bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1836: Triple negative breast cancer: Metabolomics and flux analysis to identify targets for drug development
Authors: Stewart Delisha A. , Winnike Jason , McRitchie Susan , Pathmasiri Wimal , Sumner Susan
Published: August 2015

To date, no targeted therapies are clinically available to treat triple negative breast cancer (TNBC). This highly aggressive form of breast cancer (BCa) derives its name from the absence of estrogen receptor (ER+), progesterone receptor (PR+), and HER2/neu receptor (HER2+). These receptors are expressed in other subtypes of BCa (i.e. Luminal, HER2-enriched) and have been successfully targeted with therapeutics such as Tamoxifen and Herceptin. New therapeutics are needed to improve the prognosis for patients with TNBC. Metabolomics and metabolic flux analysis (MFA) were conducted to reveal differences in TNBC cell lines compared with hormone receptor-positive cell lines (ER+ and +/-HER2+) prior to and following treatment with chemotherapy drugs. Two triple-negative (MDA-MB-231 and MDA-MB-468) and two hormone-responsive (BT474 and MCF-7) cell lines were compared before and after treatment with Taxol®/Paclitaxel. The metabolic profiles were assessed using 1H NMR broad spectrum metabolomics, MFA using [U-13C]-glucose and [U-13C]-glutamine enriched media, and cytokine profiling of the media. Broad spectrum metabolomics analysis demonstrated differences in the metabolic profiles between the TNBC and hormone-responsive cell lines in the absence of treatment, with a higher level of amino acids (e.g. alanine, glutamate, glutamine, glutathione), short chain fatty acids (isobutyrate, β-hydroxybutyrate), and nucleotides and nucleotide sugars (ADP, ATP, UDP-glucose, UDP-glucuronate) in the TNBC cell lines. Further, MFA demonstrated that glucose utilization was greater in the TNBC cells (MDA-MB-468) compared to the hormone-responsive cells (MCF-7). In addition, 13C-labeled glycine and 13C-labeled glutathione were only observed in TNBC cell lines treated in the presence of [U-13C]-glucose. In response to Taxol treatment, more metabolites were altered in the hormone-responsive cell lines compared with the TNBC cells. Profiling of 80 inflammatory cytokines (RayBiotech arrays) also demonstrated different responses in secreted inflammatory signals following treatment. We observed significant upregulation in interleukins-4 and -16 in MDA-MB-468 cells, while there was a downregulation of osteoprotegerin in all four cell lines after treatment with Taxol. This approach holds promise for identifying biomarkers which may be leveraged for development of targeted treatments.

Citation Format: Delisha A. Stewart, Jason Winnike, Susan McRitchie, Wimal Pathmasiri, Susan Sumner. Triple negative breast cancer: Metabolomics and flux analysis to identify targets for drug development. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1836. doi:10.1158/1538-7445.AM2015-1836
Abstract 1838: MUC1 serves as metabolic regulator in triple negative breast cancer
Authors: Goode Gennifer D.
Published: August 2015

Breast cancer, the second leading cause of cancer deaths in women, is the most common cancer among North American women, accounting for nearly 1 in 3 cancer cases diagnosed in the U.S. women. Triple negative breast cancer (TNBC) subtype accounts for approximately 15-25% of all breast cancer cases and has an increased incidence of metastasis, high recurrence within 1-3 years and a high mortality rate. Therefore, identifying factors that facilitate tumor growth and metastases have the potential to serve as novel molecular targets for breast cancer therapy. Mucin1 (MUC1), a glycoprotein associated with chemoresistance, is aberrantly overexpressed in TNBC and facilitates growth and metastasis of TNBC cells. This occurrence can be partially attributed to MUC1 interaction with hypoxia-inducible factor alpha (HIF1α), a key regulator of glycolysis. In the present study we examined the effect of MUC1 expression on cancer cell metabolism of TNBC cell lines under normoxic (20% Oxygen) and hypoxic (1% Oxygen) conditions. MUC1 was ectopically overexpressed in the MDA-MB-231 cell line and stably knocked down in the MDA-MB-468 cell line. Results indicate that MUC1 interacts with HIF1α and the interaction is further enhanced under hypoxic conditions. In addition, MUC1 expression enhanced glucose and glutamine uptake, enhanced lactate secretion and altered the expression of several metabolic genes. Furthermore, untargeted global metabolomic profiling identified metabolite alterations in which MUC1 expression modulates cancer cell metabolism to facilitate growth properties of TNBC cells. Thus our results support the notion that MUC1 serves as a metabolic regulator in TNBC, facilitating metabolic reprogramming that influences growth of TNBC.

Citation Format: Gennifer D. Goode. MUC1 serves as metabolic regulator in triple negative breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1838. doi:10.1158/1538-7445.AM2015-1838
Abstract 1839: Development of urinary pseudo-targeted LC-MS based metabolomics method and its application in hepatocellular carcinoma biomarker discovery
Authors: Yao Zhenzhen , Shao Yaping , Xu Guowang
Published: August 2015

Hepatocellular carcinoma (HCC) is one of the pestilent malignancies leading to cancer-related death. Discovering effective biomarkers for HCC diagnosis is an urgent demand. To identify potential metabolites biomarkers, we developed a urinary pseudo-targeted method based on liquid chromatography- triple quadrupole linear ion Q-trap 5500 mass spectrometry (LC-QTRAP- MS). Compared with non-targeted method, the pseudo-targeted method can achieve better data quality which benefits differential metabolites discovery. The established method was applied to cirrhosis (CIR) and HCC investigation. It was found that urinary nucleosides, bile acids, citric acid and several amino acids were significantly changed in liver disease groups compared with the controls, featuring the dysregulation of purine metabolism, energy metabolism and amino metabolism in liver diseases. Furthermore, some metabolites such as cycle cyclic adenosine monophosphate (AMP), glutamine and, short- and medium-chain acylcarnitines were the differential metabolites of HCC and CIR. Based on binary logistic regression, butyrylcarnitine (carnitine C4:0) and hydantoin-5-propionic acid were defined as a combinational marker to distinguish HCC from CIR. The area under curve (AUC) was 0.786 and 0.773 for discovery stage and validation stage samples, respectively. These data show that the established pseudo-targeted method is a complementary one of targeted and non-targeted methods for metabolomics study.

Note: This abstract was not presented at the meeting.

Citation Format: Zhenzhen Yao, Yaping Shao, Guowang Xu. Development of urinary pseudo-targeted LC-MS based metabolomics method and its application in hepatocellular carcinoma biomarker discovery. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1839. doi:10.1158/1538-7445.AM2015-1839
Abstract 1856: Preclinical effects of dual AKT/MAPK inhibition in PTEN-deficient prostate cancer
Authors: De Velasco Marco A. , Yamamoto Yutaka , Kura Yurie , Fukushima Emiko , Ando Naomi , Davies Barry , Hatanaka Yuji , Oki Takashi , Shimizu Nobutaka , Nozawa Masahiro , Yoshikawa Kazuhiro , Yoshimura Kazuhiro , Nishio Kazuto , Uemura Hirotsugu
Published: August 2015

Targeting signal molecules with single agent monotherapy is met with limited or transient clinical benefits. Therefore, establishing effective rational combinatorial treatment strategies provides a better option for achieving improved clinical outcomes. The AKT/PI3K and MAPK signaling pathways have been identified as key compensatory signal transduction pathways in prostate cancer. We have used genetically engineered mouse models of prostate cancer to show the treatment efficacy of monotherapy with the AKT inhibitor AZD5363 and the MAP/ERK kinase (MEK) inhibitor selumetinib (AZD6244, ARRY142886). We hypothesized that a rational combinatorial approach utilizing both of these inhibitors would improve treatment efficacy compared to single agent monotherapy. To test this hypothesis, we evaluated the treatment effects of AZD5363 and selumetinib in preclinical models of prostate cancer. In vitro experiments were carried out using a panel of human prostate cancer cell lines, and mouse prostate cancer cell lines, derived from PTEN-deficient castration-naïve prostate cancer (CNPC) and castration-resistant prostate cancer (CRPC). Treatments with AZD5363 and selumetinib demonstrated a synergistic combination effect in both human and mouse prostate cancer cell lines. We evaluated the antitumor effects in vivo by comparing the effects of AZD5363 and selumetinib alone or in combination in PTEN-deficient mice. Our studies show that combination therapy of AZD5363/selumetinib was well-tolerated by the mice and demonstrated significantly improved inhibition of tumor growth compared to single agent therapy. In the CNPC model, mice showed 16.7, 10.0 and 24.9% reductions in tumor burden after treatments with AZD5363, selumetinib and AZD5363/selumetinib, respectively (P<0.001). In the CRPC model, mice showed 15.8, 3.8 and 27.0% reductions in tumor burden after treatments with AZD5363, selumetinib and AZD5363/selumetinib, respectively (P<0.001). Target validation studies confirmed the inhibition of downstream targets. Overall, our findings suggest a potential role for targeting the PI3K/AKT and MAPK signaling pathways with AZD5363 and selumetinib for treatment of PTEN-deficient prostate cancer.

Citation Format: Marco A. De Velasco, Yutaka Yamamoto, Yurie Kura, Emiko Fukushima, Naomi Ando, Barry Davies, Yuji Hatanaka, Takashi Oki, Nobutaka Shimizu, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuhiro Yoshimura, Kazuto Nishio, Hirotsugu Uemura. Preclinical effects of dual AKT/MAPK inhibition in PTEN-deficient prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1856. doi:10.1158/1538-7445.AM2015-1856
Abstract 1858: Estrogen receptor beta as a tractable therapeutic target
Authors: Filgueira Carly S. , Benod Cindy , Lou Xiaohua , Strom Anders , Gustafsson Jan-Åke , Berkenstam Anders , Webb Paul
Published: August 2015

Nuclear receptors (NRs) regulate a variety of biological processes and are critically important in the emergence, prevention, and treatment of cancers. Many NRs represent well-validated drug targets as their ligands are used extensively in medicine. However, there still remains an active search for novel agonists and antagonists with enhanced selectivity. Estrogen receptor β (ERβ) is an attractive target for drug development. It plays different roles in gene expression from ERα, implicated in breast and uterine cancers, and displays overlapping but distinctive tissue distributions from ERα and non-redundant roles. Further, emerging evidence indicates that ERβ plays anti-proliferative and anti-apoptotic roles in a variety of cancers. These considerations have led to the suggestion that ERβ is an appealing tractable target for development of selective agonists to treat and prevent colon, breast, prostate, lung, and skin cancer and natural and synthetic estrogens for ERβ are being studied in colon, breast, and lung cancers. However, because of the high sequence similarities between ERα and ERβ, preferentially targeting one subtype can prove challenging. Repositioning of existing approved drugs with known side effect profiles can provide advantages because de novo drug design suffers from high developmental failure rates and undesirable side effects which have dramatically increased costs. Our research strategy is to identify and develop an ERβ agonist lead compound by using a systematic step-wise hit-to-lead campaign. We have identified a series of ERβ-selective lead compounds derived from screening libraries of clinically relevant compounds and have shown that some of these compounds display interesting and desirable selective modulator activities.

Citation Format: Carly S. Filgueira, Cindy Benod, Xiaohua Lou, Anders Strom, Jan-Åke Gustafsson, Anders Berkenstam, Paul Webb. Estrogen receptor beta as a tractable therapeutic target. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1858. doi:10.1158/1538-7445.AM2015-1858
Abstract 1859: Discerning the molecular basis of DNA-PK pro-tumorigenic functions and translational capacity as a therapeutic target in prostate cancer
Authors: Dylgjeri Emanuela , Goodwin Jonathan , Knudsen Karen
Published: August 2015

Prostatic adenocarcinoma (PCa) is dependent on androgen receptor (AR) signaling at all stages of disease, as AR activation induces both cell proliferation and survival. Though organ-confined disease can be treated, the response is transient and reactivation of AR results in incurable state of disease (castration resistant prostate cancer, CRPC). Thus, there is a significant need to discern the mechanisms by which aberrant AR activity arises, and to develop new means to clinically target advanced disease. Recently, AR activation was found to promote tumor cell survival and proliferation through a feed-forward loop involving the repair factor DNA-dependent protein kinase (DNA-PK). Emerging new data correlates elevated DNA-PK expression with poor prognosis and identifies DNA-PK as a driver of metastatic signaling. Although the role of DNA-PK activity in DNA repair is well characterized, the underpinning mechanisms and consequences of DNA-PK in transcriptional regulation are understudied. Data to be discussed will build on our new preliminary DNA-PK interactome data to identify sequence-specific transcription factors found in complex with DNA-PK. In order to molecularly dissect DNA-PK transcriptional events, the DNA-PK cistrome and transcriptome will be defined using clinically relevant models, resulting in a detailed understanding of DNA-PK mediated transcriptional regulation. The correlation of high expression of DNA-PK with poor prognosis in advanced PCa, as well as the known roles of DNA-PK in cancer-relevant pathways make DNA-PK a viable therapeutic target for PCa. Despite the current clinical assessment of DNA-PK inhibitors in multiple tumor types, there are no existing trials targeting PCa. Additional studies that assess the efficacy of current DNA-PK inhibitors in PCa will be presented and used to determine the best clinical space for their use.

Citation Format: Emanuela Dylgjeri, Jonathan Goodwin, Karen Knudsen. Discerning the molecular basis of DNA-PK pro-tumorigenic functions and translational capacity as a therapeutic target in prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1859. doi:10.1158/1538-7445.AM2015-1859
Abstract 186: microRNA-146a inhibits epithelial mesenchymal transition in non small cell lung cancer by targeting insulin receptor substrate 2
Authors: Son Ji Woong , Lee Soo Young , Na Moon Jun , Kwan Sun Jung , Jeon Hyo Sung
Published: August 2015

BACKGROUND: During cancer progression, some tumor cells show changes in their plasticity by morphological and phenotypical conversions, as an expression of mesenchymal markers and loss of epithelial markers, collectively referred to as epithelial-mesenchymal transition (EMT). EMT has been increasingly recognized as a critical phenomenon in lung cancer progression. The goal of this study was to identify microRNAs involved in lung cancer progression.

METHODS: A microarray and qRT-PCR were performed to investigate the miRNA expression profiles in mesenchymal like lung cancer cells. The role of miR-146a in lung cancer progression was measured by invasion and migration assays in vitro. Bioinformatics and luciferase report assays were used to identify the target of miR-146a.

RESULTS: The expression of miR-146a was reduced in mesenchymal like lung cancer cell lines. The over-expression of miR-146a induced a marked reduction of mesenchymal marker and increase epithelial marker in lung cancer cell lines. Moreover, the over-expression of miR-146a suppressed lung cancer cells migration and invasion. The expression of miR-146a was down-regulated in advanced lung cancer tissues. Insulin receptor substrate 2 (IRS2), an adaptor proteins that modulates normal growth, metabolism, survival, and differentiation, was identified as a target of miR-146a. miR-146a regulated the expression of IRS2 at the mRNA and protein levels.

CONCLUSION: These data demonstrate for the first time that miR-146a suppresses lung cancer progression by repressing IRS2 expression. This provides a new insight into the post-transcriptional regulation of lung cancer progression by miRNAs, a potential approach for the treatment of lung cancer.

Citation Format: Ji Woong Son, Soo Young Lee, Moon Jun Na, Sun Jung Kwan, Hyo Sung Jeon. microRNA-146a inhibits epithelial mesenchymal transition in non small cell lung cancer by targeting insulin receptor substrate 2. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 186. doi:10.1158/1538-7445.AM2015-186
Abstract 1860: DNA-PK-mediated transcriptional regulation drives tumor progression and metastasis
Authors: Goodwin Jonathan F. , Kothari Vishal , Drake Justin M. , Zhao Shuang , Dylgjeri Emanuela , Dean Jeffry L. , Schiewer Matthew J. , McNair Christopher , Magee Michael S. , Den Robert B. , Zhu Ziqi , Graham Nicholas A. , Vashisht Ajay A. , Wohlschlegel James A. , Graeber Thomas G. , Karnes R Jeffrey , Takhar Mandeep , Davicioni Elai , Tomlins Scott A. , Sharifi Nima , Witte Owen N. , Feng Felix Y. , Knudsen Karen E.
Published: August 2015

Emerging evidence demonstrates that the DNA repair kinase DNA-PK exerts divergent roles in transcriptional regulation of unsolved consequence. Here, in vitro and in vivo interrogation demonstrate that DNA-PK functions as a selective modulator of transcriptional networks that induce cell migration, invasion, and metastasis. Accordingly, suppression of DNA-PK inhibits tumor metastases. Clinical assessment revealed that DNA-PK is significantly elevated in advanced disease, and independently predicts for metastases, recurrence, and reduced overall survival. Further investigation demonstrated that DNA-PK in advanced tumors is highly activated, independently of DNA damage indicators. Combined, these findings put forth new paradigms for DNA-PK function, identify DNA-PK as a potent driver of tumor progression and metastases, and nominate DNA-PK as a therapeutic target for advanced malignancies.

Citation Format: Jonathan F. Goodwin, Vishal Kothari, Justin M. Drake, Shuang Zhao, Emanuela Dylgjeri, Jeffry L. Dean, Matthew J. Schiewer, Christopher McNair, Michael S. Magee, Robert B. Den, Ziqi Zhu, Nicholas A. Graham, Ajay A. Vashisht, James A. Wohlschlegel, Thomas G. Graeber, R Jeffrey Karnes, Mandeep Takhar, Elai Davicioni, Scott A. Tomlins, Nima Sharifi, Owen N. Witte, Felix Y. Feng, Karen E. Knudsen. DNA-PK-mediated transcriptional regulation drives tumor progression and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1860. doi:10.1158/1538-7445.AM2015-1860
Abstract 1862: Discerning the role of USP22 in prostate cancer development and progression
Authors: Jones Jennifer , Dean Jeffry L. , Schrecengost Randy S. , Knudsen Karen
Published: August 2015

Prostate cancer (PCa) is the second leading cause of cancer-related death for men in the United States. While organ-confined disease is manageable, advanced and disseminated PCa currently has no durable treatment options. Thus, understanding the causes and consequences of the transition from early stage to late stage castrate-resistant PCa (CRPC) are critical. While this transition has a requisite for the androgen receptor (AR) activity, the mechanisms by which AR and other oncogenes are functionally increased, even after initial treatment with androgen deprivation therapy (ADT) have not been completely characterized. USP22, a known deubiquitinase associated with the SAGA transcriptional activation complex, was originally designated in a “death from cancer” gene signature. Importantly, USP22 is significantly upregulated in PCa patients with late-stage disease, and specifically indicates for poor outcome in PCa patients. Moreover, tumor-associated USP22 increases AR levels and activity as well as the oncogene MYC function, partially defining the mechanism by which tumor-associated USP22 drives PCa progression. Thus, while certain consequences of tumor-associated USP22 expression have been elucidated, the remaining downstream effects have not been thoroughly characterized. Newly generated data begin to establish the biochemical events controlled by USP22 that confer proliferation and survival on PCa cells. Through use of novel proteomic technology, specifically Ubiscan analysis, tumor-associated USP22 will be shown to participate in driving PCa development and progression as well as therapeutic bypass through differential regulation of the ubiquitylome.

Citation Format: Jennifer Jones, Jeffry L. Dean, Randy S. Schrecengost, Karen Knudsen. Discerning the role of USP22 in prostate cancer development and progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1862. doi:10.1158/1538-7445.AM2015-1862
Abstract 1863: Using a cell based model system to characterize CYP17A1 genetic variants
Authors: Capper Cameron P. , Liu Jiayan , McIntosh Lauren , Larios José M. , Johnson Michael D. , Hollenberg Paul F. , Auchus Richard J. , Rae James M.
Published: August 2015

Androgen and estrogen steroid hormones are potent growth stimuli for both prostate and breast cancer. Cytochrome P450 17A1 (CYP17A1), a P450 monooxygenase, is required for the synthesis of these hormones and is the target of abiraterone, a drug approved for the treatment of castration-resistant prostate cancer. CYP17A1 catalyzes the 17α hydroxylation of the C21 steroids pregnenolone and progesterone and has a lyase activity that cleaves the C17-20 bond to produce the sex steroid precursors, dehydroepiandrosterone (DHEA) and androstenedione. We identified twelve nonsynonymous single nucleotide polymorphisms (SNPs) in CYP17A1 and hypothesized that the resultant amino acid changes might alter hydroxylase and lyase activity and thereby impact steroid hormone synthesis. This hypothesis was examined by transiently transfecting HEK293T cells, with either wild-type or polymorphic CYP17A1 constructs. Cells were treated for 3 hours with tritiated ([3H]) progesterone and the production of hydroxylated (OH) metabolites analyzed by HPLC. In cells transfected with wild-type CYP17A1, 8.11% and 55.5% of the [3H]-progesterone was recovered as 16-OH progesterone, and 17-OH progesterone, respectively. Using the ratio of 17-OH progesterone to 16-OH progesterone as a quantitative measure of CYP17A1 enzyme activity, wild-type CYP17A1 yields a ratio of 6.9+/- 0.05. In contrast, the SNP (rs200063521) which causes an aspartic acid to histidine amino acid change at residue 216, results in a 2.2 fold decrease in 16OH-progesterone synthesis compared to wild-type suggesting altered hydroxylase function. CYP17A1 protein expression, as determined by western blot analysis, suggests that a glycine to arginine conversion at residue 162 (rs141821705) results in decreased protein levels. Studies to further characterize the CYP17A1 variants are underway. This work could lead to a better understanding of the role of CYP17A1 SNPs in cancer progression and response to CYP17A1 targeted therapies.

Citation Format: Cameron P. Capper, Jiayan Liu, Lauren McIntosh, José M. Larios, Michael D. Johnson, Paul F. Hollenberg, Richard J. Auchus, James M. Rae. Using a cell based model system to characterize CYP17A1 genetic variants. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1863. doi:10.1158/1538-7445.AM2015-1863
Abstract 1865: TRIM24 acts as a transcriptional co-activator of the androgen receptor during prostate cancer progression
Authors: Groner Anna C. , de Tribolet-Hardy Jonas , Theurillat Jean-Philippe , Wild Peter J. , Brown Myles
Published: August 2015

The steroid hormone androgen mediates a wide range of developmental, physiological and malignant responses by acting as an agonist for the androgen receptor (AR). AR in turn functions as a nuclear receptor transcription factor and executes specific gene expression programs in an androgen-dependent manner. Since the growth of prostate cancer cells initially depends on androgen, cancer therapy uses hormone-deprivation approaches to reduce the levels of androgen. While prostate cancer (PC) initially responds to androgen-ablation, most tumors progress to a castration resistant (CR) state insensitive to treatment. Both the androgen-dependent and the CRPC state depend on AR, which is probably activated by alternative molecular pathways in CRPC in response to low androgen levels. Here, we assessed the role of the transcriptional co-activator TRIM24 in mediating AR-dependent gene expression and growth in our CRPC cell model LNCaP-abl. We observed that knock-down of TRIM24 impairs CRPC cell growth, while TRIM24 and AR physically interact in these cells in the absence of androgen. When we identified TRIM24-dependent gene expression programs by combining genome-wide binding analyses with microarray studies, we found that TRIM24 and AR co-regulated many genes in CRPC cells. TRIM24- and AR-co-activated genes often function in mitosis and their expression is higher in patients with an increased chance of prostate cancer recurrence. These findings are corroborated by IHC-analyses of prostate cancer patients, where higher TRIM24 protein levels correlated with a greater chance of biochemical recurrence, higher Gleason scores and larger tumor sizes. Importantly, TRIM24 staining was positively correlated with AR staining. This supports a model where TRIM24 acts as an important activator of AR in the castration-resistant setting and therefore may harbor therapeutic potential.

Citation Format: Anna C. Groner, Jonas de Tribolet-Hardy, Jean-Philippe Theurillat, Peter J. Wild, Myles Brown. TRIM24 acts as a transcriptional co-activator of the androgen receptor during prostate cancer progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1865. doi:10.1158/1538-7445.AM2015-1865
Abstract 1877: Age at introduction to solids is associated with the odds ratio of pediatric acute lymphoblastic leukemia
Authors: Schraw Jeremy M. , Scheurer Michael , Forman Michele R.
Published: August 2015

Introduction: Breastfeeding for > 6 months is associated with protection from acute lymphoblastic leukemia (ALL) (OR 0.81 95% CI 0.72-0.91), the most common pediatric cancer in the United States. These data are based on any breastfeeding. There is little research concerning infant formula or solid food consumption patterns and risk of pediatric ALL. We previously identified greater duration of milk formula feeding and later age at introduction to solid foods as risk factors for ALL. The purpose of this study was to determine whether the increased OR of ALL was confined to children who began receiving solids after 4 or 6 months of age.

Methods: 172 incident cases of ALL aged 0-14 years were recruited at Texas Children's Cancer Center (TCCC) and matched on sex, age at diagnosis/interview and ethnicity to 344 healthy population-based controls from Houston, San Antonio and Austin, Texas. Data were collected on sociodemographics, infant feeding patterns and known or hypothesized risk factors for ALL. Logistic regression was used to model the OR of ALL according to quartile of age at introduction of solids with the first/earliest quartile as the reference group.

Results: In adjusted models, the OR of ALL among children in quartile 3 (7-9 months) was 3.43, 95% CI 1.66 - 7.09; for children in quartile 4 (≥ 10 months) the OR was 4.19, 95% CI 2.00 - 8.75. Each additional month of milk formula feeding was associated with a 7% increase in the OR of ALL (95% CI 1.02 - 1.12). The OR of ALL among non-Hispanic compared to Hispanic children was 1.76 (95% CI 1.10 - 2.84).

Discussion: These results support our published findings that later introduction to solids is positively associated with ALL. Our finding of lower risk among Hispanic children contrasts with previous research indicating this group is at greater risk than non-Hispanics.

Citation Format: Jeremy M. Schraw, Michael Scheurer, Michele R. Forman. Age at introduction to solids is associated with the odds ratio of pediatric acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1877. doi:10.1158/1538-7445.AM2015-1877
Abstract 1886: Dietary scores and cancer incidence, mortality and survival: Epidemiologic evidence and meta-analysis
Authors: Mccullough Marjorie L. , Wang Ying , Shah Roma , Gapstur Susan
Published: August 2015

Diet scores or indices assess the degree to which an individual's reported diet conforms to dietary recommendations or culturally-defined healthful diets. Characterizing diet exposures in this way provides a more complete picture of individual dietary behavior, and potentially captures additive and/or interactive effects on disease risk. Such metrics, where higher points reflect better diets, and lower scores reflect worse diets, have been examined in relation to cancer incidence, mortality and survival. To date, the literature on the association between diet scores and cancer risk is inconsistent across and within scores. We conducted a literature search for prospective cohort studies of a priori dietary scores or indices in relation to cancer incidence, mortality or survival using PubMed. Meta-analyses were performed when at least five separate studies published results on a cancer outcome, using the same diet score. When studies on both incident and fatal outcomes were available from the same cohort for the same cancer/s, estimates from incident outcomes were used. If more than one version of the same score in the same cohort was evaluated, we used the most recent score. Estimates in men and women were included separately. Meta-analyses of four out of six diet scores and the outcome “total cancer incidence or mortality” showed significant inverse associations comparing top vs bottom quantiles, often with significant study heterogeneity: the Alternate Healthy Eating Index (AHEI) Relative Risk (RR) = 0.88, 95% confidence interval (CI) 0.84-0.93, p heterogeneity = 0.02; Healthy Eating Index (HEI) RR = 0.84, 95% CI 0.77-0.92, p heterogeneity<0.001; Mediterranean Diet Score (MDS) RR = 0.81, 95% CI 0.78-0.85, p heterogeneity = 0.35; and WCRF/AICR RR = 0.77, 95% CI 0.68-0.86, p heterogeneity = 0.046. The associations with the Recommended Food Score (RFS) and Diet Quality Index (DQI) were not statistically significant. The MDS was associated with significantly lower colorectal cancer risk (RR = 0.82, 95% CI 0.73-0.91, p heterogeneity = 0.034). In studies of cancer survivors, less than three studies were available for each diet score/outcome combination; thus no meta-analyses of cancer survival were conducted. These results suggest that following recommended dietary guidelines or the Mediterranean healthy diet pattern may significantly reduce overall cancer risk. The limited number of studies in cancer survivors indicates a need for further research.

Citation Format: Marjorie L. Mccullough, Ying Wang, Roma Shah, Susan Gapstur. Dietary scores and cancer incidence, mortality and survival: Epidemiologic evidence and meta-analysis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1886. doi:10.1158/1538-7445.AM2015-1886
Abstract 1906: CXCR4 is a novel target of cancer chemopreventative isothiocyanates in prostate cancer cells
Authors: Amjad Ali I. , Sakao Kozue , Vyas Avani R. , Chinni Sreenivasa R. , Parikh Rahul , Singh Shivendra V.
Published: August 2015

Isothiocyanates (ITCs) derived from cruciferous vegetables, including phenethyl isothiocyanate (PEITC) and sulforaphane (SFN), exhibit in vivo activity against prostate cancer in xenograft and transgenic mouse models, and thus are appealing for chemoprevention of this disease. Watercress constituent PEITC and SFN-rich broccoli sprout extract are under clinical investigation but the molecular mechanisms underlying their cancer chemopreventive effects are not fully understood. The present study demonstrates that chemokine receptor CXCR4 is a novel target of ITCs in prostate cancer cells. Exposure of prostate cancer cells (LNCaP, 22Rv1, C4-2, and PC-3) to pharmacologically applicable concentrations of PEITC, benzyl isothiocyanate (BITC), and SFN (2.5 and 5 μmol/L) resulted in downregulation of CXCR4 expression. None of the isothiocyanates affected secretion of CXCR4 ligand (stromal-derived factor-1). In vivo inhibition of PC-3 xenograft growth upon PEITC treatment was associated with a significant decrease in CXCR4 protein level. A similar trend was discernible in the tumors from SFN-treated TRAMP mice compared with those of control mice, but the difference was not significant. Stable overexpression of CXCR4 in PC-3 cells conferred significant protection against wound healing, cell migration, and cell viability inhibition by ITCs. Inhibition of cell migration resulting from PEITC and BITC exposure was significantly augmented by RNA interference of CXCR4. This study demonstrates, for the first time, that cancer chemopreventive ITCs suppress CXCR4 expression in prostate cancer cells in vitro as well as in vivo. These results suggest that CXCR4 downregulation may be an important pharmacodynamic biomarker of cancer chemopreventative ITCs in prostate adenocarcinoma.

Citation Format: Ali I. Amjad, Kozue Sakao, Avani R. Vyas, Sreenivasa R. Chinni, Rahul Parikh, Shivendra V. Singh. CXCR4 is a novel target of cancer chemopreventative isothiocyanates in prostate cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1906. doi:10.1158/1538-7445.AM2015-1906
Abstract 1908: Evaulate biomarkers in NSCLC tumors receiving neoadjuvant chemotherapy
Authors: Pataer Apar , Shao Ruping , Correa Arlene M. , Behrens Carmen , Roth Jack A. , Wistuba Ignacio I. , Swisher Stephen G.
Published: August 2015

In previous study, we have evaluated the ability of histopathologic response criteria to predict overall survival (OS) and disease free survival (DFS) and observed that the percentage of residual viable tumor cells were associated with OS and DFS in 192 NSCLC patients receiving neoadjuvant chemotherapy. In current study, we examined the selected protein biomarkers by IHC in surgical specimens of 98 NSCLC patients receiving neoadjuvant chemotherapy. All NSCLC patients treated with neoadjuvant chemotherapy received a platinum and taxane-based. For IHC analysis of protein markers, we selected candidate biomarkers (VEGFR2, EZH2, ERCC1 and Rad51) on the basis of the literature. Next, the results of VEGFR2, EZH2, ERCC1 and Rad51 expression were analyzed with regard to pathologic response and overall survival time of the patients. The surgical pathologic stage, percentage of viable tumor cells and Rad51 were associated with OS in 98 NSCLC patients receiving neoadjuvant chemotherapy in univariate and multivariate analysis. We found that patients with high-level expression of Rad51 had a poorer prognosis than did those with low-level expression. We observed that Rad51 expression was associated with pathologic response (viable tumor cells). However, there is no association between VEGFR2, EZH2, or ERCC1 expression and pathologic response (viable tumor cells). We further examined K-Ras and EGFR Mutation in these NSCLC tumors receiving neoadjuvant chemotherapy. We analyzed the hotsport mutations in K-Ras gene (Codon 12 and 13) and EGFR gene (Exon 19 and 21) by pyrosequencing and confirmed by direct sequencing method in 98 NSCLC tumors receiving neoadjuvant chemotherapy. We observed both methods had similar results on these patients’ samples. The L858R mutation results in an amino acid substitution at position 858 in EGFR, from a leucine (L) to an arginine (R). This mutation occurs within exon 21, which encodes part of the kinase domain,We found K-Ras mutation tended to be associated with OS, but not EGFR mutation. There is no association between percentage of viable tumor cells, VEGFR2, EZH2, ERCC1, Rad51 expression and K-Ras or EGFR mutation.

Citation Format: Apar Pataer, Ruping Shao, Arlene M. Correa, Carmen Behrens, Jack A. Roth, Ignacio I. Wistuba, Stephen G. Swisher. Evaulate biomarkers in NSCLC tumors receiving neoadjuvant chemotherapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1908. doi:10.1158/1538-7445.AM2015-1908
Abstract 1924: Genes associated with histopathologic features of triple negative breast tumors predict molecular subtypes
Authors: Purrington Kristen , Visscher Daniel , Yannoukakos Drakoulis , Carpenter Jane , Nevanlinna Heli , Mannerma Arto , Wang Xianshu , Giles Graham , Zheng Wei , Cox Angela , Brauch Hiltrud , Hamann Ute , Eccles Diana , Vachon Celine M. , Couch Fergus J.
Published: August 2015

Distinct subtypes of triple negative (TN) breast cancer have currently been identified by tumor expression profiling and are thought to be influenced by the tumor microenvironment. However, little is known about the relationship between distinguishing histopathologic features of TN tumors, which reflect aspects of both tumor behavior and tumor microenvironment, and molecular TN subtypes. We analyzed whole genome expression profiles of 593 TN tumors from the Triple Negative Breast Cancer Consortium (TNBCC) using the Illumina Whole Genome DASL array. Histopathologic review of mitotic index, grade, trabecular pattern, growth pattern, involution, presence of ductal carcinoma in situ, and degree of lymphoid infiltrate, necrosis, fibrosis, and lobulitis was completed for 310 of these tumors. Clusters were identified both agnostically and using genes associated with these histopathologic features, and subtypes were evaluated for enriched pathways and associations with histopathologic features. Agnostic clustering identified four stable clusters which represented a basal-like (BL) signature, a luminal signature with overexpression of androgen receptor and targets, an immune signature, and a stromal (STR) signature. Overall, the TN tumors tended to have little or no necrosis (91%), high grade (88%), a high mitotic index (80%), fibrosis (77%), an epicentric growth pattern (62%), incomplete lobular involution (55%), trabecular structures (37%), lymphocytic infiltration (25%), adjacent ductal carcinoma in situ (24%), and variable amounts of lobulitis. Using a combined total of 2,776 unique probes individually associated with the ten histopathologic features measured, we identified six distinct TN subtypes, with an additional BL and STR signature compared to the four agnostic subtypes. Our findings suggest that histopathologic features that reflect heterogeneity in tumor cell structure and behavior as well as the tumor microenvironment, particularly the presence of lymphocytic infiltrate, are critical for TN subtype identification. Further work is needed to validate these signatures and to better understand the immunologic pathways associated with each subtype.

Citation Format: Kristen Purrington, Daniel Visscher, Drakoulis Yannoukakos, Jane Carpenter, Heli Nevanlinna, Arto Mannerma, Xianshu Wang, Graham Giles, Wei Zheng, Angela Cox, Hiltrud Brauch, Ute Hamann, Diana Eccles, Celine M. Vachon, Fergus J. Couch. Genes associated with histopathologic features of triple negative breast tumors predict molecular subtypes. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1924. doi:10.1158/1538-7445.AM2015-1924
Abstract 1938: Comprehesive RNA-sequencing pipeline of cancer xenograft models
Authors: Hwang Jinha , Kim Jong-Il , Choi Jee-Soo , Kim Min Jung , Lee Charles
Published: August 2015

Since a solid pipeline for massively parallel sequencing in cancer xenograft model has not been built yet, it is necessary to establish a precise analysis pipeline. Massively parallel sequencing on xenograft samples result in mixed sequencing reads from the engrafted human tumor cells and murine tissue. It is necessary to accurately distinguish between these reads for subsequent data analysis. In the present study, we constructed an integrated RNA-Seq pipeline including genomic variation, expression profiling, and fusion gene analysis. To distinguish these reads into individual species, we used the merged reference that incorporates the two genome reference from human (hg19) and mouse (mm10). We tested our pipeline using 6 Primary and 4 metastatic gastric tumor of 6 patients and their derivative xenografts sequencing data. There were almost same amount of ambiguous reads when we aligned sequencing reads to merged reference or separate reference genome. Using merged reference, we efficiently distinguished compound sequencing reads between human and mouse without repetitive alignment processes. The proportion of sequencing reads from mouse in xenograft samples widely ranged from 2.64% to 49.25% per each sample. For expression profiling, we used HTSeq-count for measuring expression level of each sample. As expected, human-specific reads of ECM-receptor and focal adhesion related genes were specifically down-regulated in the xenograft tumors compared to primary tumor and normal tissues suggesting the depletion of human cells from the microenvironment of engrafted tumor tissues. Mouse-specific reads of those genes were highly expressed in the engrafted tissue. Differentially expressed genes between engraft tumor and paired normal tissue were shown to be enriched in the cell cycle and p53 signaling pathway and these showed that engrafted tumor tissues well represent characteristics of cancer cells.

Note: This abstract was not presented at the meeting.

Citation Format: Jinha Hwang, Jong-Il Kim, Jee-Soo Choi, Min Jung Kim, Charles Lee. Comprehesive RNA-sequencing pipeline of cancer xenograft models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1938. doi:10.1158/1538-7445.AM2015-1938
Abstract 1952: Adverse effects of erythropoietin stimulates tumor growth via EphB4
Authors: Pradeep Sunila , Huang Jie , Mora Edna , Nick Alpa , Cho Minsoon , Wu Sherry , Rupaimoole Rajesha , Xiong Chiyi , Li Chun , Lopez-Berestein Gabriel , Jackson David , Sood Anil
Published: August 2015

Abstract: While recombinant human erythropoietin (rhEpo) has been widely used to treat anemia in cancer patients, concerns about its adverse effects on patient survival have emerged. However the lack of correlation between expression of the canonical EpoR and rhEpo's effects on cancer cells prompted us to consider the existence of an alternative Epo receptor. Methods: We analyzed the membrane-bound portion of the proteome for candidate receptors possessing structural, regulatory, and functional features consistent with Epo binding and tumorigenic signaling. To examine the potential for EphB4 binding to Epo, we generated EpoR−/- MEFs from EpoR−/- mice. We injected A2780-shControl and-shEpoR cells, shEphB4 or -shEpoR/shEphB4 cells in nude mice to study rhEpo induced tumor progression. We also assessed potential effects of tumoral EpoR or EphB4 expression on the effects of ESA treatment in patients with ovarian (N = 175) or breast (N = 88) cancer. Results: Through systems-based approaches we identified EphB4 as an alternate Epo receptor. MST experiments with fluorophore labeled rhEpo demonstrated binding to EpoR with an apparent dissociation constant (KD,app) of 28.0 ± 14.0 nM and EphB4 with a KD,app value of 880.6 ± 128.6 nM. EphB4 and EphrinB2 bound with a KD,app value of 140.5 ± 28.3 nM. We showed that [I125]rhEpo can bind to these cells EpoR−/- MEFs cells.

EphB4 triggered the downstream signaling via STAT3 and promoted rhEpo-induced tumor growth and progression. rhEpo stimulated in vivo growth of A2780-shControl (p<0.01) and-shEpoR cells (p<0.01), but not -shEphB4 or -shEpoR/shEphB4 cells. In human ovarian and breast cancer samples, expression of EphB4 rather than the canonical EpoR correlated with decreased disease-specific survival in rhEpo-treated patients. Conclusion: These results identify EphB4 as a critical mediator of erythropoietin-induced tumor progression in cancer patients and further provide a novel and clinically significant dimension to the biology of erythropoietin.

Citation Format: Sunila Pradeep, Jie Huang, Edna Mora, Alpa Nick, Minsoon Cho, Sherry Wu, Rajesha Rupaimoole, Chiyi Xiong, Chun Li, Gabriel Lopez-Berestein, David Jackson, Anil Sood. Adverse effects of erythropoietin stimulates tumor growth via EphB4. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1952. doi:10.1158/1538-7445.AM2015-1952