bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 126: BRCA-positivity: Multicultural scenes from the rear view mirror .
Authors: Miller-Samuel Susan , Simone Nicole , Rosenberg Anne L.
Published: August 2014

Background: To date, BRCA1 and BRCA2 mutations have been associated with specific breast cancer subtypes and patient demographics. BRCA testing has predominantly been performed in the Caucasian population. Certain breast cancer subtypes such as Triple Negative Breast Cancer (TNBC), have previously been associated with African American (AA) women and BRCA1 positivity. Novel targeted agents such as PARP inhibitors may be beneficial for TNBC and BRCA-positive cancers. Appropriate genetic referral and testing could influence patient treatment options. We sought to determine the patterns of patient referral and outcome for BRCA testing at Thomas Jefferson University Hospital (TJUH).

Methods: After obtaining IRB approval, a retrospective case review was performed to identify women who proceeded with genetic counseling from January 2008 through May 2012. Patients with positive genetic tests were assessed. Pathology reports of mutation-positive women were reviewed. Patient characteristics such as ethnicity were confirmed from genetic counseling records.

Findings: 461 new patients were seen for genetic counseling from January 2008 through May 2012. 309 (67%) proceeded with BRCA genetic testing and 26 (9%) were found to carry a deleterious BRCA1 or BRCA2 mutation. Hormone receptor profiles, age at diagnosis, and ethnic background varied among mutation carriers. The predominant ethnic background represented in TJUH mutation carriers is now AA. In 2008-2009 (24 months), only one AA woman was diagnosed with a BRCA mutation; however, from 2010-May 2012 (29 months), 9 AA women and 2 Hispanic women were diagnosed as BRCA-mutation carriers.

Conclusions: BRCA-positive patients have diverse demographic and hormone receptor profiles. Although we found that TNBC remains associated with BRCA1 carriers and estrogen positivity with BRCA2 carriers, it is not absolute. A recent shift has noted substantially more AA women being diagnosed with BRCA mutations. While this may indicate that more diverse populations are being referred for genetic counseling, more importantly, it suggests that diverse populations are becoming more receptive to participating in genetic counseling and when appropriate, receiving genetic testing.

Citation Format: Susan Miller-Samuel, Nicole Simone, Anne L. Rosenberg. BRCA-positivity: Multicultural scenes from the rear view mirror . [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 126. doi:10.1158/1538-7445.AM2013-126

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1264: Mutated CD80 may facilitate T-cell activation by inhibiting PDL1-PD1 suppression and by costimulating.
Authors: Dalal Sonia P. , Haile Samuel , Somasundaram Preethi , Ostrand-Rosenberg Suzanne
Published: August 2014

CD80 functions as a costimulatory signal to activate T cells when it binds to CD28 or as an apoptotic signal when it binds to CTLA4. We recently identified another function for CD80 and showed that co-expression of CD80 by Program Death Ligand-1+ (PDL1+) human tumor cells prevents PDL1 binding to its receptor PD1 on activated T cells, and enhances activation of tumor-reactive T cells. To distinguish if CD80 is facilitating T cell activation by preventing PDL1-PD1 interactions and/or by functioning as a costimulatory molecule, we have generated a recombinant CD80 molecule that does not bind to CD28. Since we hypothesize that CD80 may be a therapeutic for inhibiting tumor-mediated immune suppression via PDL1-PD1 interactions, the recombinant CD80 is also designed to not interact with CTLA4. Using site directed mutagenesis we generated CD80 with the following mutations: H96A, L97A, and E99A (CD8096,97,99). CD28 and CTLA4 bind to 35.3% and 37.4%, respectively, of tumor cells expressing wild type CD80. In contrast, only 1.2% and 1.4% of tumor cells expressing CD8096,97,99 bind CD28 and CTLA4, respectively, demonstrating that the mutant CD80 does not bind to CD28 or CTLA4. Previous studies demonstrated that CD80 prevents PDL1-PD1 interactions by binding to PDL1 and obscuring its detection. Only 7.05% of CD8096,97,99 mutants have detectable PDL1 (vs. 98% of untransfected parental human tumor cells), demonstrating that the CD8096,97,99 mutant does not bind CD28 or CTLA4, but retains the ability to inhibit PDL1-PD1 interactions. We are also generating a soluble form of CD8096,97,99 (sCD8096,97,99) that could be administered to cancer patients to prevent tumor cell-mediated PDL1 immune suppression. The construct for the soluble form was generated by ligating the two extracellular domains of CD80 containing the 96,97,99 mutations to the Fc region of human IgG1, followed by incorporation into the Pet21a+ plasmid containing the T7 promoter. Recombinant protein will be isolated from transformed E. coli using a protein G affinity column, and tested in functional experiments with PDL1+ tumor cells to determine if CD80 increases T-cell activation by inhibiting PDL1-PD-1 immune suppression and/or by facilitating costimulation.

This work was funded, in part, by grants from the HHMI Precollege and Undergraduate Science Education Program (52006949), CBI (T32 GM066706), and NIH (R01CA84232 and RO1CA115880).

Citation Format: Sonia P. Dalal, Samuel Haile, Preethi Somasundaram, Suzanne Ostrand-Rosenberg. Mutated CD80 may facilitate T-cell activation by inhibiting PDL1-PD1 suppression and by costimulating. [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 1264. doi:10.1158/1538-7445.AM2013-1264
Abstract 1267: Identification of HPV-derived CD4+ T-helper epitopes for improving therapeutic anti-HPV vaccine potency.
Authors: Grabowska Agnieszka K. , Hoppe Stephanie , Blatnik Renata , Riemer Angelika B.
Published: August 2014

Persistent infection with high-risk types of human papillomavirus (HPV) can lead to malignant transformation and the development of various human malignancies, such as cervical cancer, anal cancer or a subset of oropharyngeal carcinomas. Fortunately, most HPV infections are successfully eradicated by the immune system. Accumulating evidence suggests that both CD8+ and CD4+ T cell responses play a pivotal role in the clearance and control of HPV infection. Up to now, therapeutic anti-HPV vaccines have mostly been developed to elicit CD8+T cell responses. The exclusive targeting of HLA class I-restricted HPV epitopes might be insufficient, as HLA class I surface expression may be reduced as a result of HPV immune evasion strategies. In contrast, HLA class II molecules are expressed in high-grade cervical lesions and cervical cancer. Therefore in this study, we aim to identify novel E2-, E5-, E6- and E7-derived CD4+ T cell epitopes that bind to multiple HLA class II alleles, are naturally processed and presented, and immunogenic.

We started with in silico epitope predictions using several web-based prediction algorithms. 15-mer candidate epitopes from the HPV16 E2, E5, E6 and E7 proteins were determined for seven HLA class II alleles - DRB1*0101 (DR1), DRB1*0301 (DR3), DRB1*0401 (DR4), DRB1*0701 (DR7), DRB1*1101 (DR11), DRB1*1501 (DR2b), and DRB1*0901 (DR9); together providing >90% population coverage. To identify promiscuous epitopes, the average binding score to multiple HLA-DR alleles was calculated and peptides ranked accordingly. To assess immunogenicity of HPV-derived candidate peptides, IFN-γ-ELISPOT assays were conducted with lymphocytes isolated from healthy donors. T cells isolated from the majority of healthy donors responded to at least one HPV-derived peptide. The most reactive HPV-derived peptides were further investigated in long-term T cell cultures. Significant production of IFN-γ as well as T cell proliferation was observed in nearly 100% of tested individuals against this panel of epitopes.

As the above immunoreactivities most likely represent memory responses that led to viral clearance in healthy donors, we conclude that the identified CD4+ T-helper epitopes are candidates for the development of comprehensive epitope-specific HPV immunotherapy approaches.

Citation Format: Agnieszka K. Grabowska, Stephanie Hoppe, Renata Blatnik, Angelika B. Riemer. Identification of HPV-derived CD4+ T-helper epitopes for improving therapeutic anti-HPV vaccine potency. [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 1267. doi:10.1158/1538-7445.AM2013-1267
Abstract 1273: Adaptive immunity in a zebrafish model of melanoma.
Authors: Painter Corrie A. , Ceol Craig
Published: August 2014

The recent success of the anti-CLTA-4 antibody, ipilimumab, for late stage metastatic melanoma, provides proof of principle that stimulating the immune system can have profound effects on patient outcome. However, a detailed understanding of the role of immune-modulating cells within the tumor microenvironment has been confounded by the lack of a suitable model system that would allow for direct visualization of cells within the intact tumor microenvironment. Previous work has demonstrated that the zebrafish, Danio Rerio, can be genetically manipulated to produce melanomas that recapitulate BRAFV600E;P53-/- human tumors. Due to its translucency, ease of genetic manipulation and high fecundity, the zebrafish is an attractive model system that allows for in vivo characterization of cells within intact tissue. Using this model system, we have begun investigating the adaptive immune response to melanoma in zebrafish. We have focused our initial investigations on determining whether transcript expression of key adaptive immune-modulatory proteins are present in the zebrafish and to what extent they are expressed intratumorally . Our studies have found that orthologs of CLTA-4, FOXP3, and CD8 are all expressed in wild-type zebrafish, but not in rag-/- zebrafish. Furthermore, transcript levels of CTLA-4 were greatest in the melanoma tumor relative to other regions of the zebrafish, which is consistent with the constitutive expression of CTLA-4 in human melanomas. Additionally, we have cloned the promoters for zebrafish FOXP3 and CD8 and are developing fluorescent reporter lines that will enable us to visualize and ablate subpopulations of T cells within the tumor. These lines, along with previously described reporter lines for MHC II and lck, will enable us to determine the role of adaptive immune cells during melanoma onset and progression in the zebrafish. Our research supports the notion that there is conservation between the human and zebrafish adaptive immune systems, and that similar immune evasion mechanisms may be contributing to melanoma growth in zebrafish. These data position the zebrafsh as a powerful model system for immunological investigations that focus on the adaptive immune response to melanoma.

Citation Format: Corrie A. Painter, Craig Ceol. Adaptive immunity in a zebrafish model of melanoma. [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 1273. doi:10.1158/1538-7445.AM2013-1273
Abstract 1283: Role of ERCC1 in metformin enhancing paclitaxel-induced cytotoxicity.
Authors: Lin Yun-Wei , Tseng Sheng-Chieh , Huang Yu-Ching , Chiu Hsien-Chun , Chen Huang-Jen , Weng Shao-Hsing , Huang Yi-Jhen , Wo Ting-Yu
Published: August 2014

Metformin, an extensively used and well-tolerated drug for treating individuals with type 2 diabetes, has recently gained significant attention as an anticancer drug. On the other hand, paclitaxel (taxol) is a new antineoplastic drug that has shown promise in the treatment of non-small cell lung cancer (NSCLC). High expression levels of excision repair cross-complementary 1 (ERCC1) in cancers have been positively associated with the DNA repair capacity and a poor prognosis in NSCLC patients treated with platinum-containing chemotherapy. In this current study, paclitaxel was found to increase phosphorylation of mitogen-activated protein kinase (MAPK) kinase 3/6 (MKK3/6)-p38 MAPK as well as protein and mRNA levels of ERCC1 in H1650 and H1703 cells. Moreover, paclitaxel-induced ERCC1 protein and mRNA levels significantly decreased via the downregulation of p38 activity by either a p38 MAPK inhibitor SB202190 or p38 knockdown with specific small interfering RNA (siRNA). Specific inhibition of ERCC1 with siRNA was found to enhance the paclitaxel-induced cytotoxic effect and growth inhibition. Furthermore, metformin was able to not only decrease the paclitaxel-induced p38 MAPK-mediated ERCC1 expression, but also augment the cytotoxic effect induced by paclitaxel. Finally, expression of constitutive activate MKK6 or HA-p38 MAPK vectors in lung cancer cells was able to abrogate ERCC1 downregulation by metformin and paclitaxel as well as cell viability and DNA repair capacity. Overall, our results suggest that inhibition of the p38 MAPK signaling by metformin coupled with paclitaxel therapy in human NSCLC cells may be a clinically useful combination, which however will require further validation.

Citation Format: Yun-Wei Lin, Sheng-Chieh Tseng, Yu-Ching Huang, Hsien-Chun Chiu, Huang-Jen Chen, Shao-Hsing Weng, Yi-Jhen Huang, Ting-Yu Wo. Role of ERCC1 in metformin enhancing paclitaxel-induced cytotoxicity. [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 1283. doi:10.1158/1538-7445.AM2013-1283
Abstract 1284: Combined genetic and nutritional risk models of triple negative breast cancer.
Authors: Hu Jennifer J. , Lee Eunkyung M. , Allen Glenn O. , Levine Edward A.
Published: August 2014

Purpose: Triple negative breast cancer (TNBC), which lacks treatment targets for estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), has been associated with African-American race, BRCA1/2 mutations, TP53 mutations, and poor prognosis. However, the etiology of TNBC remained unknown, which represents a critical barrier in reducing breast cancer mortality. Therefore, we designed a case-only study to test combined genetic and nutritional risk models of TNBC.

Methods: In 354 breast cancer patients diagnosed between 1998 and 2004, we evaluated 18 DNA-repair non-synonymous single nucleotide polymorphisms (nsSNPs) using the Sequenom MassARRAY system. The 110-item Block 1998 food frequency questionnaire was used to evaluate dietary/nutritional intakes. The Multivariate Adaptive Regression Splines (MARS)-logit models were used to define nutritional intakes cut-offs for their associations with TNBC.

Results: TNBC was associated with 6 DNA-repair nsSNPs, ERCC4 R415Q (rs1800067), MSH3 R940Q (rs184967), MSH6 G39E (rs1042821), POLD1 R119H (rs1726801), XRCC1 R194W (rs1799782), and XPC A499V (rs2228000) and/or 3 micronutrients (zinc, folate, and β-carotene). Combined DNA-repair risk genotypes and lower micronutrient intakes had a significant dosage-dependent association with TNBC (p<0.001): ORs=2.77 (95%CI=1.01-7.64), 9.57 (95%CI=2.87-31.84), and 20.78 (95%CI=2.46-175.71) for 2, 3, and at least 4 risk factors, respectively.

Conclusion: To the best of our knowledge, this is the first study to demonstrate multiple DNA repair SNPs and nutritional factors are associated with TNBC. Our results suggest that dietary modulation and micronutrient supplementation may have implication in preventing or reversing TNBC, particularly in genetically susceptible women.

Citation Format: Jennifer J. Hu, Eunkyung M. Lee, Glenn O. Allen, Edward A. Levine. Combined genetic and nutritional risk models of triple negative breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1284. doi:10.1158/1538-7445.AM2013-1284
Abstract 1298: The role of nuclear factor kappa B in survival of human papillomavirus-positive head and neck cancer cell lines.
Authors: Zhang Hongzheng , Hu Zhongliang , Jiang Ning , Marullo Rossella , Doetsch Paul W. , Hunt William D. , Chen Georgia Z. , Shin Dong M.
Published: August 2014

Nuclear factor kappa B (NFκB) has been implicated in the development of head and neck squamous cell carcinoma (HNSCC) from premalignant changes to tumor progression, and in treatment resistance and recurrence. However, the roles of NFκB in the sensitivity to chemoradiation of human papillomavirus (HPV)-associated HNSCC have not been well defined. Previously we reported that high expression of NFκB detected by immunohistochemistry in pretreated HPV+ oropharyngeal cancer patients was significantly associated with poor progression free survival (AACR 2012 Abstract #4489). Therefore, we hypothesized that NFκB could play a role in pro-survival in HPV+ head and neck cancer cells. We first examined the sensitivity of five HPV+ head and neck cancer cell lines (HPV16: UDSCC2, 93-VU-147T, UM-SCC47, UPCI:SCC90, HPV18: SCC1483) to radiation and cisplatin, Consistent with current models, when we knocked down expression of the HPV16E6 oncogene, SCC47 and SCC1483 cells with restored expression of p53 displayed a significant reduction in S phase entry as revealed by cell cycle analysis (SCC47: scrambled control 14.08% ± 0.55% versus siRNA-E6 6.39% ± 2.40, p=0.004; SCC1483: scrambled control 13.39% ± 1.28% versus 7.49% ± 0.83, p=0.002). In addition, all five cell lines displayed reduced sensitivity to cisplatin compared with cells treated with scrambled control siRNA. The basal level of activated NFκB in HPV+ cell lines was correlated with the sensitivity to radiation and cisplatin as assessed by colony formation and cell growth assays. Suppression of NFκBp65 by siRNA resulted in marked increases in apoptosis associated with reduction of Bcl-2 in both SCC47 and SCC1483 cells. This pattern was more pronounced with increasing doses of the proteasome inhibitor bortezomib (32.5 to 130 nM for 48 to 72 hr). Furthermore, SCC47 and SCC1483 cells treated with siRNA NFκBp65 displayed a significantly greater level of apoptosis induced by radiation or cisplatin compared to cells treated with non-targeted control siRNA, indicating a pro-survival effect for NFκB. Further studies are warranted to delineate the effects of HPV oncogene interactions with NFκB signaling pathways on chemoradiation sensitivity. (This study was supported by the V Foundation).

Citation Format: Hongzheng Zhang, Zhongliang Hu, Ning Jiang, Rossella Marullo, Paul W. Doetsch, William D. Hunt, Georgia Z. Chen, Dong M. Shin. The role of nuclear factor kappa B in survival of human papillomavirus-positive head and neck cancer cell lines. [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 1298. doi:10.1158/1538-7445.AM2013-1298
Abstract 1308: ERRβ ligand DY131 regulation of the hedgehog signaling pathway by modulation of Akt phosphorylation.
Authors: Lu Yuan , Merideth Benjamin R. , Lin Hui , Lubahn Dennis B.
Published: August 2014

Nuclear receptor family member, Estrogen Related Receptor β (ERRβ), and hedgehog (Hh)-signaling are reported to be significant for both tumorigenesis and stem cell maintenance. ERRβ itself is a crucial factor in inducing pluripotent stem cells. Here we investigate the hypothesis that ERRβ can interact with Hh signaling pathway, and modulation of ERRβ can influence Hh signaling. NIH3T3 and HEK293 cells, transfected with a Gli response element driven luciferase reporter, were treated with different concentrations of DY131, a synthetic ERRβ ligand, in the presence or absence of exogenous ERRβ. We monitored the Gli reporter activity, as well as Gli1, Akt and phosphorylated Akt via western blot.

Results: We found that DY131 inhibited Hh signaling activity when the pathway was activated by hedgehog peptide-conditioned medium, or the Smoothened Agonist (SAG), with an IC50 of 500nM. We also found that in HEK293 cells, where Gli reporter activity cannot be activated by either conditioned medium or SAG, DY131 dose dependently increased both endogenous and exogenous Gli1-induced reporter activity up to 4 fold. In addition, the transfection of ERRβ potentiated DY131’s effect on Gli reporter activity up to 7 fold. Western blot results, which are consistent with Gli reporter activity in both cell lines, showed that when Hh signaling is activated by Hedgehog peptide-conditioned medium, DY131 decreased Gli1 protein concentration; however when Hh signaling is not activated by conditioned medium, DY131 increases the concentration of Gli1 protein. Phosphorylated Akt, which is a non-canonical activator of Gli1, is up-regulated when Gli1 concentration increases, and down-regulated when the cells are treated with DY131.

Conclusion: Our results suggest a new level of regulation by ERRβ on the Hh-signaling pathway. ERRβ, induced by DY131, can modulate Akt's regulation of Gli1. Additionally, our results indicate that modulation of ERRβ, via ERRβ selective drugs like DY131, may serve as a new target for Hh-driven cancer treatment.

Citation Format: Yuan Lu, Benjamin R. Merideth, Hui Lin, Dennis B. Lubahn. ERRβ ligand DY131 regulation of the hedgehog signaling pathway by modulation of Akt phosphorylation. [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 1308. doi:10.1158/1538-7445.AM2013-1308
Abstract 1321: Castration-resistant prostate cancer: BAY 1024767 blocks function of mutated androgen receptor.
Authors: Haendler Bernard , Sugawara Tatsuo , Lejeune Pascale , Koehr Silke , Faus Hortensia , Busemann Matthias , Cleve Arwed , Luecking Ulrich , von Nussbaum Franz , Brands Michael , Mumberg Dominik
Published: August 2014

Analysis of biopsies from prostate cancer metastases and of circulating tumor cells shows that androgen receptor (AR) mutations are often found in patients developing resistance to antiandrogen therapy, i.e. at the castration-resistant prostate cancer (CRPC) stage. Here we report on preclinical studies with a novel androgen receptor suppressor compound BAY 1024767 that exhibits strong antagonism for wild-type and mutated AR, both in vitro and in vivo.

Mutations located in the AR ligand-binding domain (LBD) and potentially involved in the resistance to the standard of care bicalutamide were generated and tested in cell-based transactivation assays, using 0.1 nM R1881 for stimulation. Bicalutamide showed reduced antagonistic activity for AR E709Y and no antagonism up to 10 μM for AR W741C. In comparison, BAY 1024767 exhibited strong antagonism for these mutants.

Another mechanism leading to CRPC is the elevation of intratumoral androgen levels. To mimick this situation, the transactivation assays were performed at a higher (1 and 10 nM R1881) androgen concentration. Bicalutamide, and also the novel antiandrogen MDV3100, showed much reduced antagonism for wild-type and W741C or E709Y mutant AR. Conversely, antagonism was still observed for BAY 1024767, especially at 1 nM R1881.

The KuCaP-1 model, which is derived from a CRPC patient and harbors the W741C mutation, was obtained from the group of O. Ogawa (Kyoto University). Mice bearing the KuCaP-1 xenograft subcutaneously were treated with bicalutamide (60 mg/kg, PO, qdx20) or BAY 1024767 (50 mg/kg, PO, q2dx10). No effect on tumor growth was observed with bicalutamide (T/C=103%) whereas BAY 1024767 was able to inhibit tumor growth with a T/C of 36% on day 54 post-implantation. In another study we found that MDV 3100 (100 mg/kg, PO) and abiraterone acetate (200 mg/kg, PO) were inactive in the KuCaP-1 model (100% and 73% T/C, respectively).

Citation Format: Bernard Haendler, Tatsuo Sugawara, Pascale Lejeune, Silke Koehr, Hortensia Faus, Matthias Busemann, Arwed Cleve, Ulrich Luecking, Franz von Nussbaum, Michael Brands, Dominik Mumberg. Castration-resistant prostate cancer: BAY 1024767 blocks function of mutated androgen 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 1321. doi:10.1158/1538-7445.AM2013-1321
Abstract 1325: Chlorinated bisphenol A diglycidyl ether (EPI-001) mediates degradation of the androgen receptor in prostate cancer cells.
Authors: Brand Lucas J. , Dehm Scott M.
Published: August 2014

The androgen receptor is a critical target for the treatment of locally advanced and metastatic prostate cancer (PCa). Blocking AR signaling via castration or antiandrogens such as bicalutamide and enzalutamide results in tumor regression. However, resistance to AR-targeted therapy inevitably develops, marked by the outgrowth of castration-resistant prostate cancer (CRPC) cells. Mechanisms of CRPC include amplification of the AR gene, leading to overexpression of AR mRNA and protein, as well as expression of truncated AR splice variants. AR overexpression can convert bicalutamide to a weak agonist, thereby driving AR reactivation and ongoing cellular proliferation and survival. AR splice variants lack the COOH-terminal AR ligand binding domain (LBD) and function as constitutively-active, ligand-independent transcription factors. Recently, the chlorinated bisphenol A diglycidyl ether, EPI-001, was shown to bind to the AR NH2-terminal domain and inhibit AR signaling by interfering with coactivator recruitment. Here, we show that EPI-001 has a novel activity of mediating degradation of AR protein in various PCa cell lines, including cells driven by AR gene amplification and cells driven by AR splice variants. Cells displaying resistance to the next-generation antiandrogen enzalutamide were growth inhibited at EPI-001 concentrations corresponding to maximal AR degradation. Moreover, using VCaP cells as a model of AR overexpression, we show that EPI-001-mediated AR degradation prevents inappropriate agonist response to bicalutamide. This work provides proof-of-principle that targeting full-length and splice variant forms of the AR for degradation with EPI-001 may enhance the efficacy of conventional and next-generation antiandrogens for treatment of CRPC.

Citation Format: Lucas J. Brand, Scott M. Dehm. Chlorinated bisphenol A diglycidyl ether (EPI-001) mediates degradation of the androgen receptor in prostate 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 1325. doi:10.1158/1538-7445.AM2013-1325
Abstract 1331: Ethnic-specific polymorphisms in CHRNA3 and CHRNA9 genes.
Authors: Chikova Anna , Grando Sergei
Published: August 2014

Several recent studies demonstrated significant association of numerous common polymorphisms in the CHRNA5-CHRNA3-CHRNB4 gene cluster encoding the α5, α3 and β4 nicotinic acetylcholine receptor (nAChR) subunits with the risk of lung cancer. However, due to high linkage disequilibrium, role of each protein in the tumorigenesis remains unclear. Large number of the single nucleotide polymorphisms (SNPs) including a majority of amino acid substitutions escaped the focus of previous research. Although some ethnic variations in this cluster are known, previous research was focused on the SNPs common for various ethnic groups. Little is currently known about epidemiological role of haplotypes specific for ethnical groups. We have assessed the global frequency distributions of coding and some non-coding nAChR SNPs by direct sequencing of 3 regions of CHRNA3 and 5 regions of CHRNA9 genes in 330 individuals from anthropologically defined human populations. The results revealed statistically significant ethnic-specific differences. In CHRNA3, for 8 SNPs, including one non-synonymous amino acid substitution, and in CHRNA9 for 7, including 2 non-synonymous SNPs. Among these SNPs, some were unique to one or two ethnical groups. For example, Arg110His in CHRNA3 (rs77574318) was seen only in Russians with heterozygosity 0.06. Thus, analysis of any amino acid substitution uniquely present in a particular ethnic group may guide further studies of the role of nAChRs gene polymorphism in various types of cancer

Citation Format: Anna Chikova, Sergei Grando. Ethnic-specific polymorphisms in CHRNA3 and CHRNA9 genes. [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 1331. doi:10.1158/1538-7445.AM2013-1331

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1332: Functional killer-immunoglobulin-like receptor variation and risk of non-melanoma skin cancer.
Authors: Vineretsky Karin A. , Karagas Margaret R. , Pawlita Michael , Nelson Heather H.
Published: August 2014

Natural killer (NK) cells of the immune system are the first line of defense against transformed and infected cells. NK cell action is determined through binding of killer immunoglobulin-like receptors (KIR) with specific human leukocyte antigen (HLA) class I ligands on target cells. The KIR gene family is diverse and highly polymorphic, is poorly covered on GWAS platforms, and exhibits significant functional copy number variation. The KIR locus has been primarily studied in the context of HCV, HIV, and transplant outcomes. Here, we have tested the hypothesis that KIR genotypes are associated with Non-Melanoma Skin Cancer (NMSC) etiology. These are among the most prevalent malignancies worldwide with rising incidence. In addition, they are highly antigenic tumors, with a possible viral etiology, making NMSC an ideal model to test KIR as a cancer susceptibility trait. A random subset of a population-based case-control study in New Hampshire was selected for the study: 422 basal cell carcinomas (BCC) (1 BCC=243, 2 BCC=131, ≥3 BCC=48), 132 squamous cell carcinomas (SCC), and 222 controls. KIR genotyping was performed with Lifecodes KIR-SSO typing kit on Luminex using buffy coat derived genomic DNA, capturing KIR loci: 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 2DP1, 2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 3DL1, 3DS1, and 3DL2. Cutaneous HPV status was assessed using serum samples and established Luminex detection assays covering 16 genus beta HPV types.

The KIR2DL2/3 locus was associated with a significantly increased risk of BCC (OR=2.65, 95% CI: 1.08-6.48), which was particularly strong for cases with 3 or more tumors (OR=5.77, 95% CI: 1.30-25.66). For SCC, the KIR3DL1 gene was associated with a significantly increased risk of malignancy (OR=3.73, 95% CI: 1.51-9.22). Next we investigated the association of KIR and SCC within HPV strata. The KIR3DL1-SCC association was strongest among those with no evidence of HPV infection (OR=6.81, 95% CI: 1.78-26.12). In contrast, the non-functional KIR2DS4 variant related to an increased risk and the KIR2DL2 gene related to reduced risk of SCC among those with multiple β-HPV infections. Our results implicate a strong role for KIR in the etiology of NMSC. KIR2DL2/3 may be particularly important in the development of multiple BCC, and KIR3DL1 in the pathogenesis of SCC. Finally, we identified a pattern of KIR-SCC associations that supports a potential etiologic role of cutaneous HPV in SCC occurrence.

Citation Format: Karin A. Vineretsky, Margaret R. Karagas, Michael Pawlita, Heather H. Nelson. Functional killer-immunoglobulin-like receptor variation and risk of non-melanoma skin 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 1332. doi:10.1158/1538-7445.AM2013-1332
Abstract 137: Racial differences in physical activity after breast cancer diagnosis.
Authors: Hair Brionna , Hayes Sandi
Published: August 2014

Background: The established benefits of physical activity (PA) after breast cancer diagnosis, including reduced mortality and treatment-related side effects and improved quality of life, suggest that PA should become a component of breast cancer care. Research shows that breast cancer survivors decrease PA post-diagnosis, but these studies have been underpowered to examine racial differences or were not population-based.

Methods: The Carolina Breast Cancer Study, a population-based study of incident breast cancer cases, assesses PA levels pre- and post-diagnosis in a large sample (n=1,735) with a substantial proportion of African-Americans (21%). PA was calculated as the number of minutes spent performing moderate and/or vigorous activity per week.

Results: Women were less likely to meet PA recommendations of >150 minutes of weekly activity after their breast cancer diagnosis (35%) than before diagnosis (61%). African-American, lower income and less educated women were less likely to meet recommendations both pre-diagnosis and post-diagnosis, but the decline in meeting recommendations following diagnosis was consistent across these characteristics. Though the majority of women (63%) decreased activity post-diagnosis, 19% increased and 18% did not change their activity levels. The average decrease in weekly minutes of PA was 230 minutes (95% CI: 190, 270). On average, African-Americans reduced their PA by more minutes than whites (276 versus 218 minutes, respectively; p-value=0.14) as did lower income compared to higher income women (311 versus 199 minutes, respectively; p-value=0.002). Compared to those with decreased post-diagnosis PA, women who were white (OR: 1.3, 95% CI: 1.0, 1.7), higher income (OR: 1.6, 95% CI: 1.2, 2.1), or higher educated (OR: 1.5, 95% CI: 0.9, 2.5) were more likely to increase activity post-diagnosis.

Conclusions: After breast cancer diagnosis, cases who were African-American, had lower income, or were less educated were less likely to meet PA recommendations, less likely to increase activity, and tended to reduce PA by more minutes. Interventions should seek to increase post-diagnosis PA overall, with a focus on these subgroups.

Citation Format: Brionna Hair, Sandi Hayes. Racial differences in physical activity after breast cancer diagnosis. [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 137. doi:10.1158/1538-7445.AM2013-137
Abstract 1377: Guided imagery use in the peri-operative period of gynecologic oncology surgery.
Authors: Dickson Elizabeth L. , Gerber Matt , Jonson Amy L.
Published: August 2014

Objective: To assess the feasibility of using guided imagery therapy in the peri-operative period at the time of Gynecologic Oncology surgery.

Methods: IRB approval was obtained for this randomized control trial. Patients were recruited based on having an exploratory laparotomy surgery with the gynecologic oncology group. Research staff performed a phone interview and obtained written consent. Patients were randomized to standard of care therapy, blank CD which was to simulate noise reduction headphones, or a validated guided imagery CD. The intervention would be used in pre op waiting area, during surgery, and after surgery. VAS scores, amount of narcotics used, amount of anti-emetics used, FACT G and POMS surveys were collected.

Results: 12 patients were attempted to be recruited for the study from April 2012 to the present. Prior to study enrollment, all patients were excited about the study and willing participants. All 12 pre op nurses were a barrier to discuss the study with the patients in the pre op area. 2 patients were unable to be consented due to the pre-operative nursing staff baring the research staff from consenting. 3 of the 10 RNAs stated that the player was dysfunctional, though the players were tested prior to delivery to the pre-operative area. 7 of the 12 patients stated that they ‘forgot’ to listen to the CD prior to surgery, or that the nurses did not let them take the time to listen. Only 2 of the 12 patients actually listened to the CD prior to surgery; only 2 of the 12 had the CD played during surgery. 4 of the 12 never received the CD player and surveys after surgery on the surgical oncology floor. Only 1 of the 12 actually listened to the CD throughout the entire hospital stay. 5 patients were told that it was “too busy” to listen to the CD by the floor nursing staff. 6 CD players were lost or stolen during the course of the study. To date, only 1 patient in the control group completed all surveys. However, 3 patients voiced excitement and satisfaction when using the intervention CD during their immediate peri-operative time, and were documented by external observers to have resolution of their anxiety while using the intervention CD.

Conclusions: While subjectively beneficial for patients to decrease stress responses peri-operatively, integrative medicine techniques such as guided imagery will be difficult to implement until the culture of medicine embraces this novel modality.

Citation Format: Elizabeth L. Dickson, Matt Gerber, Amy L. Jonson. Guided imagery use in the peri-operative period of gynecologic oncology surgery. [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 1377. doi:10.1158/1538-7445.AM2013-1377
Abstract 1407: A transgenic mouse model for the Hedgehog pathway effector GLI1 induced mammary gland tumors.
Authors: Norum Jens H. , Andersson Agneta , Kasper Maria , Sørlie Therese , Toftgård Rune
Published: August 2014

Increased expression of the Hedgehog (Hh) signaling pathway effector GLI1 in breast cancer patients has been shown to correlate inversely with disease free and overall survival. Activation of the Hh signaling pathway is known to promote proliferation of breast cancer cells and stimulate mammosphere formation. In addition, the Hh signaling pathway has been reported activated in CD44 positive breast cancer stem cells. Despite the evidence supporting a role for Hh signaling in breast cancer development and maintenance, the model systems to study the Hh pathway in breast carcinogenesis are limited.

We used a transgenic mouse model to show that GLI1 expression in the mammary gland cause tumor formation. The GLI1 induced primary tumors showed different histological characteristics, suggesting that the GLI1 expression influenced the tumor initiating cells. The GLI1 induced primary tumors were orthotopically transplanted on NOD/SCID mice and we have established serial transplantable tumor lines. The different histological characteristics of the individual primary tumors have been maintained for >10 serial transplantations in NOD/SCID mice. The stem cell marker CD44 is expressed in the mouse mammary gland ducts. The CD44 positive cell population is increased in the GLI1 induced tumors. The G protein coupled receptor (GPCR) like protein Lgr5 has been suggested as a stem cell marker in several tissues, including mammary gland. We show that cells in the basal layer of the adult mammary gland ducts express Lgr5. The Lgr5 positive cell population is increased in the GLI1 induced tumors, suggesting a basal like characteristic of these tumors.

Taken together, our data show that expression of GLI1 in the mammary gland result in formation of mammary gland tumors with heterogeneous histological characteristics. The cell populations identified by the stem cell markers CD44 or Lgr5 are increased in the GLI1 induced tumors compared with the normal mammary gland. Furthermore, we have established serial transplantable GLI1 induced mammary gland tumor lines. These serial transplantable tumor lines are novel tools to address the role of GLI1 expression in breast carcinogenesis.

Citation Format: Jens H. Norum, Agneta Andersson, Maria Kasper, Therese Sørlie, Rune Toftgård. A transgenic mouse model for the Hedgehog pathway effector GLI1 induced mammary gland tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1407. doi:10.1158/1538-7445.AM2013-1407
Abstract 1428: Bone marrow adipocyte-derived CXCL1 and CXCL2 in prostate tumor progression in bone.
Authors: Hardaway Aimalie L. , Herroon Mackenzie M. , Rajagurubandara Erandi N. , Podgorski Izabela
Published: August 2014

Prostate cancer (PCa) is the second most common cause of death from cancer in men. Bone is a primary site for tumor metastasis and obese and overweight men have a three-fold higher risk of progression to metastatic disease compared to normal-weight men receiving the same treatment. Recent epidemiological evidence suggests that obesity is an independent prognostic contributor to the development of distant metastases and death from PCa. Obesity leads to adipocyte accumulation within the bone marrow, which stimulates infiltration of inflammatory cells, particularly macrophages, and promotes inflammation. Many of the inflammatory cytokines released by both adipocytes and macrophages are involved in increased tumor growth, neovascularization, and cell survival. CXCL1 and CXCL2 are two such chemokines that have been implicated in prostate, breast, and ovarian tumorigenesis. Therefore, we hypothesized that obesity-induced bone marrow adiposity and inflammation would alter prostate tumor behavior in the bone via CXCL1- and CXCL2-dependent mechanisms. Histological evaluations confirmed that mice fed high fat diet (HFD) had increased adipocyte accumulation within the bone. PC3-DsRed cells intratibially injected into HFD mice progressed strikingly faster and grew much larger than those injected into mice on normal (LFD) diet. RT PCR analyses revealed an induction of several inflammatory genes, including CXCL1 and CXCL2, in the marrow of obese mice, and these levels were further increased in tumor-bearing tibiae of HFD hosts. In vitro short-term exposure of tumor cells to media conditioned by primary bone marrow derived adipocytes (Adipo CM) led to increased growth and invasiveness. This effect was further potentiated by chronic exposure to Adipo CM. CXCL1 and CXCL2 expression and secretion increased in co-culture of tumor cells with bone marrow macrophages in the presence of adipocyte-derived factors. Studies are currently underway to determine the role of obesity-induced marrow CXCL1 and CXCL2 in osteoclastogenesis and bone resorption. We are also investigating additional inflammatory mediators that sustain the upregulation of CXC1 and CXCL2 in the tumor microenvironment. Collectively, the results of our studies demonstrate that adipocyte-induced upregulation of CXCL1 and CXCL2 may play a critical role in promoting prostate tumor aggressiveness and survival in the bone metastatic niche.

Citation Format: Aimalie L. Hardaway, Mackenzie M. Herroon, Erandi N. Rajagurubandara, Izabela Podgorski. Bone marrow adipocyte-derived CXCL1 and CXCL2 in prostate tumor progression in bone. [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 1428. doi:10.1158/1538-7445.AM2013-1428
Abstract 1431: Upregulation of CXCR2 and its ligands in KRAS oncogene-induced initiation of pancreatic cancer.
Authors: Purohit Abhilasha , Varney Michelle , Rachagani Satya , Ouellete Michelle , Batra Surinder K. , Singh Rakesh K.
Published: August 2014

Pancreatic cancer (PC) is considered the fourth leading cause of cancer-related deaths in the United States. The high mortality-to-incidence ratio for the disease can be attributed to late clinical presentation. Thus, the detection of early markers of this disease remains a major challenge in the field of PC research. Somatic mutations in oncogene KRAS are the most common and earliest events during development of PC, suggesting their crucial role in tumor initiation; however, the precise functional molecular mechanism of action remains poorly understood. CXCR2, a CXC chemokine receptor, and its ligands have also been found to be important in regulating angiogenesis and metastasis. Their enhanced expression is observed as early as the precursor lesions of PC. The objective of this study was to investigate the role of CXCR2 signaling in the KRAS(G12D)-induced PC development. To validate that mutant KRAS(G12D) is essential for this increased secretome, studies were performed using two cell line models having both exogenous and endogenous expression of mutant KRAS(G12D): Htert-HPNE-E6-E7-st/KRAS(G12D), which is an immortalized pancreatic duct-derived cell line having exogenous expression of mutant KRAS (G12D), and CD18/HPAF KRAS (G12D) -scram/Sh, which is a tumor-derived cell line. The endogenous expression of mutant KRAS (G12D) was silenced by stable expression of Sh-RNA specific to the KRAS(G12D) allele. Real time PCR, ELISA and western blot analysis of these cells revealed that mutant KRAS (G12D) regulates the expression of CXCR2 and its ligands at both transcription and protein levels. We observed an enhanced expression of the CXCR2 and its ligands in Htert-HPNE-E6-E7-st-KRAS(G12D) cells. Interestingly, stable knock-down of KRAS in CD18/HPAF-shKRAS(G12D) cells upregulated CXCR2 expression, but inhibited the expression of its ligands. In addition, immunohistochemical analysis of PC lesions from Pdx-cre-LSL-KRAS (G12D) mice showed a higher expression of both receptor and its ligands as the lesions progress to advanced stages. Taken together, these results suggest that KRAS effector pathways are essential for modulating the expression of CXCR2 and its ligand, which, in turn, can mediate cellular- responses in PC growth and progression.

Citation Format: Abhilasha Purohit, Michelle Varney, Satya Rachagani, Michelle Ouellete, Surinder K. Batra, Rakesh K. Singh. Upregulation of CXCR2 and its ligands in KRAS oncogene-induced initiation of pancreatic cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1431. doi:10.1158/1538-7445.AM2013-1431