bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1880: Role of fatty acid synthase and do novo lipogenesis in liver cancer development in mice.
Authors: Chen Xin , Li Lei , Wang Chunmei , Evert Matthias , Calvisi Diego F. , Semenkovich Clay F.
Published: August 2014

Aberrant metabolism, including increased de novo lipogenesis, is one of the hallmarks of cancer. Fatty Acid Synthase (Fasn) catalyzes the de novo synthesis of long-chain fatty acids from acetyl-CoA and malonyl-CoA. Increased Fasn expression has been reported in multiple tumor types, and inhibition of Fasn expression has been shown to have tumor-suppressing activity. However, how increased de novo lipogenesis contributes to tumor initiation and progression, especially in vivo, remains unknown.

In our previous studies, we showed that overexpression of the activated form of AKT (myr-AKT) induced de novo lipogenesis, hepatocyte proliferation and, eventually, liver cancer formation in mice. The tumorigenesis process could be significantly accelerated via co-expression of activated form of Ras. In the AKTRas tumor model, tumors were predominantly (>80%) hepatocellular carcinoma (HCC), and the remaining (<20%) were intrahepatic cholangiocarcinoma (ICC). The ICCs were induced via hepatocyte-biliary epithelial cell metaplasia in a Notch dependent manner. Intriguingly, fat droplet formation, increased expression of Fasn and other lipogenesis pathway genes were identified in HCC lesions, but not ICC lesions. To define whether Fasn-mediated de novo lipogenesis is a key metabolic event downstream of mTORC1 during AKTRas-induced hepatocarcinogenesis, we utilized Fasn flox/flox mice, and co-expressed AKTRas in the Fasn KO hepatocytes. Of note, we found that ablation of Fasn completely inhibited AKT induced lipogenesis and hepatocyte proliferation in mice. However, loss of Fasn modestly delayed AKTRas-induced liver tumor development. Histological analysis revealed that these tumor lesions were predominantly ICCs. The ICC cells were highly proliferative, and did not express Fasn or any other lipogenic pathway gene. The results suggest that Fasn-mediated de novo lipogenesis is required for AKTRas-induced HCC formation, but this process is dispensable for ICC formation.

In summary, our experiments support a critical role for Fasn as a downstream effector of mTORC1 in HCC pathogenesis. However, Fasn-mediated do novo fatty acid synthesis is not required in all tumor types. It is likely that other sources of fatty acids, presumably including those derived from the diet, can be utilized by cancer cells for membrane synthesis during cell proliferation.

Citation Format: Xin Chen, Lei Li, Chunmei Wang, Matthias Evert, Diego F. Calvisi, Clay F. Semenkovich. Role of fatty acid synthase and do novo lipogenesis in liver cancer development in mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1880. doi:10.1158/1538-7445.AM2013-1880

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1883: 2-Hydroxyglutarate: A candidate oncometabolite in breast cancer.
Authors: Mishra Prachi
Published: August 2014

Recent work in our laboratory identified a poor outcome tumor subtype with a high tissue 2HG concentration, distinct genome-wide DNA methylation, and a stem cell-like transcriptional signature. Currently, we do not know the origin of 2HG in these tumors but ruled out that it is caused by acquired isocitrate dehydrogenase mutations. Tumors with high 2HG tended to be estrogen receptor (ER)-negative and were over-represented among the African-American patients. Additional investigations revealed that 2HG can accumulate in a subset of human breast cancer cell lines to 50- to 100-fold above the baseline in non-cancerous cells, reminiscent of the difference between 2HG-high tumors and their adjacent non-cancerous tissue. The majority of the 2HG-high cell lines showed basal-like characteristics. While some cancer cells had these highly increased 2HG levels, other cell lines did not accumulate 2HG, providing a model to study 2HG treatment effects in cell lines with low endogenous 2HG and to identify the origin of increased 2HG production in those cell lines with high endogenous 2HG. Thus, we tested the hypothesis that 2HG influences breast cancer phenotypes. These experiments showed that 2HG induced phenotypic alterations, DNA methylation pattern alterations, proliferation, increased invasion and migration, and EGFR pathway activation in breast epithelial cells consistent with its oncogenic function in breast cancer. However, the observed phenotypes were partly cell line- and dose-dependent in our short term assays. Our results also indicated that different subtypes of breast cancer cells may respond differentially to 2HG, which suggests that possibly different cellular pathways are activated by 2HG in the various subtypes.

Citation Format: Prachi Mishra. 2-Hydroxyglutarate: A candidate oncometabolite in breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1883. doi:10.1158/1538-7445.AM2013-1883
Abstract 1886: Metabolic alterations associated with pancreatic cancer-induced cachexia.
Authors: Shukla Surendra K. , Singh Pankaj K.
Published: August 2014

Cachexia, a metabolic syndrome, leads to loss of muscle weight and fat tissues. Cancer-induced cachexia accounts for nearly 20% of all cancer-related deaths and is most prevalent in patients with gastric, pancreatic, colon and lung cancer. Although association of cachexia with various types of cancers has been known for a long time, the molecular mechanism of cancer-induced cachexia is still not well understood. Better understanding of the cancer-related cachexia syndrome may provide novel therapeutic targets and improved health care for cancer patients. By culturing differentiated C2C12 and 3T3-L1 cells in the presence of pancreatic adenocarcinoma cell or immortalized epithelial cell supernatants, we established cell culture-based cachexia model systems to investigate the depletion of muscle mass and fatty acid storage. Progressive physiological and biochemical alterations during muscle fibre breakdown and adipocyte fat depletion were assayed by utilizing western blotting, real-time PCR and cell imaging. We then investigated metabolomic alterations in C2C12 and 3T3-L1 cell lines in response to the cachectic factors secreted by the tumor cells. We performed differential metabolomic analyses on tumor cell supernatant-treated or control treated differentiated C2C12 muscle fibers and 3T3-L1-derived adipocytes by utilizing 2D 1H-13C HSQC NMR technique. Alterations in metabolic pathways were also investigated by evaluating the expression of metabolic genes in the corresponding pathways by performing quantitative real-time PCR. To obtain further insights into the metabolic alterations that trigger cachexia, we extended our studies to in vivo mouse orthotopic xenograft models and investigated alterations in metabolic gene expression and metabolite flux in muscle and fat tissues of tumor-bearing mice. Our results identify crucial nodes in metabolite flux that can be targeted to regulate cell growth, proliferation, and turnover in muscle and fat tissues during cancer-induced cachexia.

Citation Format: Surendra K. Shukla, Pankaj K. Singh. Metabolic alterations associated with pancreatic cancer-induced cachexia. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1886. doi:10.1158/1538-7445.AM2013-1886
Abstract 1889: Can the moonlighting glycolytic enzyme α-enolase be a therapeutic target in pancreatic cancer.
Authors: Capello Michela , Principe Moitza , Chattaragada Michelle Samuel , Riganti Chiara , Zhou Weidong , Ferri-Borgogno Sammy , Rolla Simona , Liotta Lance , Petricoin Emanuel , Cappello Paola , Novelli Francesco
Published: August 2014

Aberrant metabolism together with invasion and metastasis are hallmarks of cancer. This is particularly true for pancreatic ductal adenocarcinoma (PDAC), characterized by rapid progression, invasiveness and resistance to treatments. We have previously described α-enolase (ENOA) as a PDAC-associated antigen. It is a moonlighting protein that works both as a key metabolic enzyme and as a membrane plasminogen receptor. In order to clarify its multifunctional role in pancreatic tumorigenesis we investigated the effect of ENOA knockdown in PDAC cells. Protein expression alterations following ENOA knockdown in the human PDAC cell line CFPAC-1 were revealed by LC-MS/MS analysis. On the basis of a spectra count label-free quantitation approach a large number of proteins mainly involved in cell adhesion, metabolism and proliferation were found to be differentially expressed in ENOA silenced cells compared to the control. After ENOA silencing, PDAC cells displayed a delay in proliferation and decreased survival and colony formation capabilities, even if the pyruvate production was not affected. The growth inhibition was partially due to an increased concentration of intracellular reactive oxygen species (ROS) mainly generated through the sorbitol and NADPH oxidase pathways. Moreover in ENOA silenced cells, the in vitro plasminogen-driven invasion was abolished and the number of lung tumor masses was significantly reduced in SCID-beige mice injected with ENOA silenced cells compared to mice injected with control cells. These effects are under further confirmation in other PDAC cell lines. All together, these findings propose ENOA as a promising target for developing new therapies in pancreatic cancer management.

Citation Format: Michela Capello, Moitza Principe, Michelle Samuel Chattaragada, Chiara Riganti, Weidong Zhou, Sammy Ferri-Borgogno, Simona Rolla, Lance Liotta, Emanuel Petricoin, Paola Cappello, Francesco Novelli. Can the moonlighting glycolytic enzyme α-enolase be a therapeutic target in pancreatic cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1889. doi:10.1158/1538-7445.AM2013-1889
Abstract 1892: Integrated energetics and flux analysis reveals differential metabolic reprogramming in highly and less invasive cancer cells.
Authors: Achreja Abhinav , Yang Lifeng , Zhao Hongyun , Marini Juan C. , Nagrath Deepak
Published: August 2014

Cancer cell metabolism was investigated using systems biology tools such as metabolic flux analysis (MFA) to reveal intracellular cancer cell metabolic flux distribution. To obtain a complete role of nutrient-induced deprivation in metabolic reprogramming of cancer cells, we need to use metabolic flux analysis to determine unknown intracellular fluxes. MFA provides a framework for the estimation of intracellular fluxes and energy balance analysis (EBA) ensures the thermodynamic feasibility of the computed optimal metabolic fluxes. MFA uses stoichiometric and mass balance constraints to compute the intracellular fluxes. Here, we have developed an integrated energy balance and flux balance analysis framework. Our framework allows estimation of intracellular fluxes and reveals specific adaptations and dysregulation of metabolic pathways in ovarian cancer cells as a result of the nutrient (glucose and glutamine) deprivation conditions.

Glutamine and glucose are known to be important for cancer cell survival, growth and progression. Thus, there is a strong motivation to study metabolic alternations caused by glucose and glutamine deprivation and uncover links between various fuel sources for highly and less invasive cancer cells. Our findings indicate that NAD+/NADH ratio regulates glutamine's direct effect on lactate production irrespective of the bioenergetic substrate requirement for different kinds of cancer cells. Our findings, using quantification of metabolites pertaining to central carbon and nitrogen metabolism provided clues as to which pathways relevant to glycolysis, the pentose phosphate pathway, the TCA, urea cycles and lipid cycles, amino acid and nucleotide metabolisms may be altered by glucose and glutamine in cancer cells. This resulted in identification of dominant factors which control cancer metabolism. We believe that development of our oncosecretomics based metabolic strategy provides an unprecedented opportunity for the development of nutritional supplementation that can deregulate the adaptations and alterations induced by various nutrients in cancer metabolism.

Citation Format: Abhinav Achreja, Lifeng Yang, Hongyun Zhao, Juan C. Marini, Deepak Nagrath. Integrated energetics and flux analysis reveals differential metabolic reprogramming in highly and less invasive cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1892. doi:10.1158/1538-7445.AM2013-1892

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1893: Spectroscopic imaging unveils altered cholesterol metabolism in prostate cancer .
Authors: Yue Shuhua , Li Junjie , Lee Seung Young , Shao Tian , Song Bing , Cheng Liang , Hu Chang-Deng , Liu Xiaoqi , Ratliff Timothy L. , Cheng Ji-Xin
Published: August 2014

Metabolic reprogramming allows cancer cells to sustain high proliferative rates and resist cell death. Whereas alterations to glucose and amino acid metabolism have been extensively studied, altered lipid metabolism in cancer is increasingly recognized based on the findings of imbalanced lipid signaling network and dysregulated lipogenic enzymes. Storage of neutral lipids in lipid droplets (LDs), an essential aspect of lipid metabolism, is however long underappreciated until recent advances in LD biology. In human prostate cancer cell lines, neutral lipid accumulation was commonly seen and partially attributed to the up-regulation of fatty acid synthase, a key lipogenic enzyme catalyzing fatty acid biosynthesis and implicated in many human malignancies including prostate cancer. However, because the composition of LDs is not accessible with fluorescence microscopy, the exact role of lipid accumulation in prostate cancer progression remains elusive. By vibration-based spectroscopic imaging of 65 specimens collected from 61 prostate cancer patients and healthy donors, we found for the first time a marked shift from lipofuscin-like autofluorescent granules in normal, benign, and prostatic intraepithelial neoplasia lesions to cholesteryl ester (CE)-rich LDs in high-grade and metastatic prostate cancers. Quantitatively, CE amount was found ∼5 folds higher in high-grade relative to low-grade prostate cancers. Cellular studies demonstrate that the activated PI3K/Akt/mTOR/SREBP pathway accounts for accumulation of CEs via elevated LDL uptake and cholesterol esterification. Blockage of cholesterol esterification with an acetyl-coA cholesterol acyltransferase (ACAT) inhibitor significantly slows the growth and induces apoptosis of prostate cancer both in vitro and in vivo. These findings suggest that CE can be a quantifiable molecular marker for diagnosis of prostate cancer, especially for differentiation between low-grade and high-grade prostate cancers which currently has up to 40% inter-observer discordance. Moreover, targeting the altered cholesterol metabolism opens new opportunities for treating advanced prostate cancer in human patients.

Citation Format: Shuhua Yue, Junjie Li, Seung Young Lee, Tian Shao, Bing Song, Liang Cheng, Chang-Deng Hu, Xiaoqi Liu, Timothy L. Ratliff, Ji-Xin Cheng. Spectroscopic imaging unveils altered cholesterol metabolism in prostate cancer . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1893. doi:10.1158/1538-7445.AM2013-1893

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1897: Metformin treatment leads to the accumulation of lipid in breast cancer cell lines - a potential new biomarker of response and mechanism of action.
Authors: Lord Simon , Collins Jennifer , Fielding Barbara , Patel Neel , Gleeson Fergus , Harris Adrian , Karpe Fredrik
Published: August 2014

There is growing interest in the anticancer effect of the diabetes drug, metformin, and several clinical studies are underway worldwide. Metformin's anticancer effects are thought to be mediated by its activation of AMPK and subsequent downstream catabolic effects on protein and lipid metabolism pathways that are commonly dysregulated in cancer cells.

We have used mass spectrometry techniques and stable isotopes to investigate the effects of metformin on breast cancer cell lipid metabolism.

There was great variability in lipid metabolism response to 2mM metformin treatment in a panel breast cancer cell lines (MCF7, BT474, ZR-75-1, T47D MDA-MB-157, MDA-MB-468 and MDA-MB-231). Levels of stearic acid (p=0.019), palmitoleic acid (p=0.01), oleic acid (p=0.002), cis-vaccenic acid (p=0.003) increased 3-4 fold with metformin treatment in MCF7 cells but there was no significant change for MDA-MB-231 cells. 2mM Metformin decreased fatty acid β-oxidation of oleate 4-fold (p=0.03). Accumulation of de novo lipogenesis derived (p=0.004) and exogenous fatty acid (p=0.02) occurred in MCF7 cells but siRNA knockdown of AMPK did not abrogate these effects. A 3-fold decrease in the diglyceride to triglyceride ratio (p=0.02) suggested reduced lipolysis and inhibited adipose triglyceride lipase activity. Reduced incorporation of exogenous fatty acid into phospholipid was evident (p=0.002). Heterogeneity of lipid metabolism response correlated with effects on proliferation, oxygen consumption and mitochondrial morphology.

Our data shows that metformin has marked effects on accumulation of fatty acid in breast cancer cells most likely due to reduced lipolysis and β-oxidation and these effects cannot be explained as a result of AMPK activation. This work will contribute to developing biomarkers for metformin's metabolic anticancer effect and our understanding of how to combine it with other therapies to exploit synthetic lethality. We have initiated a multicenter, phase II, single arm clinical study, to further characterise metformin influenced, cancer relevant metabolic pathways. We will look to assess the correlation between metabolic biomarkers, including the use of mass spectrometry and gas chromatography analysis of tissue samples, and metabolic response using 18F-FDG PET-CT scans.

Citation Format: Simon Lord, Jennifer Collins, Barbara Fielding, Neel Patel, Fergus Gleeson, Adrian Harris, Fredrik Karpe. Metformin treatment leads to the accumulation of lipid in breast cancer cell lines - a potential new biomarker of response and mechanism of action. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1897. doi:10.1158/1538-7445.AM2013-1897
Abstract 190: Sulindac inhibits pancreatic carcinogenesis in LSL-KrasG12D-LSL-Trp53R172H-Pdx-1-Cre mice by suppressing aldo-keto reductase family 1B10 (AKR1B10).
Authors: Liao Jie , Chung Yeon Tae , Li Haonan , Zhang Wanying , Yang Guang-Yu
Published: August 2014

Sulindac inhibits carcinogenesis via the mechanism that appears beyond cyclooxygenase inhibition. Recently sulindac has been identified as competitive inhibitors of aldo-keto reductase family 1B10 (AKR1B10). AKR1B10 is involved in the process of carbonyls, farnesyl and geranylgeranyl as well as retinal metabolisms, and these metabolic activities are important to carcinogenesis. However, there is no evidence on if inhibition of pancreatic carcinogenesis by sulindac is via inhibition of AKR1B10 and protein prenylation (i.e., Kras protein). To determine the effect of sulindac on pancreatic carcinogenesis in LSL-KrasG12D-LSL-Trp53R172H-Pdx-1-Cre mice, five-week-old mice were fed with either AIN-93M diet or AIN-93M diet contained 200ppm sulindac (n=20 mice/group). Kaplan-Meier survival analysis with Log-Rank test was performed to analyze the effect of sulindac on animal survival from the development of pancreatic carcinoma. The results showed that average animal survival in LSL-KrasG12D-LSL-Trp53R172H-Pdx-1-Cre mice was 143.7 ± 8.8 days; and with the sulindac 200ppm treatment, animal survival significantly increased to 168.0 ± 8.8 days (p<0.005). Extensively histopathological analyses revealed that 90% of mice developed malignant pancreatic carcinomas, of which 10% invasive pancreatic adenocarcinoma had metastasis to liver and lymph nodes. With 200ppm sulindac treatment, incidence of pancreatic cancer and its metastasis were significantly reduced to 56% and 0%, respectively (P<0.01). Immunchemical analysis showed that sulindac treatment significantly decreased Ki-67-labeled cell proliferation, increased caspase-3-labeled cell apoptosis, and suppressed phosphorylated ERK1/2. In vitro mouse pancreatic carcinoma cells derived from the LSL-KrasG12D/+-LSL-Trp53R172/+-Pdx-1-Cre mice were further treated with sulindac and revealed that sulindac inhibited AKR1B10 enzyme activity, suppressed Kras and HDJ2 protein prenylation, and further inhibited phosphylation of C-raf, ERK1/2 and MEK1/2. Similar to sulindac treatment, siRNA-silencing AKR1B10 expression resulted in a significant reduction of Kras protein prenylation and its downstream signals. Our results demonstrate that sulindac inhibits pancreatic carcinogenesis, mainly via inhibition of kras protein prenylation and its activated signals by targeting AKR1B10. (supported NCI R01 CA164041)

Citation Format: Jie Liao, Yeon Tae Chung, Haonan Li, Wanying Zhang, Guang-Yu Yang. Sulindac inhibits pancreatic carcinogenesis in LSL-KrasG12D-LSL-Trp53R172H-Pdx-1-Cre mice by suppressing aldo-keto reductase family 1B10 (AKR1B10). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 190. doi:10.1158/1538-7445.AM2013-190
Abstract 1905: Influence of serum and oxygen on lipid biosynthesis and energy metabolism in murine glioblastoma.
Authors: Ta Nathan , Seyfried Thomas
Published: August 2014

Glucose and glutamine are essential energy metabolites for brain tumor growth and survival under both normoxic and hypoxic conditions. Carbons derived from either metabolite can be used for lipid synthesis. Here, we used uniformly labeled C14-glucose and C14-glutamine to examine de novo lipid synthesis in the VM-M3 murine glioblastoma cells. The major lipids synthesized included phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (SM), phosphatidic acid (PA), cholesterol (Chol), and cardiolipin (CL). De novo lipid synthesis was greater with glucose than with glutamine as the metabolic substrate under normoxia. Conversely, glutamine was the preferred substrate for lipid synthesis under anoxia. Lactate was produced largely from glucose carbons with minimal lactate derived from glutamine. Total lipid synthesis, using either glucose or glutamine, was greater in media without serum than in media containing 10% FBS under normoxia. Significant triglyceride accumulation was observed only under anoxia. Our findings show that lipid biosynthesis using glucose or glutamine is dependent on the type of growth environment, and that the VM-M3 glioblastoma cells can grow under anoxia only through acquisition of lipids, especially cholesterol, from serum. Supported in part from Boston College Res. Expense Fund.

Citation Format: Nathan Ta, Thomas Seyfried. Influence of serum and oxygen on lipid biosynthesis and energy metabolism in murine glioblastoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1905. doi:10.1158/1538-7445.AM2013-1905
Abstract 1917: Loss of steroid sulfotransferase (SULT) in human prostate cancer: Potential implications for SULT as a biomarker and therapeutic target.
Authors: Mirkheshti Nooshin , Song Chung S. , Chatterjee Bandana
Published: August 2014

Purpose: Reactivated androgen receptor, fueled by elevated tumor tissue 5α-dihydrotestosterone (DHT), plays a key role in the reemergence of prostate cancer. Primary prostate cancer almost always regresses in response to androgen deprivation therapy. The disease, however, recurs and progresses to therapy resistance and death in ∼30% patients. New-generation drugs that inhibit androgen receptor (MDV3100™) or enzymes involved in androgen biosynthesis (Zytiga™, TAK-700™, Dutasteride™) extend survival of post-chemotherapy patients for a limited period following which therapy resistance emerges– hence the challenge to develop alternate means for reducing intra-tumor androgen signaling.

Design and Results: 3β-sulfation of dehydroepiandrosterone (DHEA), mediated by the prostate-expressed sulfotransferase SULT2B1b (hereon SULT), is likely to reduce intra-prostate androgen levels since sulfated DHEA cannot be converted to androstenedione and thus to testosterone and DHT. We tested whether enzyme-catalyzed sulfation of DHEA, the obligate precursor steroid for androgen synthesis, is targetable to inhibit prostate cancer. Results show that SULT silencing increased prostate cancer cell proliferation. Clinical prostate cancer specimens showed markedly reduced SULT expression, revealed by immunohistochemistry, western blotting and mRNA quantification. 1,25-dihydroxy vitamin D3 (D3) and sterols enhanced SULT expression due to induction of the SULT2B-encoding gene by D3-activated vitamin D receptor (VDR) and sterol-activated liver X receptor (LXRα).

Conclusion: Tumor SULT levels may be a marker for predicting prostate cancer recurrence and progression. We also anticipate that enhancement of SULT activity in tumors by activating VDR and LXRα through combined therapeutic intervention with D3 and sterols would limit the tumor androgen pool and inhibit recurrent, therapy-resistant prostate cancer.

Citation Format: Nooshin Mirkheshti, Chung S. Song, Bandana Chatterjee. Loss of steroid sulfotransferase (SULT) in human prostate cancer: Potential implications for SULT as a biomarker and therapeutic target. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1917. doi:10.1158/1538-7445.AM2013-1917
Abstract 1921: Mutational and expressional analyses of DICER1 gene in ovarian stromal tumors and other common tumors.
Authors: Kim Min Sung , Je Eun Mi , Choi Youn Jin , Yoo Nam Jin , Lee Sug Hyung
Published: August 2014

Recurrent missense somatic mutations in DICER1 gene affecting exons 25 and 26 have recently been reported in non-epithelial tumors of ovary, especially in Sertoli-Leydig cell tumors (SLCT). Such a recurrent nature of the mutations strongly suggests that DICER1 mutation may play an important role in the development of ovarian non-epithelial tumors. The aim of this study was to further characterize the DICER1 mutations in human tumor tissues. We analyzed the DICER1 mutations in 2,819 tumor tissues from ovary and other organs by single-strand conformation polymorphism analysis. We found the DICER1 exon 25 and 26 hot-spot mutations in 2 of 14 SLCT (14.3%) and one of 132 non-Hodgkin lymphomas (0.8%), but none in other tumors. In SLCT, DICER1 expression was observed in Sertoli cells, but neither in Leydig nor spindle-shaped cells. We also analyzed DICER1 expression in lung, stomach, colon and prostate cancer tissues. DICER1 expression was lost in 37% of prostate cancers, but not in lung, stomach and colon cancers. Our data indicate that DICER1 recurrent mutations commonly occur in SLCT, but the incidence was significantly lowerthan that in the earlier study. Our data suggest that both DICER1 mutation and its expression loss may be important in tumor pathogenesis and that they are tumor type-specific.

Citation Format: Min Sung Kim, Eun Mi Je, Youn Jin Choi, Nam Jin Yoo, Sug Hyung Lee. Mutational and expressional analyses of DICER1 gene in ovarian stromal tumors and other common tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1921. doi:10.1158/1538-7445.AM2013-1921
Abstract 193: Effect of omega-3 fatty acid and dietary energy restriction on mammary carcinogenesis, circulating biomarkers and molecular regulators in rats.
Authors: Jiang Weiqin , Zhu Zongjian , McGinley John N. , Manni Andrea , Thompson Henry J.
Published: August 2014

Available evidence indicates that diets enriched in the ratio of n-3 fatty acids or dietary energy restriction (DER) have a broad range of health-related benefit including prevention of breast cancer. However, the number of studies in the combined effects of these intervention strategies have been investigated is limited. The objective of this investigation was to determine the effect of the combination of dietary n-3 and DER on mammary carcinogenesis, on plasma levels of cancer associated growth factors and cytokines and on molecular regulators in mammary carcinomas using a pre-clinical rat model for breast cancer. Female Sprague Dawley rats were injected with 1-methyl-1-nitrosourea (50 mg/kg) and 7 days thereafter randomized to four groups (n=30/group), 1) control (dietary n-6/n-3=1:1); 2) high n-3 (dietary n-6/n-3=1:10); 3) 20%DER (dietary n-6/n-3=1:1); 4) high n-3 + 20%DER (dietary n-6/n-3=1:10). Rats were fed a modification of the AIN-93G diet formulation that contained 30% of dietary calories from fat (currently recommended in the US Dietary guidelines). It was observed that mammary cancer incidence, cancer multiplicity and tumor burden were reduced and cancer latency was elongated for the rats in groups 2-4 compared to the rats in group 1, and the significant differences were observed between group 1 (n-6/n-3=1:1) and 4 (n-6/n-3=1:10 + 20%DER). The protective dietary combination of high n-3 and DER was associated with significant reductions in plasma IGF-1, insulin, and leptin and increased adiponectin compared to rats fed the control diet (n-6/n-3=1:1, p<0.01). Carcinomas induced in the protected animals were smaller in mass with altered levels of proteins in the PPAR and mTOR pathways on which plasma insulin had the most impact. This work was supported by grant KG081632 from the Susan G. Komen Foundation.

Citation Format: Weiqin Jiang, Zongjian Zhu, John N. McGinley, Andrea Manni, Henry J. Thompson. Effect of omega-3 fatty acid and dietary energy restriction on mammary carcinogenesis, circulating biomarkers and molecular regulators in rats. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 193. doi:10.1158/1538-7445.AM2013-193
Abstract 1930: The unknown piece of the pie: Molecular markers in triple-negative lung cancer.
Authors: Schweitzer Brock L. , Lawrence Kasey D. , Handshoe John , Skelton Rachel , Chatfield Lindsay , Xue Liquan , Morris Stephan W. , Hout David R.
Published: August 2014

Lung cancer is the main cause of cancer mortality worldwide. Approximately ∼80% of lung cancer cases are non-small cell lung cancer (NSCLC) in type and >50% of NSCLC are adenocarcinoma in histopathology. A shifting paradigm in the field of pulmonary oncology is the identification of molecular markers that are therapeutic targets and/or provide prognostic information. The recent recognition of the role biomarkers such as EGFR, KRAS, and ALK play in NSCLC adenocarcinoma patients demonstrates this trend. Although these three highly characterized molecular markers make up one-quarter to two-thirds of NSCLC adenocarcinoma patients, the remaining population, termed “Triple-Negative Lung Cancer” (TNLC), has yet to be fully defined. The complete absence of therapeutic targets for triple-negative lung cancer patients undeniably supports the need to identify up-regulated and mutated genes that better define this poor prognostic cohort. Recent genome-wide expression profiles subdivided TNLC patients into a poor prognostic group based on the overexpression of the DEP domain containing 1 gene (DEPDC1) (Okayama et al., 2011). We hypothesized that overexpression of DEPDC1 correlates with a specific sub-population of adenocarcinoma patients. Our initial studies indicate that DEPDC1 is constitutively expressed in both normal lung as well as lung cancer specimens. DEPDC1 transcripts are present as two variants (V1 and V2), with constitutive expression of the V1 variant in normal lung. We hypothesized that expression of the V2 transcript could be unique to NSCLC adenocarcinomas. Screening of FFPE NSCLC adenocarcinoma specimens using a proprietary qPCR approach demonstrated the presence of the DEPDC1 V2 variant but at levels that were not sufficient to serve as a sole biomarker. We therefore investigated whether the ratio of DEPDC1 V2 expression relative to V1 could serve as a biomarker in TNLC. Our preliminary data suggest that the ratio of V2 to V1 expression of DEPDC1 does in fact segregate TNLC into specific sub-populations. These studies have exciting diagnostic as well as therapeutic implications as Phase I/II studies using novel peptide vaccines derived against DEPDC1 have proven tolerable and efficacious for patients with bladder cancer and could potentially be extended to target these newly identified TNLC sub-populations.

Citation Format: Brock L. Schweitzer, Kasey D. Lawrence, John Handshoe, Rachel Skelton, Lindsay Chatfield, Liquan Xue, Stephan W. Morris, David R. Hout. The unknown piece of the pie: Molecular markers in triple-negative lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1930. doi:10.1158/1538-7445.AM2013-1930
Abstract 1931: Development of a functional assay for the determination of BRCA1 carrier status.
Authors: Bathurst Lauren , Uy Paolo , Feilotter Harriet , Davey Scott
Published: August 2014

Germline mutations in the breast cancer susceptibility gene BRCA1 confer an overall lifetime risk of developing breast cancer of up to 80%. Given the high risk of cancer development associated with BRCA1 mutation carriers, it is important that they be identified early and accurately. However, identification is impeded by the number of Variants of Unknown Significance (VUS), therefore there is compelling rationale for a functional assay that will identify pathogenic mutations. Previous work in our lab developed a novel functional assay to predict BRCA1 status based on gene expression profiles. That study analyzed EBV-transformed lymphoblastoid cell lines (LCLs) of BRCA1 mutation carriers (BRCA1+/−) and controls (BRCA1+/+) and found that lymphocytes from BRCA1 carriers could be distinguished with high fidelity (∼90% accuracy) from individuals with two wild type copies of the gene. Interestingly, carriers were recognized through their basal gene expression profiles. Global gene expression level changes following ionizing radiation (IR) to induce the DNA damage response (DDR) did not support the development of an accurate class predictor.

The final predictor included a set of 43 genes. Many of these genes were also markers of differentiation in blood cells. A subset of these genes have now been validated in LCLs using qRT-PCR and protein-based methods including single reaction monitoring mass spec (SRM-MS) and flow cytometry. These assays were able to distinguish BRCA1 mutation carriers from controls based on the expression levels of specific markers, including genes with known roles in lymphocyte differentiation. Using these techniques, ongoing work is being done to test the ability of these markers to discriminate BRCA1 mutation carriers from controls using lymphocytes isolated from fresh blood samples.

Citation Format: Lauren Bathurst, Paolo Uy, Harriet Feilotter, Scott Davey. Development of a functional assay for the determination of BRCA1 carrier status. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1931. doi:10.1158/1538-7445.AM2013-1931
Abstract 1932: Association of genetic variants with prostate cancer risk.
Authors: Andavolu Radhika G. , Cardenas Jean-Luc , Shore Ross , Rubin Zach , MacMurray James , Chiravuri Krishna Kanth , Andavolu Murthy VS , Rubakovic Svetlana
Published: August 2014

Background: Genetic alterations can contribute to the malignant transformation and progression of prostate cancer. We have previously shown a strong association between specific leptin gene alleles, elevated serum leptin level, and body mass index in subjects with prostate and breast cancer. These studies showing association with leptin suggest a strong link with obesity as an increased risk factor. Studies have shown that adipose tissue TNFa and leptin, which are both overexpressed in obesity, induces the production of acute phase reactants through their autocrine or endocrine role. When GSTP1 is inactivated, prostate cells appear to become more vulnerable to somatic alterations upon chronic exposure to genome-damaging stresses such as oxidants and electrophiles that are contributed by environment and lifestyle. Studies also show that the methylation status of glutathione S-transferase p1 (GSTP1) could individually distinguish prostate cancer from benign prostatic hyperplasia. Based on prior studies, it is possible that multi gene analysis can provide better diagnostic or prognostic values for prostate cancer. Methods: In this study, we screened leptin gene markers, TNFa, GSTP1 and BCCIP(TOK-1A) in 126 prostate cancer and normal healthy age & sex matched controls (Prostate cases: Age range 45-94, Mean age 73.3 Control males: Age range 65-98, Mean age 73.5). Polymerase chain reaction was used to amplify Leptin, TNF-308 and GSTP1 genes and the products are analyzed using 3130 Genetic analyzer and PCR-RFLP methods. Statistical analysis was performed using the SPSS/PC statistical program (version 10.0 for Windows; SPSS, Inc.,Chicago, IL). Relationships between two groups and genotype variables were assessed by Pearson's correlation coefficient.

Conclusions: Prostate cancer is a complex, heterogeneous disease and there a variety of gene-based biomarkers that have been associated with prostate cancer. Our results show strong association of leptin (p<0.006), GSTP1 (p<0.023) with prostate cancer. Individually TNF and BCCIP showed a modest association but TNF/BCCIP gene interaction had a significant effect on prostate cancer risk and disease progression suggesting a multigene model for prostate cancer screening.

Citation Format: Radhika G. Andavolu, Jean-Luc Cardenas, Ross Shore, Zach Rubin, James MacMurray, Krishna Kanth Chiravuri, Murthy VS Andavolu, Svetlana Rubakovic. Association of genetic variants with prostate cancer risk. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1932. doi:10.1158/1538-7445.AM2013-1932
Abstract 1933: Visfatin promotes malignant breast cancer behavior through activation of STAT3.
Authors: Yuan Shyng-Shiou F. , Chen Yun-Ju , Lee Yi-Chen , Hou Ming-Feng
Published: August 2014

Background. Obesity-associated adipocyte dysfunction and adipocytokine imbalance has become a major focus in breast cancer research. In this study, the role of circulating visfatin, a recently discovered adipocytokine, in breast cancer was explored.

Methods. Serum visfatin was analyzed by enzyme-linked immunosorbent assay and correlated with clinicopathological characteristics and survival, while the expression of phosphorylated and activated STAT3 in breast cancer tissue was analyzed by immunohistochemistry. The biologic activity of visfatin as well as the underlying mechanisms was analyzed by using XTT cell proliferation assay, flow cytometry and immunoblotting analysis in human breast cancer cells.

Results. Serum visfatin was substantially elevated in breast cancer patients. It was positively associated with tumor stage and lymph node metastasis, but negatively associated with estrogen receptor (ER) and progesterone receptor (PR) status. Breast cancer patients with high serum visfatin, especially in combination with ER− and PR− status, had a poor patient survival. On the other hand, patients with high serum visfatin had a lower incidence of disease recurrence and a better patient survival after adjuvant hormone therapy. Serum visfatin was positively associated with STAT3 tyrosine phosphorylation in breast cancer tissues. In agreement with the clinical observation, visfatin treatment in breast cancer cells activated JAK1 and SRC as well as their downstream molecule STAT3, followed by an enhanced G1/S progression and cell proliferation.

Conclusion. Serum visfatin, through activation of STAT3, leads to a malignant cancer behavior and poor survival in breast cancer patients.

Key words: visfatin, adipocytokine, breast cancer, STAT3

Citation Format: Shyng-Shiou F. Yuan, Yun-Ju Chen, Yi-Chen Lee, Ming-Feng Hou. Visfatin promotes malignant breast cancer behavior through activation of STAT3. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1933. doi:10.1158/1538-7445.AM2013-1933

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1937: Development of microRNA-based prediction model of Trastuzumab treatment response for HER2-positive breast cancer using FFPE specimens.
Authors: Sato Fumiaki , Wang Zhipeng , Ueno Takayuki , Myomoto Akira , Takizawa Satoko , Pu Feng Ling , Masuda Norikazu , Mikami Yoshiki , Mikami Yoshiki , Shimizu Kazuharu , Saji Shigehira , Toi Masakazu
Published: August 2014

[Background and Aim] Although Trastuzumab has been used for HER2(+) breast cancer, the treatment response of Trastuzumab therapy depends on unknown mechanisms among individual cases. In order to avoid unnecessary adverse events and to lighten financial burden, pre-treatment prediction of trastuzumab treatment response would be beneficial for patients. Thus, in this study, we develop a prediction algorithm using microRNA expression profile using formalin-fixed paraffin-embedded (FFPE) specimens of HER2(+) breast cancer. [Materials and Methods] Eighty-three breast cancer patients who underwent trastuzumab-chemo combined therapy before operations were enrolled with written informed consent. FFPE specimens of pre-treatment core needle biopsy samples were collected, and regions containing cancer and adjacent stromal cells were laser-microdissected. Total RNA samples extracted from the microdissected specimens were subjected for microRNA microarray (3D-Gene®, Toray, Japan) analysis. Among these 83 patients, 39 cases had pCR (definition: complete response in IDC regions regardless presence of DCIS without lymph node metastasis), and the other 44 cases did not. According to the pCR/non-pCR information, we develop a prediction model using 35 signature microRNAs by a SVM technique. Prediction accuracy assessed by Leave-one-out validation was AUROC=0.889. The 35 signature microRNAs for trastuzumab treatment response included 7 out of 8 let-7 family members and miR-125a-5p/b-5p, which were downregulated in pCR specimens. [Conclusion] microRNA profile could predict treatment response of trastuzumab-chemo combined therapy for HER2(+) breast cancer, and the developed prediction algorithm might be a useful tool for clinical decision making.

Prediction Accuracy Outcome: non-pCR Outcome: pCR total Prediction: non-sensitive 35 6 41 Prediction: sensitive 9 33 42 total 44 39 83

Citation Format: Fumiaki Sato, Zhipeng Wang, Takayuki Ueno, Akira Myomoto, Satoko Takizawa, Feng Ling Pu, Norikazu Masuda, Yoshiki Mikami, Yoshiki Mikami, Kazuharu Shimizu, Shigehira Saji, Masakazu Toi. Development of microRNA-based prediction model of Trastuzumab treatment response for HER2-positive breast cancer using FFPE specimens. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1937. doi:10.1158/1538-7445.AM2013-1937
Abstract 1942: Identification of a microRNA expression signature for radiochemosensitivity of colorectal cancer cells, involving miRNAs-320a, -224, -132 and let7g.
Authors: Salendo Junius , Spitzner Melanie , Kramer Frank , Jo Peter , Beissbarth Tim , Wolff Hendrik A. , Becker Heinz , Dobbelstein Mathias , Ghadimi Michael , Grade Marian , Gaedcke Jochen
Published: August 2014

Background: Preoperative 5-fluorouracil-based chemoradiotherapy is the standard treatment for locally advanced rectal carcinomas. However, the individual tumor response is very heterogeneous, ranging from complete resistance to regression. Therefore, ascertaining the role of microRNAs (miRNAs) in therapy response as well as identification certain miRNA as predictive markers for response remains very crucial.

Materials/Methods: Using an in-vitro model of 12 colorectal cancer cell lines, we compare the pretherapeutic miRNA expression profiles of these cell lines with the previously assessed radiochemoresistance of each cell lines, respectively. Differences in treatment sensitivity of the cell lines and miRNAs expression were then correlated. Colony formation assays in two independent cell lines were used for the functional validation in-vitro. To asses the clinical applicability miRNA expressions of 64 pretherapeutic biopsies from patients with locally advanced cancer treated with 5-FU based RCT were analyzed.

Results: We identified 36 miRNAs whose expression levels correlated significantly with the heterogeneous sensitivity of the cell lines to chemoradiotherapy (p < 0.05). Microarray measurements were independently validated using semi-quantitative real-time PCR. miR-320, miR-132, miR-429, miR-224 and let-7g were then functionally validated in-vitro by transfection of corresponding miRNA mimics. Analysis of miRNA expression in rectal cancer patients biopsies showed high correlation of miR-224 expression with poor prognosis (p=0.043).

Conclusion: We identified 36 miRNAs that associated with response to chemoradiotherapy. Functional validations of miR-320, miR-132, miR-429, miR-224 and let-7g have been undertaken in-vitro underlining the regulatory effects of the specific miRNAs. Further miR-224 found out to be significantly correlated to Disease Free Survival (DFS). An independent prospective validation in a clinical trial will be performed.

Citation Format: Junius Salendo, Melanie Spitzner, Frank Kramer, Peter Jo, Tim Beissbarth, Hendrik A. Wolff, Heinz Becker, Mathias Dobbelstein, Michael Ghadimi, Marian Grade, Jochen Gaedcke. Identification of a microRNA expression signature for radiochemosensitivity of colorectal cancer cells, involving miRNAs-320a, -224, -132 and let7g. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1942. doi:10.1158/1538-7445.AM2013-1942