bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-176: A novel high density DNA methylation array with single CpG site resolution
Authors: Bibikova Marina , Barnes Bret , Tsan Chan , Ho Vincent , Klotzle Brandy , Le Jennie M. , Delano David , Gunderson Kevin L. , Fan Jian-Bing , Shen Richard
Published: July 2012

Aberrations in DNA methylation patterns are widely recognized as a hallmark of the cancer cell. DNA methylation has emerged as a promising biomarker for the detection and classification of various types of cancer, and ability to accurately assess the DNA methylation changes plays an important role in understanding of disease onset and progression.

We developed a new generation of genome-wide DNA methylation BeadChip which allows high-throughput methylation profiling of human genome. The new high density BeadChip can assay over 480K CpG sites and analyze twelve samples in parallel. The innovative content includes coverage of 99% of RefSeq genes with multiple probes per gene, 96% of CpG islands from the UCSC database, CpG island shores and additional content selected from whole-genome bisulfite sequencing data and input from DNA methylation experts. The well-characterized Infinium® assay is used for analysis of CpG methylation using bisulfite-converted genomic DNA. In this assay, unmethylated cytosines (C) are converted to uracil (U) when treated with bisulfite, while methylated cytosines remain unchanged. The assay design is using single probe assays for CpG loci with up to two underlying CpG dinucleotides, and two-probe assays for CpG loci with multiple underlying CpG sites, with one probe querying the “unmethylated” allele and the other probe querying the “methylated” allele. The assays are designed under the assumption that methylation is regionally correlated (within 50 base pairs) and that all CpG sites underlying the probe are assumed to be “in phase” with the queried CpG site.

For the BeadChip development, we used methylation standards created by de-methylating gDNA with Phi29 whole genome amplification, methylating the amplified DNA with SssI methylase and mixing the unmethylated and methylated DNA in a 1:1 ratio to create a 50% methylation state. 500 ng gDNA input is used for bisulfite conversion and is sufficient for two assays. Reproducible DNA methylation profiles were obtained between replicates (an average R2 of 0.98).

We applied this technology to DNA methylation analyses in cancer cell lines of different tissue of origin and normal DNA samples derived from multiple tissues. Highly specific methylation signatures were obtained for each sample type.

The ability to determine genome-wide methylation patterns will rapidly advance methylation research and ultimately lead to the development of powerful tools for diagnosis, prognosis, and treatment of human diseases.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-176. doi:10.1158/1538-7445.AM2011-LB-176
Abstract LB-329: Copy number variations associated with hereditary breast and colorectal carcinomas
Authors: Abreu Francine B. , Santos Erika Maria Monteiro , Rossi Benedito Mauro , Achatz Maria Isabel , Cristina Ana , Krepischi Victorino , Kowalski Luiz Paulo , Brentani Ricardo Renzo , Rogatto Silvia Regina
Published: July 2012

Breast and colorectal cancer are the most common malignant neoplasia in the world. In Brazil, breast cancer is the first malignant neoplasia and colorectal cancer, the third. It is estimated that among 5–10% of all cancer cases are hereditary. Copy number alterations (CNVs) may affect genes associated to cancer, providing an explanation for high risk cancer families. In this study, 57 individuals with Hereditary Breast and Colorectal Carcinoma were screened for BRCA1, BRCA2, CHEK2, TP53, MLH1 and MSH2 mutations by Sanger's sequencing method. In addition, copy number alterations were evaluated by array comparative genomic hybridization (aCGH) using a platform of 4×180K (Agilent). The data were compared with the Database of Genomic Variations (DGV;http://projects.tcag.ca/variation/) and with a reference dataset obtained from 82 healthy Brazilian individuals. Genomic data were extracted and flagged with Feature Extraction software and analyzed using Genomic Workbench Standard 5.0.14, statistical algorithm ADM2 and sensitivity threshold of 6.7. Alterations identified in more than 5% of the control women were excluded from both groups. The patients were divided into two groups: 28 patients with breast or colorectal cancer (group 1) and 29 women with breast and colorectal carcinoma (group 2). Sixteen relatives from seven patients were also evaluated by aCGH. All patients were negative for BRCA1, BRCA2, CHEK2, TP53, MLH1 and MSH2 pathogenetic mutations. However, the aCGH analysis detected a total of 569 chromosomal alterations in healthy Brazilian dataset (in average 6.8/individual), 170 CNVs (in average 6.1/individual) in group 1, and 232 (in average 8.0/individual) in group 2 patients. Forty-five and 63 chromosomal regions altered were detected in group 1 and 2, respectively. Nineteen chromosomal alterations were common to both groups. Mapped on these regions altered, it was identified 28 microRNAs. Ten specific chromosomal alterations were found in patients and their relatives; seven of them were losses involving genes not previously described in breast and colorectal carcinomas patients. Our results suggest that these chromosomal regions harbor genes associated with Hereditary Breast and Colorectal Carcinomas.

Financial Support: FAPESP (2008/57887–9) and CNPq (57589/2008–9)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-329. doi:10.1158/1538-7445.AM2011-LB-329
Abstract LB-332: High-throughput proteomic analysis identifies protein S as a potential modulator of androgen-independent prostate cancer
Authors: Saraon Punit , Jarvi Keith , Diamandis Eleftherios P.
Published: July 2012

Prostate cancer is the most commonly diagnosed and second leading cause of death due to cancer in men. Androgen deprivation remains the mainstay therapy, which is efficient in shrinking androgen-dependent tumors. However, many men eventually stop responding to this treatment, progressing to a more lethal androgen-independent prostate cancer. The mechanisms mediating these transitions remain unknown, and as a result, there are no effective treatments. We hypothesize the presence of specific modulators, important for the progression to androgen-independency. To test this, we performed high-throughput proteomic analysis on the conditioned media of the androgen-dependent cell lines LNCaP and VCaP, androgen-independent cell lines LNCaP-SF, DU-145, PC3, PPC1, 22Rv1, as well as a normal prostate cell line (RWPE). This allowed us to compile a comprehensive data set, permitting us to mine for differentially secreted proteins that could serve as potential markers for androgen-independent cancers. We utilized quantitative real-time PCR, as well as in vitro wound healing assays to corroborate our proteomic analysis results. We were able to identify the anti-coagulant factor, Protein S (PROS1), as a potential modulator and marker of androgen-independence. Based on spectral counts, PROS1 was found highly secreted in all the androgen-independent cell lines, with no detectable secretion in the normal or androgen-dependent cell lines. The gene expression profile of the androgen-independent cell lines also maintained the over-expression of PROS1, when compared to the normal and androgen-dependent cell lines, while only prostate cancer tissue samples showed an upregulation of PROS1. Furthermore, recombinant PROS1-treated LNCaP cells demonstrated an increase in wound repair and invasive growth potential, implying that PROS1 may have functional roles outside of the coagulation pathway. Our results suggest that PROS1 is involved in the progression of androgen-independent prostate cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-332. doi:10.1158/1538-7445.AM2011-LB-332
Abstract LB-337: Post-transcriptional regulators of microRNA biogenesis drive glioblastoma-specific expression
Authors: Chung Wei-Jen , Sumazin Pavel , Yang Xuerui , Bansal Mukesh , Califano Andrea
Published: July 2012

MicroRNAs have emerged as key regulators of both normal and pathologic phenotypes, including cancer. Fine grain regulation of their biogenesis, however, is still a poorly understood process and only a relatively small repertoire of proteins controlling its key steps has been characterized. In order to understand the extent and specificity of microRNA biogenesis control, as well as its role in glioblastoma tumorigenesis and progression, we developed and experimentally validated miRage, an algorithm for genome-wide inference of microRNA biogenesis regulators. MiRage identifies biogenesis-regulator candidates using the conditional mutual information between the candidates, mature microRNAs precursors, and abundance to identify those proteins that are required for the maturation of microRNAs.

miRage predicted hundreds of potential biogenesis regulators that are either specific to individual microRNAs or to microRNA families. Several of the inferred regulators that are differentially expressed across glioblastoma prognosis were validated in the SNB19 cell line. Differential expression of mature microRNAs, their host genes, and their precursors were measured before and after ectopic expression or RNAi mediated silencing of predicted regulators. ACSM3, which is predicted to regulate 70% of the differentially expressed microRNAs in glioblastoma, was shown to post-transcriptionally inhibit miR-155. This microRNA is an experimentally validated regulator of CEBPβ, a master regulator of the mesenchymal signature of GBM; indeed siRNA-mediated silencing of ACSM3 dramatically activated miR-155 and down-regulated CEBPβ. In addition, several members of the DDX RNA-binding protein family were inferred as specific regulators of microRNA biogenesis. For instance, we show that DDX10 activates miR-218 and inhibits miR-9; up regulation of DDX10 may account for miR-218 overexpression in poor-prognosis tumors. We showed that RBBP4 inhibits miR-23b, WDR8 inhibits miR-18b, ACTA2 inhibits miR-218, and PRIC285 inhibits miR-196a and miR-25.

Taken together, these results suggest that microRNA biogenesis is a much more complex and finely regulated process than previously thought, and that microRNA biogenesis regulators may play a key role in tumor initiation and progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-337. doi:10.1158/1538-7445.AM2011-LB-337
Abstract LB-339: MicroRNA modulators regulate oncogenes and tumor suppressors in glioblastoma
Authors: Sumazin Pavel , Yang Xuerui , Chung Wei-Jen , Bansal Mukesh , Califano Andrea
Published: July 2012

MicroRNAs, including miR-21, MiR-26a and miR-221/2, are potential key regulators of gene expression in glioblastoma, the most common and the most aggressive type of primary human brain tumor. Their activity is modulated by post-transcriptional factors that regulate mRNA processing pathways and by the abundance of their targets. Despite recent efforts to identify microRNA modulators, this mode of regulation is still poorly understood, and the repertoire and extent that microRNA modulators regulate gene expression in tumors and affect tumorigenesis remain unknown.

We developed a genome-wide approach to identify glioblastoma-specific microRNA modulators. We identified hundreds of candidate modulators, including modulators that either activate or suppress miRISC-mediated regulation, and mRNAs that act as decoys and compete for microRNA binding in glioblastoma. Validation experiments were done in SNB19 cell line by measuring differential expression of mature microRNAs and their targets after ectopic expression or inhibition of predicted regulator proteins. 3′ UTR luciferase assays were used to establish the post-transcriptional nature of the regulation. Focusing on two known drivers of gliomagenesis and progression, we showed that PTEN is post-transcriptionally up regulated by WNT7A through miR-21 and down regulated by PALB2 through miR-106a, and that RUNX1 is post-transcriptionally down regulated by WIPF2 through miR-17–5p. Further, we showed that PTEN and RUNX1, which are widely considered functionally unrelated, act as decoys for one another by competing for a common microRNA-regulatory program: they post-transcriptionally regulate one another through their microRNA regulators. In addition, other modulators with prognosis-predictive genetic alterations act through common microRNA-regulatory programs to regulate PTEN and RUNX1 expression. For example, prognosis-predictive genetic alterations at the CTBP2 locus significantly alter both CTBP2 and PTEN expression in glioblastoma; siRNA-mediated silencing of CTBP2 resulted in a 30% reduction in the expression of PTEN 3′ UTR luciferase reporters.

Our results demonstrate that microRNA modulators regulate the expression of key glioblastoma drivers, and that genetic alterations at microRNA modulators lead to aberrant post-transcriptional regulation of these drivers. Taken together, these results suggest that microRNA modulation plays a significant role in tumorigenesis and progression of gliobalstoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-339. doi:10.1158/1538-7445.AM2011-LB-339
Abstract LB-342: MicroRNA-130b contributes to mesenchymal differentiation and leiomyosarcomagenesis
Authors: Danielson Laura , Guijarro Maria , Zavadil Jiri , Wei Jianjun , Levine Douglas , Hernando Eva
Published: July 2012

Leiomyosarcoma (LMS) is a highly aggressive cancer of smooth muscle origin for which the molecular bases of transformation and progression are poorly defined. MicroRNAs (miRNAs) are small non-coding RNAs that have been implicated in tumor development by acting as tumor suppressors or oncogenes. Therefore, identifying a mechanistic role for miRNAs in the biology of LMS will result in better understanding of this complex disease. Moreover, investigation of miRNAs that may also play a role in smooth muscle cell (SMC) differentiation might enhance our understanding of this highly undifferentiated neoplasia. MiRNA microarrays previously performed in our lab identified miR130b as upregulated in uterine LMS compared to normal myometrium. The objective of the current study is to investigate the role of this miRNA in SMC differentiation and LMS genesis and progression.

Preliminary studies have demonstrated increased proliferation, invasion, and colony formation in LMS cell lines overexpressing miR130b indicating that it may be pro-oncogenic. Moreover, miR130b overexpression in human mesenchymal stem cells (hMSCs) induced to differentiate into SMCs inhibited their maturation. Target analysis thus far has revealed that miR130b reduces the levels of its predicted targets Tuberous sclerosis 1, Dicer1, and Connexin 43.

Ongoing overexpression and knockdown studies will determine the necessity and sufficiency of miR130b on in vitro tumorigenic properties of LMS cell lines and SMC differentiation of hMSCs. Microarray expression studies and direct target analysis are being used to elucidate the molecules through which miR130b exerts its functions. Additionally, patient samples will be analyzed to determine the relevance of miR130b overexpression in LMS. Genomic amplification will be assessed as a potential mechanism for the increased levels of miR130b in LMS tissues. Human specimens will also be used to investigate correlations between miRNA expression and its direct targets.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-342. doi:10.1158/1538-7445.AM2011-LB-342
Abstract LB-343: Ultraviolet radiation modulates distinct mouse epidermal miRNAs expression profile
Authors: Jamal Mohammad S. , Verma Ajit K.
Published: July 2012

Chronic exposure to ultraviolet radiation (UVR) is the most common etiologic factor linked to the development of squamous cell carcinoma (SCC), a non-melanoma form of skin cancer that can metastasize. To find clues about the mechanism of UVR-induced development of SCC in FVB/N mice, we found that chronic UVR exposures lead to aberrant expression of several microRNAs (miRNAs). In the present experiment, mice were exposed to four (Monday, Wednesday, Friday and Monday) UVR (2 kJ/m2) treatments, emitted by Kodacel-filtered FS-40 sun lamps (approximately 60% UVB and 40% UVA). At 3 hrs post last UVR exposure, mice were sacrificed. RNAs were prepared from the scraped epidermis of these mice. To screen for unknown miRNAs, we performed the 1040 miRNAs microarray analysis. UVR treatment resulted in modulation of 15 miRNAs: six miRNAs (miR-145, miR-1937a, miR-1937b, miR-341*, miR-451, miR-720) were up-regulated while nine miRNAs (let-7g, miR-125–5p, miR-195, miR-200b, miR-29a, miR-30c, miR-429, miR-92a) were down-regulated. The microarray results were further validated by qRT-PCR. These UVR-induced over-expressed (e.g; miR-451) and down-regulated (e.g; let-7g, miR-195) miRNAs have been observed to be oncogene and tumor suppressor, respectively in various types of cancers. To determine the downstream target genes (DTG) of these miRNAs, insilico analysis was performed. We employed the DIANA LAB DNA Intelligent and Target-Scan program to reveal the various pathways and genes regulated by these miRNAs. Finally we did the IPA to decipher the existing network between these target genes. Among all the miRNAs modulated by UVR treatment, only miR-429 and miR-195 shown to have significant network. MiR-429 network consist of few important hubs. Those were TARDBP, PCBP (Poly (rC) Binding Protein) and PUF60. TARDBP is reported to bind to chromosomally integrated TAR DNA and repress HIV transcription. PCBP1 and PCBP2 are multifunctional protein reported to be translational coactivator. PUF60 is known as a transcriptional repressor. The network obtained by miR-195 DTG revealed that various cytokines exist in the downstream network. Among them, IL-10, IFN-γ and TNF-α are the most significant hub of the network. We also found ZFP36 as an important hub, which is known for recruiting the miRNA at the 3′UTR of mRNA. IPA analysis suggests that all of the above hub molecules play major roles in cell cycle and cancer initiation and progression. UVR-induced deregulated miRNAs (miR-145, miR-1937a, miR-1937b, miR-341*, miR-451, miR-720 – let-7g, miR-125–5p, miR-195, miR-200b, miR-29a, miR-30c, miR-429, miR-92a) may constitute important components of UVR carcinogenesis (Support: NIH grant CA35368).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-343. doi:10.1158/1538-7445.AM2011-LB-343
Abstract LB-428: Upregulation of serum amyloid A1 (SAA 1) gene is associated with ulcerative colitis-related colon cancer
Authors: Schaffer Michael W. , Smith Joan C. , Roy Tanusri Ghosh , Moses Harold L. , Adunyah Samuel E. , M'Koma Amosy E.
Published: July 2012

Purpose: Ulcerative colitis (UC)-associated colon cancer is frequently diagnosed in an advanced stage. SAA1 is a secreted protein made in the liver and circulates in low levels in blood. SAA1 gene is expressed in colon carcinomas and is a chemoattractant with induction of migration, adhesion, and tissue infiltration of monocytes and Polymorphnuclear-leucocytes (PMN-L).

Methods: Laser capture microdissection of colonic submucosa from UC (n=8), Crohn's colitis (CC) (n=8)), and normal (NL, n=8) samples was performed. The submucosal mRNA was extracted using the PicoPure(TM) RNA Isolation Kit. Comprehensive gene expression analysis of the pooled mRNA from each group was then performed using the Affymetrix GeneChip® Gene 1.0 ST Array System. Statistical comparisons of UC vs. CC and NL as well as CC versus UC and NL were performed (Wilcoxon Rank Test). To detect changes for UC, we compared UC to CC and UC to NL; and for CC, we compared CC to UC and CC to NL.

Results: Analysis of all UC vs. CC and NL controls showed 28,869 genes which were represented on the array by 26 probes spread across the full length of each gene. When compared UC vs. CC and NL SAA1 remained the most upregulated gene showing a 56.8-folds and 91.3-folds (p< 0.0001) over expression.

Conclusion: Microarray assessments show significantly overexpression of SAA1 in UC specimens. SAA1 has a role in local inflammation in the microenvironment of malignant tissue and is expressed in colon carcinomas. Our finding may potentially facilitate and validate the SAA1 as a new early predictive clinical marker and a unique target for designing novel selective inhibitors for therapeutic intervention of UC-associated CRC.

Acknowledgement: 3U54CA091408–09S1 (MMC-VICC partnership)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-428. doi:10.1158/1538-7445.AM2011-LB-428
Abstract LB-432: Effect of chronic nicotine consumption on NNK-induced lung tumors in the A/J mouse
Authors: Murphy Sharon E. , von Weymarn Linda B. , Schutten Melissa M. , Lewellen Mitzi , Kassie Fekadu , Modiano Jaime F.
Published: July 2012

Nicotine replacement therapy (NRT) is often used to maintain smoking cessation. Some individuals continue to use NRT for years after smoking cessation, and some NRT users continue to smoke. It is important to understand the impact of chronic nicotine use on lung cancer risk. Therefore, we determined the effect of nicotine administration on NNK-induced lung tumors in the A/J mice. P450 2A enzymes are key catalysts in NNK bioactivation, and nicotine is both an inhibitor and a mechanism based inactivator of P450 2A-catalyzed nicotine metabolism. These data suggest nicotine might protect from NNK-induced tumorigenesis. However, several investigators have reported that nicotine inhibits apoptosis, enhances angiogenesis and enhances proliferation. We hypothesized that nicotine would prevent or delay initiation of NNK-induced tumors, but accelerate tumor growth by reducing apoptosis and promoting inflammation. To test this, female A/J mice were administered a single dose of NNK (100 mg/kg i.p) and 200 ug/ml nicotine in the drinking water. Nicotine was administered 2 weeks prior to NNK, 44 weeks after NNK, throughout the experiment, or without NNK treatment. There were several key outcomes of this study: Chronic nicotine consumption did not affect P450 2A5-catalyzed metabolism in the lung of liver. Nicotine administration alone for 44 weeks did not increase lung tumor multiplicity. Nicotine given before or after NNK administration did not increase, or decrease, tumor number or tumor size. There was also no difference in the number of carcinomas per mouse or the number of adenomas with dysplasia. In conclusion, nicotine consumption in the drinking water had no effect on the tumorigenicity of NNK in the A/J mouse.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-432. doi:10.1158/1538-7445.AM2011-LB-432
Abstract LB-436: Rectal aberrant crypt foci for estimate of colorectal tumor recurrence: 3-year follow up study
Authors: Uchiyama Takashi , Takahashi Hirokazu , Sakai Eiji , Hosono Kunihiro , Endo Hiroki , Nakajima Atsushi
Published: July 2012

Background and Aims: Endoscopic screening and removal of colorectal tumor can reduce colorectal cancer, although surveillance colonoscopy is currently recommended after the initial screening and removal of colorectal tumor due to the possibility of newly developing tumor. Aberrant crypt foci (ACF) serve as surrogate biomarkers of human colorectal carcinogenesis. In this study we aimed to investigate the relationship between ACF and the recurrence of colorectal tumor after endoscopic polypectomy.

Methods: We performed high-magnification chromoscopic colonoscopy to identify lower rectal ACF on 82 subjects who underwent endoscopic polypectomy, and surveillance colonoscopy 3 years after baseline polypectomy at Yokohama City University Hospital.

Results: Marked elevation of the ACF number at baseline findings was observed in the tumor recurrence patients in comparison with the non recurrence patients (p < 0.01). ROC analysis showed high specificity of the number of ACF for diagnosis of any tumor recurrence.

Conclusion: This is the first study to investigate relation between ACF and the recurrence of colorectal tumor after endoscopic polypectomy. These results suggested that ACF is useful for predictive factors for the recurrence of tumor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-436. doi:10.1158/1538-7445.AM2011-LB-436
Abstract LB-462: Chemopreventive effects of freeze-dried strawberries in inflammation-associated mouse colon tumorigenesis
Authors: Yan Fei , Guan Huaji , Wang Yongquan , Huang Xin , Nines Ronald G. , Stoner Gary D. , Chen Tong
Published: July 2012

Colorectal cancer (CRC) is one of the most common malignant neoplasms in the United States. The risk for CRC is associated with extent and duration of inflammatory bowel disease (IBD). Patients with IBD are at increased risk of developing CRC when compared to the general population. Therefore, primary cancer chemoprevention is an important strategy to reduce the risk of CRC in IBD patients and thus, this approach has recently received more attention. Our laboratory has conducted extensive research on cancer chemoprevention in GI tract. In our previous study, we found that dietary freeze-dried black raspberries significantly inhibited adenoma and adenocarcinoma formation and decreased the level of oxidative stress in an azoxymethane (AOM)-induced rat CRC model. In a carcinogen-induced rat esophageal cancer model, we found that dietary freeze-dried strawberries significantly inhibited tumor development. However, the effects of strawberries in inflammation-associated CRC have not been investigated as yet. The principal objective of this study is to investigate the inhibitory effects of dietary freeze-dried strawberries in AOM/Dextran Sodium Sulfate (DSS)-induced mouse colon carcinogenesis. At 4–5 weeks of age, male Crj: CD-1 mice were given a single intraperitoneal injection of AOM (10mg/kg body weight), followed by 2% DSS in their drinking water for seven days, starting one week after the AOM injection. After 3 weeks, animals were fed AIN-76A diet or AIN-76A diet containing 10% freeze-dried strawberries. At week 20, mice were sacrificed for histopathological examination. We found that strawberries significantly decreased tumor incidence from 100% in mice treated with AOM/DSS only to 44.4% in mice treated with AOM/DSS plus strawberries (55.6% reduction, P = 0.02). We also found that the average animal body weight was significantly higher (P = 0.036) in animals fed strawberries (41.9 ± 5.3 g) when compared to animals fed control diet (38.7 ± 3.7 g). A large-scale investigation of the inhibitory effects of strawberries on biomarkers associated with colon carcinogenesis is under way. (Supported by California Strawberry Commissions)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-462. doi:10.1158/1538-7445.AM2011-LB-462
Abstract LB-49: The endoplasmic reticulum chaperone protein GRP94 is required for maintaining hematopoietic stem cell interactions with the adult bone marrow niche
Authors: Luo Biquan , Lam Ben S. , Lee Sung Hyung , Wey Shiuan , Zhou Hui , Wang Miao , Chen Si-Yi , Adams Gregor B. , Lee Amy S.
Published: July 2012

Hematopoietic stem cell (HSC) homeostasis in the adult bone marrow is regulated by both intrinsic gene expression products and interactions with extrinsic factors in the HSC niche. GRP94, an endoplasmic reticulum chaperone, has been reported to be essential for the expression of specific integrins and selectively regulates early T and B lymphopoiesis. Interestingly, multipotent hematopoietic progenitors persisted and even increased in GRP94 deficient bone marrow chimeras, however, the mechanism is not well understood. Here we employed a conditional knockout strategy to acutely eliminate GRP94 in the hematopoietic system. Cell cycle analysis revealed that a loss of quiescence and an increase in proliferation led to an increase in Grp94 knockout (KO) HSCs. This expansion of the HSC pool can be attributed to the impaired interaction of HSCs with the niche, evidenced by enhanced HSC mobilization and severely compromised homing and lodging ability of primitive hematopoietic cells. Transplanting wild-type (WT) hematopoietic cells into a GRP94 null microenvironment yielded a normal hematology profile and comparable numbers of HSCs as compared with WT control, suggesting a cell autonomous effect. Investigating this, we further determined that there was a near complete loss of integrin α4 expression on the cell surface of Grp94 KO HSCs, which showed impaired binding with fibronectin, an extracellular matrix molecule known to play a role in mediating HSC-niche interactions. Collectively, our studies establish GRP94 as a novel cell intrinsic factor required to maintain the interaction of HSCs with their niche, and thus regulate their physiology.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-49. doi:10.1158/1538-7445.AM2011-LB-49
Abstract LB-5: The dominant-negative effect of an insulin-like growth factor-1 mutant that is defective in binding to integrins (αvβ3 and α6β4)
Authors: Fujita Masaaki , Ieguchi Katsuaki , Cardiff Robert , Borowsky Alexander D. , Takada Yoshikazu
Published: July 2012

Insulin-like growth factor-1 (IGF1) is a major therapeutic target for cancer. Integrin αvβ3 plays a critical role in regulating IGF1 signaling, while the specifics are unclear. We recently reported that IGF1 directly binds to αvβ3. The integrin-binding defective IGF1 mutant (R36E/R37E) is defective in inducing IGF signaling and a ternary complex (αvβ3, IGF1R, and IGF1) formation, although the mutant still binds to the type 1 IGF receptor (IGF1R). These findings suggest that direct binding of IGF1 to αvβ3 is required for intracellular signaling (J. Saegusa, S. Yamaji, K. Ieguchi, C.Y. Wu, K.S. Lam, F.T. Liu, Y.K. Takada, and Y. Takada, The direct binding of insulin-like growth factor-1 (IGF-1) to integrin αvβ3 is involved in IGF-1 signaling. J Biol Chem, 2009. 284: 24106–14). Here we describe that R36E/R37E suppressed proliferation and intracellular signaling induced by WT IGF1 in NIH3T3 fibroblasts, C2C12 myoblasts, and MCF-7 breast cancer cells (a dominant negative effect). R36E/R37E suppressed anchorage-independent growth of MCF-7 cells, while WT IGF1 enhanced it. Also, R36E/R37E suppressed the tumorigenesis of MCF-7 cells in SCID mice, while wt IGF-1 markedly enhanced it. Since MCF-7 cells express little or no αvβ3, we studied which integrins are involved in IGF signaling. We discovered that α6β4, which is over-expressed in cancer and implicated in tumor progression, directly binds to WT IGF1 (but not to R36E/R37E) in MCF-7. Interestingly, WT IGF1 induced a ternary complex formation (α6β4, IGF1, and IGF1R) but R36E/R37E did not. These results suggest that α6β4 in addition to αvβ3 plays a role in IGF signaling through direct binding to IGF1, and that R36E/R37E acts as an inhibitor of IGF signaling. We propose that R36E/R37E has potential as a therapeutic agent in cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-5. doi:10.1158/1538-7445.AM2011-LB-5
Abstract LB-50: Ionizing radiation can enhance TGFß induced EMT: Investigation of signatures of cross-talk with the ATM pathway
Authors: Wang Minli , Hada Megumi , Pluth Janice , Anderson Jennifer , O'Neill Peter , Cucinotta Francis
Published: July 2012

Epithelial-mesenchymal transition (EMT), a process whereby epithelial cells undergo transition to a mesenchymal phenotype, has long been known to play a role in cellular differentiation during development and tumor invasion. EMT can be viewed as an extreme form of cell plasticity characterized by loss of epithelial markers, cytoskeletal reorganization, and transition to a spindle-shaped morphology concurrent with acquisition of mesenchymal markers. Whereas EMT can be induced by a wide array of stimuli and cytokines, TGFβ is considered to be a key player of the process. Ionizing radiation (IR), both a carcinogen and a therapeutic agent, induces TGFβ activation in vivo, and also sensitizes human epithelial cells to undergo TGFβ-mediated EMT. In order to understand the likelihood of EMT following space radiation exposures to astronauts, we investigated possible interactions between the TGFβ and ATM pathway following radiation using hTERT immortalized human esophageal epithelial cells (EPC-hTERT), and mink lung epithelial cells (Mv1Lu). The identification of EMT can be shown in changes to morphology, related gene over expression or down regulation, which can be detected by RT-PCR, immunostaining and western blotting. Our study shows EMT can be induced by TGFβ alone, and a combination of heavy ion or proton radiation and TGFβ treatment in non-transformed Mv1Lu and EPC-hTERT cells can enhance the induction of EMT, with a dose as low as 0.1Gy. Pretreatment with TGFβRI inhibitor (SD208) can efficiently attenuate the Smad signaling pathway as detected by the phosphorylation of Smad2, as well as EMT. Serum deprival can also induce EMT-like phenotype in epithelial cells, which can be minimized by the TGFβRI inhibitor. In order to detail the role of the DNA damage response in EMT, we are studying inhibition of ATM or p53 in relation to TGFβ induced EMT. Results for cells irradiated in the presence of TGFβ, and the resulting function of ATM or p53 in the TGFβ mediated EMT process will be reported.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-50. doi:10.1158/1538-7445.AM2011-LB-50
Abstract LB-65: Advocating for Cancer Patients
Authors: Cowan Belita
Published: July 2012

The Lymphoma Foundation of America (LFA) is a national, nonprofit charity dedicated to helping cancer patients and their families. The organization provides support and counseling services, research, grants, a patient hotline, referrals, second opinions, information about patients' rights, and financial assistance to low-income families.

LFA in comprised of a scientific panel of experts with backgrounds in epidemiology, genetic, and biomedical research. Peer-reviewed research reports are published by the Foundation. LFA's report entitled, “Do Pesticides Cause Lymphoma”, is a meta-analysis of the scientific literature on the association between pesticide exposures and nonhodgkin lymphoma. “Lymphoma and Solvents” is another well-known LFA-published report on the relationship between benzene and lymphoma.

Each year, LFA selects promising, young university-based researchers for their outstanding work in cancer epidemiology to receive the “Young Scientist Award”. As part of the Foundation's education and outreach, it produces webcasts and partners with Patient Power to broadcast radio talk shows. These resources help patients gain the information needed to make appropriate decisions about their treatment options.

The Foundation also takes an active role in national health policy. LFA served as the lead amicus organization in the Agent Orange case argued before the U.S. Supreme Court. Lymphoma Foundation of America also filed a precedent-setting Freedom of Information Act request with the National Cancer Institute (NCI). This FOIA request established the right of cancer patients to receive copies of their experimental treatment protocols in all federally-funded cancer clinical trials. LFA has testified at multiple congressional hearings on the environmental causes of cancer.

Lymphoma Foundation of America is an independent nonprofit organization and does not accept drug company or medical industry funding. LFA is entirely supported by individuals and charitable foundations. The Lymphoma Foundation of America was founded in 1986 by lymphoma patients who came together to help each other recover from cancer. Today, LFA serves as a leading voice for the lymphoma patient community.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-65. doi:10.1158/1538-7445.AM2011-LB-65
Abstract LB-77: The Closing of the Every Woman Counts Program in California and Our Efforts to Reopen It
Authors: McDonald Sammantha
Published: July 2012

The California Collaborative of Susan G. Komen for the Cure includes the seven California affiliates located in Sacramento, San Francisco, Fresno, Los Angeles, Inland Empire, Orange County and San Diego. The Collaborative was originally formed to share best practices and synergies, but has since included public policy issues in its initiatives.

The ‘Every Woman Counts’ program was administered by California's Department of Public Health and provided breast and cervical cancer testing, diagnosis and treatment to uninsured women 40 years and older. The Public Policy Committee of the Collaborative worked very hard with the state's elected officials to help helped draft new legislation, propose an audit of the program, and lobby Capitol Hill. Advocates worked with the governor's health staff to ask for the capitol building to be lit pink to help raise breast cancer awareness. Supporters also spoke with anyone who would listen about the importance of this program for the insured and the lives that could potentially be lost if breast cancer went undetected. As a result of their efforts, the Collaborative found several supportive members in the state's legislature. The organization's representatives managed to pass legislation in late August to reinstate the ‘Every Woman Counts’ program.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-77. doi:10.1158/1538-7445.AM2011-LB-77
Abstract LB-84: The Importance of Consumer Involvement in Design, Implemenation and Dissemination of Breast Cancer Research
Authors: Werner Kathleen
Published: July 2012

There is a growing recognition among researchers and clinicians to include consumers and patients in all stages of cancer research. Various advocacy groups and individuals are designing programs to educate and equip survivors and consumers on various aspects of the cancer research and advocacy processes. It is crucial to make these consumers valued members of the dialogue on developing research that may one day directly impact them. The consumer voice is a valued aspect of research that has been consistently overlooked by many researchers and clinicians in the past. By including consumer perspective, these researchers and clinicians are now finding it can greatly enrich the scope, design, impact and outcomes of their research. This poster follows the path of one such consumer advocate_a young, pre-menopausal breast cancer survivor_and how other such consumers can impact the research process to maximize consumer benefit. Researchers will be able to explore how they may better engage consumers for enrichment of their research impact and stated aims.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-84. doi:10.1158/1538-7445.AM2011-LB-84
Abstract LB-93: The role of GSK-3 in mammary gland development and oncogenesis
Authors: Dembowy Joanna , Woodgett Jim
Published: July 2012

Glycogen synthase kinase 3, comprised of two isoforms: α and β, is a central regulator of several signaling pathways that play key roles in development and cancer. Chemical inhibition or genetic loss of GSK-3 in a normal mouse breast cell line cultured under conditions resembling the physiological breast environment resulted in morphological defects reminiscent of early stages of disease. A similar phenotype was also observed upon expression of an activated allele of β-catenin, a major GSK-3 target protein often mutated and elevated in human cancers. These findings implicate GSK-3 as a key regulator of normal ductal development, which may be disrupted early in cancer progression. Upon genetic deletion of GSK-3 (double knock-out of all four alleles, DKO) from breast tissues of mice, dams are unable to nurse their pups. These animals develop severe mammary intraepithelial neoplasia and squamous metaplasia frequently at first pregnancy. Following 4–6 months of continuous breeding, breast adenocarcinomas are observed with elevated levels of ß-catenin and Gli, along with increased expression of a putative mammary progenitor cell marker. Increased cell growth as well as expression of smooth muscle actin, a myoepithelial marker, are also features of these tumors. Similar developmental and functional defects are also observed in mice with breast glands lacking GSK-3 as well as ß-catenin (triple knock-out, TKO). The epithelium of TKO mammary glands undergoes various degrees of squamous differentiation and keratinization, although at a lower penetrance than the DKOs. Multiparious TKOs develop tumors, albeit with a longer latency (7–9 months). Preliminary flow cytometry analysis of mammary glands from DKO and TKO animals show the ratio of luminal to basal cell populations is increased in absence of GSK-3. We are currently investigating how GSK-3 may regulate mammary stem/progenitor cell commitment to these mammary cell lineages.

Previously reported expression of stabilized ß-catenin in the mammary epithelium induces squamous metaplasia, but no adenocarcinomas were observed at 6 months. In our hands, these animals do develop small tumors at ∼10 months of age. These data suggest the observed DKO and TKO phenotypes are not solely mediated via aberrant ß-catenin signaling and that GSK-3 has ß-catenin-independent functions in mammary gland disease. Other pathways regulated by GSK-3 including Notch, Hedgehog and PI3K are being interrogated in DKO and TKO tumor-initiating cells to gain a molecular understanding of tumor development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-93. doi:10.1158/1538-7445.AM2011-LB-93
Abstract LB-95: Inhibition of CXCR4/Stat3 signaling by AMD3465 exerts major tumor- and metastasis-suppressive effects in syngenic 4T1 breast cancer model
Authors: Ling Xiaoyang , Spaeth Erika L. , Shi Yue-xi , Konopleva Marina
Published: July 2012

Stromal-derived factor-1 α (SDF-1α, also termed CXCL12) and its receptor CXCR4 have been shown to play important roles in hematopoietic stem cell homing, and high expression of CXCR4 has been associated with poor prognosis in hematologic malignancies. The CXCR4/SDF-1α axis is also functional in solid tumors, including breast cancer. It has been postulated that Stat3 may promote metastases in breast cancer through direct interaction with CXCR4, promoting tumor growth and survival. Phosphorylated Stat3 was found to trigger overexpression or activation of several potent oncoproteins. In order to evaluate the role of CXCR4 in metastases, we used the specific CXCR4 inhibitor, AMD3465, to block CXCR4 in the syngeneic 4T1 breast cancer model. Stat3, Akt, Erk1/2, Jak2 and MMP9 activity was found to be almost completely inhibited and expression levels of GSK-3β and cMyc were greatly reduced. Growth of 4T1 cells orthotopically implanted into the mammary fat pad as well as pulmonary and liver metastases were significantly (>95%) reduced in vivo when AMD3465 was given as continuous infusion at a dose of 428.57μg/mouse/day over 14 days. Furthermore, we observed that CXCR4 inhibition by AMD3465 significantly reduced lung metastases in three immunocompetent syngeneic mouse models after surgical resection of the primary breast tumors. This experimental design mimics current clinical treatment strategies. Kozin et al. (Cancer Research 70: 5679–85; 2010) recently suggested that CD11b+ myeloid/monocytic cells (a cell type implicated in supporting angiogenesis) may play a role in tumor re-growth in response to SDF-1 α. In support of this observation, we observed significantly reduced CD11b+ cells in lungs, livers and spleens (P<0.01 for all 3 organs) following treatment with AMD3465, as compared to PBS controls. Hence, our data suggest that pharmacological CXCR4 inhibition in breast cancer-bearing mice following primary tumor resection is well tolerated and potently controls the development of distant metastatic lesions. In conclusion, interruption of the SDF-1/CXCR4/Stat3 signaling axis by the CXCR4-specific inhibitor, AMD3465, effectively inhibits pulmonary and hepatic breast cancer metastasis via disruption of intracellular microenvironment mediated signaling in breast cancer cells and through effects on CD11b+ tumor infiltrating myeloid cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-95. doi:10.1158/1538-7445.AM2011-LB-95