bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1422: Validation of primary non-small cell lung cancer model: Stable isotope-resolved metabolomics (SIRM) reveals functional biochemistry
Authors: Kinslow Connor J. , Fan Teresa W-M , Tan Jin L. , Lorkiewicz Pawel K. , Balasubramaniam Ramya , Cai Yihua , Yan Jun , Lane Andrew N.
Published: May 2015

Stable isotope-resolved metabolomics (SIRM) provides a comprehensive and functional readout of cellular activity and is therefore a useful tool both for biomarker discovery and fundamental cancer biology. We have established a primary lung squamous cell carcinoma line (PSLC1), isolated from the lung tumor tissue of a non-small cell lung cancer patient, for which the metabolic profile has been established. We used SIRM to characterize the metabolic activity of the cells in mouse xenografts and in cell culture. Here we report the metabolic activities of the cell line in culture using two different tracers to evaluate central metabolism. PSLC1 cells were grown in either [U-13C]-glucose or [U-13C,15N]-glutamine labeled medium. Cell extracts and medium were prepared and analyzed by 1H-NMR, HSQC, TOCSY, GC-MS, and FTICR-MS. Cells exhibited linear glucose consumption that persisted even as proliferation plateaued. The conversion of glucose to lactate occurred at a strikingly high 89%. Anaplerotic pathways involving pyruvate carboxylation and glutaminolysis were active and appeared to be dependent on cellular proliferation. Glutamine represented an important contributor for biosynthetic and bioenergetic cellular needs, including de novo pyrimidine biosynthesis and glucose-independent TCA cycling. In addition, glutamine served as the major precursor in proline and glutathione biosynthesis. Distinct glutamine utilization and labeling patterns indicated that this cancer might be driven by C-MYC overexpression. C-MYC overexpression was observed by targeted protein analysis guided by metabolomics-edited transcriptomics analysis (Pro-META). The ability of SIRM to generate hypotheses about protein expression demonstrates its potential use as a first line biomarker detection method. Comparison with upstream in situ and xenograft data will allow us to assess model validity and the effects of tumor microenvironment. In turn, the in vitro data will help us interpret human in situ data more robustly. Supported by grant R25-CA-134283 and 1P01CA163223-01A1 from the National Cancer Institute

Citation Format: Connor J. Kinslow, Teresa W-M Fan, Jin L. Tan, Pawel K. Lorkiewicz, Ramya Balasubramaniam, Yihua Cai, Jun Yan, Andrew N. Lane. Validation of primary non-small cell lung cancer model: Stable isotope-resolved metabolomics (SIRM) reveals functional biochemistry. [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 1422. doi:10.1158/1538-7445.AM2014-1422
Abstract 1423: Resistance to LDHA inhibitors requires signaling through the AMPK/mTOR/S6K pathway leading to increased oxidative phosphorylation
Authors: Boudreau Aaron , Peterson David , Moffat John , Liu Bonnie , Kwong Mandy , Gao Min , Purkey Hans , O'Brien Thomas , Hatzivassiliou Georgia , Daemen Anneleen , Evangelista Marie
Published: May 2015

Lactate Dehydrogenase A (LDHA) is an attractive candidate for targeting glycolysis-addicted tumors. However, due to the inherent plasticity of metabolic networks in cells, there is concern that the benefit of targeting LDHA may be transient and that resistance will quickly emerge. To identify predictive features of LHDA inhibitor sensitivity and to understand how cells adapt to long-term LHDA inhibition, we screened a large panel (∼500) of tumor cell lines with GNE-140, a newly developed LHDA inhibitor. We found that approximately 15% of lines were inherently sensitive to the LDHA inhibitor, with sensitivity correlating with increased expression of glycolysis genes and inversely correlating with expression of oxidative phosphorylation genes. Despite the metabolic plasticity of cells, the timing of acquired resistance to LDHA inhibitors was comparable with other targeted agents. Under long-term LDHAi treatment, glycolytic cells acquired resistance by increased oxidative phosporylation (OX-PHOS) in a mechanism dependent on the AMPK stress response pathway; targeting either AMPK, downstream kinases, or OX-PHOS using tool compounds synergized with and prevented acquired resistance to GNE-140. Taken together, our data suggests that targeting anaerobic glycolysis may benefit a subset of patients across indications and that combinations with agents that block AMPK signaling or the mitochondria will be effective at delaying tumor relapse.

Citation Format: Aaron Boudreau, David Peterson, John Moffat, Bonnie Liu, Mandy Kwong, Min Gao, Hans Purkey, Thomas O'Brien, Georgia Hatzivassiliou, Anneleen Daemen, Marie Evangelista. Resistance to LDHA inhibitors requires signaling through the AMPK/mTOR/S6K pathway leading to increased oxidative phosphorylation. [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 1423. doi:10.1158/1538-7445.AM2014-1423
Abstract 143: Non-canonical role of TAM receptors in breast cancer
Authors: Hsieh Shelly , Nguyen Khanh Q. , Kimani Stanley , Birge Raymond B.
Published: May 2015

TAM receptors (Tyro-3, Axl, Mer) are a family of three homologous receptor tyrosine kinases that play a role in the phagocytosis of apoptotic cells (a process termed efferocytosis in recent years). TAM receptors on the surface of epithelial cells bind to soluble ligands Gas6 and Protein S, which in turn, bind to phosphatidylserine-expressing apoptotic cells. After epithelial cells phagocytose apoptotic cells, they release anti-inflammatory cytokines (e.g. IL-10, TGFβ) to attenuate the immune response. This non-canonical role of TAM receptors as efferocytosis receptors is in addition to their well-studied canonical role as tyrosine kinase receptors that activate cell proliferation signaling pathways (e.g. PI3K, Ras, ERK). Therefore, overexpression of TAM receptors on cancer cells has a dual effect on tumor progression. To date, TAM receptors have been implicated in numerous cancers, including breast cancer; and various family members have been shown to be overexpressed in mouse and human mammary tumor cells. We propose that efferocytosis is a unique and novel oncogenic event by virtue of its role in the production of immunosuppressing factors into the tumor microenvironment. To study this, we set up a unique 3D acini model, where luminal apoptotic cells are engulfed by the basal cells of the acini. Using this system along with 2D cultures, we demonstrated that overexpression of Mer increased the rate of efferocytosis, but does not act as a classical oncogene by filling in the luminal space. Additional studies such as time-lapse studies of acini stained with caspases showed that Mer increased the rate of clearance. This adapted method offers a unique system to study cancer cell efferocytosis, and the role of TAM receptors in cancer progression and immune evasion.

Citation Format: Shelly Hsieh, Khanh Q. Nguyen, Stanley Kimani, Raymond B. Birge. Non-canonical role of TAM receptors in 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 143. doi:10.1158/1538-7445.AM2014-143
Abstract 1430: Identification of distinct metabolic subtypes within pancreatic ductal adenocarcinoma through broad metabolite profiling
Authors: Daemen Anneleen , Peterson David , Sahu Nisebita , McCord Ron , Kowanetz Kaska , Hitz Anna , Du Xiangnan , Liu Bonnie , Gao Min , Moffat John , Hong Rebecca , Sampath Deepak , Merchant Mark , O'Brien Thomas , Yauch Bob , Settleman Jeff , Qing Jing , Hatzivassiliou Georgia , Evangelista Marie A.
Published: May 2015

Targeting cancer metabolism is currently being evaluated as a potential therapeutic target. Identification of tumor subtypes with specific metabolic requirements will enable the success of such therapies. Using cell line models of pancreatic ductal adenocarcinoma (PDAC), we conducted broad metabolite profiling and identified 3 metabolic subtypes through non-negative matrix factorization (NMF) consensus clustering. Among the most distinguishing features between the subtypes were levels of metabolites important for lipid synthesis, amino acid, and energy-related pathways, suggestive of distinct metabolic dependencies. Using RNAseq we show that these subtypes are defined at the transcriptional level and are associated with known subtypes of PDAC. In metabolic flux experiments, we also show that glucose and glutamine are differentially utilized by each subtype, and as a consequence, exhibit distinct sensitivity to inhibition of various metabolic pathways. Finally, through screening of a large panel of tumor cell lines with various metabolic inhibitors, we cross-validate our finding in PDAC and provide strong evidence that these same subtypes exist and can be identified in other tumor indications. Our findings highlight the utility of broad metabolite profiling to predict sensitivity to metabolic inhibitors in cancer cells.

Citation Format: Anneleen Daemen, David Peterson, Nisebita Sahu, Ron McCord, Kaska Kowanetz, Anna Hitz, Xiangnan Du, Bonnie Liu, Min Gao, John Moffat, Rebecca Hong, Deepak Sampath, Mark Merchant, Thomas O'Brien, Bob Yauch, Jeff Settleman, Jing Qing, Georgia Hatzivassiliou, Marie A. Evangelista. Identification of distinct metabolic subtypes within pancreatic ductal adenocarcinoma through broad metabolite profiling. [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 1430. doi:10.1158/1538-7445.AM2014-1430
Abstract 1433: Omental derived adipose stromal cells regulate nitric oxide homeostasis to maintain tumorigenicity in ovarian cancer cells
Authors: Rizi Bahar Salimian , Klopp Ann H. , Nagrath Deepak
Published: May 2015

The rate of obesity is increasing dramatically in the United States, resulting in a rising incidence of obesity associated diseases including colorectal, gallbladder, esophageal, renal, breast, endometrial and ovarian cancer. The location of excess adipose tissue is reportedly an important factor which influences the effect of obesity on cancer incidence. The tumor promoting effects of regional adipose may be attributed, at least in part, to the presence of mesenchymal stem cell (MSC) population in adipose tissue. Adipose derived stromal cells (ASCs) are a multi-potent population of MSCs, which support cancer progression in many models. These cells are uniquely tumor tropic, migrating into tumor stroma from regional sources where they support formation of tumor stroma elements and promote tumor progression. Thus, we hypothesize that the omentum provides a particularly hospitable microenvironment for OVCA metastasis due to the presence of omental ASCs (O-ASCs). One critically important, yet often overlooked contributor to ovarian tumor growth, progression and metastasis in obese individuals is nitric oxide (NO). Cancer cells' high affinity for NO and NO's characteristic high solubility in lipids could explain the proximity of many carcinomas to fatty tissue, and thus the high positive correlation between obesity and cancer. Here we report a novel NO mediated reciprocal coupling between O-ASCs and ovarian cancer which promotes cancer cell growth and energy metabolism. Our results show that O-ASCs supports viability of cancer cells by secreting arginine and cancer cells in turn induce adipogenesis in O-ASCs by secreting citrulline. We found that OASCs promote tumor growth and increase resistance to chemotherapy by modulating NO homeostasis. The inhibition of NO synthesis or scavenging of NO enhanced chemo-sensitivity. Using metabolic profiling we found that OASCs rewire metabolic pathways related to nitrogenous amino acids and reprogram energy metabolism. In summary, our data indicate that modulation of intrinsic NO substantially affects tumor growth and drug toxicity in ovarian cancer cells. Our study will lead to significant advances in the understanding of the omentum in altering cancer metabolism and lead to novel therapeutics.

Citation Format: Bahar Salimian Rizi, Ann H. Klopp, Deepak Nagrath. Omental derived adipose stromal cells regulate nitric oxide homeostasis to maintain tumorigenicity in ovarian cancer 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 1433. doi:10.1158/1538-7445.AM2014-1433
Abstract 1436: Investigating the NAD metabolome in Ewing sarcoma
Authors: Mutz Cornelia N. , Ban Jozef , Niedan Stephan , Kauer Maximilian O. , Aryee Dave N.T. , Fuchs Dietmar , Heitger Andreas , Kovar Heinrich
Published: May 2015

Ewing Sarcoma (ES) is the second most common bone cancer in children and adolescents with a high metastatic potential. Tumor development is driven by the specific t(11;22)(q24;q12) chromosomal translocation resulting in generation of the chimeric transcription factor EWS-FLI1.

Recently, ES has been reported to be exquisitely sensitive to inhibitors of poly(ADP-ribose) polymerase 1 (PARP1). This enzyme uses NAD+ as substrate and was demonstrated to regulate EWS-FLI1 in a feed-back mechanism. Another major mammalian NAD+ consumer is the deacetylase SIRT1 which we observed to be highly expressed in ES metastases, validated with immunohistochemistry of 250 primary tumors and 30 metastases. PARP1 and SIRT1 play pivotal roles in coupling cellular metabolism to transcriptional gene regulation as well as to stress response. Both of them regulate pro-apoptotic transcription factors that are suppressed in ES including p53, FOXO1 and 3, and NFκB. It has been demonstrated that PARP1 activity is at least partially dependent on acetylation which is counteracted by SIRT1 activity. Severe cell stress such as DNA damage leads to massive activation of PARP1, thus resulting in depletion of the cellular NAD+ pool and finally cell death. Usually, NAD+ is regenerated from nicotinamide via NAMPT or from the reduction of pyruvate via LDHA (Warburg effect), but can also be synthesized de novo from tryptophan. Interestingly, the knockdown of EWS-FLI1 in ES cells comes along with increased TDO and suppressed KMO expression, both being critical enzymes in tryptophan metabolism and NAD+ de novo synthesis. As a consequence, cellular tryptophan consumption drastically increases, which might indicate a regulatory function of EWS-FLI1 in maintaining the balance of the cellular NAD+ metabolism. In addition, we observed a striking sensitivity of ES cells to interferon gamma treatment at very low doses (5U/ml) resulting in the induction of the tryptophan metabolizing enzyme IDO and the production of the immune-suppressive metabolite kynurenine. We are currently studying how these changes in tryptophan metabolism affect cellular NAD+ pools and SIRT1 and PARP1 activities in ES. This study serves for a better understanding of EWS-FLI1 and of the microenvironment in the post-translational control of SIRT1 and PARP1 regulated gene transcription, and the consequences for PARP1 and/or SIRT1 directed therapies of ES.

Supported by the Austrian Science fund, grant I1225-B19.

Citation Format: Cornelia N. Mutz, Jozef Ban, Stephan Niedan, Maximilian O. Kauer, Dave N.T. Aryee, Dietmar Fuchs, Andreas Heitger, Heinrich Kovar. Investigating the NAD metabolome in Ewing sarcoma. [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 1436. doi:10.1158/1538-7445.AM2014-1436
Abstract 1440: Metabolic therapy reduces expression of PECAM-1/CD31 and decreases peritumoral edema in a mouse model of malignant glioma
Authors: Woolf Eric C. , Charlton Julie A. , Liu Qingwei , Turner Gregory , Preul Mark C. , Scheck Adrienne C.
Published: May 2015

Patients with malignant brain tumors have a median survival of approximately one year following diagnosis, regardless of currently available treatments which include surgery followed by radiation and chemotherapy. Improvement in the survival of brain cancer patients requires the design of new therapeutic modalities that take advantage of common phenotypes. One such phenotype is the metabolic dysregulation that is a hallmark of cancer cells. It has therefore been postulated that one approach to treating brain tumors may be by metabolic alteration such as that which occurs through the use of the ketogenic diet (KD). The KD is high-fat, low-carbohydrate diet that induces ketosis and has been utilized for the non-pharmacologic treatment of refractory epilepsy. We and others have shown that this diet enhances survival and potentiates standard therapy in mouse models of malignant gliomas, yet the anti-tumor mechanisms are not fully understood.

It has been previously shown that caloric restriction, which induces ketosis, reduces microvessel density in mouse and human brain tumor models, suggesting an anti-angiogenic effect. We now report that in animals fed KetoCal® (KC) (4:1 fat:protein/carbohydrates) ad libitum, peritumoral edema is significantly reduced early in tumor progression when compared to those fed a standard rodent diet (SD). Western blot analysis showed a reduction of platelet endothelial cell adhesion molecule 1 (PECAM1/CD31) in tumors from animals maintained on KC. These results were supported by immunohistochemical staining for CD31 which also revealed abnormal vessel structure in the tumors from animals fed SD but not in those fed KC, suggesting a normalizing effect by the KC. Furthermore gene expression profiling demonstrated that KC decreases expression of a group of genes involved in angiogenesis and vessel structuring including the genes encoding vascular endothelial growth factor B (VEGFB) and angiopoetin 1 receptor (TEK), integrin beta 1 (ITGB1), urokinase-type plasminogen activator (PLAU) and tissue inhibitor of metalloproteases 1 (TIMP1). Taken together our data suggests that KC alters the angiogenic processes involved in malignant progression of gliomas. A greater understanding of the effects of the ketogenic diet as an adjuvant therapy will allow for a more rational approach to its clinical use.

Citation Format: Eric C. Woolf, Julie A. Charlton, Qingwei Liu, Gregory Turner, Mark C. Preul, Adrienne C. Scheck. Metabolic therapy reduces expression of PECAM-1/CD31 and decreases peritumoral edema in a mouse model of malignant glioma. [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 1440. doi:10.1158/1538-7445.AM2014-1440
Abstract 1441: Induction of cancer cachexia by inflammatory molecules in directed complementation tumor models
Authors: Liu Qing , Lerner Lorena , Siddiquee Zakir , Huang Lucia , Wang Ruojie , Tao Nianjun , Krieger Brian , Chiu Isabel , Gyuris Jeno
Published: May 2015

Cancer cachexia is a complex multi-factorial syndrome characterized by anorexia, body weight loss that is primarily due to the loss of skeletal muscle and adipose tissue, as well as systemic inflammation. It affects the majority of advanced cancer patients, being the cause of death in 20% to 40% of the cases. Although inflammation is recognized as an important component of cancer cachexia, the role of individual cytokines is poorly understood. Our analyses of inflammatory markers in two independent human cancer cachexia plasma collections identified several cytokines (GDF-15, Activin-A, IL-6 and IL-8) that are consistently upregulated in cancer cachexia patients. To examine the ability of these cytokines to induce cancer cachexia, we utilized an inducible breast cancer model as the background in which to create a panel of genetically-engineered mouse tumor models that overexpress targets of interests, namely GDF15, Activin A and IL-6. Such complemented tumor models present a unique preclinical opportunity for dissecting the role of candidate markers in vivo, in part because they were generated on a genetically homogeneous background, allowing the role of each individual target to be identified and differentiated.

Mice bearing tumors that overexpress each inflammatory marker were characterized for phenotypic manifestation of cachexia. Mice bearing tumors that overexpress GDF15 (or MIC-1, macrophage inhibitory cytokine-1) developed severe cachexia with a rapid onset of body weight loss as well as changes in body composition. Overexpression of Activin A or IL-6 induced partial body weight loss with a milder cachexia phenotype at concentrations significantly higher that seen in cachexic human patients, while mice bearing control tumors without the engineered expression of an inflammatory cytokine showed no detectable changes in either body weight or body composition.

These results indicate that the elevated expression of GDF-15 and to a lesser extent Activin A, and IL-6 can result in the development of cancer cachexia and may represent attractive therapeutic targets.

Citation Format: Qing Liu, Lorena Lerner, Zakir Siddiquee, Lucia Huang, Ruojie Wang, Nianjun Tao, Brian Krieger, Isabel Chiu, Jeno Gyuris. Induction of cancer cachexia by inflammatory molecules in directed complementation tumor models. [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 1441. doi:10.1158/1538-7445.AM2014-1441
Abstract 1443: A systematic approach to the metastatically relevant microRNA landscape
Authors: Mudduluru Giridhar , Abba Mohammed , Batliner Jasmin , Patil Nitin , Lunavat Taral R. , Scharp Maike , Leupold Jörg , Oleksiuk Olga , Buchhalter Ivo , Thiele Wilko , Rothley Melanie , Benner Axel , Sleeman Jonathan , Allgayer Heike
Published: May 2015

Distant metastasis is the major determinant of patient outcome in colorectal cancer. By systematically analysing the miR expression profiles of resected metastasis-, corresponding primary tumor and normal tissues of colorectal cancer patients, we were able to delineate a miR-signature indicative of the metastatically critical microRNA landscape. This “hot list” of miRNAs, including established metastasis-related miRs like the miR-34- and let -7 families, as well as rather novel miR-candidates like miR-552, -218, -135, -210 and -654, together with their putative common targets were bioinformatically predicted to regulate the most significant metastasis-associated signaling pathways currently known. We were able to show for the first time that miR-135b, miR-210 and the loss of miR-218 constitute a novel network that regulates both E- and N-cadherin expression, which is achieved, in part, via several novel common targets such as the recently described tumor suppressors SIAH1 and SETD2, and, most importantly, FOXN3 transcription factor. This is paralleled by a significant impact on migration, invasion, in vivo intravasation and metastasis in chicken embryo and mouse models. We conclude that miR-218 as a novel metastasis suppressor, and pro-metastatic miR-135b and miR-210, constitute one important novel network critical for metastasis, resulting from the first study to systematically suggest the microRNA-driven master pathways of metatasis from solid cancer tissues.

Citation Format: Giridhar Mudduluru, Mohammed Abba, Jasmin Batliner, Nitin Patil, Taral R. Lunavat, Maike Scharp, Jörg Leupold, Olga Oleksiuk, Ivo Buchhalter, Wilko Thiele, Melanie Rothley, Axel Benner, Jonathan Sleeman, Heike Allgayer. A systematic approach to the metastatically relevant microRNA landscape. [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 1443. doi:10.1158/1538-7445.AM2014-1443
Abstract 1444: Sensitivity to trastuzumab for gastric cancer is regulated by miR-223/FBXW7 pathway
Authors: Eto Kojiro , Iwatsuki Masaaki , Ishimoto Takatsugu , Baba Hideo
Published: May 2015

Background: A recent large-scale phase III study (the ToGA trial) demonstrated the significant efficacy of trastuzumab in addition to chemotherapy in patients with HER2-positive gastric cancer (GC). Although trastuzumab has become a key drug in recent cancer treatment, resistance to trastuzumab for breast cancer is a major problem in clinical practice. However, it is unclear whether similar mechanisms of trastuzumab resistance act in GC. F-box and WD repeat domain-containing 7 (FBXW7) is the substrate recognition components of an evolutionarily conserved SCF (complex of SKP1, CUL1, and F-box protein)-type ubiquitin ligase complexe, which has been well characterized and shown to have important roles in regulating the stability of multiple oncoprotein substrates, including cyclin E, c-Myc, Notch, c-Jun, mTOR, and MCL1. Therefore, FBXW7 is recognized to be one of the major causes of drug resistance. The aim of the current study is to clarify the novel microRNA (miR)-FBXW7 pathway regulating resistance to trastuzumab. Method: Using 5 GC cell lines, we examined the following items in vitro. 1. We examined the status of FBXW7 expression by quantitative real-time reverse transcription-PCR (qRT-PCR). 2. We established the trastuzumab resistance GC cell line with continuous trastuzumab administration. Parent GC cell line and resistance GC cell line are subjected to miR PCR array. 3. We focused on miR223-FBXW7 pathway. We examined the status of HER2, miR223, FBXW7, and its target gene MCL1 expression by up or down regulation of miR-223 in GC cell lines. 4. We investigated whether the miR-223-FBXW7 pathway can affect the resistance of trastuzumab. Result: FBXW7 and miR-223 expression showed inverse correlation. Moreover, miR-223 expression was increased and FBXW7 expression was decreased whereas MCL1 expression, downstream targets of FBXW7, was increased while no significant change was observed both in HER2 and p-HER2 in GC trastuzumab resistance cells. Overexpression of miR-223 decreased FBXW7 expression persisting HER2 and p-HER2 expression and can induce the resistance of trastuzumab. The suppression of miR-223 increased FBXW7 expression persisting HER2 and p-HER2 expression and can simultaneously restore the sensitivity of trastuzumab.Conclusion: Our current study revealed for the first time that regulated the sensitivity of HER2-positive GC cell line to trastuzumab. These findings suggest that this pathway may be crucial to the mechanism of resistance to trastuzumab in GC, which may lead to the development of individualized treatment in clinical practice.

Citation Format: Kojiro Eto, Masaaki Iwatsuki, Takatsugu Ishimoto, Hideo Baba. Sensitivity to trastuzumab for gastric cancer is regulated by miR-223/FBXW7 pathway. [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 1444. doi:10.1158/1538-7445.AM2014-1444
Abstract 145: Fibulin-5 as a regulator of integrin-induced ROS production and potential target for pancreatic cancer
Authors: Topalovski Mary , Wang Miao , Brekken Rolf A.
Published: May 2015

Reactive oxygen species (ROS) function as signaling molecules in pathological and normal molecular pathways. Redox homeostasis is maintained by a balance between the ROS production and ROS scavenging capacity of the cell. Extracellular matrix (ECM) proteins such as fibronectin have been shown to induce ROS upon binding to cell surface integrins, such as α5β1 integrin. Solid tumors rich in ECM such as pancreatic ductal adenocarcinoma (PDAC) display with higher levels of ROS compared to normal tissue. An attractive strategy to improve therapeutic response in PDAC is to further increase the level of ROS, thereby producing irreversible stress to tumor cells and surrounding stromal cells. Furthermore, increased ROS production has been shown to enhance the efficacy of some chemotherapeutics and might also improve the potency of anti-angiogenic therapy.

We propose a novel function of the matricellular protein Fibulin-5 (Fbln5) is to regulate ROS levels during angiogenesis. Fbln5 is expressed highly in human PDAC tissue and in murine models of pancreatic cancer. We have seen that the genetic ablation of of Fbln5 results in decreased tumor growth in implant and genetic models of pancreatic cancer. Furthermore, this decrease in tumor growth is accompanied by a decrease in microvessel density. Our lab has discovered that Fbln5 reduces integrin-mediated ROS generation by competing with fibronectin for binding to α5β1 integrin. Fbln5 binds to α5β1 integrin via an RGD motif, mutation of which to RGE abolishes the Fbln5 - integrin interaction. Knock-in Fbln5RGE/RGE mice have been produced and crossed into the background of KIC mice (p48Cre/+; LSL- KrasG12D; Ink4a/Arflox/lox). By utilizing this in vivo model we will be able to elucidate the mechanism by which Fbln5 regulates tumor angiogenesis and specifically integrin-induced ROS production. In the present study, our goal is to investigate the molecular mechanism leading to increased ROS production in tumors in Fbln5RGE/RGE animals. These targeted experiments will advance our understanding of the biology of Fbln5 in pancreatic cancer and ultimately contribute to its development as a potential therapeutic target.

Citation Format: Mary Topalovski, Miao Wang, Rolf A. Brekken. Fibulin-5 as a regulator of integrin-induced ROS production and potential target for 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 145. doi:10.1158/1538-7445.AM2014-145
Abstract 148: Release of the TNF-family member BAFF by NK cells contributes to the resistance of chronic lymphoid leukemia cells to direct and Rituximab-induced NK reactivity
Authors: Wild Julia , Schmiedel Benjamin J. , Maurer Andreas , Raab Stefanie , Schneider Pascal , Salih Helmut R.
Published: May 2015

NK cells are cytotoxic lymphocytes that are particularly important for the immunosurveillance of leukemia. Due to their ability to mediate antibody-dependent cellular cytotoxicity (ADCC), NK cells substantially contribute to the therapeutic benefit of antitumor antibodies like Rituximab, which is a crucial component in the treatment of B cell malignancies. Available data indicate that the ability of NK cells to target malignant cells and to mediate ADCC is compromised in Chronic Lymphoid Leukemia (CLL), but the underlying mechanisms are still unclear (Jaglowski et al., Blood 2010). With the TNF family member B cell activating factor (BAFF) that regulates, among others, the proliferation and survival of B cells, we believe to have identified a factor which contributes to these clinical observations. We report that NK cells express BAFF on the mRNA level and release the protein in a soluble form with the levels depending on activation state. Notably, relevant BAFF surface expression was neither detected on resting nor activated NK cells. Using a tetrazolium salt based colorimetric assay we found that NK cell-derived BAFF enhances the metabolic activity of primary CLL cells. In addition, BAFF protects CLL cells from chemotherapy-induced cell death e.g. upon treatment with Fludarabine and Cyclophosphamide. Notably, the BAFF-specific antibody Belimumab, which is approved for the treatment of systemic lupus erythematosus, was found to block the leukemia cell-protective effects of BAFF and to restore the sensitivity of the leukemia cells to chemotherapy. Furthermore, BAFF was found to substantially reduce direct NK cell lysis and also Rituximab-mediated ADCC in cultures with primary CLL cells as measured by Europium cytotoxicity assays. Our data on the capacity of BAFF to enhance the metabolic activity of B-CLL cells and their resistance to NK cell cytotoxicity suggest its involvement in the compromised ability of NK cells to combat lymphoid as compared to myeloid leukemias (Pende et al., Blood 2005) and the reportedly impaired ability of NK cells to mediate ADCC upon Rituximab treatment in CLL. Furthermore, our findings point to a possible benefit of combinatory approaches employing Rituximab and Belimumab for immunotherapy of B cell malignancies.

Citation Format: Julia Wild, Benjamin J. Schmiedel, Andreas Maurer, Stefanie Raab, Pascal Schneider, Helmut R. Salih. Release of the TNF-family member BAFF by NK cells contributes to the resistance of chronic lymphoid leukemia cells to direct and Rituximab-induced NK reactivity. [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 148. doi:10.1158/1538-7445.AM2014-148
Abstract 1493: Specific miRNA signatures characterize different metastatic sites in clear cell renal cell carcinoma
Authors: Heinzelmann Joana , Stolzenbach Franzsika , Schneeweiss Robert , Wickmann Ulrike , Baumgart Sophie , Gajda Mieczyslaw , Stöckle Michael , Junker Kerstin
Published: May 2015

Background: MiRNAs are regulators of gene expression in tumorigenesis and progression. To identify miRNAs associated with metastasis miRNA expression in distant metastases was compared to primary clear cell renal cell carcinoma (ccRCC).

Material and methods: Total RNA of 27 primary ccRCC samples and 25 distant metastases (lung, bone and brain) was isolated from formalin-fixed paraffin-embedded (FFPE) samples Microarray analyses were performed for a global miRNA expression profiling. Results were validated by qPCR. For miRNA target identification a ccRCC cell line (786-O) was transfected with miRNAs differently expressed in metastatic primary tumors and metastases.

Results: We identified 9 miRNAs (including miR-30c and miR-126) with a similar expression in metastatic primary ccRCC and distant metastases from different metastatic sites compared to non-metastatic primary ccRCC. 11 miRNAs (including miR-10b and miR-204) were differently expressed in distant metastases compared to primary ccRCC. Furthermore, each metastatic site is characterized by a specific miRNA signature. Results were verified on selected miRNAs using qPCR. In vitro studies identified potential miRNA targets and associated metastasis related pathways.

Discussion: These data suggest that miRNAs play an important role in metastatic processes of ccRCC. Furthermore, our results regarding different metastatic sites suggest two important statements. Specific miRNAs characterize distant metastases in general. On the other hand, miRNA expression is associated with specific conditions at different metastatic sites. Thus, the data presented in this study give the base for a better understanding of the involvement of miRNAs as regulators of metastasis which opens new possibilities for new targeted therapy options.

Citation Format: Joana Heinzelmann, Franzsika Stolzenbach, Robert Schneeweiss, Ulrike Wickmann, Sophie Baumgart, Mieczyslaw Gajda, Michael Stöckle, Kerstin Junker. Specific miRNA signatures characterize different metastatic sites in clear cell renal 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 1493. doi:10.1158/1538-7445.AM2014-1493
Abstract 1495: Identification of markers for the presence of lymph nodes metastasis in patients with oral squamous cell carcinomas
Authors: Zamunér Fernando Tadeu , Maia Danielle Calheiros Campelo , Carvalho André Lopes , Vettore André Luiz
Published: May 2015

INTRODUCTION: Oral cancer is the most common subset of head and neck squamous cell carcinomas (OSCC). These tumors often have an aggressive clinical outcome hallmarked by a propensity for local invasion and regional nodal metastasis. The presence of metastatic disease in cervical lymph nodes is an important prognostic factor and it influences in the choice of the therapeutic strategy to be adopted. Thus, the identification of molecular markers able to select patients who are at high risk for lymph node metastasis and/or locoregional recurrence could be of great value. Recent studies have shown that microRNAs can be regulators of the tumor metastasis process by the activation of various signaling pathways involved in this process being useful in diagnosis, determination of disease prognosis and prediction of therapeutic response. PURPOSE: to identify microRNAs expressed in OSCC associated with the presence of lymph node metastasis. METHODS: Tumor samples were obtained from patients diagnosed with OSCC at the Head and Neck Surgery Department, Cancer Hospital of Barretos. For microRNA microarrays experiments, total RNA from 32 tumor samples (12 without lymph node metastasis and 12 with metastasis) was extracted labelled and hybridizated Analysis of microarray data was performed using GeneSpring GX 12.0 and R program. RESULTS: This analysis allowed the identification of six microRNAs differentially expressed in the comparison between metastatic and non-metastatic OCCS cases. These microRNAs will be validate and selected by qRT-PCR in an independent cohort of OSCC patients and correlate the molecular data with the clinical features of the patients.

Financial Support: FAPESP

Note: This abstract was not presented at the meeting.

Citation Format: Fernando Tadeu Zamunér, Danielle Calheiros Campelo Maia, André Lopes Carvalho, André Luiz Vettore. Identification of markers for the presence of lymph nodes metastasis in patients with oral squamous cell 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 1495. doi:10.1158/1538-7445.AM2014-1495
Abstract 1496: MicroRNA signature associated with poor outcome of glioblastoma patients
Authors: Sana Jiri , Lakomy Radek , Fadrus Pavel , Smrcka Martin , Slampa Pavel , Kren Leos , Hermanova Marketa , Svoboda Marek , Slaby Ondrej
Published: May 2015

Background: Glioblastoma multiforme (GBM) is the most frequently occurring primary malignant brain tumor with very poor prognosis, varying therapeutic response. Therefore, it is very important to find new biomarkers which can predict a clinical outcomes and help in treatment decisions. MicroRNAs (miRNAs) are small, non-coding RNAs that function as post-transcriptional regulators of gene expression and play key role in pathogenesis of wide range of cancer, including GBM. Aim of our study is to identify miRNA signature associated with the more aggressive phenotype of GBMs, which will enable sensitive prediction of clinical outcome in GBM patients under standard therapeutic protocols.

Methods: We determined the expression profile of 667 miRNAs in tumor tissues of 58 patients with primary GBM with completed concomitant chemoradiotherapy with temozolomide according to standard Stupp protocol and 12 non-malignant brain tissues obtained from patients with drug-resistant anteromedial temporal lobe epilepsy. Methylation status of MGMT promoter (methylation-specific HRM), isocitrate dehydrogenase (IDH1) status (immunohistochemistry) and co-deletion of 1p/19q (FISH) was also evaluated.

Results: We have confirmed frequencies of commonly used prognostic biomarkers in GBM patients (MGMT, IDH1, 1p/19q deletion), and observed significant correlation of MGMT methylation status, response to chemoradiotherapy with temozolomide and survival of GBM patients. We have identified specific signature of miRNAs diferentially expressed between GBM tissue and non-tumoral brain tissue from epilepsy surgeries (28 miRNAs, p<10-10). We have confirmed some previous observations (e.g. up-regulation of miR-21, miR-155, down-regulation of miR-128a, miR-23a) and also identified miRNAs deregulated in GBM tissue which were observed for the first time (e.g. miR-220, miR-328, miR-888, miR-504). More importantly we have found miRNA signature (miR-224, miR-31, miR-454, miR-672, miR-885-5p, miR-432) significantly associated short progression-free survival (p<10-6) and overall survival (p<10-6).

Conclusions: These findings indicate that miRNAs play an important role in GBM pathogenesis and may serve not only as promising predictors of therapeutic outcome but also as potential therapeutic targets in GBM patients.

This work was supported by Grant NT13549-4/2012, NT13860 4/2012, NT11214-4/2010 and NT13514-4/2012 of the Czech Ministry of Health, MH CZ - DRO (MMCI, 00209805).

Citation Format: Jiri Sana, Radek Lakomy, Pavel Fadrus, Martin Smrcka, Pavel Slampa, Leos Kren, Marketa Hermanova, Marek Svoboda, Ondrej Slaby. MicroRNA signature associated with poor outcome of glioblastoma patients. [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 1496. doi:10.1158/1538-7445.AM2014-1496
Abstract 1503: Rapid detection of somatic mutations in cancer genes
Authors: Powell Michael J. , Pastor Larry , Diaz Rachel , Chen Lily , Wu George , Li Claudia , Zhang Aiguo
Published: May 2015

In the context of personalized cancer treatment, tumor mutational analysis by DNA sequencing has been described frequently as the “gold standard”, yet DNA sequencing is one of the least sensitive methods for characterizing mutation. At least 10-20% of a DNA population must be mutant for the mutation to be detected by standard DNA sequencing. Implicit acceptance that this level of sensitivity is sufficient to characterize tumor mutations only makes sense if one assumes that tumors are monoclonal and that all the critical driver mutations will be present in the bulk of the tumor. Yet these are incorrect assumptions. In fact, it has become clear that most if not all colon tumors are polyclonal and heterogeneous. Consequently, tumor driver mutations may not be detectable using standard DNA sequencing methods. In order to rapidly detect these low frequency mutations in tumor derived DNA we have developed a new technology that allows the detection of tumor driver and drug resistance somatic mutations at <0.1% sensitivity using quantitative PCR (qPCR) without the need for digital PCR or droplet digital PCR (ddPCR) methodology. We utilize xeno-nucleic acid (XNA) clamping probes that are specific for the target gene wild-type template and allow only the selective amplification of mismatched mutant templates. Direct detection of driver and drug resistance mutations within 2 hours of obtaining samples in KRAS, NRAS, BRAF, EGFR and JAK2 utilizing real-time qPCR with interchelating fluorescence dye detection. As well as a new end-point post-PCR microtiterplate based target gene mutant amplicon capture format that utilizes colorimetric or chemiluminescence detection will be described.

Citation Format: Michael J. Powell, Larry Pastor, Rachel Diaz, Lily Chen, George Wu, Claudia Li, Aiguo Zhang. Rapid detection of somatic mutations in cancer genes. [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 1503. doi:10.1158/1538-7445.AM2014-1503
Abstract 1517: Detection of SMAD4, MAP2K4, RB1 and CDKN2A gene deletions in human tumor cells by multiplex qPCR
Authors: Woo Cora , Wilde Joyce , Patel Sunali
Published: May 2015

Gene duplications and deletions are critical to understanding cancer biology. Deletion in SMAD4, MAP2K4, RB1 and CDKN2A genes are known to play important roles in certain types of cancer or cancer states. Characterization of such mutations in different tumor cells provide insight into the study of novel biomarkers of cancer susceptibility, initiation, progression and metastases and help identify them as risk factors for various types of cancer. We demonstrated a quantitative method to detect multiple gene deletions in human tumor cells by multiplex PCR. We designed four TaqMan® Copy Number Assays to interrogate common deletion mutations in SMAD4, MAP2K4, RB1 and CDKN2A genes and performed multiplex PCR alongside with endogenous controls on representative human tumor and normal cells in different tissues. Since the detection of multiple deletion targets were performed in one single reaction, we were able to profile multiple deletion mutations in one single sample. This method can potentially be applied to the study of Copy Number Variations (CNV) on biological samples with limited quantity such as FFPE samples, as well as enables the characterization of multiple targets on samples in exactly the same condition and state.

Citation Format: Cora Woo, Joyce Wilde, Sunali Patel. Detection of SMAD4, MAP2K4, RB1 and CDKN2A gene deletions in human tumor cells by multiplex qPCR. [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 1517. doi:10.1158/1538-7445.AM2014-1517
Abstract 1526: Integrative and comparative genomic analysis of East-Asian lung squamous cell carcinomas
Authors: Kim Youngwook , Hammerman Peter S. , Kim Jaegil , Getz Gad , Meyerson Matthew , Park Keunchil
Published: May 2015

Purpose

Lung squamous cell carcinoma (SqCC) is the second most prevalent type of lung cancer. Currently, no targeted-therapeutics are approved for treatment of this cancer, largely due to a lack of systematic understanding of the molecular pathogenesis of the disease. To identify therapeutic targets and perform comparative analyses of lung SqCC, we probed somatic genome alterations of lung SqCC cases from Korean patients.

Patients and Methods

We performed whole-exome sequencing of DNA from 104 lung SqCC samples from Korean patients and matched normal DNA. In addition, copy number analysis and transcriptome analysis were conducted for a subset of these samples. Clinical association with cancer-specific somatic alterations was investigated.

Results

This cancer cohort is characterized by a very high mutational burden with an average of 261 somatic exonic mutations per tumor and a mutational spectrum showing a signature of cigarette-smoke exposure. Seven genes demonstrated statistical enrichment for mutation (TP53, RB1, PTEN, NFE2L2, KEAP1, MLL2 and PIK3CA). Comparative analysis between Korean and North American lung SqCC demonstrated similar spectrum of alterations in these two populations, in contrast to the differences seen in lung adenocarcinoma. We also uncovered recurrent occurrence of therapeutically actionable FGFR3-TACC3 fusion in lung SqCC.

Conclusion

These findings provide new steps towards the identification of genomic target candidates for precision medicine in lung SqCC, a disease with a significant unmet medical need.

Note: This abstract was not presented at the meeting.

Citation Format: Youngwook Kim, Peter S. Hammerman, Jaegil Kim, Gad Getz, Matthew Meyerson, Keunchil Park. Integrative and comparative genomic analysis of East-Asian lung squamous cell 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 1526. doi:10.1158/1538-7445.AM2014-1526