bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4663: Markers of anaerobic glycolysis as predictive factor in neoadjuvant chemoradiotherapy of rectal cancer
Authors: Shim ByoungYong , Jung Ji Han , Lee Kang-Moon , Kim Hyung Jin , Hong Sook Hee , Kim Sung Whan , Sun Der Sheng , Kim Hoon-Kyo , Cho Hyun Min
Published: February 2011

Background: Growing tumors adapt to a hypoxic environment and increase anaerobic glycolysis. This metabolic switch is related to resistance of radiotherapy and chemotherapy. We investigate the relationship between biomarkers related to anaerobic glycolytic metabolism (glucose transpoter-1(GLUT-1), lactate dehydrogenase (LDH)-5 and pyruvate dehydrogenase kinase (PDK)-1) and prognosis.

Methods: GLUT-1, LDH-5, PDK-1 expression determined by immunohistochemistry were assessed in 109 patients with rectal cancer treated with 5 flurouracil and leucovorin neoadjuvant chemoradiotherapy.

Results: This study included 74 male and 35 female patients with a median age of 63 (range 53-70 years). Stage I, II and III accounted for 7.3%, 43.1% and 49.5%. All patients had 45 Gy (1.8 Gy/day in 25 fractions) over five weeks, plus 5-fluorouracil (425 mg/m(2)/day) and leucovorin (20 mg/m(2)/day) bolus on days 1 to 5 and 29 to 33 and surgery was done on 7 to 10 weeks after completion of all therapies. The down staging rate of this neoadjuvant therapy was 53% and pathologic complete response rate is 18.3%. High expression of GLUT-1, LDH-5, PDK-1 was observed in 34 (31.2%), 62 (56.9%) and 45 (41.3%) patients, respectively. A negative GLUT-1 was associated with a significantly higher rate of pathologic complete response compared to positive GLUT-1 (25.3% vs. 0.02%, p=0.006). LDH-5, PDK-1 was not correlated with down staging and complete response of chemoradiotherapy.

Conclusions: The GLUT-1 expression is the predictive factor of pathologic complete response in rectal cancer patients treated with 5 flurouracil and leucovorin neoadjuvant chemoradiotherapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4663.
Abstract 4665: A combined IHC/Western method to measure phosphorylated histone H3 in skin biopsies as potential biomarker for anticancer drug action
Authors: Graeser Ralph , Birkle Marianne , Rentzsch Christine , Schaechtele Christoph
Published: February 2011

In recent years there has been a rapid increase in targeted tumor therapy approaches. Hand in hand with these novel approaches, the need has arisen to monitor specific target inhibition by the drug. In contrast to classic chemotherapy, which non-specifically targets proliferating cells, these novel approaches attempt to specifically interfere with some process thought to be crucial for the tumor cell survival. Nevertheless, despite extensive validation, an immediate effect of target interference on tumor growth may not necessarily be expected. Thus, in order to demonstrate activity of anti-cancer drugs, a marker known to be modified upon inhibition of the target may serve as a surrogate to tumor inhibition. Ideally, tumor biopsies would be analysed for monitoring of drug-induced effects. However, taking in consideration the difficulties to perform sequential tumor biopsies, the use of surrogate tissues like blood or skin is being explored instead. Blood is relatively easily accessible, but unlike skin tissue, blood may not reflect the fact that drugs have to penetrate multiple layers of tissue in order to reach their target in the tumor. Also, to monitor the effects of anti-mitotic drugs, the investigated specimen must exhibit proliferating features, which is given only in a very poor manner blood samples, as peripheral blood cells have typically exited the cell cycle. However cells in the dermis still proliferate, albeit to a small percentage.

Therefore, considering these issues, we set out to develop a method to analyse drug effects in skin biopsies. Traditionally, most analyses of skin biopsies have relied on immunohistochemistry (IHC) studies. However, the quantification of such results is time-consuming and error-prone. Western Blot analyses are easier to quantify, but require a considerable amount of protein lysate of sufficient concentration. We developed a protocol that reproducibly allows the detection and quantification of phosphorylated histone H3 in Western Blots from a single half of a skin biopsy. For normalization, the total amount of histone H3 present in the lysate is determined. The other half of the skin biopsy may be used for IHC analysis of phosphorylated histone H3, allowing the comparison to other studies relying solely on IHC.

We will present data on the measurement of phosphorylated histone H3 via Western Blots and IHC from skin biopsies of healthy donors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4665.
Abstract 4666: Identification of cancer stem cell-like fractions in cultured oral squamous cell carcinoma and expression of keratin 15 as a putative cancer stem cell marker
Authors: Isobe Tomohide , Yamamoto Gou , Tsunoda Yuko , Sasaki Akiko , Tachikawa Tetsuhiko
Published: February 2011

Although it is well known that one of the intermediate filament proteins cytokeratin 15 (CK15) can be a marker for not only epithelial stem cells but also cutaneous neoplasm and sebaceous tumors, the expression of CK15 in oral squamous cell carcinoma has not been reported. In this study, we investigated the expression of CK15 in different stages of epithelial dysplasia, carcinoma in situ (CIS) and squamous cell carcinoma (SCC) by immunohistochemical analysis and genetic analysis. Only basal cells in normal epithelia showed strong positive staining for CK15. Which was spread to basal cells and parabasal cells in epithelial dysplasia. In CIS and SCC, however, no reactivity was observed for CK15 except for the peripheral part of carcinoma nest in SCC. On the other hand, CK15 expression in chemoresistant SCC showed overall positive staining. In contrast to CK15 expression, transcription factor P63 which is known to putative keratinocyte stem cell marker showed different staining patterns from those of CK15. These observations seem to indicate that CK15 staining in combination with P63 could be a useful marker for pathologic diagnosis in terms of predicting malignant change and chemoresistance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4666.
Abstract 4678: KIAA1199 depletion targets the wnt/beta catenin signaling pathway and impairs migration and proliferation of human colon cancer cells
Authors: Birkenkamp-Demtroeder Karin , Maghnouj Abdel , Mansilla Francisco , Thorsen Kasper , Andersen Claus L. , Hahn Stephan , Ørntoft Torben F.
Published: February 2011

In a previous study KIAA1199 was found to be upregulated in colon adenomas. A depletion of beta-catenin resulted in silencing of KIAA1199, suggesting a potential role in wnt/beta signaling. Aim of this study was to correlate KIAA1199 expression with patient outcome, to identify KIAA1199 downstream target genes and to elucidate its function and pathway association. Here we show by transcript profiling of >400 colon adenocarcinomas that KIAA1199 is strongly increased in >75% of colon adenocarcinomas compared to normal mucosas. Immunohistochemical analyses with a monospecific antibody identified the protein to be expressed in the nucleus and the cytoplasm of adenocarcinomas, and a strong upregulation of the KIAA1199 protein compared to normal, as also seen in several other types of cancer. Cloning identified a splice isoform lacking exon 28 resulting in a truncated KIAA1199 protein. Microarray profiling of SW480 cells transiently overexpressing, or stably depleted from, KIAA1199 identified genes involved in cell cycle regulation, cell signaling, cellular movement and cell to cell interaction. In vitro functional analyses indicated that cells depleted from KIAA1199 showed reduced cellular adhesion, a decreased proliferation rate and migrated less than control cells. KIAA1199 silencing affected the expression of eighteen genes, known to be involved in the wnt/beta catenin signaling pathway. The identified target genes correlated with KIAA1199 expression in normal colon mucosa and adenocarcinomas of stage II and III (Pearson 0.7-0.9). In conclusion, our data provide evidence that KIAA1199 is a regulatory part of the wnt/beta catenin signaling, sited downstream of beta-catenin and upstream of the stem cell marker ASCL2.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4678.
Abstract 4681: Urothelial gene expression signatures aid bladder cancer detection
Authors: Rosser Charles , Sun Yijun , Urquidi Virginia , Goodison Steve
Published: February 2011

Bladder cancer is the fifth most commonly diagnosed malignancy in the United States and one of the most prevalent worldwide. It harbors a probability of recurrence of >50%, thus rigorous, long-term surveillance of patients is advocated, so improvements in non-invasive urinalysis assessment strategies would benefit patients. We applied gene expression microarray analysis to exfoliated urothelia recovered from bladder washes obtained prospectively from 102 patients with subsequently confirmed presence or absence of bladder cancer. Data from microarrays containing 56,000 targets was subjected to a panel of statistical analyses to identify bladder cancer-associated gene signatures. Hierarchical clustering and supervised learning algorithms were used to classify samples on the basis of tumor burden. A differentially expressed set of 323 gene probes was associated with the presence of bladder cancer (P<0.01). Supervised machine learning and a cross-validation approach were used to build a 55-gene molecular classifier that was able to classify patients with and without bladder cancer with an overall accuracy of >75%. Our results show that it is possible to achieve the detection of bladder cancer using molecular signatures present in exfoliated tumor urothelia. Further investigation and validation of the cancer-associated profiles may reveal important biomarkers for the non-invasive detection and surveillance of bladder cancer.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4681.
Abstract 470: The homing of prostate cancer to bone in vivo is modulated by Zoledronic acid
Authors: Reeves Kim J. , van der Pluijm Gabri , Cecchini Marco G. , Eaton Colby L. , Hamdy Freddie C. , Brown Nicola J.
Published: February 2011

Cancer metastasis to the skeleton is a major problem with prostate cancer and requires the development of specialised approaches to treatment. Zoledronic acid, a bone resorptive agent, is currently the only bisphosphonate with proven benefit in prostate cancer. However, there is minimal research investigating whether Zoledronic acid modulates the early mechanisms involved in metastasis. The aim of this study is to determine whether different treatment regimes using Zoledronic acid modulate the early events in tumour/bone interactions.

Metatarsals from newborn mice (1-3 day old) are engrafted into a Dorsal Skinfold Chamber (DSC) implanted on a SCID mouse (5-6 weeks old). Prostate (PC3-GFP) cancer cells (1 × 105) are then injected via the heart (i.c) to simulate micrometastatic spread (n=20/group). Animals are treated with either i) weekly s.c injection of Zoledronic acid (25µg/kg/inection=100μg/kg in total) or saline or ii) single s.c injection of Zoledronic acid (100μg/kg) or saline for the 4 week study duration. The groups are further subdivided, so half of the animals receive treatment either before or after injection of tumour cells. Recordings of the chamber tissue and metatarsal are made at 48hr intervals for the duration of the experiment for the presence of PC3-GFP cells within the metatarsal. At the end of the study, tissue is harvested and processed for microCT, multi-photon analysis and histology.TreatmentNumber of PC3-GFP cells homing to metatarsalDay 3Day 9Day 15 Weekly treatment before PC3-GFP cells1.1 ± 0.1*1.3 ± 0.2*2.1 ± 0.3*Weekly treatment after PC3-GFP cells3.2 ± 0.5*3.4 ± 0.8*4.4 ± 0.7*Single treatment before PC3-GFP cells1.7 ± 0.3*8.9 ± 0.9*13.6 ± 0.8Control3.2 ± 0.714.7 ± 1.015.9 ± 1.4

Table 1. Number of PC3-GFP cells present in metatarsal after i.c. injection (*p<0.05 compared to control).

Treatment with Zoledronic acid reduces tumour cell homing to bone. Weekly treatment of Zoledronic acid prior to prostate cancer cell administration reduces homing to bone to a greater extent than the other treatment regimes test.

Funded by Sixth European Framework Programme (PROMET).

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 470.
Abstract 4700: Acrolein-deoxyguanosine DNA adduct formation and lung cancer
Authors: Hu Yu , Rom William , Pass Harvey I. , Spivack Simon D , Tang Moon-shong
Published: February 2011

Cigarette smoke (CS) contains 1,000-10,000 fold more acrolein (Acr) and crotonaldehyde (Crot) than polycyclic aromatic hydrocarbons (PAHs). These aldehydes can induce propanoldeoxyguanosine (propanol-dG) DNA adducts formation in cultured normal human lung epithelial cells. Previously, we have found that the distribution of Acr-dG in the p53 gene coincides with the lung cancer p53 mutational spectrum of cigarette smokers, and that Acr-dG induces similar level of G to T transversion and G to A transition mutations as in lung cancer of cigarette smokers. Hence, we have proposed that Acr is a major etiological agent of CS related lung cancer. We developed a 32P-postlabelling method in combination with 2D TLC/HPLC method to quantify propanol-dG formation in human lung tissue. To date we have determined Acr-dG and Crot-dG in the lung tissue of cigarette smokers (n=27) and nonsmokers (n=25) with lung cancer. We found that 1) both α- and γ-Acr-dG are formed in lung tissues of smokers and nonsmokers, and the level of these two types of adduct in lung tissue varies from 10-4,510, and 0-1,900 fmole/μmole, respectively; and 2) smokers appear to have a higher level of γ-Acr-dG than nonsmokers although the differences have not reached statistical significance (p=0.1854); 3) the level of Crot-dG in smokers is significant higher than that in nonsmokers (p=0.0279). These results indicate a substantial amount of Acr-dG and Crot-dG formed in lung tissue of lung cancer.

Key words: α, β-unsaturated aldehydes, cigarette smoking, DNA adducts, lung cancer.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4700.
Abstract 4707: Clonal expansions as a biomarker of preneoplastic processes in ulcerative colitis
Authors: Salk Jesse J. , Salipante Stephen J. , Risques Rosa Ana , Crispin David A. , Li Lin , Bronner Mary P. , Rabinovitch Peter S. , Horwitz Marshall S. , Loeb Lawerence A. , Brentnall Teresa A.
Published: February 2011

Chronic inflammation predisposes to a variety of human cancers. Affected tissues slowly accumulate mutations, some of which affect growth regulation and drive successive waves of clonal evolution whereas a far greater number are functionally neutral and serve only to passively mark expanding clones. Ulcerative colitis (UC) is an inflammatory bowel disease in which up to 10% of patients eventually develop colon cancer. We have recently reported on mapping of mutations in non-coding polyguanine tracts in patients with UC to delineate the extent of clonal expansions associated with carcinogenesis. We genotyped colon biopsies for length altering mutations at 28 different polyguanine markers. In eight UC patients without neoplasia, blinded genotyping detected only two mutations in a single individual from among 37 total biopsies. In contrast, in eleven UC patients with neoplasia elsewhere in the colon, blinded genotyping identified 63 mutations in 51 nondysplastic biopsies, and every patient possessed at least one mutant clone. As a biomarker of cancer progression the presence of one or more clones was 100% sensitive and 88% specific. A subset of clones were large and extended over many square centimeters of colon. Of these, some occurred as isolated populations in non-dysplastic tissue, considerably distant from neoplastic lesions. Other large clones included regions of cancer, suggesting that the tumor arose within a pre-existing clonal field. This preliminary work indicated that neutral mutations in polyguanine tracts serve as a powerful tool for identifying clonally expanded populations, and suggested that these clones may be clinically useful for distinguishing a subset of UC patients who are at greatest risk for developing cancer. Here we provide updated results of a new and larger case-control study and discuss ongoing efforts to streamline clone detection using other types of neutral genetic markers and next-generation sequencing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4707.
Abstract 4712: Characterization of estrogen quinone-derived protein adducts and their identification in human serum albumin
Authors: Lin Po-Hsiung , Wang Tzu-Wen , Wei Tzu-Han , Yang Tsung-Chou , Chen Dar-Ren
Published: February 2011

Both estrogen-2,3-quinone (E2-2,3-Q) and estrogen-3,4-quinone (E2-3,4-Q) are reactive metabolites of estrogen that are thought to be responsible for the estrogen-induced genotoxicity. The aim of this study was to establish a methodology to analyze estrogen quinone-derived protein adducts and to measure the background levels of these adducts in human serum albumin (Alb) derived from female blood donors in Taiwan. The methodology, which employs trifluoroacetic acid anhydride and methanesulfonic acid to selectively cleave cysteinyl adducts, had previously been applied to measure naphthalene adducts in blood proteins. Results from in vitro experiments confirmed that the production of estrogen quinone-derived adducts on serum albumin increased with increased concentration of estrogen quinones (0-100 nM). The average formation of estrogen quinone-derived cysteinyl adducts per unit concentration of estrogen quinones was estimated to be 13.0, 48.2, and 36.1 (pmol/g Alb)/(nM) for E2-2,3-Q-1-S-Alb, E2-2,3-Q-4-S-Alb, and E2-3,4-Q-2-S-Alb, respectively. Time-course experiments indicated that both E2-2,3-Q- and E2-3,4-Q-derived adducts rapidly reached maximum values at 10 min mark and remained constant thereafter. Additionally, with 2,3,7,8-tetrachlorodibenzo-p-dioxin pretreatment (10 nM for 72 h), production of estrogen quinone-derived protein adducts was also detected in human MCF-7 breast cancer cells exposed to estrogen (1 μM for 9 h). Co-treatment of Ro 41-0960, a catechol-o-methyl transferase inhibitor, further enhanced the production of estrogen quinone-derived adducts in all cases. When we investigated the levels of estrogen quinone-derived adducts in human serum Alb derived from female blood donors in Taiwan, cysteinyl adducts of E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb were detected in all subjects (n=10). The median levels of E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb in human subjects were estimated to be 398 (range 240-807) and 39.0 (range 0.3-101) (pmol/g), respectively. We noticed that levels of E2-2,3-Q-4-S-Alb were significantly correlated with those of E2-3,4-Q-2-S-Alb (correlation coefficient r = 0.684, p<0.05). (This work was supported by the National Science Council, Taiwan, through grants NSC95-2314-B-005-001, and NSC97-2314-B-005-001).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4712.
Abstract 4715: Genetic susceptibility to radiation-associated colon and rectum cancers among atomic-bomb survivors with special reference to the CD14 gene
Authors: Hayashi Tomonori , Morishita Yukari , Nagamura Hiroko , Maki Mayumi , Kusunoki Yoichiro , Yoshida Kengo , Imai Kazue , Cologne John B. , Nakachi Kei
Published: February 2011

Past epidemiology studies reported that atomic-bomb (A-bomb) radiation exposure enhanced risk of colon cancer, but not rectum cancer, among the survivors. The reason for this different sensitivity to radiation remains to be elucidated. Colorectal cancer is a multi-factorial disease, the onset of which is attributed to both environmental and genetic factors, with animal experiments showing that intestinal bacterial toxins accelerate carcinogenesis. It was recently reported that single nucleotide polymorphisms (SNPs) identified in the promoter region of CD14, encoding a receptor for the lipopolysaccharide component of the outer membrane of Gram-negative bacteria, regulate CD14 gene expression and thereby affect susceptibility to inflammatory diseases such as atopic dermatitis, coronary artery disease, and hepatic diseases. This study investigated relationship between a novel CD14 gene polymorphism and the development of radiation-associated colon and rectum cancers among an A-bomb survivor cohort, in terms of a case-cohort study. On the basis of this T/G polymorphism, we determined CD14 genotypes with 210 colorectal cancer cases and with a subcohort of 2,160 individuals who were randomly selected from the cohort. Our results showed that the individuals exposed to A-bomb radiation (≥5 mGy) were at an increased risk of colon cancer, but not rectum cancer, with relative risk (RR) of 1.19/Gy (95%CI: 1.04-1.36). When we divided the study subjects into three groups for radiation dose and into two groups for CD14 genotypes, significantly high risk of colon cancer was found with RR of 2.40 (95%CI: 1.37-4.23) for the combined category of CD14-T/T homozygote and the highest dose (≥0.7Gy), compared with the baseline category (CD14-T/G or G/G and no dose). These results suggest that CD14-related inflammatory response might be involved in the development of radiation-associated colon cancer, not rectum cancer, among A-bomb survivors, and that the CD14 genotyping might be involved in individual risk of colon cancer together with radiation dose. To find a functional significance of this polymorphism, we are currently working on ELISA-based assay to measure serum levels of soluble CD14 among healthy individuals in relation to the CD14 genotyping.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4715.
Abstract 4718: Inherited variants in relapse-associated genes and risk of ovarian cancer
Authors: Peedicayil Abraham , Vierkant Robert A. , Hartmann Lynn C. , Fridley Brooke L. , Fredericksen Zachary S. , White Kristin L. , Elliott Elaine A. , Phelan Catherine M. , Tsai Ya-Yu , Berchuck Andrew , Iversen Edwin S. , Couch Fergus J. , Peethambaram Prema P. , Larson Melissa C. , Kalli Kimberly R. , Kosel Matthew L. , Shridhar Vijayalakshmi , Rider David N. , Liebow Mark , Cunningham Julie M. , Schildkraut Joellen M. , Sellers Thomas A. , Goode Ellen L.
Published: February 2011

Background: We previously identified a panel of genes associated with outcome of ovarian cancer. The purpose of the current study was to assess whether variants in these genes correlated with ovarian cancer risk.

Methods and Findings: Women with and without invasive ovarian cancer (749 cases, 1,041 controls) were genotyped at 136 single nucleotide polymorphisms (SNPs) within 13 candidate genes. Risk was estimated for each SNP and for overall variation within each gene.

At the gene-level, variation within MSL1 (male-specific lethal-1 homolog) was associated with risk of serous cancer (p=0.03); haplotypes within PRPF31 (PRP31 pre-mRNA processing factor 31 homolog) were associated with risk of invasive disease (p=0.03). MSL1 rs7211770 was associated with decreased risk of serous disease (OR 0.81, 95% CI 0.66-0.98; p=0.03). SNPs in MFSD7, BTN3A3, ZNF200, PTPRS, and CCND1A were inversely associated with risk (p<0.05), and there was increased risk at HEXIM1 rs1053578 (p=0.04, OR 1.40, 95% CI 1.02-1.91).

Conclusions: Tumor studies can reveal novel genes worthy of follow-up for cancer susceptibility. Here, we found that inherited markers in the gene encoding MSL1, part of a complex that modifies the histone H4, may decrease risk of invasive serous ovarian cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4718.
Abstract 472: Murine models mimicking different colorectal cancer liver metastasis phenotypes
Authors: Garcia-Albeniz Xabier , Nadal Cristina , Gomis Roger
Published: February 2011

Introduction

Mechanisms responsible of liver metastasis (LiM) in colorectal cancer (CRC) are largely unknown. Efficient and reliable in vivo and in vitro experimental approaches mimicking clinical scenarios to identify genes responsible of LiM in CRC should be the first step to identify effective targets with clinical relevance. The objective was to develop an animal model of CRC LiM

Experimental procedures

The following human CRC cell lines Colo 205, SW480, SW620, KM12C and KM12L4A and the murine cell line Colo 26 were selected to test the feasibility of the metastatic in vivo model. Each cell line was GTP/luciferase-labelled and injected in a cohort of 6 female balb-c nude mice (Charles River). Two million cells where injected via intrasplenic injection, and splenectomy was performed one minute thereafter. Mice were followed with a Xenogen IVIS system. LiM were disgregated, cells were selected by cell sorting and re-injected with the aim of enriching their metastatic phenotype. .

Results

The cell lines Colo 205 and SW480 did not give rise to any liver metastasis. The rest of the cell lines tested produced liver metastasis with very different patterns. After injection of Colo26, there was not any signal clearance in the liver, followed by a massive infiltration in less than 5 days in every mice. SW620, KM12C and KM12L4A injection was followed by a clearance of the signal in the liver area and recovering of the signal occurred when metastasis developed. SW620 produced metastasis in 40% of the mice after 60 days of follow up. The pattern of metastasis of SW620 was as single spots, recapitulating a very frequent phenotype in patients. KM12C and KM12L4A produced multiple spots in 50% of the mice after 35 days of follow up. Disgregation and re-injection of SW620, KM12C and KM12L4A metastatic cells resulted into an enriched metastatic phenotype, with higher number of metastasis, shorter time to metastasis development and higher percentage of mice developing metastasis.

Conclusion

We provide different animal models that mimic different clinical scenarios. Enrichment of metastatic phenotype is feasible by re-injecting metastatic cells.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 472.
Abstract 4720: Genome-wide copy number variation and breast cancer risk: Preliminary report
Authors: Lee Kyoung-Mu , Park Miey , Moon Sang-Hoon , Kim Hyung-Chol , Lee Ji-Young , Li Lian , Park Sue K. , Han Wonshik , Noh Dong-Young , Yoo Keun-Young , Ahn Sei-Hyun , Han Bokki , Kang Daehee
Published: February 2011

Background: DNA copy number variation (CNV) is a common phenomenon in human genome and some CNVs have been associated with susceptibility or resistance to disease. It has been suggested that gene copy number can be elevated in cancer cells. To evaluate potential role of CNV in breast cancer development, we conducted genome-wide association study of CNV in Seoul Breast Cancer Study.

Methods: Cases were diagnosed with breast cancer and underwent curative surgery at two teaching hospitals located in Seoul, Korea, between 2001 and 2007 (n=2,386). Control subjects were selected from multi-center based cohort study in 2007 (n=2,386). The Affymetrix SNP assay 6.0 array, which includes 946,000 probes for the detection of copy number variation, was used. Nexus Copy Number version 4.1 was used for detecting CNVs (significance threshold=10−6; minimum number of probes per segment=4), and statistical analysis. After quality control step (i.e., robust variance sample QC), total 2,315 cases and 2,039 controls were compared for the difference of each CNV. In this study, we selected the CNVs that were different between cases and controls by at least 5% (P adjusted for multiple comparison <0.01). Adjacent CNVs were grouped into CNV regions ranging 10 kb to 300 kb.

Results: Between case and controls, thirteen CNV regions including chr115q11.2, chr3q26.1, chr6q16.3, chr17q31.1 were significantly different for copy number. We note that those regions include genes such as UGT2B17 (UDP glucuronosyltransferase 2 family, polypeptide B18), OR4C11 (olfactory receptor, family 4, subfamily C, member 11), LRRC37A2 (leucine rich repeat containing 37, member A2), ARL17P1 (ADP-ribosylation factor-like 17 pseudogene 1), LOC644974, and the others. We are planning validation study for the regions by quantitative PCR to verify the association with breast cancer risk found in this study.

Conclusion: Our results suggest that common CNVs may be associated with breast cancer in Korean women. However, further variation study is essential to support our observation and underlying biological mechanisms for the associations needs to be elucidated.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4720.
Abstract 4722: Evaluation of common genetic variation in 1,300 cancer-related candidate genes and endometrial cancer risk
Authors: Yang Hannah P. , Spurdle Amanda , Gaudet Mia M. , Brinton Louise A. , Lissowska Jolanta , Peplonska Beata , Sherman Mark , Webb Penny , Ferguson Kaltin , O'Mara Tracy , ANECS ‘ , Chanock Stephen , Garcia-Closas Montserrat
Published: February 2011

Background: Endometrial cancer genetic susceptibility studies have largely focused on genes in the sex steroid hormone, checkpoint, and cell-cycle pathways, but single nucleotide polymorphisms (SNPs) in genes involved in other cancer-related pathways might also influence risk.

Methods: We investigated the association between endometrial cancer risk and ∼29,000 SNPs in over 1,300 cancer-related candidate genes in the inflammation, metabolism, and other carcinogenic pathways in the Polish Endometrial Cancer Study (PECS), a population-based investigation involving 417 cases and 407 controls. TagSNPs were located in the respective gene or within 10 kb or 5 kb downstream; had a minor allele frequency (MAF) >5% in various ethnic groups; and were not represented by a current tag SNP in linkage disequilibrium (LD; r2>0.80). We calculated odds ratios (ORs) and 95% confidence intervals (CIs) using unconditional logistic regression, adjusting for age (5-year categories). The most significant finding was followed up in the Australian National Endometrial Cancer Study (ANECS), a population-based case-control study of 815 cases and 822 controls.

Results: In PECS, we found three SNPs in two loci associated with endometrial cancer risk at P<0.0001. Two highly correlated SNPs in IQGAP1, rs2601207 (OR = 0.65; 95% CI: 0.53 - 0.81, P trend = 6.0 × 10−5) and rs6496677 (0.66; 0.54 - 0.81, P trend = 8.0 × 10−5), were associated with a decreased risk. In addition, a variant in UBE3A, rs4445877, was associated with increased risk (1.53; 1.24 - 1.90, P trend = 9.0 × 10−5). We followed up the rs2601207 association in ANECS and observed a non-significantly decreased risk (0.90; 0.77 - 1.05, P trend = 0.18). The pooled per allele OR was 0.81 (95% CI: 0.72 - 0.92), P trend = 0.001); however, there was evidence for heterogeneity by study (P-heterogeneity = 0.01).

Conclusion: Data from PECS suggests associations between endometrial cancer risk and variants in IQGAP1, a gene that is a key mediator in several cellular processes related to carcinogenesis, and UBE3A, encoding ubiquitin protein ligase 3A involved in the degradation of cancer-associated proteins such TP53. The association with IQGAP1 was not confirmed in a larger study population; and further studies are needed to follow up other potential associations between endometrial cancer risk and cancer candidate genes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4722.
Abstract 4745: A whole genome association study and identification of a locus for histiocytic sarcoma in the flat-coated retriever
Authors: Shearin Abigail L. , Schmidt Emmett V. , Parker Heidi G. , Mellersh Cathryn S. , Breen Matthew , Ostrander Elaine A.
Published: February 2011

Histiocytic sarcoma (HS) is a cancer characterized by unregulated proliferation of histiocytes. In the flat-coated retriever (FCR), the primary tumor site is frequently appendicular with a high rate of local and distant metastases. The disease occurs with a high prevalence in the FCR, accounting for approximately 40% of all cancers in the breed and 18% of disease-related deaths in a UK cohort. Breed-specific prevalence in FCRs indicates a strong genetic predisposition. Our goal is to identify the underlying loci and causal mutations associated with HS in the FCR.

A whole genome association study (GWAS) using the Illumina Canine SNP20 BeadChip was performed using 98 DNA samples from FCR collected from the United States (48 cases and 50 unaffected controls over the age of ten yrs). Of the 23,000 SNPs genotyped, 17,000 were informative in the FCR. A single-marker chi-squared analysis using EMMA to correct for population structure and kinship yielded an association with the HS phenotype (p=2.33×10-5). Including the flanking markers, the associated region spans 12 Mb and includes several candidate genes of interest. To reduce this critical interval, ABI SNPlex genotyping and candidate gene sequencing is in progress.

Our current data distinguish HS in the FCR from the related disease malignant histiocytosis (MH) found in Bernese mountain dogs (BMD). MH is a disseminated histiocytic cancer with primary tumor sites usually occurring in visceral organs. Our data suggest that HS in the FCR is not a localized form of MH. GWAS analyses for both breeds indicate that the major contributing heritable factors of MH in the BMD differ from those for HS in the FCR. Given the different behaviors of the cancers between the two breeds and the different heritable loci associated with them, our current evidence distinguishes them as separate diseases.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4745.
Abstract 4748: Identification of HLA-A*0201 and A*2402-restricted epitope peptides derived from mucin 5AC (MUC5AC) gene expressed in human pancreatic cancer as a potential target for immunotherapy
Authors: Yamazoe Sadaaki , Tanaka Hiroaki , Iwauchi Takehiko , Noda Eiji , Amano Ryosuke , Kubo Naoshi , Muguruma Kazuya , Inoue Toru , Takashima Tsutomu , Yamada Nobuya , Yashiro Masakazu , Maeda Kiyoshi , Onoda Naoyoshi , Sawada Tetsuji , Nakata Bunzo , Ohira Masaichi , Ishikawa Tetsuro , Hirakawa Kosei
Published: February 2011

MUC5AC was previously identified to be de novo expressed in the majority of pancreatic ductal adenocarcinoma. For the development of pancreatic cancer immunotherapy, we studied the significance of MUC5AC expressed in pancreatic ductal adenocarcinoma and the capability of MUC5AC as a target for immunotherapy of pancreatic cancer. By immunohistochemical analysis of 102 specimens, MUC5AC is absent from all cell types of the normal pancreas, but arise de novo in 85% of invasive ductal adenocarcinoma. Clinicopathlogically, tumors with lymph node metastasis had significantly higher expression of MUC5AC than those without node metastasis. To validate the potentiality of cancer vaccine, we have searched for possible epitope peptides that can elicit cytotoxic T lymphocytes (CTL) to MUC5AC. We screened 20 candidate peptides derived from MUC5AC gene sequence for each of human leukocyte antigen (HLA) - A*0201 and A*2402. As a result, we successfully established CTL clones stimulated by MUC5AC-A02-1398 (FLNDAGACV) and MUC5AC-A24-716 (TCQPTCRSL) peptide that have specific cytotoxic activities against each of the HLA-A*0201 and A*2402-positive target cells pulsed with the candidate peptide. Subsequent analysis of each CTL clones also revealed their cytotoxic activities against pancreatic cancer cells expressing MUC5AC. A cold target inhibition assay further confirmed that each CTL clones specifically recognized the major histocompatibility complex (MHC) class I-peptide complex. Our results suggested that MUC5AC is a novel tumor-associated antigen recognized by CTL, and MUC5AC-A02-1398 (FLNDAGACV) and MUC5AC-A24-716 (TCQPTCRSL) are HLA-A*0201 and A*2402-restricted epitope peptides that can induce potent and specific immune responses against pancreatic cancer cells expressing MUC5AC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4748.
Abstract 4752: Induction of cytotoxic T lymphocytes primed with tumor RNA-loaded dendritic cells in esophageal squamous cell carcinoma: Preliminary step for DC vaccine design
Authors: Moaven Omeed , Gholamin Mehran , Farshchian Moein , Mahmoudi Mahmoud , Sankian Mojtaba , Memar Bahram , Forghani Mohammad N. , Malekzadeh Reza , Abbaszadegan Mohammad R.
Published: February 2011

Backgrounds: Dendritic Cells (DC) are potent antigen presenting cells with the ability to prime naïve T cells and convert them to cytotoxic CTL. We evaluated the capability of autologous DCs transfected with total Tumor and normal RNA to induce cytotoxic CTL as the preliminary step to design DC-based vaccine in the esophageal squamous cell carcinoma (ESCC).

Methods: Monocytes-derived DCs were electroporated with either total tumor RNA or normal RNA. T cells were then primed with tumor RNA transfected DCs and lysing effects of the generated CTL were measured with Cytotoxicity assay and IFN-γ Release Elispot Assay.

Results: Cytotoxicity against DCs loaded with tumoral RNA was induced (%24.8±5.2 SEM) while in normal RNA-loaded DCs it was minimal (%6.1±2.4 SEM) and significantly lower (p < 0.05). INF-γ secretion was more than 2-folds higher in tumoral RNA-loaded DCs when compared with normal RNA-loaded DCs (p < 0.05).

Conclusion: Electroporating DCs with tumor RNA made tumor antigen presenting cells that enhanced cytotoxicity effects of T cells against ESCC. This may be a useful autologous ex vivo screening tool for confirming the lytic effects of primed T cells on tumor and evaluate probable further adverse effects on noncancerous tissue. These data are a crucial preliminary step to establish a total tumor RNA-pulsed DC vaccine therapy of ESCC.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4752.
Abstract 4755: Importance of an induction dose to the immunogenicity of cancer vaccines: A reverse translational study
Authors: Berd David A. , Braun Donald , Mokyr Margalit B.
Published: February 2011

We have described a human cancer vaccine consisting of autologous, irradiated melanoma cells modified with the hapten, dinitrophenyl (DNP), and mixed with BCG. Because of its immunopotentiating effect, low dose cyclophosphamide (CY) was administered 3 days before the first of a series of vaccine injections. The vaccine induced T cell immunity to both haptenized and unmodified autologous melanoma cells; clinical effectiveness was suggested by tumor regression in some stage IV patients and an unexpectedly high proportion of 5-year survivors in stage IIIB and IIIC patients. A serendipitous finding from these studies was the critical importance of what we came to call an “induction dose” of vaccine - a single injection of irradiated, autologous melanoma cells without BCG 5-7 days prior to CY. Patients who received a properly timed induction dose had more intense T cell responses (positive DTH to unmodified melanoma cells 73% vs. 20%, p<.01) and were more likely to survive 5 years (51% vs. 33%, p=.007) than those who did not. To support this finding in a controlled experimental system, we developed a mouse model using the mammary carcinoma, 410.4. After injection of 410.4 cells into the mammary fat pads, primary tumors developed and were excised. Then we assessed the anti-metastatic activity of low dose CY followed by a DNP-modified vaccine prepared and administered in a manner comparable to the human melanoma vaccine. The published study showed that this regimen significantly prolonged tumor-free survival in the mice while CY followed by a control vaccine (irradiated, non-haptenized cells) did not. To mimic the induction dose used in the clinical trial, a cohort of mice was injected with irradiated, DNP-modified tumor cells without BCG 3-7 days prior to CY. The relapse-free survival (RFS) of mice receiving the induction dose (N=41) was significantly longer (p < .01) than that of mice receiving DNP-vaccine without the induction dose (N=27) (e.g., RFS day 37: CY+irradiated vaccine=30.8%, CY+irradiated-haptenized vaccine=40.7%, CY+induction dose+ irradiated-haptenized vaccine=53.7%). In this reverse translational model, the murine experiments corroborated a serendipitous clinical observation. Taken together our results suggest a new strategy for increasing the effectiveness of cancer vaccines.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4755.
Abstract 4756: Anti-melanoma-specific humoral immune responses in vitro in patients vaccinated with DC-rIL-2-VMO
Authors: Berenji Manijeh , Alam Khorshed , Somasundaram Raj , Wallack Marc K.
Published: February 2011

Introduction: DC-rIL-2-VMO, an allogeneic cellular melanoma vaccine consisting of five melanoma cell lines, a recombinant vaccinia virus encoding for IL-2, as well as dendritic cells, has previously been shown to generate a potent cellular response in murine tumor models. We now demonstrate that this vaccine also has the capacity to generate a sustained humoral response in vitro.

Methods: To detect anti-tumor antibodies in pre- and post-vaccinated melanoma patients’ sera, a mixed hemadsorption assay (MHA) was used. The binding of human antibody from diluted patients’ sera (1:05, 1:15 and 1:45) to cultured melanoma cells (Mel-2, Mel V, GLM-2 and KFM cells) was determined as indicator cells, sheep red blood cells (SRBC) coated sequentially with mouse anti-SRBC sera, goat sera against mouse sera, and mouse anti-human IgG sera to detect binding of human anti-melanoma antibodies.

Results: Out of four patients evaluated with Stage IIIA-C and Stage IV resected with no evaluable disease (NED), three patients’ post-vaccination sera showed antibodies specifically binding to melanoma cell lines. Antibodies from two of the patients’ sera showed moderate to strong binding to three different melanoma cell lines (Mel 2, Mel V and GLM-2). Additionally, antibodies from patients’ sera and sera from two normal healthy donors did not show any binding to colorectal carcinoma (SW1116) cells (CRC), a negative control.

Conclusions: Treatment of patients with Stage IIIA-C and Stage IV melanoma resected to be NED with DC-rIL-2-VMO induced a tumor-specific antibody response. The presence of these anti-melanoma antibodies suggests that this vaccine may be effective in slowing the progression of melanoma in some patients via cellular and humoral immune mechanisms. DC-rIL-2-VMO has been FDA-approved for Phase I trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4756.