bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5716: A study on the mechanism of the chemopreventive effect of different ratios of fish oil and corn oil in experimental colon carcinogenesis
Authors: Agnihotri Navneet , Sarotra Pooja , Sharma Gayatri , Kansal Shevali
Published: February 2011

AIM: The present study was designed to study the effect of different ratios of n-3 rich fish oil and n-6 rich corn oil in the prevention of colon cancer mediated through cell signaling and apoptosis.

Methods: Male Wistar rats were divided into the following groups -

Group-I and Group-II were fed synthetic diets and Group-III and Group-IV modified synthetic diets with 1:1 and 2.5:1 ratio of fish oil and corn oil for 4 weeks. All the groups except group-I (vehicle treated) received 4 injections of dimethyl hydrazine (DMH). The DMH injections were given after initiation on the synthetic diet for a week and subsequently received one injection per week. The animals were sacrificed 48 hrs after last injection. The last 10cm of the distal colon was used for histopathological study while the rest was used for isolation of colonocytes. The differential effect of the different ratios of oil on oxidative stress by measuring reactive oxygen species’(ROS) generation by FACS analysis using dichlorofluorescin diacetate (DCFH-DA) and p53 levels using monoclonal antibody was assessed. The apoptosis was evaluated by DNA fragmentation and by using M30 cytodeath antibody. To understand the role of mitochondria in apoptosis mitochondrial membrane potential was measured using JC-1. Akt levels were also measured using a monoclonal antibody in orderto understand its effect on p53 and apoptosis.

Results: In the carcinogen administered animals as compared to the vehicle treated animals, there was- 1.mild to moderate inflammation 2. a decrease in ROS levels 3. a decrease in p53 4. an increase in apoptosis 5. an increase in mitochondrial membrane potential 6. an increase in Akt.

Administration of fish oil and corn oil led to the following- 1. Fish oil feeding in the ratio 1:1 showed severe inflammation in the animals while feeding a higher ratio showed mild inflammation with normal mucosa. 2. a decrease in ROS levels with equal ratios as compared to the higher ratio of fish oil. 3. an increase in p53 levels with both the ratios 4. a decrease in apoptosis 5. a decrease in mitochondrial membrane potential 6. a decrease in Akt in equal ratios but a significant increase in the higher ratios.

Conclusion: The results of the present study suggest that administration of fish oil in the higher ratio has a beneficial role in colon cancer as shown by histopathological examination. Fish oil derived n-3 PUFA exerts chemopreventive effect mediated by interplay among oxidative stress, apoptosis and cell survival signals depending on the dosage.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5716.
Abstract 5720: Chromosomal instability predicts the progression of premalignant oral lesions
Authors: Hamers Rinske MPT , Siebers Theke JH , Haesevoets Annick , Zwijnenberg Iris R. , Voogd Adri C. , Ramaekers Frans CS , Kremer Bernd , Merkx Thijs MAW , Slootweg Pieter J. , Speel Ernst-Jan M.
Published: February 2011

A major dilemma in the management of patients with precursor lesions of the oral mucosa lies in deciding which lesions will progress into carcinoma. Because a strong association between histological differentiation and clinical prognosis is lacking, the aim of this study was to evaluate the value of chromosomal instability (CIN) detected by fluorescence in situ hybridization (FISH) for the identification of oral premalignant lesions at risk for progression.

We examined a series of premalignant oral mucosa lesions of 106 patients by means of FISH on paraffin-embedded tissue sections using biotin- and digoxigenin-labeled probes specific for the centromeric regions of chromosomes 1 and 7. CIN was indicated by the presence of chromosome imbalances and/or polyploidization for chromosomes 1 and 7. Results were correlated with histopathological data as well as with clinical follow up data.

The 5-year progression-free survival rate was 83%. Outcome of routine histopathology did only predict malignant progression when comparing severe dysplasia with lower stage precursor lesions (hyperplasia, mild en moderate dysplasia) (p<0.001). CIN was detected in all different subgroups of histopathological differentiation. Moreover, the percentage of precursor lesions harboring CIN increased with progressing histopathological stage. CIN was significantly associated with a lower progression-free survival as compared with lesions without CIN (p<0.001).

In conclusion, tumorigenesis of the oral mucosa is associated with the development of CIN, which can reliably identify lesions at risk for progression.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5720.
Abstract 573: 18alpha- glycyrrhetinic acid (AGA) inhibits advanced prostate cancer cell proliferation and induces apoptosis by downregulating inflammation-related cytokine growth factor genes
Authors: Munirathinam Gnanasekar , Dakshinamoorthy Gajalakshmi , Olesen John Eric , Sivasakthivel Thirugnanam , Zheng Guoxing , Chen Aoshuang , Bosland Maarten , Kajdacsy-Balla André , Kalyanasundaram Ramaswamy
Published: February 2011

Glycyrrhetinic acid is an active triterpenoid metabolite of glycyrrhizin abundantly present in licorice roots. Glycyrrhetinic acid exists as alpha and beta stereo-isomeric forms. Both stereo-isomeric forms are known to have anti-inflammatory and anti-cancer activity. However, the effects and anti-cancer mechanism of alpha glycyrrhetinic acid in prostate cancer cells is not yet evaluated. Therefore in this study, we have investigated the growth inhibition, induction of apoptosis and the anti-cancer mechanisms of 18α-glycyrrhetinic acid (AGA) on androgen independent metastatic prostate cell line DU-145. Our results showed that AGA inhibited proliferation and growth of these cells in a time and dose dependent manner. Induction of apoptosis was confirmed by Annexin V-FITC and propidium iodide staining. Further analysis showed that AGA treatment activated caspase- 8 and −3 death related enzymes in DU-145 cells. Interestingly, AGA treatment also simultaneously down regulated the expression of proinflammatory cytokine/growth factor genes HMGB1, IL-6 and IL-8. This is the first report demonstrating that AGA could play a dual role as an inducer of apoptosis and suppressor of pro-inflammatory gene expression in DU-145 prostate cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 573.
Abstract 5736: Household endotoxin levels and risk of non-Hodgkin lymphoma
Authors: Wang Jun , Colt Joanne S. , Thorne Peter S. , Berhane Kiros T. , Richard Severson K. , Davis Scott , Cerhan James R. , Ward Mary H. , Bernstein Leslie , Morton Lindsay M. , Patricia Hartge , Cozen Wendy
Published: February 2011

Subclinical variation in immune response may be a risk factor for non-Hodgkin lymphoma (NHL). Endotoxin is a substance located in the cell wall of gram-negative bacteria that elicits a strong innate and inflammatory immune response. It is present in large amounts in animal and human fecal material, other organic material, and cigarette smoke. We hypothesized that higher endotoxin exposure might be a risk factor for NHL because of its effect on B-cell activation. Using a population based, multi-center (Detroit, Iowa, Los Angeles and Seattle) case-control study, we evaluated the association between endotoxin levels in carpet dust and NHL risk. During an in-person interview, we collected used vacuum cleaner bags from participants who had used their vacuum within the past year and had owned at least half of their carpets or rugs for 5 years or more. Dust samples were analyzed using the kinetic chromogenic Limulus amebocyte lysate assay in the Pulmonary Toxicology Facility, University of Iowa. Endotoxin measurements from 629 cases and 467 controls were log-transformed and evaluated as both continuous and categorical variables. In order to examine a possible threshold effect, endotoxin levels among controls were categorized into tertiles, with the highest tertile further split into two equal groups based on a preliminary model search. Multivariable logistic regression was used to estimate the effect of endotoxin on NHL risk adjusted for sex, age, race, education, study center, and last change of vacuum bag. The highest category of household endotoxin level in dust was associated with a 35% significantly decreased risk of NHL relative to the lowest category (95% confidence interval [CI] = 0.44, 0.96). The effect was stronger for the diffuse large B-cell lymphoma (DLBCL) subtype (Odds Ratio [OR]= 0.49; 95% CI= 0.26,0.91). In conclusion, contrary to our initial hypothesis, our results suggest an inverse association between high levels of household endotoxin and NHL risk. Further studies are needed to validate this finding, ideally with endotoxin measurements prior to diagnosis. Possible mechanisms of action will be discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5736.
Abstract 5737: Parity and lactation in relation to ER-/PR- breast cancer in African American women
Authors: Palmer Julie R. , Boggs Deborah A. , Adams-Campbell Lucile L. , Rosenberg Lynn
Published: February 2011

Women of African ancestry are more likely than women of European ancestry to be diagnosed with aggressive tumors characterized by negative staining for estrogen receptors (ER) and progesterone receptors (PR). Reasons for the disparity are unclear, and previous studies have lacked statistical power to investigate risk factors for the more aggressive subtypes. A recent report from the Carolina Breast Cancer Study indicated that basal-like breast cancer, a subtype of breast tumors with ER-, PR-, HER2-, HER1+ and/or CK5/6+ characteristics, was positively associated with increased parity, whereas other subtypes were inversely associated with parity. The increased risk of basal-like cancer was strongest for parous women who had never breastfed. The majority of breast tumors with negative staining for both ER and PR would likely be classified as basal-like. We examined the relation of parity and lactation to risk of ER-/PR- breast cancer among 59,000 African American women from the Black Women's Health Study in 12 years of follow-up data. Reproductive history was obtained by questionnaire at baseline in 1995 and updated every two years. ER and PR status were obtained from pathology reports and state cancer registry records. Included in this analysis were 809 incident breast cancer cases with complete data on ER and PR status, with 285 classified as ER-/PR-, 395 as ER+/PR+, 114 as ER+/PR-, and 15 as ER-/PR+. Incidence rate ratios (IRR) and 95% confidence intervals (CI) were derived from proportional hazard models for the relation of each reproductive factor to risk of specific tumor subtype, with control for all other reproductive factors, age, region, education, body mass index, waist-hip ratio, hormone use, alcohol, smoking, and physical activity. Risk of ER-/PR- breast cancer increased with increasing parity (p trend = 0.02), whereas risk of ER+/PR+ cancer decreased with increasing parity (p trend = 0.0002). For ER-/PR- breast cancer, the IRR for women who had 3+ births and never breastfed relative to nulliparous women was 1.66 (95% CI 1.09-2.54). The increased risk associated with higher parity was ameliorated by breast feeding: the comparable IRR for women who had 3+ births but breastfed at least one of the babies was 1.01 (95% CI 0.63-1.63). These results from a cohort of African American women support findings from the previous study of basal-like breast cancer and are consistent with the hypothesis that pregnancy and breastfeeding may influence risk of ER-/PR- cancer through inflammatory/immune system alterations linked to mammary gland involution.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5737.
Abstract 5742: Hand pattern and prostate cancer risk
Authors: Rahman Aneela A. , Lophatananon Artitaya , Harris Duncan , Anderson John , Easton Douglas , Key Tim , Pocock Richard , Dearnaley David , Guy Michelle , Edwards Steven , O'Brien Lynne , Wilkinson Rosemary A. , Hall Amanda L. , Sawyer Emma , Page Elizabeth , Liu Jo-Fen , Eeles Rosalind , Muir Kenneth
Published: February 2011

ABSTRACT

Background

Hand pattern (pattern of index compared to ring finger on the right hand, 2D:4D) has been studied for a number of decades as a potential predictor for disease occurrence. It is known to be associated with certain hormone related diseases developed during adult life but there have been no studies so far of 2D:4D ratio and prostate cancer risk. This study assesses the association between hand and prostate cancer.

Methods

We analysed the right hand pattern and prostate cancer risk in 1524 cases aged ≤ 80 and 3044 population-based controls. Odds ratio (OR) and 95% confidence interval (CI) were calculated using multivariate logistic regression (to allow for adjustment of confounders) estimating whether there was any association between right hand pattern and the risk of prostate cancer.

Results

The reference group was chosen to be the commonest hand pattern seen in males which is index finger shorter than ring finger. Compared to this group there was no difference in men with index finger equal to ring finger (OR 1.05, 95% CI 0.88-1.25) whereas men with index finger longer than ring finger showed a negative association indicative of a marker of a protective effect with a 33% risk reduction compared to index finger shorter than ring finger (OR 0.67, 95% CI 0.57-0.80).

Conclusion

This study showed that the pattern of finger lengths appears to be a community based surrogate marker of prostate cancer risk with length of 2D greater than 4D indicative of lower risk which has previously been shown to be also associated with lower prenatal testicular activity.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5742.
Abstract 5753: RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth
Authors: Hatzivassiliou Georgia , Song Kyung , Yen Ivana , Brandhuber Barbara J. , Anderson Daniel J. , Alvarado Ryan , Ludlam Mary JC , Stokoe David , Gloor Susan L. , Vigers Guy , Morales Tony , Aliagas Ignacio , Liu Bonnie , Sideris Steve , Hoeflich Klaus , Jaiswal Bijay S. , Seshagiri Somasekar , Koeppen Hartmut , Belvin Marcia , Friedman Lori S. , Malek Shiva
Published: February 2011

Activating mutations in KRAS or B-RAF are found in over 30% of all human tumors and targeting this pathway could have broad therapeutic impact. Small molecule ATP-competitive RAF kinase inhibitors have potent anti-tumor effects on B-RAFV600E tumors but, in contrast to MEK inhibitors, are not potent against RAS mutant tumor models, despite RAF functioning as a key effector downstream of RAS and upstream of MEK. In this study we show that ATP-competitive RAF inhibitors have two opposing mechanisms of action depending on cellular context. In B-RAFV600E tumors, RAF inhibitors effectively block the MAPK signaling pathway and decrease tumor growth. Surprisingly, in KRAS mutant and RAS/RAF wildtype tumors, RAF inhibitors activate the RAF/MEK/ERK pathway in a RAS-dependent manner, thus enhancing tumor growth in some xenograft models. Inhibitor binding activates wildtype RAF isoforms by inducing dimerization, membrane localization and interaction with RAS-GTP. These events occur independently of kinase inhibition and are, rather, linked to direct conformational effects of inhibitors on the RAF kinase domain. Based on these findings, we demonstrate that ATP-competitive kinase inhibitors can have opposing functions as inhibitors or activators of signaling pathways, depending upon cellular context. In addition, this work provides new insights into the therapeutic use of ATP-competitive RAF inhibitors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5753.
Abstract 5760: 14-3-3zeta cooperates with ErbB2 to promote ductal carcinoma in situ progression to invasive breast cancer by inducing epithelial-mesenchymal transition
Authors: Lu Jing , Li Ping , Yu Dihua
Published: February 2011

ErbB2, a metastasis-promoting oncoprotein, is overexpressed in approximately 25% of invasive/metastatic breast cancers, but in 50%-60% of noninvasive ductal carcinomas in situ (DCIS). It has been puzzling how a subset of ErbB2-overexpressing DCIS develops into invasive breast cancer (IBC). We found that co-overexpression of 14-3-3zeta in ErbB2-overexpressing DCIS conferred a higher risk of progression to IBC. ErbB2 and 14-3-3zeta overexpression, respectively, increased cell migration and decreased cell adhesion, two prerequisites of tumor cell invasion. 14-3-3zeta overexpression reduced cell adhesion by activating the TGF-beta/Smads pathway that led to ZFHX1B/SIP-1 upregulation, E-cadherin loss, and epithelial-mesenchymal transition. Importantly, patients whose breast tumors overexpressed both ErbB2 and 14-3-3zeta had higher rates of metastatic recurrence and death than those whose tumors overexpressed only one. Therefore, we discovered that 14-3-3zeta overexpression is a “second hit” or “risk factor” facilitating a subset of ErbB2-overexpressing DCIS transition into IBC and identified molecular mechanisms/pathways through which ErbB2 and 14-3-3zeta co-overexpression promotes invasion. This study identified biomarkers that allow selection of high-risk DCIS patients for more aggressive treatments at early stages of cancer development, while saving low-risk patients from ablative clinical procedures. Moreover, it provided promising targets for future intervention strategies to prevent DCIS progression to IBC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5760.
Abstract 5761: An alternative isoform of the metastasis susceptibility gene Brd4 promotes EMT and metastatic progression
Authors: Alsarraj Jude , Walker Renard C. , Crawford Nigel P.S. , Webster Joshua D. , Simpson R. Mark , Ozato Keiko , Hunter Kent W.
Published: February 2011

Recent work from our laboratory has identified the bromodomain 4 (Brd4) as a suppressor of tumor progression and metastasis. Brd4 however has an alternatively spliced isoform that truncates the protein and substitutes a different 3′-UTR. Unlike the longer isoform, expression of the short isoform (ΔC) does not impair primary tumor growth of a highly metastatic mammary tumor cell line orthotopically implanted into the mammary fat pad, although pulmonary metastasis is significantly reduced compared to control tumors. Unexpectedly, a dramatic enhancement of metastatic colonization is observed following injection of the cells directly into the circulation of mice. In vivo invasion of the cell line was not impaired by expression of the short isoform suggesting that the inability of the orthotopic tumors to metastasize was not due to the inability of the tumor cells to escape from the primary tumor. Consistent with the tail vein experiments, in vitro analyses demonstrate that ΔC-expressing cells exhibit a more malignant phenotype with an increased ability to grow in a three dimensional culture compared to control cells. Furthermore, ΔC-expressing cells are able to form tumorspheres in serum-free and non adherent conditions, suggesting a reversion to a more stem-like state. Microarray gene expression analysis demonstrates that ectopic expression of ΔC modulates the expression of a subset of EMT markers and stem cell markers. Microarray analysis further shows that ectopic expression of ΔC induces a gene expression signature that predicts poor survival in multiple human breast cancer datasets. This expression signature nearly perfectly matches a molecular classifier of high-grade tumors. Taken together, these data suggest that the short isoform may be a metastasis-promoting molecule and that metastasis susceptibility mediated by Brd4 may be the result of a complex equilibrium between the metastasis-suppressing long isoform and the metastasis-promoting shorter isoform.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5761.
Abstract 5764: Opposing roles of the COX-2 pathway receptors EP1 and EP4 in metastasis and contribution to disparities
Authors: Fulton Amy M. , Ma Xinrong , Kundu Namita , Ioffe Olga , Goloubeva Olga , Konger Raymond , Baquet Claudia , Reader Jocelyn
Published: February 2011

Cyclooxygenase-2 is highly expressed in breast and other malignancies and contributes to a poor prognosis. Our goal is to identify novel approaches to target the COX-2 pathway in order to reduce metastatic potential and improve survival. The primary COX-2 product, prostaglandin E2, exerts cellular effects by binding to four distinct G-protein-coupled receptors designated EP1, EP2, EP3 and EP4. Using pharmacologic and genetic approaches and a preclinical model of metastatic, triple-negative (ER, PR, Her-2 negative) breast cancer, we have examined the role of EP4 and EP1 in malignant behavior. Inhibition of EP4 receptor signaling with selective EP4 antagonists limits breast cancer metastasis to the same degree as achieved with a COX-2 inhibitor. Likewise, gene silencing of EP4 with specific shRNA also limits metastatic capacity by a mechanism involving Natural Killer cells. Conversely, blocking EP1 with either pharmacologic antagonists or genetic ablation promotes tumor metastasis. The expansion of mammary-gland implanted tumors was not affected by changes in EP1 expression levels, indicating that EP1 functions selectively to suppress metastasis. In primary breast tumors or cell lines, EP1 protein is detected both on the plasma membrane and in the nucleus. Immunohistochemical examination of a tissue array containing primary breast tumors reveals that the absence of nuclear EP1 in malignant cells is associated with significantly worse long term survival. Levels of cytoplasmic EP1 were not correlated with differences in outcome. Interestingly, breast tumors from African-American women express significantly less EP1 mRNA than tumors from Caucasian women, suggesting that low EP1 could contribute to worse outcome in AA women. These data support the hypothesis that EP1 acts as a metastasis suppressor but EP4 promotes metastasis and that shifting the balance towards activation of EP1 could be a novel approach to reduce breast cancer metastasis and reduce disparities in outcome.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5764.
Abstract 5766: The Hsp90 inhibitor PU-H71 is a potent suppressor of metastasis in triple-negative breast cancer models
Authors: Lopes Eloisi Caldas , Bos Paula , Peinado Hector , Cerchietti Leandro , Lavotshkin Simon , Melnick Ari , Lyden David , Chiosis Gabriela
Published: February 2011

Background: Triple-negative breast cancer (TNBC) characterized by estrogen and progesterone receptor and HER-2-negativity is a particularly aggressive tumor with currently no effective targeted therapies. Our previous findings demonstrated that Hsp90 inhibition by the purine-scaffold inhibitor PU-H71 is a promising therapeutic approach in this tumor type. In TNBC cells, a multitude of malignancy driving proteins, including EGFR, HER3, Akt, PDK1, Bcl-xL and Raf-1, were in a complex with Hsp90 and were preferentially targeted for degradation by PU-H71 (Abstract 5637, AACR-Annual Meeting 2008/2009). In TNBC xenografts, PU-H71 exhibited potent and durable anti-tumor activity inducing complete responses without toxicity to the host. PU-H71 also inhibited the invasiveness of TNBC cells through a mechanism involving downregulation and inhibition of several pro-metastatic proteins such as NF-kB, p-Akt, ERK2, TYK2 and PKC, suggesting a potential effect of PU-H71 on TNBC metastasis.

Study design: In order to determine whether in vivo PU-H71 also inhibits TNBC metastasis, we analyzed the effect of the Hsp90 inhibitor on lung colonization and metastastic body formation in immunocompromised mice. A lung tropic MDA-MB231 derivative expressing luciferase was used to establish the experimental models of metastasis. Animals were treated with 75mg/kg PU-H71 three times/week and lung metastases were monitored periodically by bioluminiscence imaging (IVIS system).

Results: At completion of treatment both bioluminescent imaging and histological analyses demonstrated significant reduction of the metastastatic burden in mice treated with PU-H71 compared to the vehicle treated group. Importantly, flow cytometry analysis demonstrated that circulating hematopoietic precursors related to metastatic spread were reduced after treatment with PU-H71 and several hematopoietic cell types recruited to metastatic sites such as CD11b+/Gr1+ were reduced after the treatment. In addition, analysis of metastatic lungs demonstrated that CD24low/CD44+ cells were attracted to metastatic lung in MDA-MB231 bearing mice and strongly reduced after PU-H71 treatment.

Conclusion: These data demonstrate that in addition to inhibition of primary tumors, PU-H71 leads to potent impairment of lung metastasis in an in vivo model of TNBC, supporting the use of PU-H71 for clinical trials involving patients with advanced and metastatic disease.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5766.
Abstract 5768: Predictive diagnostics from circulating tumor cells
Authors: Atwal Siminder K. , Savage Heidi , Spoerke Jill , Mohan Sankar , Pandita Ajay , Chen Daniel , Lackner Mark , Punnoose Elizabeth
Published: February 2011

Introduction: Tumor tissue collection is essential for biomarker assessment but can be difficult in tumor types such as non small-cell lung cancer (NSCLC) where often no surgery is performed and diagnosis is done with biopsies yielding only very limited tissue quantities. Also, in cases where primary tissue is available, the samples may not be representative of patient's current disease, i.e. in the case of hormone refractory prostate cancer (HRPC).

Rationale: Evaluation of cancer biomarkers (both predictive and pharmacodynamic) from blood and other biological material could significantly enable our ability to generate biomarkers by providing an accessible and relevant source of samples that can be analyzed. Successful assessment of biomarkers that do not require access to tumor “tissue” could improve the inclusion of biomarker evaluations in clinical development and across our oncology practice.

Results: We report on our efforts to evaluate Circulating Tumor Cells (CTC) for utility in tumor biomarker assessment. These efforts included evaluating multiple technologies for isolation and molecular analysis of CTCs from blood. By spiking tumor cells into blood as a model system for CTCs we have developed assays for biomarker evaluation in CTCs including: Fluorescence in situ hybridization (FISH), Immunofluoresecnce (IF), qRT-PCR and mutation detection. Validation of these assays in patient blood is ongoing and early data suggests that biomarker assessment is indeed possible from CTCs. Using blood samples from metastatic, HER2+ breast cancer patients; we were able to evaluate the HER2 status from CTCs using immunofluorescence and/or FISH.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5768.
Abstract 5775: A novel role for ID1 in prostate cancer chemotherapy
Authors: Geng Hao , Beer Tomasz , Nelson Pete , Qian David
Published: February 2011

To identify potential mechanisms underlying prostate cancer chemotherapy response and resistance, we compared the gene expression profiles in individual human prostate cancer specimens before and after chemotherapy collected from previously untreated patients with high-risk, localized disease. Among the molecular signatures associated with treatment, transcripts encoding Inhibitor of DNA Binding 1 (ID1) were significantly upregulated. Among patients who developed biochemical relapse 5 years after chemo-surgery, patients with the ID1 upregulation following chemotherapy had significantly longer relapse-free survival time than patients without ID1 increases. In the prostate cancer LNCaP cell line that is characterized by low ID1 expression, docetaxel dose-dependently induced ID1 expression. This docetaxel-induced ID1 transcription was mediated by ID1 promoter E-box chromatin modification and c-Myc binding. Compared to the empty-vector control, stable ID1 overexpression in LNCaP increased cell proliferation, promoted cell cycle progression through the G1 phase, and enhanced docetaxel-induced cytotoxicity. These phenotypic changes were accompanied by an increase in BCL2 phosphorylation at serine 70, caspase-3 activation and PARP cleavage. In contrast, ID1 siRNA in the LNCaP and C42B cell line reduced cell proliferation and decreased docetaxel-induced cytotoxicity. The ID1-mediated chemosensitivity enhancement was in part due to the ID1-suppression of p21. Overexpression of p21 in LNCaP-ID1 overexpressing cells restored the p21 level and reversed ID1-enhanced chemosensitivity. These data demonstrate that ID1 has a novel therapeutic role in prostate cancer treatment and ID1 upregulation after chemotherapy exposure may be a useful marker of chemotherapy efficacy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5775.
Abstract 5782: Androgen receptor antagonists: Lead optimization and preclinical pharmacology
Authors: Gavai Ashvin , Norris Derek , Balog Aaron , Austin Joel , Shan Weifang , Zhao Yufen , Han Wen-Ching , Sharma Lisa , Nation Andrew , Wu Dauh-Rurng , Mathur Arvind , Zhang Litai , Cvijic Mary Ellen , Yarde Melissa , Apanovitch Donald , Twamley Celeste , Tang Yuwei , Johnson Benjamin , Elzinga Paul , Cornelius Georgia , Obermeier Mary , Foster William , Marathe Punit , Dell-John Janet , Dito Gennaro , Masters Gregg , Rizzo Cheryl , Schweizer Liang , Jure-Kunkel Maria , Attar Ricardo , Gottardis Marco , Vite Gregory
Published: February 2011

The development and progression of prostate cancer is known to be dependent on androgens and their signaling mediated by the androgen receptor (AR). The primary therapeutic intervention involves using agents that lower serum testosterone (e.g., LHRH agonists), often in concert with an AR antagonist, such as bicalutamide. Despite a favorable initial anti-tumor response, most patients progress to the advanced hormone-refractory disease. The development of resistance to anti-androgen therapy has been shown to be associated with an increase in the levels of both AR mRNA and protein. This observation supports the concept that an AR antagonist with a significant improvement in potency as compared to bicalutamide and a broader spectrum of in vivo anti-tumor activity, including the bicalutamide-refractory human prostate tumor xenografts, may provide a significant clinical advantage in the treatment of advanced prostate cancer. This presentation will describe structure-activity relationships in a novel tetracyclic series of androgen receptor antagonists leading up to the identification of BMS-779333. It is a potent AR full antagonist, which exhibited broad spectrum efficacy in four human prostate tumor xenograft models. BMS-779333 did not exhibit agonist activity for AR mutant isoforms. Tumors that failed bicalutamide treatment were shown to retain their sensitivity to respond to BMS-779333. Transcriptomic changes in LuCaP-35 tumors treated with BMS-779333 were closer to castration than with other drug treatments. Based on its overall profile, BMS-779333 was selected for further preclinical evaluation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5782.
Abstract 580: Bone marrow cells trafficking after Salmonella typhimurium A1-R tumor targeting
Authors: Zhao Ming , Liu Fang , Zhang Lei , Hoffman Robert M.
Published: February 2011

Our laboratory has developed the tumor-targeting S. typhimurium double auxotrophic mutant termed A1-R. The A1-R mutant requires leucine and arginine, which disable it from growing in tumor tissue but the bacteria maintain high tumor virulence. In the present study, we used fluorescent imaging to observe bone marrow cells (BMC) trafficking in Lewis lung cancer-bearing nude mice after A1-R treatment. BMC were obtained from transgenic nude mice ubiquitously expressing green fluorescent protein (GFP). The GFP bone marrow cells were injected into the tail vein of red fluorescent protein (RFP)-expressing Lewis lung carcinoma-bearing nude mice. GFP-BMC trafficking to the tumor, lung, liver, and spleen were imaged at various time points after treatment with S. typhimurium A1-R. Untreated tumor-bearing mice injected with GFP-BMC were imaged as controls. Fluorescence imaging showed that there were more GFP-BMCs trafficking to tumor, lung, liver, and spleen in A1-R treated mice compared to untreated mice at 5 hours after A1-R infection. At day-2 after infection, there were significantly more GFP-BMCs in the tumor, and at day 4 after A1-R infection, BMCs appeared to die within the A1-R-treated tumors. GFP dendritic cells were observed in the liver of A1-R infected mouse at day 2 and day 4 after A1-R infection. These results suggest that immune cells in nude mice may be involved in the bacteria-induced antitumor response. The immune cells may also play a role in clearing the bacteria from normal organs in the host. In future experiments, we will identify the types of immune cells which play various roles in A1-R tumor targeting.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 580.
Abstract 582: Targeting the blood-brain barrier with a non-canonical iron-mimicry mechanism: A translational approach for the detection and treatment of human brain tumors
Authors: Staquicini Fernanda I. , Ozawa Michael G. , Moya Catherine A. , Driessen Wouter H.P. , Barbu Elena Magda , Soghomonyan Suren , Flores Leo G. , Alauddin Mian M. , Gelovani Juri G. , Liang Xiaowen , Hook Magnus , Basilion James P. , Bogler Oliver , Sidman Richard L. , Cavenee Webster K. , Pasqualini Renata , Arap Wadih
Published: February 2011

Treatment of central nervous system (CNS) diseases is limited by the blood-brain barrier (BBB), a selective vascular interface restricting passage of most molecules from blood into brain. Here we used the in vivo phage display technology to isolate peptide-targeted phage particles that can cross the BBB under normal and pathological conditions. We show that a non-canonical association of (i) transferrin, (ii) an iron-mimic ligand motif, and (iii) transferrin receptor mediates binding and transport of phage particles into the normal mouse brain through an allosteric mechanism. We also show that, in an orthotopic human glioma model, a combination of transferrin receptor overexpression plus extended vascular permeability and ligand retention results in remarkable brain tumor targeting. Such unique attributes enable molecular imaging and targeted therapy of intracranial tumors in a clinic-ready setting. Finally, we expand our data by analyzing a large panel of primary CNS tumors through comprehensive tissue microarrays. Together, our approach and results provide a translational avenue for the detection and treatment of brain tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 582.
Abstract 587: Development of brain tumor-specific targeted system by using retroviral-mediated gene therapy with SSX4 promoter
Authors: Shimizu Keiji , Ishida Eri , Kitahara Masahiro , Tamura Masakazu , Yawata Toshio
Published: February 2011

Despite many efforts to develop effective therapy, the outcome of malignant glioma remains poor. Previously, we reported the eradication of mouse glioma by retroviral-mediated gene therapy. In this study, a tumor-specific targeting system was studied to develop the effective and safe gene therapy. We first searched for genes expressing at high frequency in brain tumors but not in normal human astrocyte (NHA) among cancer testis antigen (CTA) genes by in silico and RT-PCR screening. MAGEA3 and SSX4 were identified as tumor-specific genes. The promoter activity was measured in glioma, telomerase-immortalized fibroblast and NHA cells. The SSX4 promoter but not MAGEA3 showed the tumor-specific activity. In order to define a useful tumor-specific promoter for targeting in context of retroviral-mediated gene therapy, the SSX4 promoter were used to restrict the expression of suicide gene HSVtk in retroviral vector. The glioma cell lines transduced with the retroviral vector were killed efficiently by addition of ganciclovir (GCV), but telomerase-immortalized fibroblast BJ-5ta cells were not. Mouse glioma RSV-M cells transduced with the retroviral vector were transplanted into S.C. of syngeneic mouse. The administration of GCV suppressed the tumor growth completely. These results suggest that trans-acting factors may play a role for tumor-specific activity of SSX4 promoter while the expression of most CTA genes is regulated by epigenetic factors and the retrovirus vector expressing HSVtk under control of SSX4 promoter is a promising therapeutic reagent for malignant brain tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 587.
Abstract 588: Effect of antibiotic treatment on Salmonella typhimurium A1-R tumor targeting of prostate cancer
Authors: Zhao Ming , Geller Jack , Liu Fang , Zhang Lei , Zhang Yong , Hoffman Robert M.
Published: February 2011

We have previously demonstrated that a genetically-altered Salmonella typhimurium, auxotrophic for arginine and leucine, could effectively treat metastatic PC-3 tumors in nude mice and profoundly increase survival. The auxotrophic mutations, induced by nitrosoguamidine, allow the bacteria to grow in tumors but not in normal tissue. Since human prostate cancer eventually becomes non-androgen-dependent similar to PC-3 tumors, bacterial treatment offers a novel treatment for this disease. In the current study, we observed the effect of antibiotic treatment on A1-R bacteria therapy of PC-3 tumors in nude mice. Thirty mice were implanted subcutaneously with PC-3 tumors expressing GFP. Approximately two weeks following implantation, the tumors were externally imageable. Ten of the 30 mice served as untreated control, and 10 mice were given A1 weekly intravenously as treated controls. Ten mice were given A1 intravenously weekly, combined with Gentamicin administered by intra-peritoneal injection at 100 µg daily for 10 day cycles with a two-day break between cycles. The treatment was repeated until the study end. Tumor growth was externally imaged by GFP expression of the tumors. All mice in the untreated group were sacrificed by 6 weeks after tumor implantation due to tumor size of 1.5-2.0 cm diameter. All mice in the A1-R-antibiotic combination group were sacrificed at 8 weeks due to tumor size. At the same time, all mice in the A1-R-only treatment group were alive and well. Currently, 3 treated mice were close to tumor eradication at 16 weeks following implantation and are still being injected weekly with A1-R. These results suggest that the tumor-targeting activity of A1-R bacteria was antibiotic dependent. This study gives proof of principle that A1-R can be regulated by antibiotics, an important safety feature for clinical application.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 588.
Abstract 598: EMT and nectin-1 mediate sensitivity to HSV-1 oncolysis in human HCC cell lines
Authors: Fuchs Bryan C. , Kuroda Toshihiko , Yamada Suguru , McGinn Christopher M. , Schmitt Anthony , Lanuti Michael , Tanabe Kenneth K.
Published: February 2011

Background: Oncolytic viral therapy mediated by herpes simplex virus 1 (HSV-1) is an emerging and promising cancer therapy. The entry of HSV-1 into cells is mediated in part by cell surface co-receptors. One of these co-receptors, nectin-1, is highly expressed in epithelial cells. Nectin-1 is localized to adherens junctions (AJs) where it interacts with E-cadherin through their respective peripheral membrane proteins, afadin and catenin. The loss of E-cadherin is a hallmark of epithelial-to-mesenchymal transition (EMT) and results in the disassembly of AJs. Here, we investigate the effects of EMT on cell surface nectin-1 expression, HSV infection and viral oncolysis. Methods: Western blot analysis was performed to determine E-cadherin expression in human HCC cell lines. Cytotoxicity to HSV-1 KOS strain was assessed by MTT analysis. KOS replication was assessed by a single-step burst assay, while viral infectivity was examined using the mutant bM24-BAC#3 virus which encodes GFP. Cell surface levels of nectin-1 and herpesvirus entry mediator (HVEM) were determined by flow cytometry and the role of nectin-1 in HSV-1 infectivity was further assessed using a nectin-1 neutralizing antibody. Results: Four of the human HCC cell lines were epithelial and four were mesenchymal. KOS oncolysis resulted in 50% cell death in the epithelial cell lines at multiplicities of infection (MOIs) 2-3 log orders less than the mesenchymal HCC cell lines. This difference was not attributed to cell growth rates as determined by population doubling times or to viral replication as assessed by burst assay. Instead, the epithelial cell lines had higher rates of infectivity which were attributed to greater cell surface expression of nectin-1 and not HVEM. Finally, blocking of nectin-1 with a neutralizing antibody inhibited viral oncolysis in all four epithelial HCC cell lines. Conclusions: EMT in human HCC cells results in the loss of nectin-1 cell surface expression and resistance to HSV-1 oncolysis. E-cadherin and nectin-1 are markers of sensitivity to HSV-1 oncolysis in human HCC cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 598.
Abstract 601: In vitro and in vivo inhibition of β-catenin by two novel β-catenin RNA antagonists, EZN-3889 and 3892
Authors: Zhang Yixian , Kim Steven , Shi Victoria , Qu Zhengxing , Castaneda Stephen , Wang Maoliang , Zhu Peifang , Filpula David , Greenberger Lee M. , Horak Ivan D.
Published: February 2011

Background: β-Catenin is a subunit of the cadherin protein complex and a key integral component in the Wnt signaling pathway. Therefore, it not only plays an important role in cell-cell adhesion but also in tumorgenesis. β-catenin is activated in most colon cancers due to either mutations in APC or activating mutations in β-catenin itself. Furthermore, activating β-catenin mutations have been found in a variety of other tumors such as melanomas, hepatocellular carcinomas, breast, and prostate cancer, whereas β-catenin is not activated in most normal tissues. Therefore, inhibition of β-catenin is likely to have therapeutic effects in many cancers. Unfortunately, because β-catenin is devoid of any enzymatic activity, the development of inhibitors of β-catenin has been slow. We present here both in vitro and in vivo evidence that the lack of specificity of small-molecule inhibitors may be overcome by two novel β-catenin LNA-based mRNA antagonists, EZN-3889 and EZN-3892. Material and Methods: In vitro, the ability of the compounds to inhibit mRNA, cell growth, and reporter gene were evaluated by qRT-PCR, MTS, and luciferase assays respectively, in multiple cell lines. In vivo, β-catenin mRNA down-modulation in liver and human tumors, which were grown on the flank of nude mice, was evaluated after intravenous administration of the compounds. Results: Without lipofection, we found that these two antagonists could specifically down modulate β-catenin mRNA and protein effectively in multiple cell lines with a long-lasting effect. Additionally, these two molecules effectively inhibited β-catenin transcriptional activity in SW480 colorectal and H1581 lung cancer cells stably expressing a TCF/LEF luciferase reporter gene, resulting in growth inhibition. Combination of EZN-3889 or 3892 with sulindac demonstrated improved activities in both growth and reporter assay. Moreover, EZN-3889 and 3892 inhibited the spontaneous growth of spheroids in SW480 culture and convert SW480 spheroids to monolayers. Preliminary data suggest that these spheroids were more tumorgenic than the monolayer cells. In mice, both molecules significantly down modulated β-catenin mRNA in the liver. Conclusions: Without lipofection, β-catenin antagonists potently and specifically inhibited β-catenin mRNA expression both in vitro and in vivo after intravenous injection. Target inhibition correlated with growth inhibition and phenotypical change of SW480 spheroids. Further studies will examine the significance of the inhibition of the spheroids and the effect of these antagonists on tumor growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 601.