bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4327: Mitochondrial reprogramming in triple negative breast cancer progression
Authors: Park Junhyoung , Arora Kavisha , Vithayathil Sajna A. , Donti Taraka R. , Creighton Chad , Lewis Michael T. , Sreekumar Arun , Wong Lee-Jun , Kaipparettu Benny A.
Published: May 2015

Unlike other subgroups of patients with breast cancer (BCa), for triple negative (TN) BCa, there is a current lack of understanding of driver pathways and hence are often managed using more generic therapies. Thus, it is important to identify the underlying mechanisms of triple negative breast cancer progression and its racial disparity. Mitochondria-nuclear crosstalk is a bidirectional pathway of communication between mitochondria and nucleus that influences many cellular and organismal activities. This crosstalk can regulate several oncogenic pathways that regulate cellular tumor characteristics. With the advent of transmitochondrial cybrid (cybrid) technology, it is now possible to examine the specific contribution of tumor-associated mitochondria in neoplastic growth and development under a defined common nuclear background. We used this model system to identify mitochondria regulated pathways in TN BCa. Among different oncogenic pathways, Src pathway is frequently over-expressed in TN BCa and Src inhibitors are proposed as one of their current treatments of choice. However, single drug therapy with Src inhibitors failed to control metastatic TN BCa in recent Phase-II trials. These highlight an important question on the mechanism behind the regulation of Src oncogenic signature in TN BCa. Using cybrid technology, we have discovered critical role of cancer mitochondria in the post-translational modification of Src oncogenic pathway. Interestingly we discovered that the Src autophosphorylation, which activate Src-mediated oncogenic pathway, depends on mitochondrial tumor characteristics. Specifically, our strong preliminary data suggest that mitochondrial reprogramming is a critical step in the autophosphorylation and activation of Src pathway in metastatic TN BCa cells. Our findings suggest mitochondria targeted drugs as promising combination therapy for the management of Src positive TN BCa.

Citation Format: Junhyoung Park, Kavisha Arora, Sajna A. Vithayathil, Taraka R. Donti, Chad Creighton, Michael T. Lewis, Arun Sreekumar, Lee-Jun Wong, Benny A. Kaipparettu. Mitochondrial reprogramming in triple negative breast cancer progression. [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 4327. doi:10.1158/1538-7445.AM2014-4327
Abstract 4338: The lipogenic phenotype of HER2/neu-positive breast cancer cells
Authors: Baumann Jan M. , Conklin Douglas S.
Published: May 2015

15 - 30 % of all breast cancer cases are classified as HER2/neu-positive, a very aggressive form of breast cancer that correlates with a poor prognosis, reduced survival time and increased incidence of metastasis. We have shown that the HER2-positive breast cancer cell line BT474 relies on a unique Warburg-like metabolism for survival and aggressive behavior. These cells are dependent on fatty acid synthesis, presumably as a means to regenerate NAD+ for aerobic glycolysis. The cells show markedly increased levels of stored fats compared to other breast cancer cell lines and disruption of the synthetic process results in apoptosis.

This metabolic phenotype is operating at its limits in HER2-positive cells, as addition of exogenous palmitate results in cell death, whereas HER2-normal cells are not affected by this treatment. Microarray analysis of palmitate-treated HER2/neu-positive SKBR3 cells and HER2-normal MCF7 cells suggests that palmitate induces an ER-stress response in the HER2-positive cells. This is congruent with protein markers for stress-related translational stalling, which ultimately result in CHOP-dependent cell death. Palmitate distinctly reduces the expression of the HER3 protein and combination treatments with palmitate and HER2-targeted therapeutic compounds show synergistic effects and markedly increase the cytotoxic effects of either compound alone.

In addition to these translational changes, palmitate treatment of HER2/neu-positive SKBR3 cells results in AMPK activation and subsequent inhibition of fatty acid synthesis. Transcriptome and global metabolite profiling analyses of the effect of increased palmitate on these cells indicate that palmitate alters the metabolic phenotype by re-routing the glucose carbon flux in the cells.

Supported by CDMRP Award W81XWH-11-1-0094 to Jan Baumann and NCI Award 5R01CA136658-03 to Douglas Conklin.

Citation Format: Jan M. Baumann, Douglas S. Conklin. The lipogenic phenotype of HER2/neu-positive breast 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 4338. doi:10.1158/1538-7445.AM2014-4338
Abstract 4348: TGF-β enhances mir-455-5p expression that regulates tumorigenesis through UBE2B in betel quid-associated oral cancer
Authors: Cheng Chao-Min , Shiah Shine-Gwo , Chen Ya-Wen , Chang Jang-Yang
Published: May 2015

Oral cancer is the fourth foremost case of death from cancer, and the major causes of death are local recurrence and distant metastasis. The strong associations between oral squamous cell carcinoma (OSCC) and dietary habits such as alcohol consumption (A), betel quid chewing (B) and cigarette smoking (C) and its predominance in men have been well documented. Evidences have shown that miRNAs are involved in tumorigenesis of various cancers through regulating their downstream targets. In order to identify changes in miRNAs specific for betel quid-associated oral squamous cell carcinoma (OSCC), we investigated the expression profiles of miRNAs in a panel of 40 pairs of OSCC specimens and their matched non-tumorous epithelial counterparts. Our data showed that the expression levels of mir-455-5p are significantly (P<0.0001) higher in tumor tissue than in their counterpart normal tissue. In addition, the expression levels of mir-455-5p in oral cancer cell lines are also higher than normal oral epithelial cell line. Knockdown of mir-455-5p reduces both anchorage-independent growth and proliferative ability in two head and neck cancer cells, and vice versa in mir-455-5p overexpressing cells. Collectively, these evidences showed that mir-455-5p serves as an oncomir in betel quid-associated oral cancer. Furthermore, by analyzing both the array data from cancer patients and cell lines, and then intersected with the results from website tools, we have identified ubiquitin-conjugating enzyme E2B (UBE2B) to be the target of mir-455-5p, which was further validated by 3′-UTR luciferase reporter assay and Western blot analysis. We also revealed that UBE2B suppression actually rescued both cell transformation and growth ability in mir-455-5p knockdown cells. Our data further demonstrated that the expression level of mir-455-5p is regulated by transforming growth factor-β (TGF-β) pathway. Moreover, mir-455-5p expression was suppressed by the treatment of TGF-β downstream regulators. In conclusion, our results showed that the expression of mir-455-5p is regulated by TGF-β-dependent pathway, which subsequently leads to alteration the expression of UBE2B and contribute to the tumorigenesis of betel quid-associated oral cancer. (This work was supported by following grants: National Health Research Institutes CA-101-PP-22, National Research Program for Biopharmaceuticals 100CAP015-5 and National Science Council 102-2325-B-400-007)

Citation Format: Chao-Min Cheng, Shine-Gwo Shiah, Ya-Wen Chen, Jang-Yang Chang. TGF-β enhances mir-455-5p expression that regulates tumorigenesis through UBE2B in betel quid-associated oral 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 4348. doi:10.1158/1538-7445.AM2014-4348
Abstract 4349: Regulation of proliferation and platelet-derived growth factor expression in desmoid tumors by microRNA-29
Authors: Al-Jazrawe Mushriq , Poon Raymond , Liu Jessica , Alman Benjamin
Published: May 2015

Desmoid tumors (DT), also called aggressive fibromatosis, are a rare, locally invasive soft tissue tumors. While DTs harbor mutations that activate β-catenin, a major effector molecule of canonical Wnt signaling, they also display elevated levels of both Platelet-Derived Growth Factor Receptor Beta (PDGFRβ) and its PDGF-B ligand. We hypothesize that active PDGFRβ signaling maintains DT progression.

We studied the effect of PDGFRβ inhibition on DT growth. Primary cultures of DTs treated with the PDGFRβ inhibitor Sunitinib show a decrease in proliferation rate as measured by bromodeoxyuridine (BrdU) incorporation assay. In vivo, Sunitinib treatment of Apc1638N/+ mice, a mouse model of DT, also decreased the number and size of tumors. To investigate the relationship of β-catenin activation and PDGFRβ activation, we treated human skin fibroblasts with Wnt3a to activate canonical Wnt signaling, and utilized transgenic murine skin fibroblasts harboring an activated form of β-catenin. In both systems, PDGF-B ligand expression increased both at the mRNA and protein level. However, this increase was not observed immediately, suggesting an indirect mode of regulation. The PDGF-B gene contains a conserved microRNA-29 (miR29) binding site in its 3′ untranslated region. In silico miRacts analysis of five independent DT microarray studies predicted miR29 family deregulation. As such, we studied whether miR29 deregulation in DT leads to PDGF-B overexpression and tumor proliferation. By performing quantitative PCR on human DT samples, we observed that the levels of mature forms of all miR29 family members (MIR29A, B, and C) are significantly down-regulated. Overexpressing miR29 in DT cells decreases PDGF-B ligand levels as well as BrdU incorporation rate. Finally, β-catenin-induced modulation of the PDGF-B ligand is abrogated upon inhibiting miR29.

Taken together, these data suggest that PDGFRβ signaling is a potentially important pathway in DT progression. PDGFRβ may be linked to β-catenin by the novel discovery of microRNA-29 down-regulation in DT tumors. This work will offer new perspectives on the molecular biology underlying DT development and help identify novel targets for therapy.

Citation Format: Mushriq Al-Jazrawe, Raymond Poon, Jessica Liu, Benjamin Alman. Regulation of proliferation and platelet-derived growth factor expression in desmoid tumors by microRNA-29. [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 4349. doi:10.1158/1538-7445.AM2014-4349
Abstract 4353: microRNA-196b induces epithelial-to-mesenchymal transition through targeting HOXA9 in non-small cell lung cancer cells
Authors: Son Ji Woong , Yu Seong-Lan , Lee Dong Chul , Lee Shin Yup , Park Chang Gyo , Lee Hoi Young , Kang Jaeku
Published: May 2015

MicroRNAs (miRNAs) play important roles as oncogenes or tumor suppressor through modulation of expression of target genes as posttranscriptional regulators. miR-196b has been previously reported dysregulation in multiple tumor types including gastric cancer, colorectal cancer, glioblastoma and breast cancer. However, there is unknown about the role of miR-196b in lung cancer. There has been reported reduction of invasiveness via Homeobox A9 (HOXA9) overexpression in non-small cell lung cancer (NSCLC) cells. We identified an inverse expression between miR-196b and HOXA9 in clinical samples compared to their corresponding normal counterparts. We demonstrated that HOXA9 expression was significantly regulated through alteration of miR-196b expression and miR-196b overexpression or knockdown of HOXA9 induced invasion in NSCLC cells. Moreover, the expression levels of matrix metalloproteinase (MMP) 9 and plasminogen activator, urokinase (uPA) associated with epithelial-mesenchymal transition (EMT) was induced by miR-196b overexpression or knockdown of HOXA9. In addition, the expression level of N-cadherin (Cdh2) was increased by upregulation of miR-196b and knockdown of HOXA9 decreased E-cadherin (Cdh1) expression. Therefore, our findings suggest that miR-196b may be potential therapeutic target in lung cancer.

Citation Format: Ji Woong Son, Seong-Lan Yu, Dong Chul Lee, Shin Yup Lee, Chang Gyo Park, Hoi Young Lee, Jaeku Kang. microRNA-196b induces epithelial-to-mesenchymal transition through targeting HOXA9 in non-small cell lung 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 4353. doi:10.1158/1538-7445.AM2014-4353
Abstract 4358: MicroRNA-198 inhibits non-small cell lung cancer migration and invasion through targeting OTX1 and VCP
Authors: Kim Jin-Ah , Yang Junguk , Jo Yeong Nang , Lee Chang Woo , Lee Ok-Jun , Choi Song-Yi , Kang Jong Soon , Yun Jieun
Published: May 2015

Lung cancer is the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) accounting for the majority of cases. In previous studies, microRNA-198 (miR-198) was shown to be significantly downregulated in lung adenocarcinoma-associated malignant pleural effusion and suggested as a diagnostic biomarker. Herein, we show that miR-198 regulates migration and invasion of NSCLS cells by directly targeting orthodenticle homeobox 1 (OTX1) and valosin containing protein (VCP). Expression of miR-198 suppressed migration and invasion of NSCLS cells, whereas inhibition of miR-198 expression increased the invasive efficiency of the cancer cells. MiR-198 directly targets 3′UTR of OTX1 and VCP, thus regulates their protein level. Furthermore, we found reduced levels of miR-198 in human NSCLS tissue samples compared to adjacent normal tissues. Our findings indicate that miR-198 regulates NSCLC cell migration and invasion, partially via the down-regulation of OTX1 and VCP. Thus, miR-198 may represent a potential therapeutic target for NSCLC intervention.

Citation Format: Jin-Ah Kim, Junguk Yang, Yeong Nang Jo, Chang Woo Lee, Ok-Jun Lee, Song-Yi Choi, Jong Soon Kang, Jieun Yun. MicroRNA-198 inhibits non-small cell lung cancer migration and invasion through targeting OTX1 and VCP. [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 4358. doi:10.1158/1538-7445.AM2014-4358
Abstract 4367: Tumor suppressor miR-30a inhibits LOX expression and progression in anaplastic thyroid cancer
Authors: Boufraqech Myriem , Nilubol Naris , Zhang Lisa , Sadowski Samira , Gara Sudheer Kumar , He Mei , Davis Sean , Quezado Martha , Kebebew Electron
Published: May 2015

one of the most aggressive human malignancies with a median survival time of less than one year. A better understanding of the molecular events involved in ATC initiation and progression that could be targeted for therapy is gravely needed. There is limited data on the role of miRNAs in ATC. Screening of miRNA expression profile in thyroid cancer samples showed significant downregulation of miR-30a in ATC compared to differentiated thyroid cancer and normal tissue. Ectopic expression of miR-30a in vitro decreased invasion and migration of ATC cells with downregulation of epithelial-mesenchymal transition (EMT) markers and Lysyl Oxidase (LOX) expression, a predicted target of miR-30a. Using an in vivo mouse model of ATC metastases, miR-30a significantly decreased lung metastasis with decreased vimentin, CD44 and LOX expression in lung metastases tumor tissue. We also found that LOX and its downstream target TWIST-1 are highly expressed in ATC, and the expression of both were significantly, inversely correlated with miR-30a expression (p= 0.0026). In addition, the analysis of 454 primary thyroid cancer samples from The Cancer Genome Atlas dataset demonstrated upregulation of LOX and downregulation of miR-30a in high-risk thyroid cancer. These findings suggest that LOX and miR-30a are prognostic markers for aggressive thyroid cancer.

To determine whether LOX is a direct effector of miR-30a, the 3′UTR region of LOX was cloned into a luciferase reporter plasmid. Transient transfection of 3′UTR-LOX along with miR-30a in ATC cell lines led to a decrease in luciferase activity when compared to the co-transfection of 3′UTR-LOX with negative control miRNA, demonstrating that LOX is a direct target of miR-30a.

In vitro treatment of 4 ATC cell lines with β-aminoproprionitrile (BAPN), an irreversible inhibitor of LOX, had no effect on cell proliferation but inhibited cell invasion and migration and significantly decreased EMT markers and CD44 expression. Moreover, BAPN-treated mice with metastatic human ATC tumors had significantly fewer tumors and smaller size of lung metastases. Our data taken together support a critical role of miR-30a in ATC progression, which is mediated by LOX, and suggests that miR-30a and LOX is possible therapeutic target in ATC.

Note: This abstract was not presented at the meeting.

Citation Format: Myriem Boufraqech, Naris Nilubol, Lisa Zhang, Samira Sadowski, Sudheer Kumar Gara, Mei He, Sean Davis, Martha Quezado, Electron Kebebew. Tumor suppressor miR-30a inhibits LOX expression and progression in anaplastic thyroid 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 4367. doi:10.1158/1538-7445.AM2014-4367
Abstract 4369: The microRNA-218-survivin axis regulates cervical cancer cell migration and invasion
Authors: Kogo Ryunosuke , How Christine , Bruce Jeff , Shi Willa , Yip Kenneth W. , Ailles Laurie , Liu Fei-Fei
Published: May 2015

MicroRNA (miR)-218 down-regulation has been reported in numerous human malignancies. In cervical cancer, we identified that lower miR-218 expression was significantly associated with poorer overall survival, disease-free survival, and pelvic/para-aortic lymph node recurrence. Further analyses of cervical cancer data from The Cancer Genome Atlas (TCGA) identified that this down-regulation was associated with a genomic locus loss (hsa-mir-218-1:4p15.31, hsa-mir-218-2:5q34, n=105). The objective of the current study was to elucidate the cellular and molecular functions of miR-218. MiR-218 transfection into cervical cancer cells (SiHa and ME-180) significantly reduced cell migration (by 66% and 89%, respectively), invasion (by 49% and 67%, respectively), and clonogenic capacity (by 42% and 53%, respectively), relative to control-transfected cells (P<0.05). In order to identify clinically relevant miR-218 target genes, we used an integrated trimodal approach, incorporating DNA microarray (Affymetrix Human Genome U133 Plus 2.0) analyses of 79 clinical samples, miR-218 transfection, and miRDB target prediction. The most significant target was survivin (BIRC5); miR-218 transfection confirmed a reduction in survivin mRNA and protein expression in both SiHa and ME-180 cells. Furthermore, a direct interaction between the survivin-3′UTR and miR-218 was validated using a luciferase reporter assay. siRNA knockdown of survivin in SiHa and ME-180 significantly reduced cell migration (by 76% and 83%, respectively), invasion (by 79% and 88%, respectively), and clonogenic capacity (by 98% and 97%, respectively), relative to control cells (P<0.05). YM155, a small-molecule survivin suppressant, effectively reduced survivin mRNA and protein levels in a concentration- and time-dependent manner. This compound correspondingly decreased cervical cancer cell proliferation and clonogenic survival. Our results suggest that the miR-218-survivin axis plays an important role in cervical cancer progression.

Citation Format: Ryunosuke Kogo, Christine How, Jeff Bruce, Willa Shi, Kenneth W. Yip, Laurie Ailles, Fei-Fei Liu. The microRNA-218-survivin axis regulates cervical cancer cell migration and invasion. [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 4369. doi:10.1158/1538-7445.AM2014-4369
Abstract 4376: Upregulation of miR-21 in cisplatin-resistant ovarian cancer via JNK-1/c-Jun pathway
Authors: Echevarria Ileabett M. , Encarnacion Joel , Valiyeva Fatma , Vivas Pablo
Published: May 2015

Cisplatin has been the most accepted drug for the treatment of ovarian cancer for almost 40 years. Although the majority of patients with ovarian cancer respond to front-line platinum combination chemotherapy, many patients will develop cisplatin-resistance disease, which is extremely rapid and fatal. Although various mechanisms of cisplatin resistance have been postulated, the key molecules involved in such resistance have not been identified. MiRNAs are endogenously expressed small non-coding RNAs, which are evolutionarily conserved and function as post-transcriptional regulators of gene expression. Dysregulation of miRNAs have been associated with cancer initiation, progression and drug resistance. The oncogenic miRNA-21, one of the best-studied miRNAs, is upregulated in almost all human cancers. However, the regulation of miR-21 in cisplatin resistant ovarian cancer cells has not been assessed. In this study, we measured the miR-21 expression by real-time PCR and found upregulation of miR-21 in cisplatin resistant compared with cisplatin sensitive ovarian cancer cells. Chromatin immunoprecipitation studies demonstrated the association of the c-Jun transcription factor to the mir-21 DNA promoter regions. Blocking the JNK-1, the major activator of c-Jun phosphorylation, reduced the expression of miR-21 and increased the expression of its well-known target gene, PDCD4. Overexpression of miR-21 in cisplatin sensitive cells decreased PDCD4 levels and increased cell proliferation. Finally, targeting miR-21 reduced cell growth, proliferation and invasion of cisplatin resistant ovarian cancer cells. These results suggest that the JNK-1/c-Jun/miR-21 pathway contributes to the cisplatin resistance of ovarian cancer cells and demonstrated that miR-21 is a plausible target to overcome cisplatin resistance.

Citation Format: Ileabett M. Echevarria, Joel Encarnacion, Fatma Valiyeva, Pablo Vivas. Upregulation of miR-21 in cisplatin-resistant ovarian cancer via JNK-1/c-Jun 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 4376. doi:10.1158/1538-7445.AM2014-4376
Abstract 4378: miR-100 mediates resistance to paclitaxel in cervical cancer cells
Authors: Kato Rina , Nishi Hirotaka , Nagamitsu Yuzo , Sasaki Toru , Isaka Keiichi
Published: May 2015

Objectives: Hypoxia occurs during development of cervical cancer and is considered to correlate with its resistance to chemotherapy. We found that hypoxia induced miR-100 whose expression was low in cervical cancer. We sought to determine the regulators of the paclitaxel response during hypoxia in cervical cancer. Methods and Results: We found that the expression of miR-100 was significantly higher under hypoxic condition (1% O2) by realtime RT-PCR. According to the results of in silico analysis, miR-100 targeted human ubiquitin-specific protease (USP) 15. Hypoxia and overexpression of miR-100 respectively decreased the activity of luciferase-reporter containing the 3′-untranslated region (UTR) of USP15 with the predicted miR-100-binding site in cervical cancer cell lines, SiHa and HeLa. By western blot analysis, hypoxia enhanced the endogenous USP15 protein expression. USP15 mRNA level was also up-regulated by hypoxia using realtime RT-PCR. Overexpression of miR-100 induced paclitaxel resistance reducing apoptosis using MTT assays and annexin V staining. Conclusions: Our results provide evidence that regulation of USP15 expression by miR-100 represents a mechanism for paclitaxel resistance in cervical cancer. Our results suggest that miR-100 and USP15 could be a therapeutic target in paclitaxel-resistant cervical cancer.

Citation Format: Rina Kato, Hirotaka Nishi, Yuzo Nagamitsu, Toru Sasaki, Keiichi Isaka. miR-100 mediates resistance to paclitaxel in cervical 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 4378. doi:10.1158/1538-7445.AM2014-4378
Abstract 4406: Role of KRAS in modulating the metabolomic profile and the response of NSCLC cells to PI3K/mTOR and AMPK interfering agents
Authors: Caiola Elisa , Brunelli Laura , Marabese Mirko , Garassino Marina Chiara , Farina Gabriella , Broggini Massimo , Pastorelli Roberta
Published: May 2015

Background: Non-small-cell lung cancers (NSCLC) constitute the majority of lung cancer cases, whose risk is associated with environmental factors, such as air pollution, and smoking habits. KRAS mutations are present in about 20% of NSCLC patients, mostly at codon 12 and have been associated to tumor progression and resistance to chemotherapy in many types of cancer. Increasingly evidences seem to highlight that the definition of KRAS status is not sufficient to determine its predictive and/or prognostic role in NSCLC, and indicate that different KRAS mutations could differently impact on crucial cellular processes. An issue still to be addressed is whether different KRAS mutations may modulate cellular metabolism with possible implications on different response to anticancer treatments which specifically inhibit metabolism modulating pathways, such as the PI3K/AKT/mTOR axis.

Methods: We generated from the human NSCLC cell line NCI-H1299 clones overexpressing WT KRAS, and the three most common NSCLC KRAS mutations (G12C, G12V and G12D). The clones were used to determine in vitro response to PI3K, mTOR and AMPK interfering agents. We applied an explorative mass spectrometry-based untargeted metabolomics strategy to characterize the largest possible number of metabolites that might distinguish the different KRAS mutated clones.

Results: KRAS overexpressing clones resulted sensitive to the PI3K inhibitors BKM-120 and PIK-75 and to the PI3K/mTOR dual inhibitor BEZ-235. Metformin (acting on AMPK) did not display a cytotoxic activity on the clones at concentrations below or equal to 1mM. Metabolomics analyses revealed that clones harboring different KRAS mutations at codon 12 had different metabolic remodellings such as different redox buffering system and different glutamine-dependency.

Conclusion: The preliminary findings reported herein indicate that specific KRAS mutations at codon 12 are associated to different metabolomics profiles that might have an impact to the response to anticancer treatments. Combination studies including metformin and PI3K/mTOR interfering agents and more detailed metabolomics analyses are ongoing in order to clarify whether metabolic adaptations dependent on KRAS mutants might account for the behaviour of KRAS overexpressing clones treated with different drugs.

Citation Format: Elisa Caiola, Laura Brunelli, Mirko Marabese, Marina Chiara Garassino, Gabriella Farina, Massimo Broggini, Roberta Pastorelli. Role of KRAS in modulating the metabolomic profile and the response of NSCLC cells to PI3K/mTOR and AMPK interfering agents. [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 4406. doi:10.1158/1538-7445.AM2014-4406
Abstract 441: Functional characterization of recurrent CD74-NRG1 fusions in lung adenocarcinoma
Authors: Plenker Dennis , Fernandez-Cuesta Lynnette , Osada Hirotaka , Sun Ruping , Bos Marc , Daßler Juliane , Wright Gavin , Brambilla Elisabeth , Büttner Reinhard , Ansen Sascha , Haas Stefan , Yatabe Yasushi , Thomas Roman K.
Published: May 2015

In nearly 50% of all lung adenocarcinomas the oncogenic driver is still unknown. We performed transcriptome sequencing on 25 pan-negative lung adenocarcinomas of never smokers leading to the identification of the novel CD74-NRG1 gene fusion. In an additional cohort of 102 adenocarcinomas we could identify and validate four additional cases bearing the CD74-NRG1 fusion gene - all of the invasive mucinous subtype.

We then transduced NIH-3T3 cells with the fusion and found it localized on the cell surface in FACS experiments. As we found the identical breakpoint in CD74 in our fusion as in CD74-ROS1, we hypothesized that the functional active part needs to be NRG1. (Soda et al. 2012) The NRG1 isoform III-β3 that we found is generally not expressed in lung tissue - therefore the CD74 part leads to the expresseion of NRG1 III-β3 in the lung. As Neuregulins are known interaction partners of HER receptor family members we transduced the human adenocarcinoma cell lines H322 and H1568, that are HER receptor and KRAS wildtype, with the fusion and found phosphorylation of HER2 and HER3 in the transduced cells under starvation conditions compared to empty vector controls. (Hobbs et al. 2002) This holds true as well in the patient situation in which phospho-HER3 was only found in tumors expressing the fusion (p<0.0001).

We could further show that the CD74-NRG1 fusion is associated with activation of the phosphoinsitide-3-kinase pathway activation and the mitogen-activated protein (MAP) kinase pathway . As functional consequence we could observe in several human adenocarcinoma cell lines increased colony forming ability in soft-agar assays showing the CD74-NRG1 fusion being a new oncogenic driver in lung adenocarcinoma. This finding may lead to new treatment options for patients harboring this fusion gene who do not benefit from any effective treatment option up to now.

Citation Format: Dennis Plenker, Lynnette Fernandez-Cuesta, Hirotaka Osada, Ruping Sun, Marc Bos, Juliane Daßler, Gavin Wright, Elisabeth Brambilla, Reinhard Büttner, Sascha Ansen, Stefan Haas, Yasushi Yatabe, Roman K. Thomas. Functional characterization of recurrent CD74-NRG1 fusions in lung adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 441. doi:10.1158/1538-7445.AM2014-441
Abstract 4411: DRAK1 overexpressed in head and neck cancer suppresses the TGF-β1 tumor suppressor activity in head and neck cancer cells
Authors: Yang Kyung-Min , Kim WonJoo , Lee Jeong-Mi , Kim Seong-Jin
Published: May 2015

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide and a highly malignant cancer associated with poor prognosis and low overall survival. Here, we describe a role of DRAK1 (DAP kinase-related apoptosis-inducing kinase 1) in head and neck cancer cells. DRAK1 is significantly overexpressed in primary head and neck tumors and cell lines. Overexpression of DRAK1 decreased the TGF-β1-induced transcriptional activity and expression of tumor suppressor target genes, including p15, p21, whereas DRAK1 silencing enhanced. Furthermore, endogenous depletion of DRAK1 enhanced the TGF-β1-induced cell growth inhibition and suppressed the tumorigenic ability of head and neck cancer cells in vivo. Interestingly, the gain- or loss-of-function of DRAK1 did not change the TGF-β1-induced phosphorylation of Smad2/3. DRAK1 specifically binds to Smad3 through its kinase domain, thereby blocking recruitment of Smad4 and translocation of Smad3 into nucleus. Thus, our findings suggest that aberrant expression of DRAK1 increases tumorigenic potential through the inhibition of TGF-β1 tumor suppressor activity in head and neck cancer cells.

*This work was supported by a National Research Foundation grant of Korea (2009-0081756) funded by the Korea government.

Citation Format: Kyung-Min Yang, WonJoo Kim, Jeong-Mi Lee, Seong-Jin Kim. DRAK1 overexpressed in head and neck cancer suppresses the TGF-β1 tumor suppressor activity in head and neck 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 4411. doi:10.1158/1538-7445.AM2014-4411
Abstract 4413: Mnk activity in diffuse large b-cell lymphoma: targeting translation
Authors: Landon Ari , Muniandy Parameswary A. , Gartenhaus Ronald B.
Published: May 2015

Augmentation of protein translation has been observed in a number of human malignancies. We have previously demonstrated that cap-dependent protein translation is dysregulated in diffuse large B-cell lymphoma (DLBCL). This cancer-associated phenomenon may result in the deregulation of many proto-oncogenes and tumor suppressors leading to multiple cellular abnormalities. A major rate-limiting step in cap-dependent translation is the binding of eIF4E to the 7-methyl guanosine cap in the 5′ untranslated region of mRNA thus, bridging the eIF4F pre-initiation complex with the ribosome and initiating translation. eIF4E has been widely implicated as a proto-oncogene contributing to cancer development in numerous published reports. In addition to the mTOR signaling network, there is a potential for eIF4E to be modulated by the mitogen-activated protein kinases via map kinase interacting kinases (MNK). MNK proteins (MNK1 or MNK2) exert their effect on eIF4E through phosphorylation of eIF4E at serine 209. This phosphorylation is believed to promote B-cell lymphomagenesis. However, the physiologically relevant regulatory mechanisms and potential for targeting eIF4E phosphorylation in human B-cell lymphoma is largely unexplored. This study aims to elucidate the mechanistic and therapeutic effects of MNK regulation of eIF4E phosphorylation in DLBCL.

Counter-intuitively, we have found differential expression of MNK2 over MNK1 in activated B-cell (ABC) lymphoma versus germinal center B-cell (GCB) lymphoma subtypes even though both are known to converge to phosphorylate eIF4E. Interestingly, ERK does not play a significant role in activating MNK1/2 or affect the downstream phosphorylation of eIF4E in DLBCL, however, p38 plays an important role in eFI4E phosphorylation via MNK1. We show that phosphorylation of eIF4E by MNK is critical for malignant cell survival in both ABC and GCB DLBCL. The inhibition of p38 resulted in a significant and selective killing of DLBCL via inactivation of MNK1 and down-regulation of MCL1. Therefore, p38 inhibition may be a useful targeted approach in DLBCL through its suppression of MNK mediated eIF4E phosphorylation.

Citation Format: Ari Landon, Parameswary A. Muniandy, Ronald B. Gartenhaus. Mnk activity in diffuse large b-cell lymphoma: targeting translation. [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 4413. doi:10.1158/1538-7445.AM2014-4413
Abstract 4414: Neuropilin2 is a novel target gene of Wnt signaling pathway regulating angiogenesis, metastasis and growth of osteosarcoma
Authors: Ji Tao , Guo Yi , Lin Carol , Chen Cheng-Fong , Zi Xiaolin , Hoang Bang H.
Published: May 2015

Objective: Multivascularity of osteosarcoma (OS) rendered the anti-angiogenesis is a promising therapeutics. Neuropilin 2 (NRP2), a coreceptor of vascular endothelial growth factor (VEGF), has been implicated in tumor angiogenesis, growth, and metastasis of many cancers. Its act in osteosarcoma still remains poorly understood. In order to explore the role of NRP2 in modulating tumor growth, angiogenesis, invasion, and lung metastasis of osteosarcoma, we conduct the current study.

Methods: Cell culture, RNA interference, stable transfection, real time PCR, western blot, motility and invasion assay, in vivo tumorigenesis model, tube formation assay, ChIP.

Results: By microtissue array assay, the expression of NRP2 is implicated in metastasis and poor prognosis of osteosarcoma patients. The in vitro data showed NRP2 knockdown has an inhibitory effect on OS tumor cell growth, both anchoragedependent and anchorageindependent way. Xenograft model showed silencing of NRP2 expression in OS cells has an extraordinary inhibitory effect. Both migration and invasion were significantly impaired in two NRP2 knockdown OS cell lines in vitro. Again, a lung metastasis mice model was used to verify the antimetastasis effect after NRP2 silencing and the results showed metastasis was inhibited. The blood vessels and capillaries in the xenograft samples were evaluated by immunohistochemistry staining for CD31. The capillaries in NRP2 knockdown tumors were rarely seen compared with the intact capillaries in control tumors. In order to explore the mechanism underlying the anti angiogenesis effect, we conducted the coculture of tumor cells and HUVEC cells in both contact and noncontact way. The ability of recruiting HUVEC cells by OS cells were dramatically inhibited by NRP2 knockdown. Also NRP2 silencing caused the tumor cells to behave less involved in tube formation. ChIP assay revealed TCF4, wnt effector, bound all the five binding sites in NRP2 promoter region.

Conclusion: We demonstrated that NRP2 plays an important role in modulating tumor growth, angiogenesis, invasion, and lung metastasis of osteosarcoma.

Citation Format: Tao Ji, Yi Guo, Carol Lin, Cheng-Fong Chen, Xiaolin Zi, Bang H. Hoang. Neuropilin2 is a novel target gene of Wnt signaling pathway regulating angiogenesis, metastasis and growth of osteosarcoma. [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 4414. doi:10.1158/1538-7445.AM2014-4414
Abstract 4494: A clinical candidate anti-mesothelin-MMAE antibody-drug conjugate (ADC) for therapy of mesothelin-expressing cancers
Authors: Scales Suzie J. , Gupta Nidhi , Pacheco Glenn , Firestein Ron , French Dorothy M. , Chuh Josefa , Zhang Yin , Berry Leanne , Bostrom Jenny , Luis Elizabeth , O'Donohue Aimee Fourie , Kozak Katherine R. , Ross Sarajane , Dennis Mark S. , Tibbitts Jay , Spencer Susan D.
Published: May 2015

Mesothelin (MSLN) is a cell surface glycoprotein widely expressed in a variety of cancers, with prevalence levels by IHC of 85% in ovarian, 75% in pancreatic and 45% in mesothelioma, and normal expression limited to the mesothelia, suggesting it could be an ideal target for antibody-drug conjugate therapy of these cancers. We have generated a high affinity (subnanomolar), humanized antibody to MSLN and conjugated it to auristatin anti-mitotic drugs (monomethylauristatin E and F, Seattle Genetics) via an uncleavable linker (anti-MSLN-mc-MMAF) or a cathepsin-cleavable valine-citrulline linker (anti-MSLN-mc-vc-PAB-MMAE/F) for comparison. The in vivo efficacy obtained in an ovarian transplant model was superior with the anti-MSLN-MMAE ADC. Anti-MSLN-MMAE was specifically internalized by MSLN-expressing cells in vitro, resulting in cell death compared to control ADCs. Pancreatic, ovarian and mesothelioma tumor cell lines endogenously expressing physiological levels of MSLN were identified and established as xenografts in mice. A single dose of anti-MSLN ADC was sufficient to inhibit or shrink tumor growth in models of each of the three indications in vivo, as well as inducing complete regressions in primary human pancreatic models, even those expressing low levels of MSLN typical of most human pancreatic tumors. Additionally, anti-MSLN-MMAE (at suboptimal doses) appeared to synergize with gemcitabine at clinically relevant doses in an HPAC xenograft model. Furthermore, anti-MSLN-MMAE was well tolerated in non-clinical toxicity studies (see accompanying abstract by Gupta et al.). Our data suggest that anti-MSLN-vc-MMAE is a promising clinical candidate for the treatment of several types of mesothelin-positive cancers.

Citation Format: Suzie J. Scales, Nidhi Gupta, Glenn Pacheco, Ron Firestein, Dorothy M. French, Josefa Chuh, Yin Zhang, Leanne Berry, Jenny Bostrom, Elizabeth Luis, Aimee Fourie O'Donohue, Katherine R. Kozak, Sarajane Ross, Mark S. Dennis, Jay Tibbitts, Susan D. Spencer. A clinical candidate anti-mesothelin-MMAE antibody-drug conjugate (ADC) for therapy of mesothelin-expressing cancers. [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 4494. doi:10.1158/1538-7445.AM2014-4494
Abstract 4526: BIBF1120, an investigational triple angiokinase inhibitor, in combination with inhibitors of mTOR signaling shows potent antitumor activity in preclinical models of sarcoma
Authors: Patwardhan Parag P. , Musi Elgilda , Ivy Kathryn S. , Giovino Natalie , Schwartz Gary K.
Published: May 2015

BIBF1120 (Nintedanib), an investigational potent small molecule angiokinase inhibitor, selectively binds to and blocks receptor tyrosine kinases (RTKs) such as vascular endothelial growth factor receptor, (VEGFR), fibroblast growth factor receptor (FGFR) and platelet derived growth factor receptor (PDGFR). Nintedanib has shown promising results in preclinical as well as clinical studies of non-small cell lung cancer, colorectal, ovarian cancer and other solid tumors. We tested the in vitro anti-proliferative activity of BIBF1120 in a panel of sarcoma cell lines including malignant peripheral nerve sheath tumor (MPNST), synovial (SYO-I), Ewing's (CHP100), osteo (SaOS2) and liposarcoma (LS141). Our results indicate that when compared to other potent RTK inhibitors such as Imatinib and Pazopanib, BIBF1120 is highly efficient at nanomolar to low micromolar concentrations in blocking proliferative activity of sarcoma cell lines, specifically, SYO-I and MPNST, which are highly dependent on PDGFR signaling. Western blot analysis to test the efficacy of BIBF1120 confirmed significant blockade of RTK signaling pathways. Ras activation and/or activation of mitogen activated protein kinase (MAPK) pathway has been shown to be critical in sarcomas including MPNST and synovial sarcomas. Unlike Imatinib and Pazopanib, BIBF1120 also blocked MAPK signaling pathway as demonstrated by p-ERK1/2 blockade. In addition to the RTKs, mTOR (mammalian Target of Rapamycin) protein plays a key role in AKT activation and downstream survival signaling. Combination of RTK blockade by BIBF1120 and mTOR blockade by TORC1 inhibitor rapamycin resulted in significantly stronger inhibition of in vitro cellular proliferation compared to single agent treatments. Taken together, our data strongly supports further clinical research of BIBF1120 in sarcoma. Combining potent inhibitors of tyrosine kinase signaling, MAPK signaling and inhibitors of mTOR pathway such as rapamycin could provide a new therapeutic approach in this disease.

Citation Format: Parag P. Patwardhan, Elgilda Musi, Kathryn S. Ivy, Natalie Giovino, Gary K. Schwartz. BIBF1120, an investigational triple angiokinase inhibitor, in combination with inhibitors of mTOR signaling shows potent antitumor activity in preclinical models of 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 4526. doi:10.1158/1538-7445.AM2014-4526
Abstract 4528: Comparison of anti-proliferation activity of BEZ 235, a multi-step inhibitor of mTOR pathway, vs. BKM120, a pan PI3K inhibitor, in lung cancer cell lines (LCCL)
Authors: Huang Chao H. , Beaudoin Christopher , Veldhuizen Peter Van , Farassati Faris
Published: May 2015

Background: PI3K/AKT/mTOR(Mammalian target of rapamycin) signal transduction pathway is over-activated in many cancers including lung cancers. The activation of cell surface receptors i.e.EGFR (Epidermal Growth Factor Receptor) signals through PI3K/AKT/mTOR pathway which then results in increase cell proliferation, angiogenesis and anti-apoptosis process. BEZ 235 is a multi-step inhibitor of PI3K and mTORC 1 / 2. BKM 120 is a novel pan class I PI3K inhibitor of all class I PI3K isoforms (α, β, Υ, δ). Inhibition of multiple steps of PI3K/AKT/mTOR pathway would be advantageous and could yield higher activity and overcome over activation of PI3K/AKT, a potential mechanism of resistance to mTOR inhibitors. Docetaxel (D) is an approved agent used in the treatment of lung cancer. We previously demonstrated that the sequence of D followed by mTOR inhibitor temsirolimus (T) in lung cancer cell lines (LCCL) had increased suppression of cell proliferation as compared to reverse sequence of T→D due to suppression of over-activation of PI3K/AKT resistance mechanism. We studied the anti-proliferation effect of each compound in LCCL and also compared the activity of sequential docetaxel followed by BKM 120 vs. D followed by BEK235. Methods: Adenocarcinoma LCCL H2122 and A549 were treated with increasing concentrations of BEZ 235 (50nM, 75 nM, 100 nM) and BKM 120 at same concentrations. Both agents were provided by Novartis.We assessed the cell proliferation by optical density. In the sequential treatment study, we incubated LCCL with D 100nM for 24h then added BKM 120 or BEZ235 at 50nM, 75nM and 100nM. We then assessed the anti-proliferation effect at 48h and 72h.

Results: BEZ 235 single had greater inhibitory effect in LCCL proliferation as compared to BKM120. The sequential treatment of D followed by BEZ235 or BKM120 did not yield higher anti-proliferation effect compared to single agent treatment in both LCCL.

Conclusion: Single agent BEZ235 had greater anti-proliferation effect than BKM120 in both LCCL. The sequential treatment of D followed by BEZ235 or BKM120 did not lead to increased anti-proliferation effect. The potential explanation of lack of synergistic effect may due to suppression of PI3K by pretreatment with D.

Note: This abstract was not presented at the meeting.

Citation Format: Chao H. Huang, Christopher Beaudoin, Peter Van Veldhuizen, Faris Farassati. Comparison of anti-proliferation activity of BEZ 235, a multi-step inhibitor of mTOR pathway, vs. BKM120, a pan PI3K inhibitor, in lung cancer cell lines (LCCL). [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 4528. doi:10.1158/1538-7445.AM2014-4528