bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1345: Evidence for genetic mediation of lung cancer through hay fever.
Authors: D'Amelio Anthony , Nguyen Chi H. , El-Zein Randa , Spitz Margaret R. , Wu Xifeng , Etzel Carol J.
Published: August 2014

Introduction: In the past decade, advances in genetics have led to the discovery of numerous lung cancer susceptibility variants. The majority of these variants have been found to influence lung cancer susceptibility via tobacco exposure or nicotine addiction; however, recently, researchers have observed lung cancer susceptibility variants that mediate through Chronic Obstructive Pulmonary Disease (COPD). Studies involving potential genetic factors related to other lung-related conditions, such as pneumonia and/or hay fever have been limited. The genetic variants related to underlying mechanism of hay fever on the development of lung cancer are interesting to pursue as hay fever has been showed to be protective against lung cancer.

Methods: Cases included 1154 histological-confirmed Caucasian lung cases from MD Anderson Cancer Center in Houston, Texas, and controls included 1137 individuals recruited through the Kelsey-Seybold Clinics in Houston, Texas. These cases and controls were a subset of participants from a lung cancer case-control study conducted that has available Genome-wide association study (GWAS) data (317,498 SNPs). We first conducted an association analysis in PLINK to determine the association of each SNP with both lung cancer and hay fever and find SNPs that showed a joint significance for both hay fever and lung cancer in opposite directions (p-value < 0.05 for both). We then performed mediation analyses with the SNPs that showed significance for both hay fever and lung cancer. Finally, we inferred SNPs in regions that contained a collection of tag SNPs that mediate lung cancer risk through hay fever.

Results: Two hundred and forty six SNPs were found to be statistically significant (p-value < 0.05) for both hay fever (protection) and lung cancer (risk), and 76 of these SNPs maintained their significance level after mediation analysis. SNP rs7159751 on chromosome 14 had the highest mediation effect of 19.83%. A collection of SNPs on the Neuregulin 3 gene (NRG3) that mediate lung cancer risk through hay fever was found on Chromosome 10 which has been shown to activate the JAK-STAT signal transduction pathway and stimulate lung epithelial cell proliferation.

Conclusion: This is the first study to have investigated the mediation effects of hay fever on lung cancer risk. Our data supports the mediation role of certain SNPs in lung cancer through hay fever and points to specific transduction pathway. Future studies are needed to validate these results in external populations.

Citation Format: Anthony D'Amelio, Chi H. Nguyen, Randa El-Zein, Margaret R. Spitz, Xifeng Wu, Carol J. Etzel. Evidence for genetic mediation of lung cancer through hay fever. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1345. doi:10.1158/1538-7445.AM2013-1345
Abstract 1349: Genetic variants in DNA repair pathway genes are associated with the risk of esophageal adenocarcinoma.
Authors: Wang Yan , Ajani Jaffer A. , Ye Yuanqing , Huang Maosheng , Wu Xifeng
Published: August 2014

Esophageal Cancer (EC) is one of the leading causes of cancer death in the world. Over 90% of ECs are either esophageal squamous cell carcinomas (ESCC) or adenocarcinomas (EAC). EAC is the fastest growing malignancy in the US in the past three decades. In this large case/control study, we aim to assess genetic susceptibility to EAC by investigating single nucleotide polymorphisms (SNPs) in DNA repair related pathways, including nuclear excision repair (NER), base excision repair (BER) and double-strand-break repair (DSBR). The study population included 513 Caucasian EAC cases 1499 Caucasian controls. Genotyping was performed by candidate gene approach and obtained from genome-wide association study (GWAS). A total of 464 SNPs (254 SNPs in 26 NER genes, 173 SNPs in 22 BER genes and 37 SNPs in 26 DSBR genes) were included in the final analysis. Among the top significant SNPs, SNP rs10258834 in RPA3 from the NER pathway contributed to over 30% increased EAC risk (OR=1.30, 95% CI=1.12-1.51, P=5.26x10−4), SNP rs3827392 in SMC1B from the DSBR pathway was associated with a 25% increased EAC risk (OR=1.25, 95% CI=1.07-1.47, P=4.80x10−3); whereas rs3135998 in LIG3 from the BER pathway was associated with a decreased risk of EAC (OR=0.73, 95% CI=0.59-0.90, P=3.87x10−3). Unfavorable genotype analysis was performed among those SNPs showing significant association (P<0.05) in individual SNP analysis. We found a strong cumulative effect overall or stratified by DNA repair sub-pathway. For example, individuals carrying 10-13 unfavorable genotypes in BER pathway exhibited a nearly four-fold increased EAC risk compared to those in the reference group with fewer than 5 unfavorable genotypes. Our results suggest that genetic variants in DNA repair pathways significantly modify EAC risk individually and jointly. The results from this study, if validated in independent populations, could potentially categorize population into different risk groups and identify high-risk individuals for cost-effective screening and early detection of this deadly disease.

Citation Format: Yan Wang, Jaffer A. Ajani, Yuanqing Ye, Maosheng Huang, Xifeng Wu. Genetic variants in DNA repair pathway genes are associated with the risk of esophageal adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1349. doi:10.1158/1538-7445.AM2013-1349
Abstract 1400: Obesity induced leptin-notch signaling axis in breast cancer.
Authors: Battle Monica , Gillespie Corey , McGlothen Tanisha , Torroella-Kouri Marta , Gonzalez-Perez Ruben R.
Published: August 2014

Obesity leads to increased leptin levels and signaling which has been linked to the growth of breast cancer (BC). We early identified leptin as an inducer of Notch in BC cells. However, whether leptin-induction of Notch is instrumental to the development of BC is unknown. Objective: Therefore, we examined the impact of obesity and inhibition of leptin signaling (via LPrA2 antagonist) on the growth of syngeneic E0771-derived BC and tumor expression of Notch in lean and obese mice and in E0771 cell cultures. Methods: C57BL/6J female mice were fed either high-fat diet (HFD, 60% Kcal from fat) or normal diet (10% Kcal from fat). Obesity was defined as BW ≥ 20% BW Lean-control at week 5. Lean and diet-induced-obesity (DIO) mice were orthotopically inoculated with mouse E0771 cells in the lower left region of the mammary pad. One week later the mice were treated with either (1) pegylated leptin peptide receptor antagonist 2, PEG-LPrA2 or (2) PEG-Scrambled peptide (as control; 50μl i.v. tail vein/ 0.1 mM) following the two-arm design: (a) one and (b) two doses/weekly. Treatment lasted for 4 weeks. Tumor onset and growth was determined overtime and expression of Notch, its ligands, and crosstalk factors were assessed using western blot, immunohistochemistry, and ELISA. Leptin's effects on Notch and crosstalk factors were also investigated in E0771 cell cultures. Results: Obesity induced E0771-BC onset and growth and, increased the expression of Notch in vivo. Inhibition of leptin signaling had a higher impact on DIO mice than in lean mice by delaying onset and reducing tumor growth and the expression of Notch. Therefore, high levels of leptin in DIO-mice played a positive role in tumorigenesis. Similarly, leptin induced and PEG-LPrA2 decreased Notch expression in E0771 cells in vitro. Present data further validate the idea that the consumption of a high-fat diet leads to early BC onset and growth, which was linked to the leptin-Notch axis. Our results reinforce the potential use of inhibition of leptin signaling as a novel approach to fight breast cancer, especially in obese patients showing higher levels of leptin and incidence of BC.

Funding: This work was supported by NIH/NCI 5SC1CA138658-03 and the Georgia Cancer Coalition Distinguished Cancer Scholar Award (to RRGP), NIH/2G12RR003034-26; U54 MSM/TU/UAB Cancer Center Partnership; and 1R21CA153172-01A1 (to MTK). This research was also supported, in part, facilities and support services at Morehouse School of Medicine (NIH 1C06 RR18386).

Citation Format: Monica Battle, Corey Gillespie, Tanisha McGlothen, Marta Torroella-Kouri, Ruben R. Gonzalez-Perez. Obesity induced leptin-notch signaling axis in breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1400. doi:10.1158/1538-7445.AM2013-1400
Abstract 1408: Opposing roles of p53 and autophagy in a model of PALB2-associated hereditary breast cancer.
Authors: Huo Yanying , Cai Hong , Teplova Irina R , Bowman-Colin Christian , Chen Guanghua , Pricea Sandy , Karantzaa Vassiliki , White Eileen , Xia Bing
Published: August 2014

Hereditary breast cancers stem from germline mutations in susceptibility genes such as BRCA1, BRCA2 and PALB2. These and the majority of other known susceptibility genes all encode products that function in the repair and response to DNA damage, particularly double strand breaks. The p53 tumor suppressor is frequently mutated in hereditary breast cancer, and results from mouse models also suggest that its inactivation is critical for tumorigenesis induced by loss of BRCA1 and BRCA2. Autophagy is an intracellular waste disposal/recycling and stress mitigation mechanism that that can impact both cancer initiation and progression, but its role in breast cancer is unclear. Here we show that, similar to Brca1 and Brca2, ablation of Palb2 in mouse mammary gland resulted in tumor development with long latency, the majority of the tumors harbored mutations in Trp53, and combined deletion of Palb2 and Trp53 led to efficient tumor formation. Interestingly, partial loss of autophagy, due to mono-allelic loss of the essential autophagy gene Becn1, delayed and reduced Palb2-associated mammary tumorigenesis in mice with Palb2 single deletion but not Palb2;Trp53 double deletion in the mammary gland. These results indicate that, in the face of DNA damage and genome alterations elicited by PALB2 loss, p53 is a barrier to cancer development whereas autophagy facilitates cell survival and tumorigenesis. Our findings also underscore the similarity between PALB2- and BRCA-associated cancers and further suggest that inhibition of autophagy may represent a new avenue to the prevention and/or treatment of hereditary breast cancers.

Citation Format: Yanying Huo, Hong Cai, Irina R Teplova, Christian Bowman-Colin, Guanghua Chen, Sandy Pricea, Vassiliki Karantzaa, Eileen White, Bing Xia. Opposing roles of p53 and autophagy in a model of PALB2-associated hereditary breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1408. doi:10.1158/1538-7445.AM2013-1408
Abstract 1440: Mechanisms of TGF-β1-induced myofibroblast/CAF differentiation in human prostate-derived mesenchymal stem cells.
Authors: Kim Woosook , Barron David , San Martin Rebeca , Ressler Steven , Rowley David
Published: August 2014

Reactive stroma in prostate cancer is typified by the co-evolution of myofibroblasts/CAFs. This reactive stroma is associated with most human carcinoma and is predictive of progression. TGF-β1 is a key factor in regulating reactive stroma biology. However, the origin of myofibroblasts and the mechanisms of how TGF-β1 recruits, activates and induces their differentiation are essentially unknown. We have identified mesenchymal stem cells from normal human prostate gland and evaluated their biology in a novel 3D co-culture system. Human prostate-derived mesenchymal stem cells (hpMSCs) were CD44+/CD90+, expressed other mesenchymal stem cell genes, and exhibited multipotent differentiation patterns. When co-cultured with LNCaP cells in Millicell-CM inserts or co-inoculated in nude mice, self-organizing organoids formed with a core of stromal cells and a peripheral mantel of LNCaP cells. To investigate the role of TGF-β1, the hpMSCs were co-cultured with LNCaP cells engineered to overexpress active TGF-β1. Significantly, LNCaP cells engineered to overexpress TGF-β1 induced hpMSC differentiation to prototypical reactive stroma myofibroblasts. Microarray analysis of stroma revealed 1617 gene probes with more than a 2 fold-change in the presence of TGF-β1, showing that expression of TGF-β1 in LNCaP cells drives differential gene expression in hpMSCs. Of those gene expression profiles, RUNX1 and ID1 were identified as key transcription factors in hpMSCs that mediate TGF-β1-induced myofibroblast differentiation. Knockdown of RUNX1 in hpMSCs significantly promoted differentiation to myofibroblasts. Conversely, overexpression of RUNX1 inhibited a myofibroblast gene expression signature. Together, these data implicate RUNX1 as a major regulator of the stem cell state of tissue-derived myofibroblast stem/progenitor cells and therefore a modulator of the co-evolution of reactive stroma. Furthermore, ID1 may function as a negative regulator of reactive stroma formation by promoting proliferation of transiently amplifying cells and inhibiting myofibroblast differentiation. Our data show that reactive stroma in prostate cancer initiates from activation and differentiation of CD44+/CD90+ endogenous hpMSCs through TGF-β1/RUNX1/ID1 pathway. Understanding these mechanisms is important for developing new strategies to target the microenvironment niche.

Citation Format: Woosook Kim, David Barron, Rebeca San Martin, Steven Ressler, David Rowley. Mechanisms of TGF-β1-induced myofibroblast/CAF differentiation in human prostate-derived mesenchymal stem cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1440. doi:10.1158/1538-7445.AM2013-1440
Abstract 1498: Alpha-Lipoic acid suppresses migration and invasiveness of thyroid cancer cells through inhibition of epithelial-mesenchymal transition .
Authors: Han Ji Min , Choi Hyun-Jeung , Yim Ji Hye , Kim Won Gu , Kim Tae Yong , Shong Young Kee , Kim Eui Young , Kim Won Bae
Published: August 2014

Reactive oxygen species (ROS) are recently known as a contributor to tumor progression and metastasis through several mechanisms including induction of epithelial-mesenchymal transition (EMT), invasion, and tumor angiogenesis. In this study, we investigated the role of alpha-Lipoic acid (ALA), a potential antioxidant, on EMT in thyroid cancer cell lines. Human thyroid cancer-derived cell lines, BCPAP (BRAF V600E), HTH83 (HRAS Q61R), CAL62 (KRAS G12R), BHT101 (BRAF V600E), and FTC133 (PTEN null) were treated with ALA, and changes in ROS generation were measured by DCFH-DA. The expression of E-cadherin, vimentin and Twist were determined by immunoblot analysis. Cell migration and invasion were evaluated by scratch assay and by Boyden chamber assay. ALA significantly decreased intracellular ROS generation in most cell lines. The total amount of E-cadherin increased and those of vimentin and Twist decreased after ALA treatment in 4 thyroid cancer cell lines (BCPAP, HTH83, BHT101 and CAL62) in a dose-dependent manner (0.2∼1.0 mM). But, the changes in E-cadherin, vimentin and Twist were opposite in FTC133 cells. The cell migration and invasion ability decreased after treatment with ALA in all cell lines except FTC133 (PTEN null cell). These results suggest that ALA is a potential agent to inhibit EMT resulting in suppression of migration and of invasiveness in thyroid cancer cell lines with RAS or BRAF mutation, probably through reducing cellular amount of Twist. We suggest that it might be caused by antioxidant effects of ALA, but the exact mechanism of action of ALA is not clear yet. ALA could be a novel, potential agent to suppress metastasis of tumors in patients with thyroid cancer.

Citation Format: Ji Min Han, Hyun-Jeung Choi, Ji Hye Yim, Won Gu Kim, Tae Yong Kim, Young Kee Shong, Eui Young Kim, Won Bae Kim. Alpha-Lipoic acid suppresses migration and invasiveness of thyroid cancer cells through inhibition of epithelial-mesenchymal transition . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1498. doi:10.1158/1538-7445.AM2013-1498
Abstract 1499: Role of Twist for maintenance of EMT in colon cancer cell .
Authors: Kim Hye Ryeon , Kim SoYoung , Baik Young Ae , Hong Hye Kyung , Lee Woo Yong , Cho Yong Beom
Published: August 2014

Epithelial-mesenchymal transition or transformation (EMT) is a process characterized by loss of cell adhesion, repression of E-cadherin expression, and increased cell motility. Epithelial-mesenchymal transition (EMT) confers tumor cells with metastatic dissemination, cancer stem cell-like characters and drug resistance. However, the molecular mechanism for metastatic dissemination remains unclear. Initiation of metastasis involves invasion, which has many phenotypic similarities to EMT, including a loss of cell-cell adhesion mediated by E-cadherin repression and an increase in cell mobility. In this study, we induced EMT in colon cancer cell line LoVo with expression of Twist, a key transcriptional factor of EMT. The metastasis associated with EMT was determined E-cadherin, vimentin and β-catenin protein expression by western blot and morphological changes associated with EMT in these cells. We found that expression of Twist induced a morphological change associated with EMT in these cells. We also found that the E-cadherin was significantly decreased in Twist-overexpressing cells LoVo. Interestingly, we showed that β-catenin was activated in these Twist-overexpressing cells. Our results indicate that activation of β-catenin is required for the sustention of EMT-associated metastasis dissemination.

Citation Format: Hye Ryeon Kim, SoYoung Kim, Young Ae Baik, Hye Kyung Hong, Woo Yong Lee, Yong Beom Cho. Role of Twist for maintenance of EMT in colon cancer cell . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1499. doi:10.1158/1538-7445.AM2013-1499
Abstract 1508: Gene expression analysis of perfusable nodules grown on ex vivo 3D model shows effects on extracellular matrix, cell signaling and immune response.
Authors: Mishra Dhruva K. , Creighton Chad , Zhang Yiqun , Gibbons Don L. , Kurie Jonathan M. , Kim Min P.
Published: August 2014

Tumor microenvironment plays a very important role in regulating the cell behavior within a tissue, largely by interacting with the extracellular matrix (ECM). Recently, our lab has developed a novel ex vivo lung model that forms perfusable tumor nodules, which mimic human lung cancer histopathology and protease secretion patterns better than tumor cells grown on a petri dish. In this study, we have compared the genome-wide gene expression profiles (Affymetrix 430_2) and protein arrays (Reverse phase protein array or RPPA) of 2D (monolayer) culture versus our ex vivo 3D cell culture using 393p cells, derived from mice lung cancer. Expression array data showed a set 1314 gene probes differentially expressed between monolayer 2D culture and 3D ex vivo cell culture. Gene Ontology (GO) analysis of differentially expressed genes showed the upregulation of several genes associated with extracellular matrix (ECM1, BMP15, FMOD, cathepsin, MMP9, MMP10), cell signaling (VEGFA, TGFB2, FGF20), and immune response (C1, C3, MHC class II, IL1α, IL1RA, IL13, TNF receptor superfamily, CXCL2). Selected genes were also validated using real time PCR. We found 46 proteins with significant (p<0.01) differential levels in 2D vs 3D cell cultures of 393p. The proteins associated with extracellular matrix (Collagen VI, Caveolin1, Fibronectin) and cell signaling (mTOR, p38 (phosphorylated), Caspase 7 (Cleaved), BAD (phosphorylated), PI3K) were upregulated in 3D ex vivo cell culture compared to conventional 2D culture. Thus, our results showed the insightful influence of the 3D ex vivo model and its three-dimensional perfusable environment over cell growth and its behavior in terms of significant differences in gene expression and protein patterns as compared to the same cells grown on conventional 2D cell cultures.

Citation Format: Dhruva K. Mishra, Chad Creighton, Yiqun Zhang, Don L. Gibbons, Jonathan M. Kurie, Min P. Kim. Gene expression analysis of perfusable nodules grown on ex vivo 3D model shows effects on extracellular matrix, cell signaling and immune response. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1508. doi:10.1158/1538-7445.AM2013-1508
Abstract 1521: Role of the guanylate-binding-protein 1 (GBP-1) in immunoediting of colorectal carcinoma.
Authors: Britzen-Laurent Nathalie , Lipnik Karoline , Ocker Matthias , Naschberger Elisabeth , Schellerer Vera , Croner Roland , Vieth Michael , Waldner Maximilian , Steinberg Pablo , Hohenadl Christine , Stürzl Michael
Published: August 2014

The human guanylate-binding protein 1 (GBP-1) is among the proteins the most highly induced by IFN-γ in every cell type investigated as yet. In vivo, GBP-1 expression is associated with the presence of inflammation and has been observed in autoimmune diseases, inflammatory bowel diseases (IBD) and cancer. In colorectal carcinoma (CRC), we previously reported that the expression of GBP-1 in the cells of the desmoplastic stroma correlates with the presence of an IFN-γ-dominated Th1 microenvironment and with an increased cancer-related 5-year survival. Moreover, GBP-1 expression in the blood vessels of CRC tissue was associated with a reduced angiogenesis. However, the expression and the role of GBP-1 in the tumor cells of CRC remained unknown.

In the present study, the analysis of GBP-1 expression in a series of 185 CRCs by immunohistochemistry confirmed that GBP-1 is expressed in stroma cells of CRCs and revealed a significantly less frequent expression in tumor cells, which was contradictory with the broad inducibility of GBP-1. Furthermore, three of six CRC cell lines treated with IFN-γ were unable to express GBP-1 indicating a reduced inducibility in colorectal tumor cells. On the contrary, non-transformed colon epithelial cells strongly expressed GBP-1 in vitro in presence of IFN-γ and in vivo in IBD. Reconstitution of GBP-1 expression in a negative CRC cell line inhibited cell proliferation, migration, invasion and anchorage-independent growth. Using RNA interference, we showed that GBP-1 mediates the anti-proliferative, anti-migratory, anti-invasive and anti-clonogenic effects of IFN-γ in CRC cells. In addition, GBP-1 was able to inhibit tumor growth in vivo. Altogether these results suggested that GBP-1 acts directly as a tumor suppressor in CRC by mediating the effects of IFN-γ. In addition, the loss of GBP-1 expression in tumor cells indicated that tumor evasion from the IFN-γ-dominated Th1 immune response might have occured. This was confirmed by showing, first, that the absence of GBP-1 inducibility in CRC cell lines correlates with resistance to the anti-proliferative and pro-apoptotic effects of IFN-γ and, second, that the resistant cell lines exhibit defects in the IFN-γ signaling pathway.

Citation Format: Nathalie Britzen-Laurent, Karoline Lipnik, Matthias Ocker, Elisabeth Naschberger, Vera Schellerer, Roland Croner, Michael Vieth, Maximilian Waldner, Pablo Steinberg, Christine Hohenadl, Michael Stürzl. Role of the guanylate-binding-protein 1 (GBP-1) in immunoediting of colorectal carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1521. doi:10.1158/1538-7445.AM2013-1521
Abstract 1523: MALDI mass spectrometry imaging for on-tissue spatial profiling of tumor-specific N-linked glycans and lipids in renal carcinomas.
Authors: Drake Richard R. , Powers Thomas , Jones Ellen , Lance Raymond S. , Mehta Anand S. , Troyer Dean A.
Published: August 2014

Pathologic staging of renal clear cell carcinoma (ccRCC) provides useful but limited prognostic information suggesting that there are underlying molecular processes not readily identified through conventional histopathological techniques in the primary tumor. At the molecular level, transcript, metabolomic and protein expression patterns have indicated a striking Warburg Effect profile in ccRCC tissues, with major affects on sugar and lipid metabolism. Our group has been applying MALDI-MS imaging approaches to ccRCC tissues to profile lipids and glycans associated with disease progression. As both lipids and glycans can act as signaling molecules and have been associated with cancer progression, ccRCC tissues were selected by a pathologist to contain substantial tumor, non-tumor and tumor margin regions to characterize the differential localization of lipid and glycan species within the tissue. Lipid profiles from fresh-frozen tissue slides coated in DHB matrix were obtained on a dual source Bruker Solarix 70 FTICR mass spectrometer. Glycans were imaged in a similar fashion with a method developed by our lab using protein N-glycanase F to release protein bound N-glycans. There are clear patterns of specific lipid and glycan species that are only present along the margin of the tumor and non-tumor interface, or those primarily localized in non-tumor or tumor regions. Off-tissue extraction of lipids and glycans from similarly processed tissues and further mass spectrometry analysis was done to confirm structural designations. Linkage of differentially expressed lipid and glycan species with clinical data and other molecular markers of ccRCC is ongoing. This approach has the potential to not only improve prognostic assessment and enhance post-operative surveillance, but also to better define the underlying biology of ccRCC aggressiveness and identify new rational targets for therapeutic intervention.

Citation Format: Richard R. Drake, Thomas Powers, Ellen Jones, Raymond S. Lance, Anand S. Mehta, Dean A. Troyer. MALDI mass spectrometry imaging for on-tissue spatial profiling of tumor-specific N-linked glycans and lipids in renal carcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1523. doi:10.1158/1538-7445.AM2013-1523
Abstract 1529: Endoneurial macrophages induce perineural invasion of pancreatic cancer cells by secretion of GDNF and activation of RET tyrosine kinase receptor.
Authors: Cavel Oren , Wong Richard J. , Shomron Olga , Amit Moran , Gil Ziv
Published: August 2014

Perineural invasion of cancer cells (CPNI) is found in most patients with pancreatic adenocarcinomas (PDAs), prostate or head and neck cancers. These patients undergo palliative rather than curative treatment due to dissemination of cancer along nerves, well beyond the extent of any local invasion. Although CPNI is a common source of distant tumor spread and a cause of significant morbidity, its exact mechanism is undefined. Immunohistochemical analysis of specimens excised from patients with PDAs showed a significant increase in the number of endoneurial macrophages (EMΦs) which lie around nerves invaded by cancer compared to normal nerves. Video microscopy and time-lapse analysis revealed that EMΦs are recruited by the tumor cells in response to colony stimulated factor-1 secreted by invading cancer cells. Conditioned medium (CM) of tumor-activated EMΦs (tEMΦs) induced a 5-fold increase in migration of PDA cells compared to controls.

Compared to resting EMΦs, tEMΦs secreted higher levels of glial-derived neurotrophic factor (GDNF), inducing phosphorylation of RET and downstream activation of extracellular signal-regulated kinases (ERK) in PDA cells. Genetic and pharmacologic inhibition of the GDNF receptors GFRA1 and RET abrogated the migratory effect of EMΦ-CM and reduced ERK phosphorylation. In an in-vivo CPNI model, CCR2-deficient mice which have reduced macrophages recruitment and activation showed minimal nerve invasion, while wild-type mice developed complete sciatic nerve paralysis due to massive CPNI. Taken together, our results identify a paracrine response between EMΦs and PDA cells which orchestrates the formation of cancer nerve invasion.

Citation Format: Oren Cavel, Richard J. Wong, Olga Shomron, Moran Amit, Ziv Gil. Endoneurial macrophages induce perineural invasion of pancreatic cancer cells by secretion of GDNF and activation of RET tyrosine kinase receptor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1529. doi:10.1158/1538-7445.AM2013-1529

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1601: Paclitaxel potentiates212Pb-radioimmunotherapy-induced cell killing efficacy by perturbing mitotic spindle checkpoint.
Authors: Yong Kwon Joong , Milenic Diane E. , Baidoo Kwamena E. , Brechbiel Martin W.
Published: August 2014

We have recently reported on the potentiation of high-LET radiation 212Pb-TCMC-trastuzumab targeting HER2 in combination with paclitaxel. To elucidate mechanisms associated with this therapy, mice bearing human colon cancer LS-174T i.p. xenografts were pre-treated with paclitaxel, followed by treatment with 212Pb-TCMC-trastuzumab and compared to groups treated with paclitaxel alone, 212Pb-TCMC-HuIgG, 212Pb-TCMC-trastuzumab and 212Pb-TCMC-HuIgG with paclitaxel pre-treatment. 212Pb-TCMC-trastuzumab with paclitaxel given 24-30 h earlier, induced mitotic catastrophe and increased apoptosis. The combined modality of paclitaxel and 212Pb-TCMC-trastuzumab markedly reduced DNA content in the S-phase of the cell cycle with a concomitant increase observed in the G2/M-phase. This treatment regimen also resulted in reduced phosphorylation of histone H3, accompanied by an increase in multimicronuclei, or mitotic catastrophe in nuclear profiles and positive stained γH2AX foci. This data suggests possible effects on the mitotic spindle checkpoint by the paclitaxel and 212Pb-TCMC-trastuzumab treatment. Consistent with this hypothesis, 212Pb-TCMC-trastuzumab treatment in response to paclitaxel reduced expression and phosphorylation of BubR1, which is likely attributable to disruption in a functional survivin/Aurora B complex, leading to impairment of the mitotic spindle checkpoint. In addition, the reduction of BubR1 expression may be mediated by the association of a repressive transcription factor, E2F4, on the promoter region of BubR1 gene. Taken together, these findings suggest that paclitaxel pre-treatment sensitizes 212Pb-TCMC-trastuzumab induced cell killing, leading to cell death by mitotic catastrophe followed by apoptosis.

Citation Format: Kwon Joong Yong, Diane E. Milenic, Kwamena E. Baidoo, Martin W. Brechbiel. Paclitaxel potentiates 212Pb-radioimmunotherapy-induced cell killing efficacy by perturbing mitotic spindle checkpoint. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1601. doi:10.1158/1538-7445.AM2013-1601
Abstract 1617: Chebulagic acid can inhibit vascular endothelial growth factor-A mediated angiogenesis.
Authors: Lu Kai , Basu Sujit
Published: August 2014

Background: Angiogenesis or new blood vessel formation is essential for the growth, progression and metastasis of malignant tumors. Among the several pro-angiogenic factors, vascular endothelial growth factor-A (VEGF-A) plays a critical role in this process of angiogenesis. However, the current anti-VEGF-A agents in the clinics are not only expensive, but also have serious side effects. Therefore it is essential to identify newer, effective, less toxic and inexpensive anti-VEGF-A agents for the treatment of cancer. We have recently demonstrated the anti-VEGF-A actions of Triphala churna (THL), a natural medicinal product prepared from three myrobalan fruits. Because chebulagic acid (CA), a bioactive tannin present in THL has been reported to inhibit tumor growth, we therefore investigated the effect of CA on VEGF-A mediated angiogenesis.

Method: Human umbilical vein endothelium cells (HUVECs) were cultured in vitro, and pre-treated with different doses of CA for 24hrs. Thereafter, we examined the proliferation migration and tube formation abilities of these cells. In addition we also determined the effects of CA on HUVEC cell permeability and on new blood vessel formation utilizing CAM assay. Finally, the effect of CA on VEGFR-2 phosphorylation was investigated.

Results: 5uM CA can significantly inhibit VEGF-A induced cell proliferation, migration and permeability of HUVECs. Besides it can also inhibit formation of tube-like structures by these cells. In vivo CAM assay also suggested that 5uM CA can suppress VEGF-A stimulated new vessel formation. Western blot confirmed that CA inhibits VEGF-A actions by suppressing phosphorylation of VEGFR-2.

Conclusion: Our findings suggest that CA can significantly inhibit VEGF induced angiogenesis.

Citation Format: Kai Lu, Sujit Basu. Chebulagic acid can inhibit vascular endothelial growth factor-A mediated angiogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1617. doi:10.1158/1538-7445.AM2013-1617
Abstract 2053: F-XA7, a novel derivative of vinca alkaloid with potent antitumor activity, efficiently overrides P-glycoprotein-mediated drug resistance both in vitro and in vivo.
Authors: Quan Haitian , Xu Yongping , Hu Lihong , Lou Liguang
Published: August 2014

Vinorelbine, a semi-synthetic vinca alkaloid, is widely used for the treatment of non-small cell lung cancer, metastatic breast cancer and ovarian cancer by inhibiting microtubule assembly. However, drug resistance mediated by P-glycoprotein (P-gp) and neurotoxic side effect possibly caused by microtubule paracrystal are major obstacles to vinorelbine application in clinic. Here, we describe a vinorelbine derivative F-XA7 as a novel microtubule-disrupting agent. F-XA7 inhibits microtubule assembly by binding to tubulin at the same site as vinorelbine and thus inhibiting the proliferation of cultured tumor cells. In vivo, F-XA7 exhibits potent antitumor activity against human tumor xenografts in nude mice. More importantly, F-XA7 is active against vinorelbine-resistant tumor cells mediated by P-gp-overexpression and significantly suppresses the growth of P-gp-overexpressing tumor xenograft in vivo, indicating that F-XA7 efficiently overrides P-gp-mediated drug resistance. In addition, the activity of F-XA7 to induce microtubule paracrystal is much weaker than that of vinorelbine in cultured cells, suggesting it may have less neurotoxic side effect. Collectively, these results indicate that F-XA7 is a novel and promising microtubule-disrupting agent for cancer therapy.

Citation Format: Haitian Quan, Yongping Xu, Lihong Hu, Liguang Lou. F-XA7, a novel derivative of vinca alkaloid with potent antitumor activity, efficiently overrides P-glycoprotein-mediated drug resistance both in vitro and in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2053. doi:10.1158/1538-7445.AM2013-2053
Abstract 2129: Aplidin triggers the activation of molecular components of the UPR as part of its pro-apoptotic program in tumor cells.
Authors: Losada Alejandro , Martínez-Leal Juan F. , Bejarano Alberto , Cuevas Carmen , Garcia-Fernández Luis F. , Galmarini Carlos M.
Published: August 2014

Aplidin (APL), a cyclic depsipeptide originally isolated from the marine tunicate Aplidium albicans and currently under phase II/III clinical trials for cancer therapy, induces oxidative stress, activation of Rac1 and phosphorylation of JNK1, which together trigger a rapid apoptotic program in tumor cells. A previously generated APL-resistant HeLa cell line (APL-R) allowed us to find out differences in protein expression with the parental HeLa cells. Among the proteins differentially expressed, several endoplasmic reticulum (ER) stress-related proteins were observed. We found decreased basal levels of BiP in HeLa APL-R, while the levels of Ero1a and phospho-eIF2a were slightly increased when compared to parental HeLa cells, indicative of a higher basal ER stress. In this work, we investigated if APL was inducing a bona fide ER stress in HeLa cells and whether this process was essential in the mechanism of action of the compound. Similarly to that observed with thapsigargin and tunicamycin, two well known ER stress inducing agents, APL triggered the activation of several key molecular components of a classical ER stress induced unfolded protein response (UPR), including the phosphorylation of eIF2a and JNK1, the proteolitic processing of ATF6 and the alternative splicing of XBP1, but not others, such as the accumulation of noxa or the proteolitic activation of caspase 4. Interestingly, although the eIF2a phosphorylation induced by APL in HeLa wt cells correlated with a strong inhibition of protein synthesis, it did not elicit an increased expression of CHOP, a transcription factor involved in the launch of UPR mediated apoptosis. Actually, it was observed a clear reduction of CHOP protein levels after the treatment of HeLa cells with APL, most probably due to its rapid degradation by the proteasome machinery. In summary, as a part of the proapoptotic program triggered by APL in HeLa cells, we observed the induction of ER stress and the activation of several endpoint executioners of the UPR apoptotic branch.

Citation Format: Alejandro Losada, Juan F. Martínez-Leal, Alberto Bejarano, Carmen Cuevas, Luis F. Garcia-Fernández, Carlos M. Galmarini. Aplidin triggers the activation of molecular components of the UPR as part of its pro-apoptotic program in tumor cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2129. doi:10.1158/1538-7445.AM2013-2129
Abstract 2326: S6K activation is essential for the development of follicular thyroid cancer.
Authors: Antico Arciuch Valeria Gabriela , Di Cristofano Antonio
Published: August 2014

The PI3K signaling cascade is frequently activated in follicular and anaplastic thyroid tumors. Mice in which the PI3K pathway is constitutively activated through loss of Pten (PtenL/L;TPO-Cre mice) develop thyroid hyperplasia that progresses to thyroid follicular carcinoma.

The wide range of downstream targets of this pathway raises the question of the relative role and relevance of its multiple effectors. This issue has important therapeutic implications, since it would be ideal to only target molecules directly involved in the transformation process, thus reducing the toxicity associated with wide-range inhibitors.

PDK1 plays a central role in the PI3K cascade, acting as a master regulator of both AKT and S6K. Notably, PDK1 possesses a substrate docking site named “PIF pocket” that is required to phosphorylate and activate S6K but not AKT.

In order to define the relative contribution of Akt and S6k to the development of thyroid lesions and to their progression to follicular carcinoma, we have generated a compound mouse strain in which, in addition to thyroid-specific Pten loss, the PIF-pocket site of Pdk1 can be conditionally disrupted by the L155E mutation, resulting in constitutive activation of Akt and mTor, but not of S6k.

We have found that young double mutant mice display a significant reduction in thyroid size, compared to PtenL/L;TPO-Cre mice, and that such rescue is associated with a marked decrease in the thyrocytes proliferation rate. More strikingly, while 60% of Pten mutants develop adenomas and carcinomas by one year of age, none of the age-matched double mutant mice displays neoplastic lesions, indicating that Akt activation, in the absence of S6k activation, is completely unable to drive transformation.

Furthermore, we have identified STAT3 as a molecule that is selectively activated downstream of S6k, and that is inactive in double mutants. Pharmacological STAT3 inhibition drastically reduces the proliferation rate of thyroid cancer cell lines, suggesting that STAT3 may be one key conduit of S6k oncogenic activity.

Citation Format: Valeria Gabriela Antico Arciuch, Antonio Di Cristofano. S6K activation is essential for the development of follicular thyroid cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2326. doi:10.1158/1538-7445.AM2013-2326
Abstract 2561: MiRNA-related SNPs and risk of esophageal adenocarcinoma and Barrett's esophagus: post genome-wide association analysis in the BEACON consortium.
Authors: Buas Matthew F. , Onstad Lynn , Levine David , Vaughan Thomas L. , Whiteman David C. ,   BEACON Consortium Investigators
Published: August 2014

The incidence of esophageal adenocarcinoma (EA) has increased significantly in recent decades. Although several major risk factors have been identified for EA and its precursor, Barrett's esophagus (BE), including reflux, Caucasian race, male gender, obesity, and smoking, only limited knowledge exists regarding the role of inherited genetic variation and its interplay with environmental factors. The Barrett's and Esophageal Adenocarcinoma Consortium (BEACON) recently completed a genome-wide association study (GWAS) of 1,517 EA cases, 2,416 BE cases, and 2,187 controls. Using this dataset, we examined single nucleotide polymorphisms (SNPs) that potentially affect the biogenesis or biological activity of microRNAs (miRNAs), a class of small non-coding RNAs that regulate post-transcriptional gene expression and are deregulated in many cancers, including EA. Polymorphisms in three classes of genes were evaluated for their association with risk of EA or BE: 1) miRNA biogenesis genes (157 SNPs), 2) miRNA gene loci (234 SNPs), and 3) miRNA-targeted mRNAs (179 SNPs). 28 SNPs were significantly associated (P<0.05) with EA, and 34 with BE. A polymorphism located in a predicted miRNA-138 binding site of the DNA repair gene XRCC1 (rs1799782) was associated with >30% reduced risk of EA (OR 0.68, 95% CI 0.55-0.83), which remained significant after correction for multiple comparisons (adjusted P=0.04). This inverse association became more pronounced with increasing pack-years (P-interaction=0.02). Alterations in disease risk associated with other polymorphisms ranged from 10-40%. This study provides the most extensive assessment to date of miRNA-related SNPs in relation to risk of EA and BE and suggests that genetic variation in this pathway may influence disease susceptibility and interact with known risk factors.

Citation Format: Matthew F. Buas, Lynn Onstad, David Levine, Thomas L. Vaughan, David C. Whiteman, BEACON Consortium Investigators. MiRNA-related SNPs and risk of esophageal adenocarcinoma and Barrett's esophagus: post genome-wide association analysis in the BEACON consortium. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2561. doi:10.1158/1538-7445.AM2013-2561
Abstract 2575: Differential anti-tumor activities of curcumin against apoptosis and cell Cycle progression in Src- and Ras-activated human gallbladder carcinoma cells.
Authors: Ono Misaki , Higuchi Takako , Takeshima Mikako , Chen Chen , Nakano Shuji
Published: August 2014

Curcumin, a phytochemical in turmeric, has been studied as a potential anticancer drug targeting multiple signaling molecules. However, the role of Src and Ras signaling pathways in curcumin sensitivity remains unknown. We investigated therefore the molecular mechanism of anti-proliferative and apoptosis-inducing activities of curcumin in HAG-1 human gallbladder adenocarcinoma cells transfected with either activated Src (HAG/src3-1) or Ras (HAG/ras5-1). Effects of curcumin on proliferation, apoptosis, cell cycle perturbation, and signal proteins for survival, proliferation, and apoptosis were evaluated by WST-1 assay, FACS analyses and Western blotting. Activation of either Src or Ras did not confer resistance to curcumin compared to HAG/neo3-5 vehicle-transfected cells, producing similar IC50 concentrations of curcumin among these 3 cell lines. Upon treatment with curcumin, constitutive activities of Erk1/2 were enhanced, but those of Akt were significantly inhibited in these three cell lines, with subsequent reductions of activities of downstream mTOR and S6K1. The sub-G0/G1 apoptotic cell populations were substantially increased in all cell lines with demonstrable cleavage of PARP, but this increase was most prominent in Src-activated cells. Reduced expression and phosphorylation of Bcl-2 and enhanced expression of Bax were demonstrated to have a causative role for the curcumin-induced apoptosis in Src-driven, but not Ras-driven cells. By contrast, drastic increases of the proportion of cells in a G2/M phase were seen in Ras-activated cells, suggesting a potential role of Ras/Erk1/2 activation in curcumin-induced G2/M cell cycle arrest. These data indicate that curcumin-induced growth declines would be mediated mainly by G2/M arrest of the cell cycle in Ras-driven cells but by apoptosis induction in Src-driven cells. Taken together, the results indicate that curcumin exhibits differential activities against apoptosis and cell cycle progression, depending on activation mechanisms of signal transduction, and that curcumin can overcome Src- and Ras-driven activation of downstream signaling pathways, thus providing evidence of potential application of curcumin for the treatment of human cancers with activated Src or Ras.

Citation Format: Misaki Ono, Takako Higuchi, Mikako Takeshima, Chen Chen, Shuji Nakano. Differential anti-tumor activities of curcumin against apoptosis and cell Cycle progression in Src- and Ras-activated human gallbladder carcinoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2575. doi:10.1158/1538-7445.AM2013-2575