bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1353: KIR copy-number variation and cutaneous melanoma risk.
Authors: Nelson Heather H. , Vineretsky Karin , Lazovich DeAnn
Published: August 2014

Prevention of cutaneous malignant melanoma requires the identification and clearance of abnormal, pre-cancerous cells by the immune system. Central to this immune surveillance is the interplay between KIR receptors present on natural killer cells and their HLA ligands expressed on target cells. Like HLA, the KIR genes are highly polymorphic. They encode both activating receptors (that trigger NK-cell mediated killing), and inhibitory receptors (important in NK cell education and self-tolerance). Many of the KIR genes occur as copy number variants, and we hypothesize that inter- individual variability in KIR copy number is associated with melanoma risk. A random sample of cases and controls within the large, population-based Minnesota Skin Health Study were selected for examination (84 cases and 87 controls). Using Luminex technology and genomic DNA isolated from subject buccal cells we tested for the presence or absence of specific KIR genes (KIR3DL1, KIR2DS1, KIR2DL2, KIR2DL3, KIR2DL5, KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS4 {truncating allele}, KIR2DS5, KIR2DS1). In addition to investigating each gene as a melanoma risk factor, we created an integrated locus score for 2DL2/DL3 and 3DL1/DS1, reflecting the typical mutual exclusivity of these genes at their respective loci. Results: Homozygous deletion of the activating KIR gene 2DS1 was associated with both an increasing number of nevi (p<0.04), and increased melanoma risk (OR 2.2, 95% CI 1.2 - 4.3). Similarly, at the shared 3DL1/3DS1 locus, absence of the activating KIR3DS1 gene was significantly associated with increased melanoma risk (OR 2.0, 95% CI 1.0 - 3.3), and a borderline significant increase in the number of nevi (p<0.07). In contrast, at the KIR2DL2/2DL3 locus, which encodes two inhibitory KIR alleles, our data suggest that a lack of diversity at this locus may be more important than the presence or absence of either gene; individuals homozygous for either 2DL2 or 2DL3 were at increased risk for both nevi (p<0.03), and melanoma (OR 1.8, 95% CI 1.0 - 3.4) relative to those who had both 2DL2 and 2DL3. These novel analyses at the complex KIR locus indicate that absence of certain activating KIR receptors, and balance in diversity of KIR inhibitory receptors are important determinants of cutaneous melanoma risk.

Citation Format: Heather H. Nelson, Karin Vineretsky, DeAnn Lazovich. KIR copy-number variation and cutaneous melanoma risk. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1353. doi:10.1158/1538-7445.AM2013-1353
Abstract 1355: Biospecimen preferences of African-American cancer survivors for cancer genetic research.
Authors: Ewing Altovise T. , Erby Lori AH , Ricks-Santi Luisel J. , Kalu Nnenna , Scott Denise M.
Published: August 2014

Background: Research based on large population-based biobanks has potential to ameliorate health disparities attributed to biological and genetic differences. Study goals were to identify associations of cancer survivors’ sample preferences with sociodemographic characteristics and familiarity with genetic research and testing.

Methods: Information was analyzed on a convenience sample of 158 adult African-American breast, colon or prostate cancer survivors who expressed a willingness to provide a biospecimen for cancer genetic research. Information was collected from a self-directed instrument that measured: 1) demographics, 2)familiarity with genetic research and testing, and 3)willingness to provide a variety of sample types. Descriptive statistics and bivariate chi-square analyses were utilized.

Results: Sample preference variation (N=158) consisted of: 65%, 62% and 18% willing to donate blood, saliva and tissue, respectively. One third (n= 53) endorsed willingness to provide only one form of biological sample (blood, saliva, or tissue). Significant associations were detected between willingness to provide tissue and individuals < 50 years of age (p=0.04). Sample preferences were not otherwise related to age at cancer diagnosis, gender, income, educational status, or familiarity with genetic testing or genetic research.

Conclusions/Impact: Providing options of acceptable biological sample types may yield increased donation from minorities. Inadvertently, this may introduce quality control challenges in obtaining sufficient or multiple forms of biological samples. Identifying additional predictive factors for samples preferences may lead to improvements in methods used to obtain biospecimens from minority populations.

Citation Format: Altovise T. Ewing, Lori AH Erby, Luisel J. Ricks-Santi, Nnenna Kalu, Denise M. Scott. Biospecimen preferences of African-American cancer survivors for cancer genetic research. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1355. doi:10.1158/1538-7445.AM2013-1355

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1358: Are physician recommendations for BRCA 1/2 testing among breast cancer patients appropriate? A population based study.
Authors: McCarthy Anne Marie , Demeter Mirar Bristol , Fredricks Tracey , Liao Kaijun , Shea Judy A. , Groeneveld Peter , Domchek Susan M. , Armstrong Katrina
Published: August 2014

Purpose: To evaluate the rates and predictors of physician recommendation for BRCA 1/2 testing in breast cancer patients.

Patients and Methods: Women aged 18-64 diagnosed with invasive breast cancer in 2007 in Pennsylvania completed a survey on family history of cancer, physician treatment recommendations, and BRCA 1/2 testing (N=2258). Based on age at diagnosis and family history, women were categorized as high, moderate, or low risk for BRCA 1/2 mutations. Multivariate logistic regression was used to assess the odds of physician recommendation for BRCA 1/2 testing by risk group, age at diagnosis, demographics, and tumor characteristics.

Results: Nearly 25% of participants were at high risk for carrying a BRCA 1/2 mutation based on age at breast cancer diagnosis and family history of breast and/or ovarian cancer. Physician recommendations for BRCA 1/2 testing were strongly associated with risk of carrying a BRCA 1/2 mutation, with 53% of women at high risk reporting a testing recommendation compared to 9% of women at low risk. In addition, physician recommendations were strongly correlated with use of testing in all risk groups. Among women at high risk of mutation, lack of a recommendation for BRCA 1/2 testing was more common among women who were older, had lower income, and were employed.

Conclusion: Although BRCA 1/2 testing recommendations appear to be appropriately correlated with mutation risk among women with breast cancer, a significant proportion of breast cancer patients who meet criteria for BRCA 1/2 testing may not be receiving recommendations for such testing from their providers.

Citation Format: Anne Marie McCarthy, Mirar Bristol Demeter, Tracey Fredricks, Kaijun Liao, Judy A. Shea, Peter Groeneveld, Susan M. Domchek, Katrina Armstrong. Are physician recommendations for BRCA 1/2 testing among breast cancer patients appropriate? A population based study. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1358. doi:10.1158/1538-7445.AM2013-1358
Abstract 1360: Reaching minority and low-income women with breast cancer prevention mobile messages: Implications for health promotion and clinical office collaboration.
Authors: Kratzke Cynthia , Vilchis Hugo
Published: August 2014

Background: While growing literature for breast cancer information supports the use of the Internet, little is known about breast cancer prevention and the use of mobile messages especially for women of different ages, race/ethnicity, income, and education levels. Minority and low-income women are more likely to be diagnosed with breast cancer at a later stage. Purpose: We examined differences for breast cancer risk factor knowledge, current channels, and mobile message preferences among Hispanic and non-Hispanic women in southern New Mexico. The study was guided using a conceptual framework of McGuire's Input-Output Persuasion Model. Methods: Women ages 40 and older at an imaging center completed a survey assessing their knowledge of breast cancer risk factors, channels of information, mobile message preferences, and demographics. In bivariate analysis, relationships between age, income, race/ethnicity and education of women and their mobile message preferences were examined. Results: A total of 157 women ages 40 to 91 (mean = 61, SD = 12.07) completed the survey. Breast cancer risk factor knowledge was significantly higher for non-Hispanic (p=.035), women with higher education levels (p=.023) and higher income levels (p=.000). The most common channels were television, magazines, Internet, and brochures. Overall, 87% used cell phones. Cell phone use was more likely among younger women ages 40-59 (p=.017), non-Hispanic women (p=.013), and women with incomes $20,000 or higher (p=.032). Of the cell phone users, 20% preferred breast cancer prevention messages. Breast cancer prevention cell message preferences were more likely among younger women ages 40-59 (p=.001), Hispanic women (p=.000), and women with lower incomes (p=.011). Nearly 47% used text messaging. Of the text users, 36% preferred breast cancer prevention text messages and 37% preferred mammogram appointment text reminders. Text message preferences were more likely among Hispanic women (p=.001). Conclusions: Traditional and mobile messages for breast cancer information are preferred by women. Targeting lower income, Hispanic, and younger women with mobile messages may help to reduce health disparities. Further research is needed for mobile technology use in health promotion. Health promotion specialists may partner with clinical offices to develop and target breast cancer prevention mobile messages for improved mammography adherence, patient education, and best practices in managing screening appointments.

Citation Format: Cynthia Kratzke, Hugo Vilchis. Reaching minority and low-income women with breast cancer prevention mobile messages: Implications for health promotion and clinical office collaboration. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1360. doi:10.1158/1538-7445.AM2013-1360
Abstract 1380: In vivo evidence for prior manipulation therapy-induced metastasis of primary osteosarcoma.
Authors: Wang Jir-You , Wu Po-Kuei , Chen Chih-Hsueh , Yen Chuen-Chuan , Hung Giun-Yi , Hung Shih-Chieh , Chen Wei-Ming , Liu Chien-Lin , Chen Tain-Hsiung
Published: August 2014

Our previous clinical study demonstrated that manipulation therapy (MT) on osteosarcoma (OS) patients prior to diagnosis resulted in poor prognosis after surgical treatment. This study was aimed to provide in vivo evidence of MT-induced metastasis in primary osterosarcoma. Eight-week-old male GFP-labeled human OS cells-transferred nude mice were randomly allocated into 2 groups, namely, MT (+) and MT (-) groups. MT was conducted with repeated massage on tumor site twice a week for 7 or 15 weeks. The parameters evaluated were x-ray diagnosis, micro-PET/CT scan, histopathology and serum metalloproteinase 9 (MMP9) level. The results showed that MT (+) mice showed a decreased body weight (30.5±0.65g) and an increased tumor volume (8.3±1.18 mm3) compared to MT (-) group with body weight (35.8±0.40g, p<0.0001) and tumor volume (3.9±1.34mm3, p=0.038), respectively. There was an increased signal intensity over lymph node region of hind limb by micro-PET/CT and the GFP-labeled human OS cells were detected in the lung and bilateral lymph nodes in MT (+) group, while there were no such findings in MT (-) group. The serum MMP9 level was higher in MT (+) group (27.1±1.29 ng/ml) than in MT (-) group (17.8±1.97 ng/ml, p=0.048). Taken previous clinical observation and the present in vivo evidence together, we conclude that physicians should pay more attention on those patients who seek MT before diagnosis or during treatment for osteosarcoma.

Citation Format: Jir-You Wang, Po-Kuei Wu, Chih-Hsueh Chen, Chuen-Chuan Yen, Giun-Yi Hung, Shih-Chieh Hung, Wei-Ming Chen, Chien-Lin Liu, Tain-Hsiung Chen. In vivo evidence for prior manipulation therapy-induced metastasis of primary osteosarcoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1380. doi:10.1158/1538-7445.AM2013-1380
Abstract 1385: PPARδ promotes mammary tumorigenesis through mTOR activation.
Authors: Glazer Robert I. , Yuan Hongyan , Upadhyay Geeta , Yin Yuzhi , Kopelovich Levy
Published: August 2014

Background: Peroxisome proliferator-activated receptorδ (PPARδ) regulates a multitude of physiological processes associated with glucose and lipid metabolism, inflammation and proliferation. One or more of these processes are potential risk factors for the ability of PPARδ agonists to promote tumorigenesis in the mammary gland. In the present study, we describe a new transgenic mouse model in which activation of PPARδ by endogenous or synthetic ligands results in mammary neoplasia.

Methods: Transgenic mice were generated in which expression of PPARδ was directed to the mammary gland under the control of the MMTV long terminal repeat.

Results: Multiparous MMTV-PPARδ mice presented with infiltrating ductal carcinomas after a latency of 12 months. When transgenic animals were maintained on a diet supplemented with the PPARδ ligand GW501516, rapid progression to ductal and alveolar hyperplasia, dysplasia and ductal carcinoma in situ occurred within three months, and multifocal ductal carcinomas occurred after 5 months. Histopathological changes occurred in concert with a marked increase in phosphatidylcholine (PC) and lysoPC levels. Gene array profiling revealed an inflammatory, invasive, metabolic and proliferative signature denoted by enhanced expression of Saa1/2, S100a8, Klk6/7, Acsl4 and Plac1, and increased mTOR signaling. Treatment with the mTOR inhibitor, everolimus, dramatically reversed many of the histopathological and gene expression abnormalities.

Conclusion: MMTV-PPARδ mice demonstrate a direct role for PPARδ in breast tumorigenesis that is linked to several previously unknown signaling processes. These findings suggest a rationale for new therapeutic approaches to prevent and treat this disease.

Supported by 1N01CN43302-WA19 and 1P30CA51008

1st Choice Category: TB1 Cell Culture and Animal Models of Cancer

Citation Format: Robert I. Glazer, Hongyan Yuan, Geeta Upadhyay, Yuzhi Yin, Levy Kopelovich. PPARδ promotes mammary tumorigenesis through mTOR activation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1385. doi:10.1158/1538-7445.AM2013-1385
Abstract 1386: Development and application of a syngeneic breast tumor preclinical model to study anticancer efficacy and cardiac safety of doxorubicin and dexrazoxane.
Authors: Gonzalez-Berrios Yanira , Dickey Jennifer S. , Aryal Baikuntha P. , Rosen Elliot T. , Herman Eugene H. , Mog Steven , Nakamura Asako J. , Redon Christophe E. , Baxa Ulrich , Parekh Palak , Mason Karen P. , Bonner William , Rao Ashutosh
Published: August 2014

Doxorubicin is a chemotherapeutic drug that causes oxidative stress-mediated cardiotoxicity. To prevent oxidative stress and help to mitigate the cardiotoxic effects of doxorubicin, the iron chelator dexrazoxane is clinically used. However, iron chelators and other antioxidants have not completely succeeded in mitigating this effect and do not protect against iron-independent oxidative damage. One limitation to the development of new cardioprotectants is the lack of physiologically-relevant animal models to simultaneously study antitumor activity and cardioprotection. Therefore, we optimized a syngeneic rat model and examined the mechanisms by which oxidative stress affects the safety and efficacy outcome. Immune-competent spontaneously hypertensive rats (SHRs) were implanted with the SHR-derived, breast tumor cell line, SST-2. Tumor growth and cytokine responses (IL-1A, MCP-1, TNF-α) were observed two weeks post-implantation. In order to demonstrate the utility of SHR/SST-2 model for monitoring both anticancer efficacy and acute cardiotoxicity, we studied cardiotoxic doxorubicin alone and in combination with dexrazoxane. As predicted, significant tumor reduction, DNA damage, and cardiomyopathy were demonstrated by doxorubicin. Treatment with the cardioprotectant dexrazoxane increased the pro-survival autophagy marker LC3-II and decreased caspase-3 in the heart, as a single agent and in combination with doxorubicin. Histopathology and transmission electron microscopy demonstrated apoptosis, autophagy, and necrosis in tumor and heart. Our results demonstrate the advantage of the immune-proficient SHR/SST-2 model to study anticancer agents and simultaneously address anticancer efficacy and cardiac safety.

Citation Format: Yanira Gonzalez-Berrios, Jennifer S. Dickey, Baikuntha P. Aryal, Elliot T. Rosen, Eugene H. Herman, Steven Mog, Asako J. Nakamura, Christophe E. Redon, Ulrich Baxa, Palak Parekh, Karen P. Mason, William Bonner, Ashutosh Rao. Development and application of a syngeneic breast tumor preclinical model to study anticancer efficacy and cardiac safety of doxorubicin and dexrazoxane. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1386. doi:10.1158/1538-7445.AM2013-1386
Abstract 139: Alcohol and breast cancer subtypes in a Spanish cohort.
Authors: Gago-Dominguez Manuela , Redondo Carmen M. , Enguix Manuel , Fernandez Maite Peña , Martinez Maria Elena , Munoz Victor , Carracedo Angel , Gude Francisco , Castelao Jose Esteban
Published: August 2014

Background Although alcohol intake is an established risk factor for overall breast cancer, few studies have looked at the relationship between alcohol use and breast cancer risk by subtype of breast cancer.

Materials and Methods A population-based study was conducted in Galicia, Spain. A total of 1211 Spanish women diagnosed with operable invasive breast cancer between 1997 and 2010 and 550 controls participated in the study. Data on demographics, breast cancer risk factors, and clinico-pathological characteristics were collected. The four major breast cancer subtypes [hormone-receptor-positive, HER2-negative (luminal A); hormone-receptor-positive, HER2-positive (luminal B); hormone-receptor-negative, HER2-negative (triple-negative); and hormone-receptor-negative, HER2-positive (HER2 over-expressing)] were compared on their clinico-pathological characteristics and risk factor profiles, particularly alcohol intake and reproductive variables.

Results With the exception of Her2 overexpressing, the risk of all subtypes of breast cancer increased with increasing alcohol intake (luminal A: OR=2.32, 95%CI:1.43-3.77; luminal B: OR=2.03, 95%CI 1.01-4.06, and triple negative: OR=2.36, 95%CI 1.11-5.00). The association of alcohol with breast cancer was similar among women who drank wine exclusively and other drinkers. The alcohol-breast cancer association appeared to be more pronounced among lobular breast cancers (OR= 2.70, 95% CI:1.33-5.52) than among ductal type breast cancers (OR= 2.09, 95%CI:1.33-3-01). It has been suggested that the association of alcohol intake with breast cancer may be stronger in women who are current or past users of hormone replacement therapy compared with never users, or among non-ovariectomized compared to ovariectomized women, we found no such differences.

Conclusions To our knowledge, no prior study has examined the association of breast cancer with alcohol intake according to breast cancer subtypes and the type of wine consumed in Spanish women. Our findings indicate that breast cancer risk increased with increasing alcohol intakes for three out of the four major subtypes of breast cancer and that the risk was similar in women who drank wine, beer and/or liquor exclusively or in mixture.

Citation Format: Manuela Gago-Dominguez, Carmen M. Redondo, Manuel Enguix, Maite Peña Fernandez, Maria Elena Martinez, Victor Munoz, Angel Carracedo, Francisco Gude, Jose Esteban Castelao. Alcohol and breast cancer subtypes in a Spanish cohort. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 139. doi:10.1158/1538-7445.AM2013-139
Abstract 1391: Cellular and molecular bases of susceptibility to estrogen-induced mammary cancer in the rat.
Authors: Ding Lina , Zhao Yang , Warren Christopher , Shull James
Published: August 2014

Known breast cancer risk factors include a family history of breast cancer, cumulative exposure to endogenous and exogenous estrogens and breast mammographic density. Although several genes have been identified that significantly impact risk when mutated or aberrantly expressed, the genetic determinants of responsiveness to estrogens and mammographic density remain poorly defined. We are using the ACI rat model of 17β-estradiol (E2)-induced mammary cancer to identify genetic and environmental determinants of susceptibility to mammary cancer. When treated continuously with physiologic levels of E2, female ACI rats develop mammary carcinoma at an incidence approaching 100%. The mammary cancers that develop in E2 treated ACI rats express estrogen receptor-α (ERα) and progesterone receptor (PgR) and are dependent upon E2 for continued growth. By contrast, Brown Norway (BN) rats are highly resistant to E2-induced mammary cancer. The purpose of this study was to compare the impact of E2 treatment on mammary gland biology in susceptible ACI and resistant BN rats. Dramatic differences in numerous cellular and molecular responses to E2 were observed when these two inbred rat strains were compared. Whereas the mammary epithelium of susceptible ACI rats exhibited a marked proliferative response to E2, the epithelium of resistant BN rats exhibited responses associated with differentiation to secretory epithelium. Strain differences in expression of multiple genes associated with mammary epithelial cell proliferation and differentiation were observed. Specifically, Areg, Pgr, Wnt4 and Tnfsf11 were highly expressed in mammary glands of E2 treated ACI rats, relative to treated BN rats, whereas Lcn2, Spp1, Cd44, Mmp7 and Mmp9 were highly expressed in mammary glands of BN rats, relative to ACI rats. This study provides novel insight into the molecular bases of inherited susceptibility to E2-induced mammary cancer. We further propose that the observed strain differences in mammary gland responsiveness to E2 represent phenotypes that underlie variation in mammographic density in humans and may have the potential to serve as biomarkers that predict breast cancer risk.

Citation Format: Lina Ding, Yang Zhao, Christopher Warren, James Shull. Cellular and molecular bases of susceptibility to estrogen-induced mammary cancer in the rat. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1391. doi:10.1158/1538-7445.AM2013-1391
Abstract 1392: Role of mutant p53 for anti-TGF-β therapy in isogenic basal-like breast cancer models.
Authors: Lugo Fernando P. , Lin Shu , Bandyopadhyay Abhik , Yang Junhua , Sun Lu-Zhe
Published: August 2014

Breast cancer remains as one of the most common cancers worldwide accounting for 450,000 deaths. During cancer progression, transforming growth factor-β (TGF-β) signaling in tumor cells is often converted from tumor suppression to tumor promotion. However, how this oncogenic switch occurs is not well understood. Tripe-negative breast cancer (TNBC) is a breast cancer subtype, which is frequently associated with p53 mutation and lacks specific molecular targets for therapeutic intervention. TGF-β signaling has been shown to be oncogenic in TNBC models. Therefore, we tested the hypothesis that mutation of p53 may cause the oncogenic switch of TGF-β and thus serve as a biomarker for anti-TGF-β therapy.

In an effort to create isogenic models resembling the transformation of normal breast epithelial cells into TNBC, oncogenic H-Ras V12, human telomerase reverse transcriptase (hTERT), and mutant p53 R175H (mP53) or vector control were sequentially introduced into untransformed human breast epithelial MCF-10A cells. These generated cell lines were orthotopically introduced into the mammary fat pads of female nude mice and tumor burden was monitored. Mice were divided into two groups receiving a TGF-β type I receptor (RI) kinase inhibitor or PBS as control. Harvested lungs were analyzed for metastasis with GFP fluorescent imaging and quantitative PCR for tumor cell specific genes.

Mice inoculated with the transformed MCF-10A cells containing mp53 (10A/mp53) displayed greater tumor burden and lung metastasis incidence than the vector control counterparts. Systemic treatment with the TGF-β inhibitor resulted in a decrease in metastasis in the mice inoculated with 10A/mp53 representing disruption of TGF-β’s oncogenic role. In contrast, inhibition of TGF-β signaling in mice inoculated with 10A/pp290 vector control displayed greater lung metastasis incidence, likely due to the disruption of TGF-β tumor suppressive effects. Our results indicate that the mp53 may serve as a biomarker for the oncogenic switch of TGF-β pathway and anti-TGF-β therapy for the inhibition of TNBC-induced metastasis.

Citation Format: Fernando P. Lugo, Shu Lin, Abhik Bandyopadhyay, Junhua Yang, Lu-Zhe Sun. Role of mutant p53 for anti-TGF-β therapy in isogenic basal-like breast cancer models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1392. doi:10.1158/1538-7445.AM2013-1392
Abstract 1394: Stable knockdown of osteoprotegerin increases cell proliferation and metastasis in breast cancer cells.
Authors: Weichhaus Michael , Segaran Prabu , Le Andrew , Geerts Dirk , Connelly Linda
Published: August 2014

Human breast cancer cells express and secrete osteoprotegerin (OPG, also called TNFRSF11B), a negative regulator for osteoclast differentiation. Previous in vitro data indicated that OPG expression promotes survival of breast cancer cells by inhibiting the apoptotic effect of TRAIL, through direct interaction of OPG with TRAIL. We generated stably transfected OPG-knockdown (KD) MDA-MB-231 breast cancer cells, using OPG shRNA clones, to examine the effect of OPG expression on primary tumor growth and metastasis in vivo. Positive selection of successfully transfected clones revealed that OPG KD cells expressed 60% less OPG (mRNA and protein) than control shRNA cells. OPG expression in control shRNA cells was not different from wild-type (wt) cells. Cell proliferation showed a small decrease after TRAIL challenge of KD cells compared to control and wt cells, assessed by MTT-assay, suggesting this was not a significant interaction. We decided to investigate the impact of OPG KD on tumor growth and metastasis using the chick embryo in vivo model. In brief, fertilized white leghorn chicken eggs were incubated for 10 days, after which the chorioallantoic membrane (CAM) was inoculated with 3 million KD or control cells. After an additional 7 days incubation, the primary tumor and a portion of the distant CAM were excised and measured for weight and the presence of human DNA, respectively. Primary tumor weight of KD cells was 31% higher than primary tumors from control cells. Moreover detection of human DNA in the distant CAM was 58% higher in eggs incubated with KD cells than in eggs incubated with control cells. Cell growth experiments in vitro showed that cell proliferation of KD cells was 40% higher compared to control and wt cells, as measured by trypan exclusion method. For validation in human cancer patients we mined a public microarray database, detailing gene expression in primary breast tumors. In agreement with our experimental data, high OPG gene expression in breast tumors was correlated with better prognosis. Contrary to the current understanding of OPG expression in breast cancer cells, our data indicates that OPG expression in breast cancer cells inhibits growth, the ability to metastasize and may promote survival in breast cancer patients.

Citation Format: Michael Weichhaus, Prabu Segaran, Andrew Le, Dirk Geerts, Linda Connelly. Stable knockdown of osteoprotegerin increases cell proliferation and metastasis in breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1394. doi:10.1158/1538-7445.AM2013-1394
Abstract 1405: Deficiency of Fes tyrosine kinase expression correlates with delayed mammary tumor onset in a HER2/Neu over-expressing transgenic mouse model.
Authors: Zhang Connie S. , Greer Peter A.
Published: August 2014

Fes (a.k.a. Fps) is a cytoplasmic protein tyrosine kinase that is highly expressed in breast epithelial cells during lactation, as well as in vascular endothelial and myeloid cells. Retrovially encoded oncogenic alleles of Fes are associated with tumors in chickens and cats and cause tumors in transgenic mice; however, a role for Fes in human cancer has not been established.

Using an orthotopic mouse mammary engraftment model, we previously found that Fes deficiency correlated with reduction in engrafted tumor growth rates, lung metastasis, and circulating tumor cells. Reduced vascularity and fewer infiltrating macrophages suggested the tumor microenvironment of Fes-deficient mice was less supportive of tumor growth and metastasis. In co-culture with tumor cells, Fes-deficiency in macrophages correlated with a reduced ability to promote tumor cell invasive behavior. These results suggested a tumor promoting role of Fes kinase in breast cancer through roles in cells of the tumor microenvironment.

In a transgenic mouse model of breast cancer driven by an activated HER2/Neu allele expressed in the mammary epithelium, we observed delayed tumor onset in Fes-deficient mice compared to wild-type mice. However, there was no difference in tumor growth rates.

Taken together, these observations argue that Fes inhibition might provide therapeutic benefits in breast cancer, by attenuating tumor-associated angiogenesis and the metastasis-promoting functions of tumor-associated macrophages, or by delaying breast tumor onset in women with HER2 overexpression.

Citation Format: Connie S. Zhang, Peter A. Greer. Deficiency of Fes tyrosine kinase expression correlates with delayed mammary tumor onset in a HER2/Neu over-expressing transgenic mouse model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1405. doi:10.1158/1538-7445.AM2013-1405

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1410: Forced expression of EWS/ATF1 induces sarcomas in mice.
Authors: Yamada Kazunari , Ohno Takatoshi , Hara Akira , Yamada Yasuhiro , Shimizu Katsuji
Published: August 2014

Clear cell sarcoma (CCS) is an aggressive soft tissue malignant tumor characterized by a unique t(12; 22) translocation, leading to the expression of a chimeric EWS/ATF1 fusion gene. However, little is known about the mechanisms underlying how EWS/ATF1 is involved in the development of CCSs. In addition, the cells of origin for CCSs remain to be determined. We generated EWS/ATF1-inducible mice, and examined the effects of EWS/ATF1 expression in adult somatic cells. We show that the forced expression of EWS/ATF1 results in the development of EWS/ATF1-dependent sarcomas in mice. The histology of EWS/ATF1-induced sarcomas resembles that of CCSs and EWS/ATF1-induced tumor cells express CCS-markers, such as S100, Sox10, and Mitf. Lineage tracing experiments using Wnt1-Cre and P0-Cre alleles indicated that neural crest-lineage cells are permissive to EWS-ATF1-driven transformation.

Citation Format: Kazunari Yamada, Takatoshi Ohno, Akira Hara, Yasuhiro Yamada, Katsuji Shimizu. Forced expression of EWS/ATF1 induces sarcomas in mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1410. doi:10.1158/1538-7445.AM2013-1410
Abstract 1415: Role of IL-8 in uveal melanoma induced angiogenesis.
Authors: Lattanzio Laura , Torta Ilaria , Tonissi Federica , Monteverde Martino , Merlano Marco , Milano Gerard , Lo Nigro Cristiana
Published: August 2014

Background: The pro-angiogenic IL-8 pathway has already been associated with resistance to VEGF-targeted therapies in various tumour types, particularly in melanoma. IL-8 expression is higher in tumour as compared to normal tissues and both tumour and endothelial cells express this chemokine and its receptors, CXCR1 and CXCR2. Uveal melanoma (UM) is a highly vascularised tumour that is generally treated with radiations (RT). In a previous pre-clinical study (Sudaka A. et al., Invest. New Drugs, 2012, Jun 20), we reported that RT-associated anti-angiogenic therapy has more than additive effects by modulating VEGF levels and allowing to deliver lower radiation doses for identical cytotoxic effects. Aim: In order to better understand the microenvironment contribution to treatment efficacy in UM we investigated the release of IL-8 in response to the anti-angiogenic drug Bevacizumab, and the cross-talk between IL-8 and VEGF signalling. Materials and methods: OCM-1 uveal melanoma cell line and Huvec cells were grown in transwell plates. Bevacizumab was used at a final concentration of 2.5 mg/ml and cells were irradiated with a 6Gy dose. Il-8 concentrations were determined by ELISA assay. Results: Bevacizumab alone or combined with RT induced a 6 fold decrease in IL-8 concentration in culture medium from OCM-1 cells grown alone (Bev vs control/CT p<0,001%; Bev vs CT p<0,01%), but this effect was completely abolished when OCM-1 cells were cultured in Huvec-conditioned medium. When cells were co-cultured on transwell plates and treated with Bevacizumab IL-8 level increased by 30% in OCM-1 culture supernatant (Bev vs CT p=0,12) whereas it was 35% lower in Huvec culture medium (Bev vs CT p<0,05). Conclusions: These data indicate that the UM microenvironment, beside VEGF, can activate IL-8 signalling as an alternative pro-angiogenic pathway. The latter seems to be upregulated in UM cells by anti-VEGF therapy only when they are co-cultured with endothelial cells, thus suggesting that this effect may be dependent on a crosstalk between the two cell types. The decrease in IL-8 concentration, observed in the Huvec chamber of the transwell plates after incubation with Bevacizumab, may be due to a more efficient Il-8 binding by CXCR1 and/or CXCR2 (hypothesis currently tested). Therefore, although preliminary, our results suggest a possible synergic effect for a therapeutic approach combining IL-8 inhibitors with anti-VEGF compounds in UM.

Citation Format: Laura Lattanzio, Ilaria Torta, Federica Tonissi, Martino Monteverde, Marco Merlano, Gerard Milano, Cristiana Lo Nigro. Role of IL-8 in uveal melanoma induced angiogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1415. doi:10.1158/1538-7445.AM2013-1415
Abstract 1420: Mechanisms of stroma-derived CCL7 in promoting lung cancer.
Authors: Choi Hyejin , Durrans Anna , Gao Dingcheng , Ryu Seongho , Altorki Nasser , Mittal Vivek
Published: August 2014

Lung cancer is a leading cause of cancer related deaths worldwide, with non-small cell lung cancer (NSCLC) being the most prevalent form. Considerable attention has been directed to cancer cell intrinsic abnormalities (K-ras, p53, EGFR, AML-ALK4 fusions etc.), but little is known about the contribution of stromal cells to NSCLC progression.

We have undertaken a comprehensive analysis to determine the contribution and biological function of the stromal cells in NSCLC. We show increased recruitment of BM hematopoietic cells in the tumor beds compared to matched adjacent non-neoplastic lung tissue in NSCLC patients. Genome-wide transcriptome analysis on specific cell populations from fresh samples of NSCLC patients identified differentially regulated genes and mRNA isoforms in NSCLC stromal cells. Genes that were successfully validated by independent RT-PCR on sorted stromal cells and tumor cells were used for further analysis. Many genes encoded secreted proteins, suggesting the activation of aberrant autocrine and paracrine regulatory loops likely to support stroma-mediated tumor progression.

Among these genes, CCL7 was upregulated in myeloid progenitor cells and was selected for further investigation. Immunostaining of lung tumor and adjacent non-neoplastic tissue of patients shows highest expression of CCL7 in BM-derived cells (CD45+ cells) in the tumor.

Cross species analysis showed that, consistent with the observation in human tissue, CCL7 was differentially regulated in BM derived cells in a mouse model of NSCLC generated in KrasG12D and p53-/- transgenic mice. We have genetically ablated CCL7 in the BM cells using lentiviral shRNA and BMT approaches and will discuss consequences of this ablation in CCL7-mediated crosstalk between tumor cells and stromal cells that regulate NSCLC progression.

Citation Format: Hyejin Choi, Anna Durrans, Dingcheng Gao, Seongho Ryu, Nasser Altorki, Vivek Mittal. Mechanisms of stroma-derived CCL7 in promoting lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1420. doi:10.1158/1538-7445.AM2013-1420
Abstract 1422: Interleukin-17A activates tumor progression locus 2 to induce neoplastic cell transformation and breast carcinogenesis.
Authors: Kim Garam , Choi Hong Seok
Published: August 2014

Interleukin-17 (IL-17) is a pro-inflammatory cytokine secreted by activated CD4+ memory T cells. Although the role of T cells in cancer promotion has been examined, little is known about the underlying molecular mechanisms of interleukin-17 A (IL-17A) in carcinogenesis. In the present study, we first found that IL-17A induces neoplastic transformation of JB6 Cl41 cells through activation of tumor progression locus 2 (TPL2). IL-17A activates MEKs-ERKs, JNKs-c-Jun, and STAT3 signaling pathways, which are inhibited by a TPL2 kinase inhibitor (TKI). Furthermore, IL-17A activates c-fos and c-jun promoter activity, resulting in increased activator protein-1 (AP-1) activity. When small interfering RNA (siRNA) of IL-17A receptor (IL-17R), IL-17A, and TPL2 were introduced into JB6 Cl41 cells, respectively, IL-17A-induced AP-1 activity was significantly decreased compared to control cells. Similarly, TPL2 inhibition suppressed AP-1 activity induced by IL-17A. The knockdown of IL-17R and TKI treatment in JB6 Cl41 cells resulted in decreased IL-17A-induced cell transformation. The in vivo chorioallantoic membrane (CAM) assay also showed that IL-17A increased tumor formation of JB6 Cl41 cells, whereas TKI inhibited the tumorigenesis promoted by IL-17A. Consistent with these observations, knockdown of IL-17A and/or inhibition of TPL2 attenuated tumorigenecity of human breast cancer MCF7 cells. Moreover, immunohistochemical staining showed that both expressions of IL-17A and TPL2 were highly upregulated in human breast cancers. Together, our findings point to a critical role for the IL-17A-induced TPL2 signaling pathway in supporting cancer-associated inflammation in the tumor microenvironment. Therapeutic approaches that target this pathway may therefore effectively inhibit carcinogenesis.

Citation Format: Garam Kim, Hong Seok Choi. Interleukin-17A activates tumor progression locus 2 to induce neoplastic cell transformation and breast carcinogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1422. doi:10.1158/1538-7445.AM2013-1422

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1423: Proinflammatory activity of type II interleukin 1 receptor, a decoy receptor.
Authors: Mar Ai-Chung , Lee Hui-Ju , Lee Te-Chang
Published: August 2014

We have previously demonstrated that type II interleukin 1 receptor (IL1R2), an IL-1 decoy receptor, plays a dual role on regulation of inflammatory signaling, i.e., IL1R2 inhibits the exogenous interleukin 1 (IL-1) signaling on the one hand but activates intracellular IL-1α and its downstream proinflammatory cytokines. To have a better understanding of the proinflammatory activity of IL1R2, we modulated the intracellular IL1R2 levels by ectopic expression or siRNA silencing techniques. In the present study, our results showed that ectopic expression of IL1R2 in HaCaT cells, an immortalized cell line of human keratinocyte, significantly enhanced the expression of IL-6, MMP-1, and VEGF-A. Since T4R2 cells, a tumorigenic cell line derived from HaCaT cells by long-term exposure to inorganic arsenic, expressed a significant amount of IL1R2, we therefore silenced the expression of IL1R2 in T4R2 cells by RNAi technique. Our results consistently showed that silencing of IL1R2 in T4R2 cells decreased expression of IL-6, MMP-1, and VEGF-A. Meanwhile, our results demonstrated that the intracellular levels of IL1R2 are closely associated with migration ability of HaCaT cells. Moreover, IL1R2 silencing reduced the growth rate of T4R2 cells in nude mice. The conditioned medium harvested from T4R2 cells was able to stimulate migration, proliferation, and tube formation of cultured endothelial cells, whereas the angiogenesis activity of conditioned medium harvested from IL1R2 silenced T4R2 cells was significantly suppressed. Furthermore, the enhanced migration ability of endothelial cells culturing in T4R2 conditioned medium was suppressed by antibodies against VEGF-A and IL-6. We also demonstrated that tumors derived from T4R2 cells contained higher levels of CD31 positive cells as compared to those derived from IL1R2 silenced T4R2 cells, indicating more blood vessels formed in T4R2 tumor tissues. Mechanistically, our present studies showed that IL1R2 may work together with cFos and bind to Ap-1 site of promoters of IL-6 and VEGF-A. Taken together, we revealed that intracellular IL1R2 acts as a proinflammatory factor and hence enhances the expression of several inflammatory cytokines involving in angiogenesis and tumor malignancy.

Citation Format: Ai-Chung Mar, Hui-Ju Lee, Te-Chang Lee. Proinflammatory activity of type II interleukin 1 receptor, a decoy receptor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1423. doi:10.1158/1538-7445.AM2013-1423
Abstract 1424: Inhibition of M-CSF or MCP-1 expression in tumor cells reduces tumor growth and lung metastasis in a murine mammary cancer model.
Authors: Cho Han Jin , Park Jung Han Yoon
Published: August 2014

Tumor-associated macrophages, which are derived from the infiltration of circulating bone marrow-derived monocytes, consist primarily of a polarized M2 macrophage population and are associated with poor prognosis in various cancers. We previously observed that solid tumor growth and lung metastasis were enhanced when 4T1 mammary carcinoma cells were injected with M2 macrophages into the mammary fat pad of BALB/c mice. Cancer cell proliferation and macrophage infiltration were markedly increased when M2 macrophages were co-injected with 4T1 cells. In the present study, we explored the mechanisms underlying the crosstalk between 4T1 cells and M2 macrophages in tumor microenvironment. The results of in vitro experiments revealed that the migration of monocytes and the secretion of many cytokines including macrophage-colony stimulating factor (M-CSF) and monocyte chemotactic protein-1 (MCP-1) were increased when 4T1 cells were co-cultured with M2 macrophages, as compared to when the 4T1 cells were cultured alone. The mRNA expression of M-CSF and MCP-1 increased in 4T1 cells when 4T1 cells were co-cultured with M2 macrophages. To investigate the potential roles of M-CSF and MCP-1 in tumor growth and metastasis, 4T1 cells were infected with M-CSF, MCP-1, or control shRNA lentivirus particles and selected with puromycin. 4T1 cells infected with M-CSF shRNA did not grow or metastasize to the lung when injected into the mammary fat pads of BALB/c mice. Inhibition of MCP-1 expression in tumor cells also suppressed 4T1 tumor growth and metastasis, however, the degree of inhibition was much smaller than that of M-CSF inhibition. M-CSF knockdown decreased M2 macrophage (mouse mannose receptor+) population, whereas MCP-1 knockdown decreased the number of both M1 and M2 macrophages in tumor tissues. Taken together, these results indicate that M-CSF and MCP-1 are produced by the crosstalk between 4T1 cells and M2 macrophages, and these cytokines play important roles in mammary tumor progression.

Citation Format: Han Jin Cho, Jung Han Yoon Park. Inhibition of M-CSF or MCP-1 expression in tumor cells reduces tumor growth and lung metastasis in a murine mammary cancer model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1424. doi:10.1158/1538-7445.AM2013-1424