In the absence of PI3K-AKT pro-survival signal, cells became sensitized to mitochondria dependent cell death such as Bcl2 antagonists ABT-263 in many epithelial cells and Fas and TRAIL in type II cells. In mice carrying human cancer cells, the addition of HPβCD (a derivative of βCD) to the 2-deoxyglucose & ABT-263 combination therapy, and to the TRAIL therapy improved their efficacies.

Citation Format: Ryuji Yamaguchi. Targeting cholesterol for increased chemotherapy efficacy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3808. doi:10.1158/1538-7445.AM2015-3808
Abstract 3816: The DNA methylation inhibitor induces telomere dysfunction and apoptosis of leukemia cells that is attenuated by telomerase over-expression
Authors: Li Bingnan , Zhang Xiaolu , Jonge Nick de , Björkholm Magnus , Xu Dawei
Published: August 2015

DNA methyltransferase inhibitors (DNMTIs) such as 5-azacytidine (5-AZA) have been used for the treatment of acute myeloid leukemia (AML) and other malignancies, and inhibition of global/gene-specific DNA methylation is widely accepted as a key mechanism behind their anti-tumor activity. It is currently unclear whether other mechanisms are involved in DNMTI's action. Because telomerase or telomerase reverse transcriptase (TERT) plays a key role in malignant transformation and progression through telomere elongation and other telomere lengthening-independent activities, and TERT or telomerase has been shown to confer chemo- or radio-resistance to cancer cells, we determine whether DNMTIs affect telomere function and whether TERT/telomerase interferes with their anti-cancer efficacy. We showed that 5-AZA induced DNA damage and telomere dysfunction in AML cell lines by demonstrating the presence of 53-BP1 foci and the co-localization of 53-BP1 foci with telomere signals, respectively. Telomere dysfunction was coupled with diminished TERT expression, shorter telomere and apoptosis in 5-AZA-treated cells. However, 5-AZA treatment did not lead to changes in the methylation status of subtelomere regions. Downregulation of TERT expression similarly occurred in primary leukemic cells derived from AML patients exposed to 5-AZA. TERT over-expression significantly attenuated 5-AZAmediated DNA damage, telomere dysfunction and apoptosis of AML cells. Taken together, 5-AZA mediates the down-regulation of TERT expression, and induces telomere dysfunction,which consequently exerts an anti-tumor activity. These findings may have implications in DNMTI and telomerase-based strategies for cancer therapeutics.

Citation Format: Bingnan Li, Xiaolu Zhang, Nick de Jonge, Magnus Björkholm, Dawei Xu. The DNA methylation inhibitor induces telomere dysfunction and apoptosis of leukemia cells that is attenuated by telomerase over-expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3816. doi:10.1158/1538-7445.AM2015-3816
Abstract 3821: Understanding the functional role of retinoic acid receptor beta2 in the development of oral cancer
Authors: Radhakrishnan Raghu , Hunter Keith
Published: August 2015

Background: Retinoids are the best studied chemopreventive agents in oral cancer patients, whose effects are mediated by retinoic acid receptors (RAR). The loss of RARβ2 expression early in the development of oral cancer is attributed to acquisition of cellular immortalization or abrogated senescence. Aim: To determine the functional effects of RARβ2 dysfunction (constitutive expression or loss) in precancerous oral keratinocytes. Methods: Characterization of a panel or mortal (D6, D30) and immortal precancerous oral keratinocytes (D19, D20, D34 and D38), and tert immortalized normal cells (FNB-6) for baseline RARβ2 expression and their role in senescence, differentiation, apoptosis, and methylation. Re-express RARβ2 in immortal cells by treatment with all trans retinoic acid, 5-aza-2′- deoxycytidine, administered alone and in combination and asses their effect on senescence and promoter methylation. Results: Mortal cells showed constitutive expression of RARβ2 in senescing cultures and loss of expression in immortal cells. This was reflected in tissue engineered 3D composites as well as in the corresponding biopsies, when stained with RARβ antibody. Concomitant expression of CDKN2A and CDKN1A in senescing cultures and their absence in immortals suggest that the loss of RARβ2 strongly correlate with transition from senescence to immortalization. Re-expression of RARβ2 in immortal cells by treatment with retinoic acid (RA), 5-aza-2′-deoxycytidine (Aza-C) and combination of RA + Aza-C with simultaneous upregulation of CDKN2A and CDKN1A confirm RARβ2 as tumor suppressor gene, which is silenced due to hypermethylation. Increase in the Histone Repressor A (HIRA) foci and Senescence associated β- galactosidase (SA-β gal) activity in treated cell confirmed the restoration of senescence program in an immortal cell. Promoter methylation analysis confirmed that the silencing of RARβ2 in immortal cell was due to hypermethylation. Conclusion: The role of demethylating agent in growth inhibition and apoptosis highlights the importance of RAR beta serving as an intermediate biomarker in chemoprevention. Identification of methylation status may prove useful as a selective biomarker prior to combined demethylation and retinoid treatment in oral precancerous lesions.

Citation Format: Raghu Radhakrishnan, Keith Hunter. Understanding the functional role of retinoic acid receptor beta2 in the development of oral cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3821. doi:10.1158/1538-7445.AM2015-3821
Abstract 3836: Identification of key survival-correlating microRNAs and Piwi-interacting RNAs dysregulated in head and neck squamous cell carcinoma
Authors: Ku Jonjei , Zou Angela E. , Honda Thomas K. , Zheng Hao , Saad Maarouf A. , Yu Vicky , Xuan Yinan , Singh Pranav , Rahimy Mehran , Kuo Selena Z. , Ongkeko Weg M. , Wang-Rodriguez Jessica
Published: August 2015

Aberrant expression of microRNAs and piwi-interacting RNAs, two distinct classes of non-coding RNA (ncRNA) and important regulators of gene expression, is known to be implicated in human diseases including cancer. The exact roles of microRNAs and piwi-interacting RNAs in the pathogenesis of head and neck squamous cell carcinoma (HNSCC), however, remain poorly understood. Utilizing next generation RNA sequencing analysis of raw data obtained from The Cancer Genome Atlas, we discovered significant dysregulation (fold change ≥ 2) in 231 microRNAs and 60 piwi-interacting RNAs between forty-three matched samples of tumor and adjacent normal tissues, including down-regulation of mirs 30e and let7-c, and up-regulation of piR34736 and mir-196a-1 in tumor tissue compared to adjacent, non-tumor tissue. Further statistical survival modeling using patient survival data and cox proportional hazard regression analysis revealed that miRs 30e, 196a-1, and let7-c, and piR34736 correlated significantly with survival in patients with HNSCC. To further investigate these microRNAs and piwi-interacting RNAs, quantitative reverse transcriptase PCR (qRT-PCR) and MTT cell proliferation assays were performed in-vitro to assess their isolated effects on cancer gene expression in established HNSCC cell lines. Endogenous expression of these ncRNA was compared between a set of three normal oral keratinocytes and five HNSCC cell lines using qRT-PCR which confirmed miR-30e and mir-let7-c down-regulation and piR34736 and mir-196a-1 up-regulation in cancer. MTT cell proliferation assays upon knock-down of miR-196a-1 and ectopically expressed mir-let7-c in HNSCC revealed significant decreases in cell proliferation compared to cells transfected with their respective negative control. Identification of key ncRNAs differentially expressed between cancer tissue and adjacent normal tissue may reveal important steps in the initiation and progression of HNSCC as they are strongly correlated with patient survival. These key microRNAs and piwi-interacting RNAs may serve as potential biomarkers and therapeutic targets for future epigenetic-based cancer therapies.

Citation Format: Jonjei Ku, Angela E. Zou, Thomas K. Honda, Hao Zheng, Maarouf A. Saad, Vicky Yu, Yinan Xuan, Pranav Singh, Mehran Rahimy, Selena Z. Kuo, Weg M. Ongkeko, Jessica Wang-Rodriguez. Identification of key survival-correlating microRNAs and Piwi-interacting RNAs dysregulated in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3836. doi:10.1158/1538-7445.AM2015-3836
Abstract 3845: Smarce1 facilitates survival of metastatic breast cancer cells in circulation
Authors: Sethuraman Aarti , Fan Meiyun , Brown Martin M. , Wu Zhaohui
Published: August 2015

Elevated BAF57/SMARCE1 protein expression has been linked to high metastatic potential of prostate cancer, while its relevance in metastatic breast cancer is not completely understood. The objective of this study is to investigate the role of SMARCE1 in regulating metastatic potential of basal-like breast tumor cells. SMARCE1 is a core subunit shared by all mammalian SWI/SNF complexes and is predicted to mediate the interaction between the SWI/SNF complex and transcriptional activators/repressors, as well as DNA binding. SWI/SNF are nucleosome modifying complexes that destabilize the histone-DNA interaction in an ATP-dependent manner to facilitate recruitment of transcription regulators to DNA. We found that SMARCE1 knockdown (SMARCE1-kd) in basal-like breast cancer cell line MDA-MB-231 reduced spontaneous lung metastasis in an orthotopic xenograft model, without affecting primary tumor growth. Through tail-vein injection of tumor cells we identified that SMARCE1-kd significantly reduced the number of metastatic foci in lungs of inoculated mice, implicating a role of SMARCE1 in promoting metastatic cell survival in later stages of metastasis occurring post-dissemination from primary tumors. In-vitro assays showed that SMARCE1-kd had no effect on cell migration and invasion, but substantially sensitized cells to anoikis. Consistently, we observed reduced activation/phosphorylation of the pro-survival proteins PTK2/FAK and MAPK/ERK and increased expression of apoptotic protein BCL2L11 in SMARCE1-kd cells compared to wild-type cells grown under suspension. We further confirmed using specific chemical inhibitors, that FAK and ERK phosphorylation are required for SMARCE1-mediated survival of tumor cells under suspension culture. Collectively, these results suggest a role for SMARCE1 in promoting metastasis by protecting cells against anoikis. The molecular mechanism underling SMARCE1-mediated regulation of FAK/ERK phosphorylation in response to loss of contact with extracellular matrix is under investigation.

Citation Format: Aarti Sethuraman, Meiyun Fan, Martin M. Brown, Zhaohui Wu. Smarce1 facilitates survival of metastatic breast cancer cells in circulation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3845. doi:10.1158/1538-7445.AM2015-3845
Abstract 3846: DNA double-strand breaks in hypoxic but not in normoxic conditions determine different radiosensitivity of tumors
Authors: Ogi Kazuhiro , Kawata Mayumi , Nakagaki Takafumi , Takeda Kousuke , Akihiro Miyazaki , Hiroyoshi Hiratsuka
Published: August 2015

Purpose:

While only few biomarkers like tumor histology and stage are used in routine clinical practice to predict tumor cure probability after chemo-radiotherapy, sustained efforts are made to establish more precise predictors for individual treatment. Recently chemotherapy corresponded with radiotherapy (CCR) to oral cancer has developed treatment efficiency of killing tumor cells. However we can't distinguish the number of tumor cells that may cause recurrences after radiotherapy. Then we hypothesized that hypoxic condition may have some influence of cell death by irradiation, and we tried to demonstrate how the correlation of phosphorylated H2AX (γH2AX) was influenced by oxygenation status of the tumor regions to predict radiation treatment response.

Results: We used oral squamous cell carcinoma (OSCC) cell lines and divided them into two groups that showed hypoxic response or not. HSC-2 cells increased the expression level of HIF-1 alpha under hypoxia. No difference was observed in G1 arrest to irradiation of 6Gy in these cell lines. While the expression revel of γH2AX decreased in HSC-2, its revel didn't change under hypoxic condition in HSC-3 and SAS. From these findings, we postulate that one of the mechanism by which hypoxic cells become resistant to irradiation may involve the expression of HIF-1 alpha.

Conclusion:

The γH2AX is highly dependent on tumor cell oxygenation. Further studied are ongoing to establish the potentially important role of γH2AX under hypoxia.

Citation Format: Kazuhiro Ogi, Mayumi Kawata, Takafumi Nakagaki, Kousuke Takeda, Miyazaki Akihiro, Hiratsuka Hiroyoshi. DNA double-strand breaks in hypoxic but not in normoxic conditions determine different radiosensitivity of tumors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3846. doi:10.1158/1538-7445.AM2015-3846
Abstract 3871: Telomere length and cancer mortality and all-cause mortality
Authors: Bojesen Stig E. , Nordestgaard Børge G. , Rode Line
Published: August 2015

Introduction:

Short telomeres are associated with increased age and age-related diseases. We tested the hypothesis that short telomeres are associated with increased cancer mortality and all-cause mortality.

Methods:

We studied 64637 individuals recruited from 1991 and onwards from two prospective cohort studies; the Copenhagen City Heart Study and the Copenhagen General Population Study. All had telomere length measured by Q-PCR in DNA from peripheral blood and had the telomere length-associated genotypes of rs1317082 (TERC), rs7726159 (TERT), and rs2487999 (OBFC1) determined. We calculated the sum of telomere length shortening alleles from the three SNPs combined. We conducted Cox regression, and instrumental variable analyses in a mendelian randomization design, using the allele sum as a genetic instrument for telomere length.

Results:

Telomere length shortened by 17 (P<10E-300) and 69 (P = 110E-104) base pairs per increase of one year of age, and one allele increase of allele sum, respectively. Of the 7607 deaths during up to 22 years of follow-up, 2420 were due to cancer. Shortening of telomeres (200 base pairs) was associated with increasing cancer mortality and all-cause mortality with hazard ratios of 1.02(95% CI;1.01-1.03) and 1.02(95% CI;1.02-1.03) in multifactorially adjusted models. In contrast, in the genetic analyses, the corresponding odds ratios were 0.86(95% CI;0.76-0.96) and 0.98(95% CI;0.91-1.05).

Conclusion:

In an observational design, short telomeres were associated with increased cancer mortality and all-cause mortality. In contrast and importantly; in a mendelian randomization design, short telomeres were associated with decreased cancer mortality, suggesting that short telomeres are likely causative in protecting against cancer mortality.

Citation Format: Stig E. Bojesen, Børge G. Nordestgaard, Line Rode. Telomere length and cancer mortality and all-cause mortality. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3871. doi:10.1158/1538-7445.AM2015-3871
Abstract 388: EGR1 is a mediator of TWEAK-Fn14 pathway induced prostate cancer bone metastasis
Authors: Ameri Amir H. , Yin JuanJuan Ivy , Jansen Keith , Wang Simeng , Snyder Jessica , Hynes Paul , Kelly Kathleen
Published: August 2015

Patients with advanced or castration-resistant prostate cancer have a high probability of developing bone metastasis. However, the molecular mechanisms driving bone metastasis in prostate cancer remain poorly understood. We have previously demonstrated that Fn14 expression is increased significantly in metastatic lesions from prostate cancer patients. Fn14 is a member of the TNF receptor family, and our in vivo study using DU145/RasB1 cells indicated that the TWEAK-Fn14 pathway promotes prostate cancer bone metastasis via the canonical NFκB pathway. However, the downstream targets of the TWEAK-Fn14 pathway have yet to be characterized in prostate cancer cells. To elucidate the precise mechanism underlying Fn14-promoted metastasis, we analyzed the gene expression profile of highly metastatic DU145/RasB1 and low metastatic DU145/RasB1 Fn14depleted (Fn14KD) cells. Fn14 depletion significantly altered expression of 147 genes, including ECM components, cell adhesives, and cytoskeletal regulators. In addition, DU145/RasB1 and DU145/RasB1 Fn14KD cells responded differently to TWEAK treatment. We found increased expression of early growth response 1, (EGR1), in the TWEAK-treated DU145/RasB1 cells relative to DU145/RasB1 Fn14KD cells. To determine if EGR1 plays a role in prostate cancer metastasis, we depleted EGR1 expression with two shRNAs in highly metastatic DU145/RasB1 cells. Our results show that both EGR1 shRNAs significantly inhibited bone metastasis in vivo. We also found that EGR1 expression was decreased when the NFκB pathways were inhibited by an IκB super repressor in DU145/RasB1 cells. Interestingly, both Fn14 depletion and EGR1 depletion increased TFPI2 expression and inhibited adhesion to fibrin. Fibrin has been shown to facilitate metastatic spread by assisting platelet adhesion to circulating tumor cells. These results suggest one scenario whereby activation of Fn14 signaling may promote prostate cancer bone metastasis through enhancing tumor-platelet interaction via an EGR1-dependent mechanism.

Citation Format: Amir H. Ameri, JuanJuan Ivy Yin, Keith Jansen, Simeng Wang, Jessica Snyder, Paul Hynes, Kathleen Kelly. EGR1 is a mediator of TWEAK-Fn14 pathway induced prostate cancer bone metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 388. doi:10.1158/1538-7445.AM2015-388
Abstract 3883: Clonal evolution of a lethal prostate cancer: Integrated whole genome analysis case study
Authors: Kallio Heini M.L. , Annala Matti , Kivinummi Kati , Högnäs Gunilla , Gundem Gunes , Wedge David C. , Van Loo Peter , Heyn Holger , Emmert-Buck Michael R. , Isaacs William B. , Esteller Manel , McDermott Ultan , Nykter Matti , Visakorpi Tapio , Bova G. Steven
Published: August 2015

Prostate cancer (PC) is prevalent in both indolent and lethal forms. Although many PC patients diagnosed today have organ-confined disease curable by prostatectomy or radiation therapy, 20-30% of PCs will relapse to lethal disease within 5-years of treatment. Relatively little attention has been paid to distinguishing the molecular characteristics of proven lethal metastatic PC from non-lethal cancers. A better understanding of the origins and evolution of lethal cancers should allow screening and treatment to be better tailored to the needs of each patient. To this end, we performed an integrative molecular profiling of a lethal PC from one patient (A21). High-coverage whole genome sequence, transcriptome sequence, and methylation analysis was performed on 9 anatomically separate metastases obtained by autopsy, and targeted sequencing was performed in multiple cancerous and noncancerous foci within the radical prostatectomy specimen removed 5 years prior to death. Molecular results were analyzed in relation to detailed clinical data. Integrated whole genome sequence analysis revealed convergent evolution of AR gene amplification events, and inception of p.L702H mutation in the AR present only in liver metastases. In addition, the analysis showed parallel increases in AR regulated transcripts in the liver metastases, suggesting a dominant effect by the mutation. Mutation of PI3/PI4 kinase member PIK3CG was found in all metastases but in no primary tumor foci studied. The study demonstrated the power of an integrated approach to interrogate clonal evolution of cancer suggesting that such studies are valuable on the individual level and could lead the way towards personalized treatment. In the individual studied, cessation of corticosteroid treatment and/or therapeutic manipulation of PI3K/AKT/mTOR activity theoretically could have provided a personalized benefit. These findings suggest that similar integrated analysis in large cohorts of patients with metastatic cancer could accelerate progress in establishing effective personalized cancer medicine.

Citation Format: Heini M.L. Kallio, Matti Annala, Kati Kivinummi, Gunilla Högnäs, Gunes Gundem, David C. Wedge, Peter Van Loo, Holger Heyn, Michael R. Emmert-Buck, William B. Isaacs, Manel Esteller, Ultan McDermott, Matti Nykter, Tapio Visakorpi, G. Steven Bova. Clonal evolution of a lethal prostate cancer: Integrated whole genome analysis case study. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3883. doi:10.1158/1538-7445.AM2015-3883