bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 989: Mild ER stress increases human ABC transporter expression and drug transport function in polarized cell monolayers.
Authors: Fung King Leung , Pixley Jessica , Talbert Darrell J. , Kapoor Khyati , Ambudkar Suresh , Gottesman Michael M.
Published: August 2014

Expression of ATP-binding cassette (ABC) transporter genes such as MDR1 (ABCB1/P-gp) is associated with poor outcome in chemotherapy and a multidrug resistant phenotype in cancer. Endoplasmic reticulum (ER) stress has been implicated in tumor growth but its role in drug resistance remains unclear. Here, we hypothesize that ER stress could increase expression of drug efflux transporters. To test this hypothesis, we developed a transient expression system in which human ABC transporters were ectopically expressed in a cell line monolayer by the BacMam baculovirus. The baculovirus is able to infect several mammalian cell lines including LLC-PK1, T-84, and MA104 without affecting the tightness of the host cell monolayers. In LLC-PK1 cells infected by BacMam vectors, induction of mild ER stress by pre-incubation (pre-conditioning) with thapsigargin (0.5 μg/ml) or tunicamycin (1 μg/ml), which are well-characterized ER-stress inducers, increased ABCB1 (P-gp) and ABCG2 (BCRP) levels. Drug transport assays showed that P-gp expressing cells pre-conditioned with thapsigargin showed increased directional transport of [3H] paclitaxel across the cell monolayer. The effect of thapsigargin was inhibited by the presence of the Erk inhibitor PD098059 but not the P38(MAPK) inhibitor SB203580, or the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor LY294002, suggesting that expression of ABC transporter is influenced by mild ER stress response and, at least in part, by the ERK/MAPK signaling pathway. Although pre-conditioning with tunicamycin can increase P-gp expression, it also inhibits N-glycosylation of P-gp. Collectively, our findings indicate that mild ER stress responses up-regulate drug transporters introduced into cells by BacMam vectors.

Citation Format: King Leung Fung, Jessica Pixley, Darrell J. Talbert, Khyati Kapoor, Suresh Ambudkar, Michael M. Gottesman. Mild ER stress increases human ABC transporter expression and drug transport function in polarized cell monolayers. [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 989. doi:10.1158/1538-7445.AM2013-989
Abstract 996: Receptor tyrosine kinase AXL is upregulated in metformin resistant prostate cancer cells.
Authors: Bansal Nitu , Mishra Prasun , Cruz Kathleen , Bertino Joseph
Published: August 2014

Metformin is a biguanine derivative, widely used as an antidiabetic drug. Epidemiological studies on several types of cancers including breast, prostate, ovarian and pancreatic cancer show that patients treated with metformin have a lower incidence of cancer. Mechanistic studies have shown that metformin inhibits cancer cell viability by activating the AMP activated protein kinase (AMPK) pathway and inhibiting mTOR. In this study we developed a metformin resistance LnCap prostate cancer cells by exposing the cells continuously increasing concentrations of metformin. Micro-array data analysis comparing the resistant cells (4-fold) to parental cells showed that genes associated with receptor binding and with cell proliferation are primarily affected. EDIL3 (EGF-like repeats and discoidin I-like domains 3), EREG (eEpiregulin) and AXL (Axl receptor tyrosine kinase) are top three up regulated genes (∼ 35 fold). CALB2 (cCalbindin 2) and TPTE (transmembrane phosphatase with tensin homology) were the top down regulated genes (10 fold down regulated). AXL overexpression has earlier been demonstrated in Imatinib resistant CML cell lines and in CML cells from patients. We are currently investigating the role of Axl receptor tyrosine kinase in metformin resistant prostate cancer cells.

Citation Format: Nitu Bansal, Prasun Mishra, Kathleen Cruz, Joseph Bertino. Receptor tyrosine kinase AXL is upregulated in metformin resistant 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 996. doi:10.1158/1538-7445.AM2013-996
Abstract 998: Poly(ADP-Ribose) polymerase inhibitor overcomes Rad51- mediated prostate cancer cells’ resistance to Topoisomerase I inhibitor under hypoxia.
Authors: Wu Minghui , Zhang Jingsong , Wang Xue
Published: August 2014

Intra-tumoral hypoxia has been proposed to create a “mutator” phenotype through down-regulation of DNA repair, leading to increased genomic instability and drug resistance. There is increasing interest to target hypoxic cancer cells with DNA-damaging agents and inhibitors of DNA repair. Here, we showed that hypoxia did not sensitize castration-resistant prostate cancer cells to the poly(ADP-ribose) polymerase (PARP) inhibitor veliparib or the DNA-damaging topoisomerase I inhibitor: Irinotecan or SN38. Although DNA repair proteins like Rad51 were decreased under hypoxia, up-regulation of Rad51 and Rad51 nuclear foci formation was observed soon after treatment with veliparib and SN-38 under hypoxia. This Rad51 up-regulation occurred at the transcription level and was mediated at least in part by E2F1. Inhibiting Rad51 expression with siRNA overcame prostate cancer cells’ resistance to SN-38 under hypoxia. Combining veliparib with Irinotecan significantly enhanced DNA damage and apoptosis under both hypoxic and normoxic culture conditions. Such enhanced DNA damage and anti-tumor activities were seen in the presence of Rad51 up-regulation and confirmed in vivo with PC3 mouse xenografts. These data illustrate a dynamic regulation of Rad51 by prostate cancer cells in response to hypoxia and DNA damage. The veliparib and Irinotecan combination can be further explored as a treatment for metastatic castration-resistant prostate cancer patients who have progressed through standard treatments.

Citation Format: Minghui Wu, Jingsong Zhang, Xue Wang. Poly(ADP-Ribose) polymerase inhibitor overcomes Rad51- mediated prostate cancer cells’ resistance to Topoisomerase I inhibitor under hypoxia. [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 998. doi:10.1158/1538-7445.AM2013-998
Abstract ADV06: The power of social media in patient advocacy.
Authors: Ciccarella Annemarie
Published: August 2014

Patient advocates are harnessing the power of social media by developing relationships with like-minded advocates from around the globe. Worldwide collaboration is now achieved seamlessly thanks to outlets like twitter with messages appended using a hash tag

Of greatest interest to me is bringing the voice of the patient to every aspect along the research continuum. Our unique perspective as stakeholders whose lives are directly impacted brings invaluable input to the table. From assisting in the writing of lay abstracts for funding applications to the review of grant applications, the voice of the patient is crucial. No longer will content sit on the sidelines, advocates want to be active participants in their treatment and many of us are especially interested in playing a role in the determining what constitutes worthwhile research. If it doesn't matter to the vast majority of the patient population, is it worth pursuing? While each individual story is important, the collective voice of the community is the most important voice of all.

Through social media interaction, a "connected" advocate has their finger on the pulse of what is important within their community. We sit in major conferences, sharing presentations in real time with others who are not in attendance. Active social media advocates are sharing stories as they hit the medical journals, discussing their relevance and oftentimes have doctors participating those discussions. Patients as partners, as collaborators whose voices are making a difference represent a powerful and positive paradigm shift in medical treatment and in the research landscape.

Citation Format: Annemarie Ciccarella. The power of social media in patient advocacy. [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 ADV06. doi:10.1158/1538-7445.AM2013-ADV06
Abstract ADV07: Cancer simplified: A web-based learning system for newly-diagnosed patients.
Authors: Darrow Duncan
Published: August 2014

Mission Statement:

Improving the quality of care for cancer patients on the East End of Long Island by establishing the Nations’ largest free cancer counseling center with a regional focus.

Initiatives:

We provide psycho-social counseling to approximately 300 newly-diagnosed cancer patients each year with a professional staff that includes two oncology social workers, a clinical psychologist with a PhD and a registered nurse. We also author a unique book - Coping with Cancer on the East End - which is a “where-to-find-it” guide to regional cancer care resources.

Poster Description:

It will focus on a relatively new initiative at Fighting Chance which is called “Cancer Simplified.” This is, in essence, a “sister website.” It serves as a web-based learning system about the “ABCs” of cancer, using a PowerPoint platform and is intended for newly-diagnosed patients with a high school reading level.

Cancer simplified aims to demystify the basic concepts of “what is cancer” using depiction instead of jargon-laden explanation. A deck of power point slides is augmented by a set of videos portraying Fighting Chance patients discussing their “cancer journeys.” Finally, a third element, known as “Easy Read,” drills into some of the largest cancer-helpful websites (like the ACS and NCCN) and takes the visitor directly to the most user-friendly pamphlets embedded in that site.

Citation Format: Duncan Darrow. Cancer simplified: A web-based learning system for newly-diagnosed patients. [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 ADV07. doi:10.1158/1538-7445.AM2013-ADV07
Abstract ADV13: Kure It Cancer Research.
Authors: Jones Karen
Published: August 2014

Kure It funds researchers conducting innovative projects in the fields of kidney cancer and other underfunded malignancies. Kure It raises funds inseveral ways: through annual special events; contributions from family and community foundations; and partnerships with the self storage industry, through which we've created tailored programs and events aimed at raising funding and awareness. Since its inception in 2007, Kure It has raised over $1.6 million for research. Our goal is to raise $1 million annually going forward. In 2012, Kure It awarded grants totaling $900,000, including two $250,000 awards given through Kure It's partnership with the AACR. Kure It funds support the work of researchers at major cancer centers across the United States.

Citation Format: Karen Jones. Kure It Cancer 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 ADV13. doi:10.1158/1538-7445.AM2013-ADV13
Abstract ADV14: One advocate's journey: I went from fighting for my life to now fighting for others.
Authors: Keesen Kristen
Published: August 2014

Kristin Keesen, 45 year old colorectal cancer survivor from Colorado, describes her personal journey with colorectal cancer, and how she became inspired to work with Fight Colorectal Cancer to help others.

Fight Colorectal Cancer demands a cure for colon and rectal cancer. We educate and support patients, push for changes in policy that will increase and improve research, and empower survivors to raise their voices against the status quo.

Fight Colorectal Cancer believes that a cure means accessible, accurate population-based screening, and personalized treatment. To that end, we connect with people touched by colorectal cancer, provide information to help them cope with their current situation and then, if they choose, empower them to raise their voices and training advocates around the country. Fight Colorectal Cancer and our advocates educate lawmakers and regulators, pushing them for better policies. Our research advocates serve on research committees and help focus on meaningful outcomes. They also collaborate with a variety of prestigious academic institution on research projects.

In 2013, our One Million Strong initiative will help our advocates engage the general public by asking them to take the pledge to get screened if appropriate, and spread the word without screening.

Our programs include:

- Research Advocacy Training and Support (RATS): training lay advocates to participate effectively in the research process.

- Patient Information Services: providing information in multiple channels.

- Lisa Fund: career development funding for young researchers performing translational research relevant to patients with metastatic colorectal cancer.

Our seminal effort is Call-on-Congress, our annual advocacy training and lobbying event which is based on our regulatory and legislative priorities.

Citation Format: Kristen Keesen. One advocate's journey: I went from fighting for my life to now fighting for others. [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 ADV14. doi:10.1158/1538-7445.AM2013-ADV14
Abstract ADV15: Survivors teaching students: Saving women's lives program.
Authors: Leighton Susan L.
Published: August 2014

Since 2002, the Ovarian Cancer National Alliance has been working to save women's lives by helping health care practitioners learn to diagnose ovarian cancer earlier through the Survivors Teaching Students: Saving Women's Lives® (STS) Program. The program brings ovarian cancer survivors into the classrooms of medical schools, nursing schools, nurse practitioners and physician assistant programs to inform students about real women's experiences with the disease. Each woman who presents has a unique story. These women_many diagnosed at an advanced stage_are in a distinct position to help students become more sensitive to the disease's symptoms and risk factors that, in turn, can result in an earlier diagnosis. The STS program is currently presented in 74 medical schools nationwide, 3 nurse practitioner programs, 12 physician assistant programs, 42 nursing schools and 13 other allied health professional schools across the country. Awareness of symptoms and risk factors is one of the only tools available to women and their physicians which may lead to earlier diagnosis. Evaluation of the Survivors Teaching Students program has revealed that the students who have attended an STS presentation have an increased awareness of the symptoms and risk factors of ovarian cancer and when asked about the program after they entered medical practice, the respondents remembered the information they had learned in these presentations and felt it had impacted the ways in which they treated women presenting with these symptoms.

Citation Format: Susan L. Leighton. Survivors teaching students: Saving women's lives program. [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 ADV15. doi:10.1158/1538-7445.AM2013-ADV15
Abstract ADV22: Susan G. Komen story map.
Authors: Seago-Coyle Rebecca
Published: August 2014

The Susan G. Komen Foundation began as a promise between two sisters to end breast cancer forever. Susan Goodman Komen was diagnosed with breast cancer at the age of 33 and died three years later. Ms. Komen's younger sister, Nancy Goodman Brinker, created the foundation in 1982. The Komen Puget Sound Affiliate was founded in 1993 to forward the mission of our national organization - to save lives and end breast cancer forever by empowering people, ensuring quality of care for all and energizing science to find the cures. Since its inception, the Komen Puget Sound Affiliate has invested nearly $23 million in local nonprofit, tribal and government agencies that provide breast health and breast cancer services to medically underserved residents of our 16-county service area (Service Area).

The Puget Sound affiliate as a community that identify disparities, gaps in service and barriers to care that prevent women who live in our community from accessing, early detecting and surviving breast cancer.

Citation Format: Rebecca Seago-Coyle. Susan G. Komen story map. [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 ADV22. doi:10.1158/1538-7445.AM2013-ADV22
Abstract LB-104: Gadolinium-based nanoparticles sensitize head and neck carcinoma stem and nonstem cells to low and high LET radiation.
Authors: Aloy Marie-Thérèse , Miladi Imen , Armandy Emma , Billotey Claire , Tillement Olivier , Lux François , Perriat Pascal , Janier Marc , Rodriguez-Lafrasse Claire
Published: August 2014

Head and neck squamous cell carcinoma (HNSCC) is an aggressive and recurrent malignancy owing to intrinsic radioresistance and lack of induction of apoptosis. Several strategies aiming at radiosensitizing these tumors are currently being developed, one of these relying on the use of nanoparticles including high Z elements such as gadolinium (Z=64). Once delivered to the tumor, gadolinium-based nanoparticles (GBNs) should amplify the efficacy of radiotherapy through the generation of electron Auger cascades and secondary electrons.

This study aims at demonstrating the in vitro and in vivo radiosensitizing effect of sub-5 nm GBNs (composed of a core of gadolinium oxide, a shell of polysiloxane, and functionalized by diethylene triaminepentaacetic acid) in a SQ20B radioresistant HNSCC model. Owing to the crucial role of cancer stem cells in tumor initiation, disease recurrence and radioresistance, the radiosensitizing effect of GBNs was also tested in SQ20B stem-like cells.

The association of 0,6 mM GBNs with a photon or a 33.6 keV/µm carbon ion irradiation decrease significantly SQ20B cell survival. Radiosensitization goes through the increase in non-reparable DNA double-strand breaks, the shortening of G2/M phase blockage, the inhibition of cell proliferation, each contributing to the commitment into apoptosis. The combined treatment of GBNs with irradiation can also overcome the resistance of SQ20B stem-like cells.

Using an SQ20B tumor-bearing mouse model, we also demonstrate that GBNs in conjunction with photon irradiation significantly retard tumor growth compared with the radiation alone, with complete remission in some mice. Significantly, an increase in apoptosis and decrease in cell proliferation are also detected inside tumors in the combined treatment group.

These results suggest the interesting potential of GBNs in sensitizing resistant HNSCC tumors to radiotherapy and their possible contribution towards overcoming limitations of current cancer treatment.

This project was conducted under the framework of LANTHARAD (PDC019 CLARA 2010), PRRH-ETOILE and LABEX PRIMES (2012)

Citation Format: Marie-Thérèse Aloy, Imen Miladi, Emma Armandy, Claire Billotey, Olivier Tillement, François Lux, Pascal Perriat, Marc Janier, Claire Rodriguez-Lafrasse. Gadolinium-based nanoparticles sensitize head and neck carcinoma stem and nonstem cells to low and high LET radiation. [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 LB-104. doi:10.1158/1538-7445.AM2013-LB-104
Abstract LB-110: Overexpression of EIF5A2 induced by gene amplification or hypoxia plays a pivotal role in tumor metastasis and angiogenesis.
Authors: CAO Tingting , Li Yan , Fu Li , GUAN Xinyuan
Published: August 2014

Hypoxia is often happened in solid tumors, which activates hypoxia response genes such as hypoxia-inducible transcription factors to keep tumor growth under the stress. Here we report that expression of EIF5A2, a frequently overexpressed oncogene in esophageal squamous cell carcinoma (ESCC), can be induced by hypoxia. Expression of EIF5A2 increased significantly in all five tested ESCC cell lines under hypoxic stress (P<0.05), compared with normaxic condition. Besides induced by hypoxia, overexpression of EIF5A2 can be induced by gene amplification. Amplification of EIF5A2 at 3q26 has been frequently detected in various solid malignancies including ESCC. In the present study, overexpression of EIF5A2 was detected in over 40% of ESCC cases, which was significantly associated with lymph node metastasis (P=0.037), tumor invasion (P=0.044) and poor outcome (P=0.09) of ESCC patients. Functional study found that EIF5A2 promoted tumor metastasis and angiogenesis, two typical phenotypic changes of a tumor under hypoxic stress. Molecular study found that EIF5A2 was able to bind to the promoter region of HIF1α and up-regulate its expression, and subsequently promote tumor invasion and metastasis via epithelial-mesenchymal transition (EMT). EIF5A2 could also promote angiogenesis via up-regulating VEGF in a HIF1α-mediated manner. Both in vitro and in vivo assays showed that silencing EIF5A2 expression by RNAi with shRNA-eIF5A2 could effectively inhibit tumor growth, especially when combined with chemotherapeutic agents. Taken together, these findings demonstrated that overexpression of eIF5A2 induced by gene amplification or hypoxia plays critical roles in cancer metastasis and angiogenesis via HIF1α-mediated signaling pathway under normaxic or hypoxic condition, respectively.

Citation Format: Tingting CAO, Yan Li, Li Fu, Xinyuan GUAN. Overexpression of EIF5A2 induced by gene amplification or hypoxia plays a pivotal role in tumor metastasis and 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 LB-110. doi:10.1158/1538-7445.AM2013-LB-110
Abstract LB-111: A stable transcription factor complex nucleated by dimeric AML1-ETO controls leukemogenesis.
Authors: Sun Xiao-Jian , Wang Zhanxin , Wang Lan , Jiang Yanwen , Soong T. David , Kost Nils , Chen Wei-Yi , Elemento Olivier , Fischle Wolfgang , Melnick Ari , Patel Dinshaw J. , Nimer Stephen D. , Roeder Robert G.
Published: August 2014

Transcription factors are frequently altered in leukemia through chromosomal translocation, mutation or aberrant expression. AML1-ETO, a fusion protein generated by the t(8;21) translocation in acute myeloid leukemia (AML), is a transcription factor implicated in both gene repression and activation. AML1-ETO exists as dimer/oligomer through its NHR2 dimerization domain. Given that the AML1-ETO dimerization has been shown to be critical for its leukemogenic activity, it is important to identify coregulatory factors that “read” the NHR2 dimerization and contribute to leukemogenesis. We now show that, in leukemic cells, AML1-ETO resides in and functions through a stable protein complex (AETFC) that contains several hematopoietic transcription factors and cofactors. These AETFC components stabilize the complex through multivalent interactions, provide multiple DNA-binding domains for diverse target genes, colocalize genome-wide, cooperatively regulate gene expression, and contribute to leukemogenesis. Within the AETFC complex, the AML1-ETO dimerization is required for a specific interaction between the dimerized NHR2 domain and a novel NHR2-binding (N2B) motif in E proteins. Crystallographic study of the NHR2-N2B complex reveals a unique interaction pattern in which an N2B peptide makes direct contact with side chains of two NHR2 domains as a dimer, providing a novel model of how dimeric transcription factors create a new protein-binding interface through dimerization. Intriguingly, disruption of this interaction by point mutations abrogates AML1-ETO-induced hematopoietic stem cell self-renewal and leukemogenesis. These results reveal new mechanisms of action of AML1-ETO in leukemogenesis and provide potential therapeutic possibilities for targeting t(8;21)+ AML.

Citation Format: Xiao-Jian Sun, Zhanxin Wang, Lan Wang, Yanwen Jiang, T. David Soong, Nils Kost, Wei-Yi Chen, Olivier Elemento, Wolfgang Fischle, Ari Melnick, Dinshaw J. Patel, Stephen D. Nimer, Robert G. Roeder. A stable transcription factor complex nucleated by dimeric AML1-ETO controls leukemogenesis. [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 LB-111. doi:10.1158/1538-7445.AM2013-LB-111
Abstract LB-113: Genome-wide mapping of TFE3-fusion target genes.
Authors: Huang Ying , Baba Masaya , Hasumi Hisashi , Hasumi Yukiko , Schmidt Laura , Wakabayashi Yoshi , Zhu Jun , Yang Wenjing , Linehan Marston
Published: August 2014

Xp11.2 translocation renal cell carcinoma (RCC), also known as TFE3-fusion associated RCC, is a recently classified distinct subtype of RCC. It is defined by several different translocations involving chromosome Xp11.2, all resulting in gene fusions involving the Transcription Factor Binding to IGHM Enhancer 3 (TFE3) gene. However, the detailed function of TFE3-fusion proteins in TFE3-fusion associated RCC still remains unknown, and there is no effective treatment for patients with this RCC subtype. In this study we sought to investigate the detailed regulatory network of TFE3-fusion proteins by using doxycycline-inducible HA-tagged TFE3-fusion cell lines and chromatin immunoprecipitation (ChIP)-sequencing. Doxycycline-inducible cell lines that express HA-tagged TFE3-fusion (PRCC-TFE3, PSF-TFE3 and NONO-TFE3) and TFE3 wild-type proteins were established in the HK2 cell line. Genome-wide screening of the binding characteristics of TFE3-fusion proteins and TFE3 wild-type protein in doxycycline-induced cell lines was performed using a chromatin immunoprecipitation sequencing (ChIP-seq) assay. Hierarchical clustering was used to analyze the genes regulated by each type of TFE3-fusion protein compared with TFE3 wild-type protein. The total number of CHIP-seq reads in TFE3 wild-type cell line was 18552874 and enrichment of the promoter region was 1.2. In contrast, the total number of CHIP-seq reads in PRCC-TFE3, PSF-TFE3 and NONO-TFE3 cell lines were 15332969, 20426159 and 17639467, respectively. Enrichment of promoter region in each of the TFE3-fusion cell line was 1.5, 2.4 and 2.2, respectively. There were 80 motifs enriched (Z score >1.5) by expression of PRCC-TFE3 protein compared to TFE3-wild type protein, 104 motifs enriched by PSF-TFE3 protein and 39 motifs by NONO-TFE3 protein expression. Work is ongoing to identify the target genes that are co-regulated in TFE3-fusion cell lines relative to the TFE3 wild-type cell line. Our aim is to identify activated pathways in common among the different TFE3-fusion cell lines compared to the TFE3 wild-type cell line. We anticipate that this study will further our understanding of mechanisms that activate TFE3-fusion proteins facilitating the identification of therapeutic targets for effective treatment of TFE3-fusion associated RCC.

Citation Format: Ying Huang, Masaya Baba, Hisashi Hasumi, Yukiko Hasumi, Laura Schmidt, Yoshi Wakabayashi, Jun Zhu, Wenjing Yang, Marston Linehan. Genome-wide mapping of TFE3-fusion target 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 LB-113. doi:10.1158/1538-7445.AM2013-LB-113
Abstract LB-114: Genome wide analysis reveals a "liability" in the mutant p53 transcriptome.
Authors: Martinez Luis A. , Kollareddy Madhusudhan , Varanasi Lakshman
Published: August 2014

One of the mechanisms by which the oncogene mutant p53 (mtp53) suppresses chemotherapy induced apoptosis is by directly regulating gene expression. We speculated that within the mtp53 transcriptome, there may be transcriptional targets that constitute "liabilities" that can be exploited to kill mtp53 bearing cancer cells. CHIP-Seq analysis of mtp53 revealed that the key enzyme required for activation of nucleoside analogs, deoxycytidine kinase (DCK), is a direct target of mtp53. We demonstrate that mtp53 associates with the promoter of the DCK gene and that knockdown of mtp53 in multiple cancer cell lines results in reduced DCK mRNA and protein levels. Furthermore, we show that mtp53 cooperates with the ETS transcription factors to induce DCK expression. Knockdown of mtp53 or ETS proteins reduces the activity of a DCK promoter-luciferase construct, thus reinforcing that DCK is a transcriptional target of an ETS/mtp53 complex. In contrast, knockdown of wildtype p53 has no effect on the expression of DCK, indicating that the ability of mtp53 to regulate DCK expression is a gain-of-function activity. Based on our findings, we reasoned that by upregulating DCK expression, mtp53 may increase the sensitivity of cancer cells to nucleoside analogs. Indeed, shRNA knockdown of DCK reduces the sensitivity of cancer cells to gemcitabine. Importantly, suppression of mtp53 or ETS proteins also renders cancer cells less sensitive to gemcitabine. Thus, these data support the notion that mtp53 can sensitize cells to nucleoside analogs via DCK. Our data reveal the presence of a transcriptional "liability" in the mtp53 transcriptome that may help predict which patients will have a better therapeutic response to nucleoside analogs.

Citation Format: Luis A. Martinez, Madhusudhan Kollareddy, Lakshman Varanasi. Genome wide analysis reveals a "liability" in the mutant p53 transcriptome. [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 LB-114. doi:10.1158/1538-7445.AM2013-LB-114
Abstract LB-117: Myc phosphorylation suppresses transformation through novel regulatory regions.
Authors: Dingar Dharmendra , Wasylishen Amanda R. , Chan-Seng-Yue Michelle , Huang Ling , Bros Christina , Tu William , Meyer Natalie , Boutros Paul C. , Penn Linda Z.
Published: August 2014

Myc is a proto-oncogenic transcription factor, whose expression is deregulated in numerous human tumours and over-expression in animal models can cause tumorigenesis. Myc is important in cancer initiation and progression and therefore to develop successful approaches targeting Myc, a better understanding of both the mechanisms of Myc regulation and subsequent deregulation in cancer is important. Identification of Myc post-translational modifications (PTMs) that are essential for its oncogenic activity remains largely unknown. Therefore, detecting those key PTMs and there effect on gene expression would shed more light on the mechanisms of Myc dependent tumorigenesis. In our study, we screened a panel of Myc phosphorylation mutants in MCF10A cells for their ability to promote anchorage-independent colony growth and we identified two previously uncharacterized mutants that increased Myc-dependent transformation: S71A/S81A and 340Cl-A (T343A, S344A, S347A, and S348A). We further confirmed that gain-of-function serine and threonine mutants potentiated transformation in SH-EP neuroblastoma cells and three-dimensional MCF10A acini. Additionally, to investigate the mechanism of increased transformation and mutants' selectivity for gene transcription, we conducted genome-wide mRNA expression analysis of MCF10A acini and identified 112 genes regulated by all MYC alleles, and 158 genes regulated by all three transforming phosphorylation mutants but not wild-type MYC. We performed genome-wide ChIP in MCF10A cells, integrated this with gene transcription data, and showed that the majority of genes with altered expression also have MYC binding in their promoter regions, further suggesting that these gene expression changes are directly regulated by Myc. In conclusion, phosphorylation of S71/S81 and 340Cl amino acid residues negatively regulates Myc dependent transformation, suggesting that better understanding of Myc regulation through the phosphorylation of these residues may provide opportunities to therapeutically target Myc.

Citation Format: Dharmendra Dingar, Amanda R. Wasylishen, Michelle Chan-Seng-Yue, Ling Huang, Christina Bros, William Tu, Natalie Meyer, Paul C. Boutros, Linda Z. Penn. Myc phosphorylation suppresses transformation through novel regulatory regions. [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 LB-117. doi:10.1158/1538-7445.AM2013-LB-117
Abstract LB-127: HTLV-1 Tax inflicts severe DNA damage through the activation of nitric oxide production: A credible pathway for transformation of HTLV-1 infected cells.
Authors: Baydoun Hicham , Ratner Lee
Published: August 2014

Adult T-cell Leukemia-Lymphoma (ATLL) is an aggressive and fatal malignancy of CD4+ T-lymphocytes associated with HTLV-1 infection, and an effective treatment is not yet available. The exact molecular mechanism underlying ATLL has not been fully elucidated. Although the activation of cell proliferation and inhibition of tumor suppressors are considered key contributors to the leukemogenesis process, the accumulation of DNA damage and genomic instability are believed to be the driving force for tumorigenesis and are known to be a hallmark of cancer cells. How genomic instability accumulates in cancer remains an elusive question. One of the most serious forms of genomic instability is DNA Double Strand Breaks (DDSB) because they pose problems for transcription, replication and chromosome segregation. We have recently shown that the HTLV-1 oncoprotein Tax induces DDSB. In addition, Tax has been found to constitutively activate the key T-cell inflammatory pathway, NF-kB, and this activation is critical for the leukemogenic process associated with HTLV-1 infection. In the present study, we investigated whether any of the NF-kB target genes is critical in inducing DDSB. Of note, we found that the inducible nitric oxide synthase (iNOS), the enzyme that catalyzes the production of nitric oxide (NO) in T-cells, is a target of the NF-kB pathway and is highly expressed in HTLV-1 and Tax expressing cells. Nitric oxide is an important cellular signaling molecule involved in many physiological and pathological processes and is a part of the network of reactive species that inflict DNA damage as a defense mechanism after infection. Interestingly, our results show a correlation between the number of DDSB and the production of NO in tumors isolated from Tax transgenic mice. We also observed a dramatic reduction of DDSB when NO production was inhibited, indicating that Tax induces DNA damage through the activation of NO. Determination of the impact of NO on tumors in an ATLL mouse model will open a new area in the development of alternative strategies for the treatment/prevention of ATLL, and the conclusions will be directly relevant to a better understanding of many types of cancer.

Citation Format: Hicham Baydoun, Lee Ratner. HTLV-1 Tax inflicts severe DNA damage through the activation of nitric oxide production: A credible pathway for transformation of HTLV-1 infected 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 LB-127. doi:10.1158/1538-7445.AM2013-LB-127
Abstract LB-128: High throughput screening kinase activators of Atg4B for cancer therapy.
Authors: Shu Chih-Wen , Liu Pei-Feng , Reed John C.
Published: August 2014

Autophagy is an evolutionarily conserved process whereby cells catabolize damaged proteins and organelles for purposes of generating substrates for sustaining ATP production during times of nutrient deprivation. This self-eating process is associated with many diseases, particular in cancers. The theory remains controversial due to limitation of the assays about autophagy, which requires specific assays and further studies to reevaluate the role of autophagy on cancer cells. Autophagins/Atg4 are a class of cytosolic cysteine proteases and play dual roles in autophagy machinery, suggesting tightly control of Atg4 is critical for regulation of autophagy. There are four human genes encoding Atg4 (Atg4A, Atg4B, Atg4C, Atg4D). Among them, Atg4B appears functional dominance in regulation of autophagy. Knocking down Atg4B or overexpression of catalytic mutant of Atg4B inhibited cell growth in various cancer cells. Moreover, Atg4B reveals three phosphorylation sites at Ser 316, 383, 392 in cells through large-scale phosphoproteomics approach. Inhibiting phosphorylation of Atg4B diminished proteolysis activity both in vitro and in cells, suggesting kinases may serve as activators for Atg4B. Further, we employed mammalian cell-based Atg4B assay to screen potential kinases activators of Atg4B with kinome siRNA library, which consists 704 kinase related genes. Twelve hits were reproducible and reconfirmed with GFP-LC3 stably expressed cells as a secondary assay. Four kinase hits were interacted with Atg4B and showed same effect on GFP-LC3 cleavage. Knocking down kinase hits diminished Atg4B proteolysis activity and cell growth in cancer cells, demonstrating novel function of the kinase hits on Atg4B-mediated autophagy and cancer growth. Our finding may provide new kinases as biomarkers or drug targets for cancer therapy.

Citation Format: Chih-Wen Shu, Pei-Feng Liu, John C. Reed. High throughput screening kinase activators of Atg4B for cancer therapy. [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 LB-128. doi:10.1158/1538-7445.AM2013-LB-128

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract LB-130: The metabolic function of RB tumor suppressor gene.
Authors: Kohno Susumu , Kitajima Shunsuke , Sasaki Nobunari , Muranaka Hayato , Takahashi Chiaki
Published: August 2014

The RB tumor suppressor gene has been implicated primarily in the control of cell cycle and terminal differentiation. However recent findings indicate that pRB may possess much more roles than previously thought (Viatour & Sage, Dis. Model Mech., 4: 581, 2011; Takahashi et al., Cancer Sci., 103: 1182, 2012). We have been exploring unique roles of pRB using mice with various genetic backgrounds (Takahashi et al., Nat. Genet., 38: 118, 2006). A seminal observation in nematode prompted us to address how pRB controls Ras activation status. We previously reported that pRB regulates mevalonate (MVA) pathway through SREBPs thus affects isoprenylation of Ras proteins (Shamma et al., Cancer Cell 15:255, 2009). We are currently extending this observation to the investigation of pRB roles in controlling cellular metabolism. We identified a number of metabolic pathways that are affected by the status of RB in various cellular contexts and genetic backgrounds. Furthermore, a recent report from another group pointed out that the product of mutated p53 tumor suppressor gene also targets SREBP although in a different fashion (Freed-Pastor et al., Cell 148: 244, 2012). We therefore hypothesized that these two tumor suppressors are redundant in the control of lipid synthesis. As expected, depending on the context, mouse cells deficient of both Rb and p53 genes showed evidence of metabolic reprogramming and moreover features of cancer stem cells. In addition, mice deficient of both genes generated tumors with unique undifferentiated phenotypes associated with many kinds of cancer stem cell-like activities. We further analyzed such cells or tumor cells by transcriptomics, metabolomics, flux analyzer and high-throughput drug screening methods: these unveiled previously unexpected functions of pRB in controlling cell metabolism and stemness (Kohno et al., in preparation for submission).

Citation Format: Susumu Kohno, Shunsuke Kitajima, Nobunari Sasaki, Hayato Muranaka, Chiaki Takahashi. The metabolic function of RB tumor suppressor gene. [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 LB-130. doi:10.1158/1538-7445.AM2013-LB-130
Abstract LB-14: High-risk cervical human papillomavirus infections among human immunodeficiency virus-positive women in the Bahamas.
Authors: Blackman Elizabeth , Dames Dionne N. , Butler Raleigh , Taioli Emanuela , Eckstein Stacy , Devarajan Karthik , Griffith-Bowe Andrea , Gomez Perry , Ragin Camille
Published: August 2014

Background: It has been demonstrated in immunocompromised females that HR HPV genotypes other than 16 and 18 have been detected in a significant proportion of these individuals. We aim to evaluate the frequency of HR HPV genotypes in a population of HIV-positive Caribbean women, detect characteristics associated with HR HPV infections other than HPV16 and 18, and investigate the clustering of HPV genotypes in this population.

Methods: One hundred and seventy-six non-pregnant, HIV-positive females ≥18 years were recruited. Each participant received a vaginal examination and completed a questionnaire.

Results: Mean age was 38.7 years for women positive for HR HPV and 44.4 years for women negative for HR HPV (P value = 0.0018). The majority of the HR HPV-positive women were on HAART for 2-4 years. Non-negative matrix factorization identified HPV 16, 18, 52 and 58 as the most influential HPV high-risk types. Among the women who were HIV+ for more than 2 years, secondary CART analysis revealed those aged ≤29 had statistically significant different cervical abnormality profile compared to women >29 years; specifically, younger women were more likely to have dysplasia, while older women had normal cervical cytology. Also, the mean number of years on HAART for ≤29 years old was only 1 year (std dev. ±1.3) while those >29 years old the mean was 3.6 years (std. dev. ± 2.5) (p = 0.009).

Conclusions: HR HPV in the immunocompromised female consists of more than just HPV 16 and 18, but also types HPV 52 and 58, which are likely to influence the course of HPV infections and the development of cervical cytological abnormalities. Targeted interventions to boost treatment compliance among HIV- positive women are needed to reduce the risk of cervical abnormalities.

Citation Format: Elizabeth Blackman, Dionne N. Dames, Raleigh Butler, Emanuela Taioli, Stacy Eckstein, Karthik Devarajan, Andrea Griffith-Bowe, Perry Gomez, Camille Ragin. High-risk cervical human papillomavirus infections among human immunodeficiency virus-positive women in the Bahamas. [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 LB-14. doi:10.1158/1538-7445.AM2013-LB-14
Abstract LB-149: Sustained activity of the mTOR pathway is required for DNA damage-induced up-regulation of RRM2 in cancer cells.
Authors: Shen Changxian , Houghton Peter J.
Published: August 2014

Balanced deoxyribonucleotides (dNTPs) are essential for the accurate and efficient replication and repair DNA synthesis. Increased or imbalanced dNTP pool leads to a mutator phenotype, while decreased dNTP level impairs DNA replication and repair, and hence cell survival. Ribonucleotide reductase (RNR) is the rate-limiting enzyme for the production of dNTPs from ribonucleotides and is under tight control during cell cycle and in response to stress. Increase of RNR is required for yeast cells to survive DNA damage and replication stress. However, the regulation and function of RNR in response to DNA damage remains unclear and controversial in mammalian cells. We report here that the mTOR pathway controls both RRM1 and RRM2 but not p53R2. mTOR signaling is required for mitomycin C (MMC) induced up-regulation of RRM2 in rhabdomyosarcoma Rh30 cells. Pharmacological inhibition of mTOR kinase with AZD8055 or mTORC1 inhibitor rapamycin resulted in the decrease of both RRM1 and RRM2, but not p53R2. 4E-BP1 and 4E-BP2 double knockout mouse embryonic fibroblast cells (4E-BP1/2-/- MEF) showed increased basal expression of RRM1 and RRM2, indicating that mTORC1 controls RRM1 and RRM2 via cap-dependent protein translation. In addition, pharmacological inhibition of either mTORC2-Akt signaling or CDK4/6 led to decrease of the mRNAs of RRM1 and RRM2, accompanied with the suppression of pRb-S780 signaling, suggesting that mTORC2-Akt signaling controls the gene transcription of RRM1 and RRM2 via regulating G1 cyclin-dependent kinases (CDK4/6). Both rapamycin and AZD8055 impaired the induction of endogenous RRM2 by MMC, but not ectopically expressed RRM2, which is not controlled by mTOR signaling. Our data demonstrate that mTOR controls both the protein translation and gene transcription of RRM1 and RRM2. DNA damage induced up-regulation of RRM2 depends on sustained mTOR signaling, suggesting that control of RNR by mTOR signaling may be one of the mechanisms by which cell growth-promoting signaling pathways enhance the survival and genome instability of cancer cells under constant DNA replication and damage induced stress. Supported by UPHS award CA77776.

Citation Format: Changxian Shen, Peter J. Houghton. Sustained activity of the mTOR pathway is required for DNA damage-induced up-regulation of RRM2 in 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 LB-149. doi:10.1158/1538-7445.AM2013-LB-149