bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract IA10: Signaling through individual Ras proteins in the absence of the Nf1-RASGAP
Authors: Ratner Nancy 
Published: May 2015

Abstract
Neurofibromatosis type 1 (NF1) patients are predisposed to develop neurofibromas, benign peripheral nerve tumors resulting from loss of function mutations in the NF1 gene. NF1 (neurofibromin), the protein product of the NF1 gene, is a GTPase activating protein (GAP) that inactivates the Ras proteins H-, N-, K-, M-, and R-Ras, and R-Ras2/TC21 (TC21). We hypothesized that relevant Ras proteins would be activated in vivo by deletion of NF1, so that removing single Ras proteins in Nf1 mutant cells would reveal specific functions of individual Ras proteins. We found that activation of TC21 accounts for migration defects in NF1 mutant Schwann cells (Huang et al, 2004), through AKT and not ERK. Removing TC21 in Nf1 mutant Schwann cell precursors identified an Nf1/TGFβ/TGFβRII/AKT autocrine survival loop (Patmore et al, 2012). Implying that delayed benign tumor formation resulted from effects on tumor initiation, benign tumor formation was delayed in mice lacking Nf1 and TC21. In Schwann cell precursors, activation of P-ERK, not P-AKT, was blocked by dnH-Ras. To define the role of ERK signaling in neurofibroma, we used a DhhCre;Nf1fl/fl mouse model of neurofibroma formation. Preclinical testing of specific signaling pathways with Phase I drug candidates in the neurofibroma mouse model was read out neurofibroma growth by 7T MRI imaging, in conjunction with pharmacokinetic and pharmacodynamic readouts (Jessen et al., 2013). MEK inhibition was highly effective although not curative in this model; we established minimum effective doses of the MEK inhibitor PD0325901 and effects of early MEK inhibition on peripheral nerve tumor growth. The data support the idea that NF1 GAP loss can be used to identify roles of individual Ras proteins and downstream signaling pathways in specific cell types, and that combinations of Ras proteins can collaborate to drive tumorigenesis. Supported by R01-NS28840, P50-NS057531 and the Children's Tumor Foundation/ Neurofibromatosis Therapeutic Acceleration Program.
Citation Format: Nancy Ratner. Signaling through individual Ras proteins in the absence of the Nf1-RASGAP. [abstract]. In: Proceedings of the AACR Special Conference on RAS Oncogenes: From Biology to Therapy; Feb 24-27, 2014; Lake Buena Vista, FL. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(12 Suppl):Abstract nr IA10. doi: 10.1158/1557-3125.RASONC14-IA10
Abstract PR01: Yap1 activation enables bypass of oncogenic Kras addiction in pancreatic cancer
Authors: Kapoor Avnish , Yao Wantong , Ying Haoqiang , Hua Sujun , Liewen Alison , Wang Qiuyun , Zhong Yi , Wu Chang-Jiun , Sadanandam Anguraj , Hu Baoli , Chang Qing , Chu Gerald , Al-Khalil Ramsey , Jiang Shan , Xia Hongai , Fletcher-Sananikone Eliot , Lim Carol , Horwitz Gillian , Viale Andrea , Pettazzoni Piergiorgio , Sanchez Nora , Wang Huamin , Protopopov Alexei , Zhang Jianhua , Heffernan Timothy , Johnson Randy , Chin Lynda , Wang Alan , Draetta Giulio , DePinho Ronald 
Published: May 2015

Abstract
Activating mutations in KRAS are among the most frequent events in diverse human carcinomas and particularly prominent in human pancreatic ductal adenocarcinoma (PDAC). An inducible KrasG12D-driven mouse model of PDAC has established a critical role for sustained KrasG12D expression in tumor maintenance, providing a model to determine the potential for, and underlying mechanisms of, KrasG12D–independent PDAC recurrence. Here we show that majority tumors will undergo spontaneous regrowth following Kras oncogene extinction and, importantly, a subset of relapsed tumors remained devoid of KrasG12D expression and canonical MAPK signaling and acquired amplification and over-expression of Hippo effector, the Yap1 transcriptional co-activator. Functional studies established Yap1 and the Tead2 transcriptional factor in driving KrasG12D–independent tumor maintenance. The Yap1/Tead2 complex acts cooperatively with E2F transcription factors to promote a cell cycle and DNA replication program and a quasimesenchymal phenotype with increased metastatic potential. Our studies, along with corroborating evidence from human PDAC models, portend a novel mechanism of escape from oncogenic Kras addiction in PDAC.
This abstract is also presented as Poster B21.
Citation Format: Avnish Kapoor, Wantong Yao, Haoqiang Ying, Sujun Hua, Alison Liewen, Qiuyun Wang, Yi Zhong, Chang-Jiun Wu, Anguraj Sadanandam, Baoli Hu, Qing Chang, Gerald Chu, Ramsey Al-Khalil, Shan Jiang, Hongai Xia, Eliot Fletcher-Sananikone, Carol Lim, Gillian Horwitz, Andrea Viale, Piergiorgio Pettazzoni, Nora Sanchez, Huamin Wang, Alexei Protopopov, Jianhua Zhang, Timothy Heffernan, Randy Johnson, Lynda Chin, Alan Wang, Giulio Draetta, Ronald DePinho. Yap1 activation enables bypass of oncogenic Kras addiction in pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on RAS Oncogenes: From Biology to Therapy; Feb 24-27, 2014; Lake Buena Vista, FL. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(12 Suppl):Abstract nr PR01. doi: 10.1158/1557-3125.RASONC14-PR01
Abstract PR03: Global quantitative proteomic and phosphoproteomic analysis of oncogenic Kras-driven mouse pancreatic cancer
Authors: Taguchi Ayumu , Kapoor Avnish , Momin Amin , Katayama Hiroyuki , Yao Wantong , Wang Hong , Ying Haoqiang , DePinho Ronald , Hanash Samir 
Published: May 2015

Abstract
Background: The most common genetic abnormality in pancreatic cancer is oncogenic mutation of KRAS, which is an initial key event in pancreatic carcinogenesis. Oncogenic KRAS activates downstream signaling pathways, such as PI3K and MAPK pathways, indicating the essential role of phosphorylation in pancreatic cancer. However, the effect of KRAS on the global phosphoproteome in pancreatic cancer has not been fully elucidated. We investigated proteomic and phosphoproteomic alterations induced by oncogenic Kras using mouse pancreatic cancer cell lines established from doxycycline-inducible KrasG12D pancreatic cancer mouse model.
Materials and Methods: Three mouse pancreatic cancer cell lines were established from triple transgenic inducible KrasG12D pancreatic cancer mouse model (tetO_LSL- KrasG12D; ROSA26A-LSL-rtTA-IRES-GFP; p48-Cre). To identify direct downstream pathways of Kras, we performed two different experiments for each cell line: (1) 20 hours Kras-off followed by 5 hours Kras-on (designated “Re-on”) vs. 25 hours Kras-off, and (2) Continuous Kras-on (designated “On”) vs. 24 hours Kras-off. Whole cell lysates were fractionated and subjected to analysis with using mass spectrometry.
Results and Conclusion: Overall 4,133 phosphorylation sites were identified. Phosphorylation levels of 2,670 sites were quantified in at least one sample, including 2,347 (87.9%) at serine, 293 (11.0%) threonine, and 30 (1.1%) tyrosine residues. Phosphorylation levels were more than 1.25 fold higher at 386 sites (139 proteins) and 558 sites (256 proteins) in “Re-on” and “On” samples, respectively. 190 sites were common between “Re-on” and “On”, 196 sites were “Re-on” unique, and 368 sites were “On” unique. mTOR and MAPK pathways were activated in both “Re-on” and “On”, while Rho kinase, Phospholipase C, and Rac signaling pathways were predominantly associated with “On”, suggestive of differential downstream pathway activation at different phases of Kras signaling.
This abstract is also presented as Poster A21.
Citation Format: Ayumu Taguchi, Avnish Kapoor, Amin Momin, Hiroyuki Katayama, Wantong Yao, Hong Wang, Haoqiang Ying, Ronald DePinho, Samir Hanash. Global quantitative proteomic and phosphoproteomic analysis of oncogenic Kras-driven mouse pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on RAS Oncogenes: From Biology to Therapy; Feb 24-27, 2014; Lake Buena Vista, FL. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(12 Suppl):Abstract nr PR03. doi: 10.1158/1557-3125.RASONC14-PR03
Abstract A20: Sphingosine 1-phosphate promotes tumor progression in a survivin-dependent manner
Authors: Samadi Nasser , Nezhad Maryam Tabasi , Ghanbari Parisa , Mohseni Mahsa 
Published: May 2015

Abstract
Introduction: sphingosine 1-phosphate (S1P) is a bioactive lipid molecule that involves in tumor progressions, metastasis and chemo-resistance. We investigated the effects of S1P and its signaling mechanism in induction of tumor progression in chronic lymphoblastic leukemia SKW3 cells and non-small cell lung cancer H1299 cells.
Methods: We investigated the effects of S1P on proliferation with MTT assay, invasion by soft-agar colony forming assay and cell migration by applying trans-well migration assay. The expression of survivin gene was assessed by Real-time PCR.
Results: We showed that low concentrations of S1P induced proliferation and migration in both cell lines. S1P also stimulated invasiveness in H1299 cancer cells. We showed that the effects of S1P in tumor progression are S1P receptor-dependent. Survivin also plays a key role in S1P tumorogenic functions.
Conclusion: Our results suggest that identifying cancer patients with higher levels of S1P and survivin and inhibiting their activities can be considered as an important strategy to increase the efficacy of chemotherapeutic agents.
Note: This abstract was not presented at the conference.
Citation Format: Nasser Samadi, Maryam Tabasi Nezhad, Parisa Ghanbari, Mahsa Mohseni. Sphingosine 1-phosphate promotes tumor progression in a survivin-dependent manner. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A20.
Abstract A23: eIF4F is a key and targetable convergence nexus of multiple mechanisms of resistance to anti-BRAF and anti-MEK cancer therapies
Authors: Boussemart Lise , Malka-Mahieu Helene , Girault Isabelle , Allard Delphine , Hemmingson Oskar , Kamsu-Kom Nyam , Agoussi Sandrine , Eggermont Alexander , Desaubry Laurent , Robert Caroline , Vagner Stephan 
Published: May 2015

Abstract
The V600E mutation in BRAF is the most frequent oncogenic mutation in human cancers, and can be targeted by specific anti-BRAF agents. However, several mechanisms of resistance have been identified, leading to reactivation of the initially blocked MAP-kinase pathway (known as ERK-dependent mechanisms) or reactivation of alternative ones, like the PI3K-AKT-mTOR pathway, or inhibition of the apoptotic cascade (ERK-independent). These pathways converge to regulate the formation of the eIF4F translation initiation complex that binds to the 7-methylguanosine cap at the 5′ end of mRNAs, thereby modulating mRNA translation of specific mRNAs. We addressed the potential role of the eIF4F eukaryotic translation initiation complex in resistance or sensitivity to anti-BRAF and anti-MEK agents in several BRAF mutant cells, xenografts and tumors. Our data demonstrate that the formation of the eIF4F complex is associated with most resistance mechanisms to these targeted therapies, independently of their capacity to reactivate or not the MAP-kinase pathway. Formation of this complex can be explored in tumor samples using a new in situ technology. Inhibitors of the eIF4F complex synergize with anti-BRAF agents and can thus reverse resistance.
Citation Format: Lise Boussemart, Helene Malka-Mahieu, Isabelle Girault, Delphine Allard, Oskar Hemmingson, Nyam Kamsu-Kom, Sandrine Agoussi, Alexander Eggermont, Laurent Desaubry, Caroline Robert, Stephan Vagner. eIF4F is a key and targetable convergence nexus of multiple mechanisms of resistance to anti-BRAF and anti-MEK cancer therapies. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A23.
Abstract A38: Optimization of whole-genome amplification for analysis of single cells using next-generation sequencing
Authors: Amiss Terry J. , Tong Frances , Snowden Eileen , Kelly Richard , Blaesius Rainer , Herrmann Nick , Hahn Friedrich , Porter Warren , Ferguson Mitchell , Chang Chen , Clancy Daphne , Jurgensen Stewart 
Published: May 2015

Abstract
The accurate analysis of both genotypic variation in tumors and tumor evolution due to selective pressure is important in determining treatment options in the clinic. Next generation sequencing (NGS) of single cells from solid tumors and circulating tumor cells has the potential to dramatically increase the amount of genotypic detail obtained from these samples. For the analysis of single cells the challenge is to employ whole genome amplification (WGA) to obtain enough DNA for analysis while minimizing amplification bias and maintaining copy accuracy. We are developing tools for single cell sequencing to improve these parameters while also increasing sample throughput. Initially, three commercially available WGA methods were tested on 5-10 genome quantities of HCT-15 genomic DNA. Using the Ion Torrent AmpliSeq Cancer Hotspot Panel, the amplification accuracy, amplicon coverage and DNA yield were determined. When compared to controls significant differences were observed in the three WGA methods ability to accurately amplify DNA. The most accurate kit produced a specificity and sensitivity of 98.5% and 100% respectively. This protocol also produced a substantial DNA yield of 25 µgs and although, the average amplicon coverage indicated some bias, greater than 99% of the amplicons had at least 100-fold coverage. This WGA kit was then tested on single cells from a human breast cancer; an ER/PR/Her2 negative spindle cell metaplastic carcinoma. To obtain single cells the tumor was grown in a PDX mouse model and index sorted using a BD FACSAria II flow cytometer. The WGA produced an average DNA yield of 2.7± 0.8 µg, with 100-fold amplicon coverage of 92±7%. For the variants identified, the amplification bias differed by as much as 3,000 reads/variant. We are now taking steps to improve WGA of single cells and will report on these efforts in detail. To date when monitoring variant allele frequency and coverage depth in the triple negative tumor, KRAS and MET mutations were identified in single cells, as well as in the bulk sample. Interestingly, a TP53 mutation was identified only in single cells. This data supports the utility of WGA and single cell analysis to identify mutations, some having clinically relevant role for targeted therapy.
Citation Format: Terry J. Amiss, Frances Tong, Eileen Snowden, Richard Kelly, Rainer Blaesius, Nick Herrmann, Friedrich Hahn, Warren Porter, Mitchell Ferguson, Chen Chang, Daphne Clancy, Stewart Jurgensen. Optimization of whole-genome amplification for analysis of single cells using next-generation sequencing. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A38.
Abstract A42: Exogenous normal mammary epithelial mitochondria increase the drug sensitivity of human breast cancer MCF-7 cells
Authors: Jiang Xian-Peng , Philips Jeff , Elliott Robert L. , Head Jonathan F. 
Published: May 2015

Abstract
The mitochondrial dysfunction of cancer cells leads to the suppression of glucose oxidation, increased glycolysis, proliferated advantage, resistance to apoptosis and drug resistance. We hypothesize that the introduction of normal mitochondria into cancer cells will restore the mitochondrial function of cancer cells and increase drug sensitivity. We stained the mitochondria of the normal human mammary epithelial cell line MCF-12A with JC-1 dye, and found orange fluorescence (JC-1 aggregates in mitochondria due to high mitochondrial membrane potential) in the MCF-12A cells. We then isolated the JC-1 stained-mitochondria of the MCF-12A cells and co-cultured them with the cells of human breast cancer MCF-7 cell line. At 30 minutes after co-culture of MCF-7 cells with the isolated JC-1-stained mitochondria, the orange fluorescence was observed in the MCF-7 cells. The fluorescence continued to increase for at least 6 hours and was still detected at 24 hours. This demonstrated that exogenous normal mitochondria of MCF-12A can enter into MCF-7 breast cancer cells. In order to determine whether mitochondria of normal MCF-12A cells improve the sensitivity of breast cancer cells to chemotherapeutic drugs, we co-cultured MCF-7 cells for 24 hours with normal mitochondria of MCF-12A, aspirated off the media and treated the MCF-7 cells with the drugs doxorubicin, abraxane or carboplatin. We found that MCF-7 cells, treated with the normal mitochondria of MCF-12A, were more sensitive to doxorubicin, abraxane and carboplatin than non-treated MCF-7 cells. We also have RT-PCR results demonstrating that uptake of normal mitochondria by cancer cells down-regulates the expression of membrane glucose transporter GLUT3 in MCF-7 cells. In conclusion, we suggest that the introduction of normal cell mitochondria into cancer cells could restore cancer cell mitochondrial function and significantly impact cancer therapy.
Citation Format: Xian-Peng Jiang, Jeff Philips, Robert L. Elliott, Jonathan F. Head. Exogenous normal mammary epithelial mitochondria increase the drug sensitivity of human breast cancer MCF-7 cells. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A42.
Abstract A44: S100A4 drives the invasive potential of non-small cell lung cancer cells and is targeted by FDA approved drugs
Authors: Chen Min , Stewart Rachel , Knifley Teresa , Carpenter Brittany L. 
Published: May 2015

Abstract
The metastatic nature of advanced non-small cell lung cancer (NSCLC), often associated with therapeutic resistance, accounts for the majority of cancer death with the 5-year survival rate less than 10%. This dismal outcome will remain the same until we gain better understanding of the crucial drivers of the metastatic process and gain the power to effectively target them. S100A4 is a tumor metastasis associated protein and an epithelial to mesenchymal transition (EMT) marker. S100A4 contributes to several hallmarks of cancer such as anti-apoptosis, proliferation and therapeutic resistance, and has been implicated in the progression of different types of cancer, including breast, colon and pancreatic cancer. However, the impact of S100A4 signaling in lung cancer progression and its potential use as a target for therapy in lung cancer has not been properly explored. Here, by using established lung cancer cell lines, we demonstrate that S100A4 is upregulated in a subset of lung cancer cell lines both at the mRNA and protein levels. We further found that inhibition of S100A4 by shRNA in A549 and H460 lung cancer cell lines reduced cell proliferation and decreased 3D invasive growth, while exogenous overexpression of S100A4 in H1299 cells increased invasive potential. Interestingly, we showed that Niclosamide, an FDA-approved anti-tapeworm agent, suppressed S100A4 expression in A549 and H358 cells and led to decreased cell proliferation, invasion and invasive growth. Similarly, we found that a CDC25 inhibitor, NSC95397, disrupts the interaction of S100A4 and non-muscle myosin IIA and also decreased A549 cell invasion. Collectively, these data highlight the importance of S100A4 in lung cancer invasion and metastasis and provide the rationale for targeting S100A4 as a potential agent to combat lung cancer.
Citation Format: Min Chen, Rachel Stewart, Teresa Knifley, Brittany L. Carpenter. S100A4 drives the invasive potential of non-small cell lung cancer cells and is targeted by FDA approved drugs. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A44.
Abstract B11: Stromal response to prostate cancer therapeutics
Authors: Gales Dominique Nicole , Fadlalla Khalda , Manne Upender , Yates Clayton , Samuel Temesgen 
Published: May 2015

Abstract
Docetaxel (D) is a clinically used front-line chemotherapeutic agent for castration resistant advanced prostate cancer (Pca). However, its long-term benefits are limited, with most patients eventually progressing because of inherent or acquired drug resistance. The treatment of advanced Pca is a significant challenge and there are no effective treatments that stably suppress the disease. The mechanisms behind the resistance to Docetaxel are not fully understood. It is well accepted that tumor microenvironment is essential for tumor cells survival, cancer progression and metastasis. However, the mechanisms by which tumor cells interact with their surrounding at different stages of cancer development or during chemotherapy are largely unidentified. The central goal is to identify the cellular and biochemical responses of stromal cells in tumor microenvironment with which prostate cancer cells interact. Specifically, the interaction of prostate cancer cells with stromal cells of the prostate or the bone microenvironment in the presence of clinically used cancer therapeutics will be examined. Key regulators of these interactions will also be identified. Such regulators include cytokines, growth factors, proteins, and their receptors. By identifying these regulators and their contribution to tumor drug response, novel therapeutics targeted to the microenvironment can be developed. Therefore, we hypothesize that the response of prostate stromal cells exposed to chemotherapeutic agents could modify drug response or therapeutic outcome of prostate cancer. Cellular proliferation was analyzed by MTT assay. The IC50 of Docetaxel for WPMY-1 (normal stromal), LNCaP (androgen dependent), DU-145 (androgen independent), and HS27A (bone stromal) showed inhibitory effects between 2-10nM. Cell cycle profiles were assessed by flow cytometry. The analysis showed that Docetaxel (2-50nM) induced prominent G2-M arrest in prostate and stromal cells within 24 hours. Gene expression profiling of prostate cancer and stromal cells lines were evaluated. To obtain the expression profiles, we conducted a human cytokines and chemokines array, which consisted of 84 specific genes. Analysis of the prostate cancer and stromal cells treated with Docetaxel revealed differential expression patterns. These results suggest Docetaxel exhibits strong anti-proliferative effects and analysis of the gene array suggest regulation of genes associated with, cell growth, survival, proliferation, metastasis, angiogenesis, and apoptosis.
Note: This abstract was not presented at the conference.
Citation Format: Dominique Nicole Gales, Khalda Fadlalla, Upender Manne, Clayton Yates, Temesgen Samuel. Stromal response to prostate cancer therapeutics. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr B11.