bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2927: Rapamycin decreases expression of thymidylate synthase and enhances the response to pemetrexed in preclinical studies and a Phase I/II clinical study of subjects with non-small cell lung cancer
Authors: Kawabata Shigeru , Chiang Chun-Te , Memmott Regan M. , Komiya Takefumi , Gills Joell J. , Dennis Phillip A. 
Published: May 2015

Abstract
Non-small-cell lung cancer (NSCLC) accounts for 80-85% of lung cancers and almost half of patients with newly diagnosed NSCLC have metastatic disease. Platinum-based chemotherapy regimens given as first-line treatment for advanced NSCLC patients with metastasis are standard but have modest response rates. Pemetrexed is well tolerated and inhibits several folate-dependent enzymes including thymidylate synthase, TS. Increased expression of TS confers resistance to pemetrexed in vitro and is a predictive factor for poor response to pemetrexed. Rapamycin is a mTOR inhibitor and suppresses protein synthesis. Here, we show that rapamycin decreases endogenous expression of TS in NSCLC cells. The combination of rapamycin and pemetrexed synergistically inhibits NSCLC cell proliferation and enhances the inhibition of 4E-binding protein 1 (4E-BP1) activation that is regulated by mTOR. Although pemetrexed as a single agent upregulated expression of TS, pretreatment with rapamycin suppressed pemetrexed-upregulated TS expression. In vivo, the combination of rapamycin and pemetrexed inhibited the growth of NSCLC xenografts, which correlated with decreased mTOR activity and inhibition of pemetrexed-induced upregulated expression of TS. Based on these preclinical data, we have conducted a Phase I/II clinical trial combining rapamycin and pemetrexed in subjects with relapsed NSCLC. This regimen was well tolerated, and resulted in a 22% partial response rate that is above what is expected for response to single agent pemetrexed (9%). In biomarker analyses, pretreatment with rapamycin decreased the expression of TS in peripheral blood mononuclear cells, which correlated with increased progression free survival. Collectively, these studies identify rapamycin and pemetrexed as an effective combination in NSCLC, and suggest that rapamycin enhances the efficacy of pemetrexed by suppressing TS expression.
Citation Format: Shigeru Kawabata, Chun-Te Chiang, Regan M. Memmott, Takefumi Komiya, Joell J. Gills, Phillip A. Dennis. Rapamycin decreases expression of thymidylate synthase and enhances the response to pemetrexed in preclinical studies and a Phase I/II clinical study of subjects with non-small cell lung 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 2927. doi:10.1158/1538-7445.AM2014-2927
Abstract 293: High-throughput methylation sequencing of targeted genes in breast cancer specimens using nanofluidic PCR prepared libraries
Authors: Wang Jun , Li Zibo , Wu Yepeng , Guo Xinwu , Li Shengyun , Xiao Zhi , Chen Feiyu , Deng Zhongping , Dai Lizhong , Yi Wenjun , Tang Lili 
Published: May 2015

Abstract
Breast cancer is a global public health issue as it is the most frequently diagnosed malignancy in women in the Western world with over a million new cases every year worldwide. Previous epigenetic analyses have identified aberrant DNA methylation signatures associated with breast cancer. Methylation alterations resulting in aberrant gene expression are key contributors to breast tumorigenesis. There are more than 100 candidate genes reported throughout the literatures as promoter hypermethylated in breast cancers (Pubmeth web resource). More recently, with the application of genome wide array-based methylation profile analysis and next generation sequencing, hundreds of new methylation candidate genes have been reported to be associated with breast cancer. It is crucial to develop an approach for targeted methylaton resequencing to validate these candidate genes in hundreds of patient samples and develop a breast cancer detection panel for early diagnostics.
We have developed a targeted gene bisulfite-sequencing approach using a nanofluidic platform, the Access Array™ system, which enables simultaneous amplification of 48 genetic regions (amplicon) from 48 samples in parallel. We have selected 48 breast cancer associated methylation candidate genes, each commonly reported by multiple literatures. Bisulfite-specific-sequencing (BSP) primers were designed for the promoter regions of these genes. In addition, we have applied a primer design strategy that incorporates sample-specific barcodes and Illumina (Miseq) sequencing adaptors into each amplicon, removing the need for additional library preparation before sequencing. We will present methylation sequencing data collected from amplicon libraries generated using the Access Array system. The 48 promoter regions have been amplified in paired tumor/normal tissues and plasma DNA from 48 breast cancer patients. Our results indicate the excellent utilization of this approach in the validation of methylation biomarkers and selection of detection panel for breast cancer.
Citation Format: Jun Wang, Zibo Li, Yepeng Wu, Xinwu Guo, Shengyun Li, Zhi Xiao, Feiyu Chen, Zhongping Deng, Lizhong Dai, Wenjun Yi, Lili Tang. High-throughput methylation sequencing of targeted genes in breast cancer specimens using nanofluidic PCR prepared libraries. [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 293. doi:10.1158/1538-7445.AM2014-293
Abstract 2935: RNAi screen identifies therapeutic targets in hepatocellular carcinoma
Authors: Huang Chun-Hao , Lujambio Amaia , Zuber Johannes , Kitzing Thomas , Tschaharganeh Darjus F. , Stanchina Elisa D. , Lowe Scott W. 
Published: May 2015

Abstract
Liver cancer is the third leading cause of cancer related mortality worldwide. Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is a very aggressive type of cancer that lacks effective treatment, in part, due to the undruggable nature of its main genetic drivers, such as oncogene MYC amplification or loss of tumor suppressor TP53. The introduction of the multikinase inhibitor Sorafenib represents the biggest therapeutic advance in the past decade, though its activity only increases the life expectancy of liver cancer patients from 8 to 11 months. To identify new therapeutic targets for HCC, we employ RNA interference (RNAi) screens as a non-biased means to identify genes required for tumor maintenance in genetically defined tumors. In the current study, we applied an optimized shRNA platform to perform a negative selection screen in genetically-defined murine hepatocellular carcinoma cell in vitro. Specifically, we have designed a library that contains 2,245 shRNAs targeting 442 “drugged” genes, including genes encoding proteins targeted by FDA-approved drugs, small molecules in clinical trials, or compounds under preclinical development. Countersceening and validation was performed in normal murine cells and human HCC cell lines and xenograft using shRNAs and corresponding small molecular drugs. Pharmacological or shRNA-mediated inhibition of certain targets led to robust antitumor effects, both in vitro and in vivo, and suggest new strategies to inhibit Myc function therapeutically. Our results highlight the utility of RNAi screening for studying cancer vulnerabilities that can be exploited for direct pharmacological intervention.
Citation Format: Chun-Hao Huang, Amaia Lujambio, Johannes Zuber, Thomas Kitzing, Darjus F. Tschaharganeh, Elisa De Stanchina, Scott W. Lowe. RNAi screen identifies therapeutic targets in hepatocellular 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 2935. doi:10.1158/1538-7445.AM2014-2935
Abstract 2936: JNJ-47117096, a selective small molecule inhibitor of the MELK oncogene decreases DNA damage tolerance in highly proliferating cancer cells
Authors: Beke Lijs , Linders Joannes T.m. , Kig Cenk , Boeckx An , Heerde Erika van , Wuyts Dirk , Parade Marc , Meerpoel Lieven , Johnson Chris , Beullens Monique , Bollen Mathieu , Brehmer Dirk 
Published: May 2015

Abstract
Maternal embryonic leucine zipper kinase (MELK, hMP38, pEG3), a Ser/Thr protein kinase, is highly overexpressed in stem cells and cancer cells. The oncogenic role of MELK is attributed to disabling critical cell cycle check points as well as enhancing replication. Most functional studies have relied on the use of siRNA/shRNA mediated gene silencing, but this often can be associated with off-target effects.
Here we want to present a novel, potent and selective small molecule inhibitor JNJ-47117096 that has enabled us to validate the biological function of MELK kinase.
Outcome: MELK inhibition in cancer cells with an intact p53 signaling pathway is linked to a replicative senescence phenotype. This phenotype correlates with a rapid ATM activation and phosphorylation of CHK2 without any effects on the alternative ATR/ CHK1 DNA damage pathways. Furthermore, JNJ-47117096 induces strong phosphorylation of p53 and prolonged up-regulation of p21 without the induction of apoptosis. Strikingly, MELK inhibition in several p53 disabled cancer cells showed induction of a mitotic catastrophe phenotype followed by prominent cell killing.
Finally, JNJ-47117096 triggers a rapid degradation of cellular MELK protein, independent of the cell cycle phase and its regulation by the E2F pathway. This observation can clearly be linked to a direct binding effect of JNJ-47117096 to cellular MELK protein confirmed by chemical proteomics.
Conclusion: Our data generated with JNJ-47117096, confirmed by selective siRNAs, indicate MELK as a key stimulator of proliferation and replication by its ability to increase the threshold for DNA damage tolerance. Targeting MELK function by selective small molecule inhibitors might sensitise tumors to DNA-damaging agents or radiation therapy.
Citation Format: Lijs Beke, Joannes T.m. Linders, Cenk Kig, An Boeckx, Erika van Heerde, Dirk Wuyts, Marc Parade, Lieven Meerpoel, Chris Johnson, Monique Beullens, Mathieu Bollen, Dirk Brehmer. JNJ-47117096, a selective small molecule inhibitor of the MELK oncogene decreases DNA damage tolerance in highly proliferating 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 2936. doi:10.1158/1538-7445.AM2014-2936
Abstract 3020: Patient-derived xenograft (PDX) models to study the role of breast cancer stem cells in metastasis formation
Authors: Ginestier Christophe , Lartigue Danae , Cabaud Olivier , Malzac Aurelie , Wicinski Julien , Josselin Emmanuelle , Finetti Pascal , Adelaide Jose , Bertucci Francois , Chaffanet Max , Birnbaum Daniel , Charafe-Jauffret Emmanuelle 
Published: May 2015

Abstract
Although advances have been made in the treatment of localized breast cancer, there has been less progress in the treatment of advanced metastatic disease. This disease is an oncoming therapeutic challenge for breast cancer since it is always fatal. Conventional therapies may fail to target cancer stem cells (CSC). Metastatic breast cancer may thus benefit from current intensive research in the CSC field. Several studies suggest that CSCs are the only cancer cells responsible for metastasis formation. CSCs may have the ability to disseminate in blood circulation as circulating tumor cells (CTCs) and to colonize new sites. To better understand the biological processes that link CSCs and metastasis, we have generated a xenobank of 50 patient-derived xenografts (PDXs) of primary tumors grafted in the humanized mammary fat pad of immunodeficient mice. These models reflect the heterogeneity of primary tumors both at molecular and cellular level. Using a lentiviral vector, we infected our PDXs to obtain GFP-luciferase-positive models designed to monitor metastasis development from the orthotopic site. To mimick the patient therapeutic management, metastasis formation is monitored during tumor growth but also after tumor surgical removal. Metastases are collected and analyzed for histology and phenotype. The different metastatic patterns are correlated with the mutational profile of PDX. CTCs are also monitored at different time of the metastatic process using a specific filter (ISET) for cell characterization and phenotype. PDXs can be classified in tumor groups with similar metastatic tropisms. To depict the role of CSC from primary tumors in the tropism of their metastases, we collected and genomic profiled the ALDEFLUOR-positive CSCs in orthotopic tumors from different models with distinct metastatic pattern. These different PDX models lead to a better understanding of the role of breast CSCs in metastasis formation from CTCs to well-constituted metastasis and their study may identify targets able to design future therapeutic strategies.
Citation Format: Christophe Ginestier, Danae Lartigue, Olivier Cabaud, Aurelie Malzac, Julien Wicinski, Emmanuelle Josselin, Pascal Finetti, Jose Adelaide, Francois Bertucci, Max Chaffanet, Daniel Birnbaum, Emmanuelle Charafe-Jauffret. Patient-derived xenograft (PDX) models to study the role of breast cancer stem cells in metastasis formation. [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 3020. doi:10.1158/1538-7445.AM2014-3020
Abstract 3035: Cripto/GRP78 signaling promotes the stem cell phenotype in normal and neoplastic mammary epithelial cells
Authors: Spike Benjamin T. , Kelber Jonathan A. , Booker Evan , Kalathur Madhuri , Rodewald Rose , Lipianskaya Julia , La Justin , He Marielle , Wright Tracy , Klemke Richard , Wahl Geoffrey , Gray Peter C. 
Published: May 2015

Abstract
Microenvironmental factors are critical regulators of stem cell and tumor cell behavior. However, few stem cell factors have been identified and targeted in breast and other cancers. Using a highly enriched stem cell pool isolated from fetal mammary rudiments and single cell analyses, we have identified the onco-fetal protein Cripto as a potent soluble factor that regulates the mammary stem cell state. Cripto is a GPI-anchored/secreted signaling protein and a known regulator of PI3K/AKT and TGF-beta pathways. Here, we develop a novel Cripto antagonist, ALK4L75A-Fc, which selectively blocks the growth factor-like effects of soluble Cripto. We show that this antagonist promotes the differentiation of mammary stem cells cultured ex vivo while Cripto treatment maintains the stem cell phenotype and yields colonies with enhanced mammary gland reconstitution capacity. We previously discovered that Cripto signaling requires binding to cell surface Glucose Response Protein 78kDa (GRP78). GRP78 is an HSP70 family member that is induced by stresses including hypoxia and glucose deprivation and it is highly expressed in tumors where these conditions prevail. We show here that cell surface GRP78 marks fetal mouse mammary stem cells and a population of bipotent adult mammary epithelial cells that are selectively responsive to soluble Cripto in vitro. GRP78high mammary epithelial cells also harbor substantially higher stem cell activity upon transplantation than GRP78low cells. Consistently, we further show that deletion of GRP78 from adult mammary epithelial cells ex vivo blocks mammary gland reconstitution. Finally, we find that Cripto antagonism with ALK4L75A-Fc inhibits the proliferation of triple negative breast cancer cell lines in vitro and that this antagonist can also inhibit tumor growth in vivo. Since Cripto and GRP78 are both induced by hypoxia and other stressful conditions found in tumors, our data raise the possibility that Cripto/GRP78 signaling may exacerbate breast and other cancers by increasing the number of stem cell-like tumor cells in tumor microenvironments where these conditions predominate.
Citation Format: Benjamin T. Spike, Jonathan A. Kelber, Evan Booker, Madhuri Kalathur, Rose Rodewald, Julia Lipianskaya, Justin La, Marielle He, Tracy Wright, Richard Klemke, Geoffrey Wahl, Peter C. Gray. Cripto/GRP78 signaling promotes the stem cell phenotype in normal and neoplastic mammary epithelial 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 3035. doi:10.1158/1538-7445.AM2014-3035
Abstract 3046: Lin28B mediated IKK-β sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling pathway
Authors: Chen Chong , Bai Lipeng , Cao Fengqi , Liu Yan , Xie Junling , Wang Wei , Si Qin , Yang Jian , Chang Antao , Xiang Rong , Luo Yunping 
Published: May 2015

Abstract
Substantial evidence indicates that many human cancers are driven by a subpopulation of cells that display stem cell properties. These cancer stem cells (CSCs) may also contribute to tumor metastasis and treatment resistance. It has been reported that inflammatory factors in tumor microenvironment activate NF-kB/IKK-β pathway in tumor cells which in turn drive CSC self-renewal, but their molecular basis remain unclear. In this study, we demonstrated a novel mechanism regarding that Lin28B, a RNA binding protein regulated by IKK-β, sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling pathway. First, we found that all stemness properties of MDA-MB-231 human breast cancer cells including ALDH+ population, Sphere-forming ability and the expression of stemness genes SOX2, OCT4, NANOG and Lin28B were decreased by IKK-β inhibition, meanwhile, the down regulated expression of Lin28B decrease the stemness properties of MDA-MB-231 cells as well. Furthermore, it was found that Wnt/TCF4 signaling pathway was inhibited by Lin28B down-regulation via reducing the expression of TCF4. Interestingly, TCF4 could also regulate the expression of Lin28B by directly binding to the intron 1 of Lin28B. Moreover, we observed that microRNA-34a was down regulated by Lin28B at transcriptionally level, and increased miR34a expression inhibited the activity of Wnt/LEF1 pathway by directly targeting LEF1. Most importantly, we found that down regulated expression of Lin28B or IKK-β inhibition combined with chemo-drug (IKK-β selective inhibitor IMD-0354 combine with paclitaxel) could markedly suppressed tumor growth and metastasis of MDA-MB-231 breast tumor cells in NOD/SCID mice model. Taken together, our study demonstrated that Lin28B, which was activated by IKK-β, could sustain the stemness of CSC via the TCF4 and miR-34a/LEF1 signaling regulation, thus, establish the Lin28B as a central regulator in govern inflammation to maintain breast cancer stemness phenotype, and could serve as a therapeutical target.
Citation Format: Chong Chen, Lipeng Bai, Fengqi Cao, Yan Liu, Junling Xie, Wei Wang, Qin Si, Jian Yang, Antao Chang, Rong Xiang, Yunping Luo. Lin28B mediated IKK-β sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling 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 3046. doi:10.1158/1538-7445.AM2014-3046
Abstract 3055: Potential role of cyclin D1 in regulation of liver cancer stem cells
Authors: Xia Wei , Wang Xiao Qi 
Published: May 2015

Abstract
Cancer stem cells (CSCs) are capable of self-renewal, multipotency, in vivo tumorigenicity, and driving metastasis. CSCs are more resistant to conventional therapy, allowing recurrence of tumor. Thus, novel therapies capable of eliminating CSCs are needed. Cyclin D1 is deregulated in many types of cancers. Beyond being a cell cycle regulator and oncogene, cyclin D1 also enhances stem cell (SC) self-renewal and promotes induced pluripotent stem cells (iPSCs) reprogramming efficiency. Yet, the role of cyclin D1 in regulation of CSC is not well defined.
Here we investigated whether and how cyclin D1 could regulate liver cancer CSC properties. We overexpressed cyclin D1 in two lines of HCC cells, Huh7 and 97H and found that the sphere formation capacity was enriched in Huh7 cyclin D1-expressing cells than Huh7 cells. From dissected single spherical cells, the secondary sphere formation capacity was significantly higher in cyclin D1-expressing cells than both parental Huh7 (p=0.00) and 97H (p=0.02) cells. Anchorage-independent spherical colonies (spheres) are composed of a small number of cancer cells with stem-like properties. Thus, the results suggest that overexpression of cyclin D1 enhanced self-renewal capacity of liver CSCs. The cyclin D1-expressing spheres displayed an increased expression of stem cell-associated genes Nanog, Oct4, and Nordal/Activin. Smad2/3, a TGF-β signaling effector, was highly phosphorylated in cyclin D1-expressing spheres compared to monolayer or spheres without cyclin D1 expression.
More importantly, TGF-β/Smad inhibitor (SB431542) inhibited cyclin D1-expressing CSC self-renewal shown by significantly reduction of sphere formation in two lines of cyclin D1-expressing HCC lines, whereas it had no effects on sphere formation in parental cells without cyclin D1 overexpression. In addition, Erk inhibitor (UO126) showed no inhibiting effects, a further proof of the role of cyclin D1 mediated activation of Smad2/3. These results indicate that TGF-β/Smad inhibitor might have therapeutic potential for targeting liver CSCs.
HCC cells are highly resistance chemotherapy resulting in enriched CSC population. We further investigated the sphere formation capacity of D1-expressing cells under the treatment of chemotherapeutic agents in combination with Smad inhibitor. We found that SB431542 could inhibit cyclin-D1 expressing spherical cancer cell proliferation therefore sensitized the cells to chemo drug cisplatin. In summary, our results suggest that cyclin D1 might promote liver CSC proliferation and self-renewal via activation of TGF-β/Smad signaling, with underlining mechanism is under investigation.
Citation Format: Wei Xia, Xiao Qi Wang. Potential role of cyclin D1 in regulation of liver cancer stem 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 3055. doi:10.1158/1538-7445.AM2014-3055