bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-166: Risk of anal cancer in women with human papillomavirus-related gynecological neoplasm: Puerto Rico 1987-2013
Authors: Acevedo-Fontanez Adrianna Isabel , Suarez Erick , Torres-Cintron Carlos R. , Ortiz Ana Patricia
Published: July 2017

Objective: This study aimed to: 1) describe age-adjusted incidences for invasive cervical, vulvar, vaginal, and anal cancer cases reported from 1987 to 2013 in Puerto Rico (PR); 2) to estimate the Standardized Incidence Ratio (SIR) of secondary anal cancer among women with HPV-related gynecological neoplasms (cervical, vulvar or vaginal) during 1987-2013; and 3) to estimate the magnitude of these associations, adjusting for age.

Methods: We identified 9,489 women who had been diagnosed with a primary cervical, vaginal or vulvar tumor during 1987-2013 in PR. To describe the trends of cervical, vulvar, vaginal, and anal cancer, the age-standardized incidences were estimated using the direct method (2000 US Std Pop). SIRs (observed/expected) were computed using the indirect standardization method; expected cases were calculated using age-specific rates of anal cancer in women from the general population of PR. The ratio of SIRs was estimated using the Poisson regression model to estimate the magnitude of the association between secondary anal cancer and HPV-related gynecological neoplasms.

Results: A significant increase in the incidence trend for anal cancer was observed from 1987 to 2013 (APC=1.1; p-value<0.05), whereas from 2004 to 2013, a significant increase was observed for cervical cancer incidence (APC=3.3; p-value<0.05). Women with HPV-related gynecological cancers had three times the risk of developing anal cancer in comparison with women with non-HPV-related gynecological cancers (RR: 3.27; 95%CI: 1.37, 7.79).

Conclusion: Anal cancer is increasing among women in PR. Women with gynecological HPV-related tumors are at increased risk of anal cancer as compared to women from the general population and to those with various other primary tumors. Further investigation on the public health implications of this association is needed, as well as appropriate screening guidelines for anal cancer in these populations.

Citation Format: Adrianna Isabel Acevedo-Fontanez, Erick Suarez, Carlos R. Torres-Cintron, Ana Patricia Ortiz. Risk of anal cancer in women with human papillomavirus-related gynecological neoplasm: Puerto Rico 1987-2013 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-166. doi:10.1158/1538-7445.AM2017-LB-166
Abstract LB-169: Targeted tumor prevention in Li-Fraumeni syndrome
Authors: Soragni Alice , Phan Nhan , Eisenberg David
Published: July 2017

Li-Fraumeni syndrome (LFS) is a rare, dominant cancer predisposition disorder. The majority of LFS cases are brought about by germline p53 mutations. Individuals affected develop a spectrum of cancers occurring since childhood or early adulthood (Mai et al, 2012). Tumors associated with LFS include but are not limited to sarcomas, lymphomas and leukemias, breast cancers, brain tumors, etc (Olivier et al, 2010; Mai et al, 2012). p53 is a tumor suppressor gene that regulates many cellular processes and prevents cancer formation, growth and dissemination. When inactivated by mutation, it not only loses its tumor suppressive function, but also acquires oncogenic properties. Mutations of p53 promote destabilization of its three-dimensional structure, which can cause p53 to partially unfold, aggregate and accumulate in cells (Xu et al, 2011). We developed a peptide, ReACp53, to arrest p53 clumping and accumulation, chaperone p53 to a functional state and promote its degradation, hence arresting tumor proliferation (Soragni et al, 2016). ReACp53 can also target p63 and p73 (Zheng et al, 2016). It has shown good tolerability and a favorable safety profile in vivo. In LFS patients and LFS animal models, p53 is undetectable in normal tissues. However, the protein starts to accumulate in tumors and pre-cancerous lesions (Lang et al, 2004; Soragni et al, unpublished data). As p53 accumulation is correlated with aggregation (Xu et al, 2011; Soragni et al, 2016), we postulated that, by periodically administering ReACp53, we could target this initial event early on, and prevent p53 accumulation and/or push the p53 aggregates-ridden cells into apoptosis (Soragni et al, 2016). Here, we tested ReACp53 as a tumor preventive agent, by administering it twice a week from weaning to LFS mice carrying the aggregation-prone R172H mutation (R175H in humans). ReACp53 delayed cancer onset resulting in a significant increase in overall survival (OS). The homozygous cohort (n=16/group) showed a 50% increase in median survival in ReACp53-treated mice (median OS 15.79 vs 23.93 weeks for vehicle vs ReACp53, p=0.005; HR=2.6). Heterozygous mice (n=8/group) showed a 36% increase in OS with a median of 58.86 vs 80.50 weeks for vehicle vs ReACp53 (p=0.03, HR=2.6). No long-term side effect was observed upon ReACp53 administration. The tumor spectrum reported differed between the two groups. Activation of a functional p53 response in mouse cells and LFS patient cells treated with ReACp53 is undergoing. Overall, our study supports ReACp53 therapy as a potential approach to tumor prevention in LFS.

Citation Format: Alice Soragni, Nhan Phan, David Eisenberg. Targeted tumor prevention in Li-Fraumeni syndrome [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-169. doi:10.1158/1538-7445.AM2017-LB-169
Abstract LB-174: Patient navigation in Appalachia: results from a survey of current practices
Authors: Dignan Mark B. , Dwyer Sharon , White Carol , Rohan Elizabeth A. , White Dana , Castro Georgina , Wilson Reda , Boyd Mary , Stockton Eric
Published: July 2017

Purpose: The Appalachian region of the US includes 420 counties in 13 states that begin with New York, follow the Appalachian Mountains and extend to Mississippi. The region has a heterogeneous population that shares distinct cultural characteristics, high rates of unemployment, low income, and high rates of chronic disease, including cancer. Patient navigators (PN) provide one-on-one guidance and assistance through the health care continuum from prevention to end-of-life and address barriers and enhance access to healthcare through the use of individual navigators who are sensitive to the needs, interests and values of the community. Although there are numerous PN training programs available, there are no known programs that are designed for the Appalachian population. As a first step in developing such a program, we designed and conducted a survey of PN training needs in Appalachia.

Methods: With guidance from formative research involving a variety of stakeholders, we identified key individuals, professional groups, health care systems, and public health agencies serving the Appalachian region. We developed a 26-item survey to collect information on current patient navigation programs, sources of current training, and training needs. After pretesting, the survey was distributed online. Data were collected from June, 2015 through May, 2016.

Results: A total of 152 responses were received. The responses were from community based organizations (29.3%), academic medical centers (15.5%), hospitals (35.8%), and community cancer centers (19.5%). PN was currently provided by individuals from a variety of professional backgrounds, chiefly nursing (32.8%) and social work (8.4%), and 15.3% were community navigators. Only 63.8% of respondents reported having received formal PN training. About half (47.3%) indicated that they were interested in attending PN training and suggested specific content and format.

Conclusions: Survey respondents identified a need for PN training that focuses on Appalachia. These results supported the development of an Appalachia Patient Navigation Training Program, currently underway.

Note: This abstract was not presented at the meeting.

Citation Format: Mark B. Dignan, Sharon Dwyer, Carol White, Elizabeth A. Rohan, Dana White, Georgina Castro, Reda Wilson, Mary Boyd, Eric Stockton. Patient navigation in Appalachia: results from a survey of current practices [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-174. doi:10.1158/1538-7445.AM2017-LB-174
Abstract LB-175: Gender and racial disparities in incidence trends of tobacco-related cancers from 2010 to 2013
Authors: Wilson Danyell S. , Parish Alice , Guo Yi
Published: July 2017

Over one in three cancer-related deaths are associated with tobacco use. Although the overall incidence trend of tobacco-related cancers (TRCs) has been decreasing in the past a few decades, evidence suggests that not all TRCs have decreasing incidence and there exists gender and racial disparities in the TRC trends. Prior studies of TRC incidence have used the Surveillance, Epidemiology, and End Results Program (SEER) database. However, the SEER database includes limited information regarding the United States southeastern region and has been shown not to be representative of the US. In this study, we analyzed TRC incidence trends from 2010 to 2013 using the Florida cancer registry data, collected from a large ethno-racially diverse state. We computed annual age-adjusted incidence rates and annual percentage change (APC) to characterized the incidence trends of the following cancers: oral cavity, cervical, respiratory (lung, bronchitis, and larynx), urinary (bladder, kidney, and renal pelvis), digestive (esophagus, stomach, and pancreas), non-Hodgkin’s lymphoma, and pharyngeal. All incidence trends were also evaluated stratifying by gender (men and women) and race (Non-Hispanic (NH) Whites, NH Blacks, and Hispanics). Overall, all TRCs had a significant decrease in age-adjusted incidence rates for the study period, except for non-Hodgkin’s lymphoma and pharyngeal cancer for which the incidence remained unchanged. Oral cavity cancer had the biggest decline in age-adjusted incidence rates (APC= −2.5; 95% CI= −3.4, −1.6), while cancers of the digestive system had the smallest decline in incidence rates (APC= −0.6; 95% CI= −1.0, −0.2). When broken down by gender, males have a greater decrease in incidence rates than women for urinary and respiratory cancers. In addition, while pharyngeal cancer incidence rate stayed unchanged overall, there was a significant increase in the rate among male (APC= 1.0; 95% CI= 0.3, 1.7), but not female. When broken down by race and ethnic groups, NH Blacks and Hispanics had greater decline in cancer incidence for cervical cancer, oral cavity cancer, and respiratory cancer. For pharyngeal cancer, there was a significant decrease in incidence among NH Blacks (APC= −3.7; 95% CI= −5.5, −1.9), while the incidence significantly increased among NH Whites (APC= 1.6; 95% CI= 0.7, 2.5). In summary, there was an overall decreasing trend of TRC incidence. However, increasing incidence of pharyngeal cancer was observed for male and NH Whites.

Citation Format: Danyell S. Wilson, Alice Parish, Yi Guo. Gender and racial disparities in incidence trends of tobacco-related cancers from 2010 to 2013 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-175. doi:10.1158/1538-7445.AM2017-LB-175
Abstract LB-182: Differential modulation of the PD-1 pathway impacts the anti-tumor activity of CAR T cells
Authors: Lin Regina J. , Gautron Anne-Sophie , Poirot Laurent , Wong Oi Kwan , Sasu Barbra , Boldajipour Bijan
Published: July 2017

Adoptive T cell therapy using ex vivo expanded tumor-infiltrating lymphocytes (TILs) or T cells engineered with chimeric antigen receptors (CAR T) has shown tremendous success in many hematological diseases, however treatments of solid tumors have been less successful so far. It is presumed that adoptive cell therapy is limited by the presence of immune-suppressive mechanisms within the solid tumor microenvironment. To enhance T cell activity against solid tumors, we seek to overcome these immune barriers and one approach is to do this by blocking checkpoints such as the PD-1 receptor. Using CAR T cells targeting the glioblastoma (GBM) tumor-specific antigen EGFRvIII, we investigated 3 approaches of inhibiting PD-1: (i) shRNA-mediated knockdown, (ii) TALEN®-mediated knockout, and (iii) antibody-mediated blockade. In contrast to the PD-1 antibody that augmented CAR T cell activity in vivo, the PD-1 shRNA impaired the anti-tumor activity of CAR T cells in vitro and in vivo. Detrimental effects of the PD-1 shRNA were associated with impaired survival of CAR T cells in vitro and in vivo. Interestingly, TALEN®-mediated PD-1 knockout generated CAR T cells with superior cytotoxic potency and prolonged functional persistence in vitro. Further characterization of PD-1 TALEN® CAR T cells, and how they compare in activity against the CAR T/anti-PD-1 combination in both subcutaneous and intracranial GBM models, is underway. These studies will not only elucidate the role of PD-1 in CAR T cell biology, but also inform CAR T cell combination therapy approaches for solid tumor indications.

Citation Format: Regina J. Lin, Anne-Sophie Gautron, Laurent Poirot, Oi Kwan Wong, Barbra Sasu, Bijan Boldajipour. Differential modulation of the PD-1 pathway impacts the anti-tumor activity of CAR T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-182. doi:10.1158/1538-7445.AM2017-LB-182
Abstract LB-185: A PSMA-directed natural killer cell approach for prostate cancer immunotherapy
Authors: Brand Lucas J. , Zaslavsky Alexander B. , Palapattu Ganesh S. , Knudsen Karen E.
Published: July 2017

Prostate Cancer (PCa) is the third leading cause of cancer mortality in men in the United States. Locally advanced or metastatic PCa is clinically managed with androgen deprivation therapy (ADT), androgen receptor antagonists, and/or taxane chemotherapy. While initially effective, patients eventually develop resistance, leading to onset of a lethal castration-resistant disease state for which no curative interventions currently exist. Cell-based immunotherapy—which is centered on infusion of modified, tumor-directed immune cells intended to drive a more potent anti-tumor response—could potentially provide a new approach to treating advanced PCa. In this study, we describe the modification of an allogeneic, immortalized human natural killer cell line, NK-92, with a chimeric antigen receptor (CAR) directed against prostate-specific membrane antigen (PSMA), a cell-surface antigen expressed by most prostate tumors. We observed that both unmodified and CAR-bearing NK-92 cells display potent cytolytic activity against the majority of PCa cell lines tested in vitro. Expression of PSMA on PCa cell lines affected the activity of CAR-modified, but not unmodified, NK-92. Combination of NK-92 immunotherapy with ADT and taxanes is also discussed, as well as ongoing in vivo preclinical modeling. Overall, CAR-modified NK-92 cells show promise as a novel immunotherapeutic agent for the treatment of PCa.

Citation Format: Lucas J. Brand, Alexander B. Zaslavsky, Ganesh S. Palapattu, Karen E. Knudsen. A PSMA-directed natural killer cell approach for prostate cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-185. doi:10.1158/1538-7445.AM2017-LB-185
Abstract LB-203: VXL100, a MUC1 signal peptide based immunotherapy, acts through a neoantigen-like mechanism, and shows promising effects in hematological cancers
Authors: Chen Limor , Kovjazin Riva , Shapira Michael
Published: July 2017

VXL100 (ImMucin®), a 21mer peptide, pan-HLA vaccine derived from the MUC1 signal peptide (SP) domain, has been shown to be safe and to elicit a robust and specific T and B-cell responses in stable and progressive multiple myeloma patients in a phase I/II clinical trial. It is now becoming recognized that the advantages of using the signal peptide domain for immunization stem from its unique B and T cells epitope restriction density and TAP independent presentation. Additionally, the low avidity of signal peptides as antigens, is believed to result in escaping negative selection in the thymus during development, and preservation of active clones in the peripheral blood. This may allow signal peptides to be presented as true neoantigens during tumor development and generate a robust immune response. To prove the neoantigenic properties of MUC1 SP, we have recently tested endogenous levels of specific anti-VXL100 T-cells by staining for VXL100-specific or control multimers. We also performed proliferation assays for patient TIL and normal donors PBMCs using cells stimulated with VXL100 or other MUC1 antigenic control peptides. VXL100 specific T-cell were found in 100% of patients (1.3-6%), VS. 0% in normal donors. We have found a direct correlation between the MUC1 signal peptide expression levels on tumor cells and levels of endogenous VXL100 specific T-cells. Positive signals were observed for multimers of other MUC1 peptides as well, but these were also seen in normal donors,

suggesting a non-specific binding. Moreover, we were able to show that VXL100 specific exhausted CD8 cells exist at a non-significant levels as compared to the total exhausted CD8. When stimulated in-vitro with VXL100, PBMCs from 70% of naive normal donors and 94% of cancer patients showed a strong proliferative response (average of 8.9% and 16.5% proliferating cells, respectively). When stimulated with control peptides 0.5% - 5.2% of patients and 0 - 13% of donors showed positive proliferation response.

Conclusions: VXL100, a novel immunotherapy agent shows the capacity to elicit a specific and robust T-cell response in healthy individuals and hematological cancer patients. These features suggest that MUC1 SP is a native sequence functioning as a cancer neoantigen and preserved through normal and tumor development. It presents an attractive opportunity for the development of an effective cancer immunotherapy.

Citation Format: Limor Chen, Riva Kovjazin, Michael Shapira. VXL100, a MUC1 signal peptide based immunotherapy, acts through a neoantigen-like mechanism, and shows promising effects in hematological cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-203. doi:10.1158/1538-7445.AM2017-LB-203
Abstract LB-217: Microenvironment mediated mechanisms of resistance to HER2 inhibitors differ between HER2+ breast cancer subtypes
Authors: Watson Spencer , Gray Joe , Korkola James , Dane Mark , Hesier Laura , Chin Koei , Mills Gordon
Published: July 2017

Clinical responses of HER2+ breast cancers to tyrosine kinase inhibitors (TKIs) are often limited by de novo and acquired resistance. Recent studies have shown that the tumor microenvironment has a substantial capacity to generate signals and create niches that enable otherwise sensitive cells to avoid therapeutic control. We describe here the use of MicroEnvironment MicroArrays (MEMAs) to identify specific signals from the tumor microenvironment that reduce sensitivity to small molecule HER2 targeted TKIs. We show here that responses to these signals differ between HER2+ cells that express transcriptional signatures associated with basal (HER2E) or luminal (L-HER2+) HER2+ subtypes. Specifically, we show that HER2E cell lines are rescued from the TKIs lapatinib and neratinib by hepatocyte growth factor (HGF), while L-HER2+ cell lines are rescued by neuregulin1-β1 (NRG1β). We further show that these divergent responses are due to differential use of the PI3K and MAPK pathways, and expression of receptors HER3 and MET. This is supported by transcriptomics of human tumors performed by The Cancer Genome Atlas. HGF and NRG1β are expressed in the stroma of primary mammary tumors, but can also be highly expressed in common sites of both HER2E and L-HER2+ metastasis. We show that NRG1β mediated resistance in L-HER2+ lines is reversed by inhibiting HER2-HER3 dimers with pertuzumab, while HGF mediated resistance in HER2E lines is reversed with the MET inhibitor crizotinib. These studies suggest that L-HER2+ and HER2E tumors should be treated as separate diseases and that treatments should be tailored to counter microenvironment mediated resistance mechanisms that may differ between metastatic locations based on local production and action of growth factors.

Citation Format: Spencer Watson, Joe Gray, James Korkola, Mark Dane, Laura Hesier, Koei Chin, Gordon Mills. Microenvironment mediated mechanisms of resistance to HER2 inhibitors differ between HER2+ breast cancer subtypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-217. doi:10.1158/1538-7445.AM2017-LB-217
Abstract LB-220: A microfluidic model for real-time monitoring of tumor-lymphocyte interactions
Authors: Moore Nathan , Doty Daniel , Santos Jose , Kratchman Louis B. , Angione Stephanie , Mott Vienna , Zielstroff Mark , Chen Hongmin , Gimbel Alla , Borenstein Jeffrey
Published: July 2017

The emergence of immune checkpoint inhibitors and the promise of increased patient survival reinforces the potential for targeting the tumor microenvironment as a drug development strategy. The tumor microenvironment consists of a heterogeneous composition of cancer cells, vascular cells, fibroblasts, and immune cells that are regulated by both intrinsic and extrinsic chemical and physical stimuli. Within this microenvironment, aberrant blood vessel formation and extra cellular matrix (ECM) restructuring lead to increased interstitial fluid flow and shear stress that stimulate differential gene expression, reorganization of tumor cell structure, and suppression of local cytotoxic T-cell activity. Elevated levels of fluid pressure and shear stress have also been demonstrated to alter cell sensitivity and response to therapeutic agents. Traditional static 2D in vitro models fail to adequately address tumor-immune cell interactions and flow mediated change in sensitivity, contributing to inaccurate predictions of in vivo therapeutic response. The development of 3D microfluidic systems have greatly enhanced the ability to model components of the tumor microenvironment, such as fluid flow, and capture the dynamics of tumor and immune cell responses to emerging therapeutic compounds. Here, we have developed a medium throughput microfluidic device which is constructed using bioinert materials that are also compatible with real-time confocal imaging, to model the effects of therapeutic treatment on tumor viability. In addition to exposing tumors to a dynamic flow environment, this model system enables the introduction of flowing lymphocytes, permitting tracking of tumor-lymphocyte interactions in real-time. As a test case, we demonstrate the ex vivo binding and cytotoxic activity of MC38-derived tumor infiltrating lymphocytes (TILs) on tumor fragments. These results highlight the power of a dynamic microfluidic flow system to model the tumor-lymphocyte microenvironment and the potential of this system to more accurately evaluate novel therapeutic agents.

Citation Format: Nathan Moore, Daniel Doty, Jose Santos, Louis B. Kratchman, Stephanie Angione, Vienna Mott, Mark Zielstroff, Hongmin Chen, Alla Gimbel, Jeffrey Borenstein. A microfluidic model for real-time monitoring of tumor-lymphocyte interactions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-220. doi:10.1158/1538-7445.AM2017-LB-220
Abstract LB-223: Whole genome sequencing of rhabdomyosarcoma germline cohort identifies low frequency of pathogenic mutations
Authors: Light Nicholas , Lupo Philip , Khan Javed , Schiffman Joshua , Hawkins Douglas , Shlien Adam , Malkin David
Published: July 2017

Introduction: Rhabdomyosarcoma (RMS) is among the most frequently occurring tumors in Li-Fraumeni syndrome patients and is often observed in patients with a strong family history of cancer. Recent studies suggest that at least 10% of children with cancer harbor an underlying pathogenic germline mutation, although the frequency in children with RMS is not well characterized. In this study we sought to determine the rate of likely pathogenic germline mutations in rhabdomyosarcoma patients, unselected for family history.

Methods: Of 275 RMS patients enrolled on one clinical trial (COG ARST0531), whole-genome sequencing was performed on blood-derived DNA of an initial set of 50 using the Illumina HiseqX to an average sequencing depth of 44.9X [34.2-58.4]. Single nucleotide variants (SNVs) and Indels were called and filtered to select only those occurring in exons or splicing regions. Variants were further filtered to include only those with a frequency of <0.5% in the ExAC database and lying in one of 99 genes identified as a cancer predisposition gene in the COSMIC Cancer Gene census. Nonsynonymous SNVs were required to be predicted as deleterious by one or more of Polyphen, mutation assessor or sift.

Results: Across the 50 initial samples, a total of 290 variants ([0-17], median of 5) passed the described filters. Notably, no point mutations were identified in TP53 in the initial 50 samples. Two samples contained variants annotated as pathogenic in ClinVar - a frameshift deletion in CHEK2 and a rare SNP in MUTYH. Also notable was a potentially pathogenic frameshift deletion in MSH6 that lacked previous annotation.

Conclusions: Our preliminary analysis suggests a lower incidence of causative TP53 mutations in RMS than previously suggested. Whole genome sequencing of the remaining samples in the cohort along with analysis of copy number and structural variants is ongoing. Statistical analysis of the entire cohort of 275 patient samples will allow for the most complete characterization of the germline genomic landscape of rhabdomyosarcoma to date. This information will better inform clinical surveillance and management parameters for RMS patients and families.

Citation Format: Nicholas Light, Philip Lupo, Javed Khan, Joshua Schiffman, Douglas Hawkins, Adam Shlien, David Malkin. Whole genome sequencing of rhabdomyosarcoma germline cohort identifies low frequency of pathogenic mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-223. doi:10.1158/1538-7445.AM2017-LB-223
Abstract LB-226: Dual inhibition of CD47 and HER2 to radiosensitize breast cancer cells
Authors: Candas Demet , Zhang Lu , Menaa Cheikh , Fan Ming , Zhang Yanhong , Liu Lin , Zhou Weibing , Azghadi Soheila , Sweeney Colleen , Shen Rulong , Lin Tzu-yin , Pan Chong-xian , Vaughan Andrew T. , Wang Ji Ming , Chen Hong-Wu , Lam Kit , Sun Lun-Quan , Monjazeb Arta M. , Murphy William J. , Li Jian Jian
Published: July 2017

Accumulating studies are aimed to combine immunoregulation with the standard of treatment such as radiotherapy in anti-cancer therapy. A potential cross resistance of tumor cells will be a barrier in furthering the long-term control by such combined treatment. CD47, an immunoglobulin-like transmembrane protein, enables cancer cells to escape the immune surveillance by flagging them with a ‘don’t eat me’ signal, which is recognized by the signal regulatory protein α (SIRPα) on macrophages. Here, we report that CD47 expression is enhanced by radiation in breast cancer cells and in vivo irradiated xenograft tumors via NF-κB regulation. In addition, CD47 and HER2 are co-induced by radiation and CD47 expression is specially enhanced by radiation in HER2-expressing cells. Contrasted with primary tumors, co-expression of CD47 and HER2 is elevated in the recurrent breast tumors. Anti-HER2 treatment inhibits CD47 transcription and protein expression with increased macrophage phagocytosis. Likewise, anti-CD47 not only augments macrophage phagocytosis but also reduces the capacity of tumorsphere formation. Radiation with anti-CD47 synergistically suppresses clonogenic survival and the maximal inhibition was achieved by dual inhibition of CD47 and HER2 with radiation. The synergy of in vivo tumor inhibition is established in mouse breast tumor model via local tumor radiotherapy combined with anti-CD47 treatment. These results demonstrate that the HER2-NF-κB-CD47 loop is responsible the cross-resistance of breast tumor against immune surveillance and radiation. Thus dual inhibition of CD47 and HER2 is a potential target to enhance tumor response to immune-radiotherapy. Funding: This study was partially supported by National Cancer Institute RO1 grant R01CA213830-01 (JJL).

Note: This abstract was not presented at the meeting.

Citation Format: Demet Candas, Lu Zhang, Cheikh Menaa, Ming Fan, Yanhong Zhang, Lin Liu, Weibing Zhou, Soheila Azghadi, Colleen Sweeney, Rulong Shen, Tzu-yin Lin, Chong-xian Pan, Andrew T. Vaughan, Ji Ming Wang, Hong-Wu Chen, Kit Lam, Lun-Quan Sun, Arta M. Monjazeb, William J. Murphy, Jian Jian Li. Dual inhibition of CD47 and HER2 to radiosensitize breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-226. doi:10.1158/1538-7445.AM2017-LB-226
Abstract LB-230: Oncogenic PKC epsilon and Pten loss converge on activating CXCL13 via atypical NF-κB transcriptional signaling in prostate cancer progression
Authors: Garg Rachana , Blando Jorge , Perez Carlos , Abba Martin , Benavides Fernando , Kazanietz Marcelo
Published: July 2017

Protein Kinase C epsilon (PKCϵ), a pro-oncogenic member of the PKC family of phorbol ester/diacylglycerol receptors, has been shown to be up-regulated in most epithelial cancers, including prostate cancer. PKCϵ transgenic overexpression in the mouse prostate causes preneoplastic lesions, however when intercrossed with mice haploinsufficient for Pten, the resulting compound mice (PB-PKCϵ;Pten+/-) develops invasive adenocarcinoma. We found that Pten-deficient prostate cell lines engineered to overexpress PKCϵ (CaP8-PKCϵ) cells become highly invasive and tumorigenic. Microarray gene expression profiling in this cell line revealed marked changes in the expression of genes implicated in proliferation, migration, adhesion, EMT, angiogenesis and metabolism. In particular, the chemokine CXCL13 was identified as the top-most up-regulated gene in CaP8-PKCϵ cells. Measurements of either CXCL13 mRNA, protein release to the culture medium (CM), or Cxcl13 promoter activity showed clear enhancements in cells overexpressing PKCϵ or Pten depleted, and these effects were enriched synergistically in CaP8-PKCϵ cells. CaP8-PKCϵ subjected to CXCL13 RNAi depletion showed reduced motility and tumorigenic growth. Similar results were observed upon silencing of CXCR5, the CXCL13 receptor. CXCL13 expression and promoter activity in CaP8-PKCϵ cells were significantly reduced by GF109203X (PKC inhibitor), IKK16, wedelolactone (NF-κB inhibitors), and BKM120 (PI3K inhibitor), but were insensitive to BX795 (PDK1 inhibitor). RNAi-mediated depletion of NIK or IKKα significantly blunted Cxcl13 luciferase promoter activity in CaP8-PKCϵ cells, but the effect was partially inhibited by IKKβ RNAi, suggesting the role of the atypical NF-κB pathway. Mutation of a responsive element for atypical NF-κB in the CXCL13 promoter abolished CXCL13 reporter activity in CaP8-PKCϵ cells. Taken together, our study identifies a novel paradigm depicting atypical NF-κB as the transcriptional signaling pathway by which PKCϵ overexpression/Pten loss regulates overproduction of the chemokine CXCL13 in prostate cancer cells.

Citation Format: Rachana Garg, Jorge Blando, Carlos Perez, Martin Abba, Fernando Benavides, Marcelo Kazanietz. Oncogenic PKC epsilon and Pten loss converge on activating CXCL13 via atypical NF-κB transcriptional signaling in prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-230. doi:10.1158/1538-7445.AM2017-LB-230
Abstract LB-241: Quantification of EGFR allele frequency predicts tumor response to EGFR tyrosine kinase inhibitors
Authors: Kim Jin-Soo , Kim Mi Young , Kim Sung-Su , Choi Hyun-jeung , Lee Yunsu , Lee In-Seon , Park Jin-Hyun , Kim Ki Hwan , Kim Hoon Gu , Kim Bong Seog , Shin Young Kee
Published: July 2017

Background: EGFR tyrosine kinase inhibitors (TKIs) are current standard treatments for non-small cell lung cancers (NSCLCs) with EGFR mutation. Current diagnostic tests for EGFR mutation include direct sequencing, pyrosequencing or real-time PCR based technology. We have developed a digital droplet PCR based analysis, GenesWell™ ddEGFR Mutation Test (IUO version 5.2. Gencurix Inc., Seoul, Korea). Methods: We have analyzed a total of 325 archived tissue from 233 lung cancer patients treated with gefitinib or erlotinib in the participating 3 hospitals from 2008 to 2013. Samples were analyzed with GenesWell™ ddEGFR Mutation Test and the results were compared with in-house results recorded in each institutional EMR. Results: Since we have analyzed left-over archived tissue, the amount of DNA was inappropriate for analysis in few cases (n=5, 1.5%), which had less than 2ng/uL concentration of DNA. ddEGFR test detected additional EGFR mutations in patients with EGFR wild-type (WT) (N=4), exon 19 deletion (19del) (N=4) and exon 21 L858R (L858R) (N=3) by in-house EGFR test. ddEGFR test detected EGFR mutations in cytology-base samples, such as pleural effusion or ascites. Small amount of sample from CSF and plasma was also detectable with ddEGFR test. Based on in-house EGFR mutation results, response rates (RRs) and disease control rates (DCRs) were as follows: 15.8% and 26.9% in WT, 71.4% and 91.8% in 19del, and 55.9% and 91.2% in L858R. Similarly, RRs and DCRs were as follows, based on ddEGFR results: 13.4% and 43.9% in wild-type (WT), 77.6% and 93.9% in 19del and 54.1% and 86.5% in exon 21 L858R (L858R). Mean allele frequencies were 48.6 (0 - 180.5) in PR, 27.4 (0 - 287.5) in SD, and 9.5 (0-159.6) in PD (P-value, PR vs SD: .0078, PR vs PD: <.000001, SD vs. PD: .029, respectively, Student T-test) Conclusion: The GenesWell™ ddEGFR Mutation demonstrate a high sensitivity for EGFR mutation detection. RRs and DCRs were comparable between in-house EGFR test and ddEGFR. Quantitative analysis of EGFR mutant allele could predict responses to EGFR TKIs. Plasma-based detection of EGFR mutations with ddEGFR are being tested in patients with EGFR mutant lung cancers. Supported by the R&D Program for Society of the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (Grant number : 2013M3C8A1078499)

Citation Format: Jin-Soo Kim, Mi Young Kim, Sung-Su Kim, Hyun-jeung Choi, Yunsu Lee, In-Seon Lee, Jin-Hyun Park, Ki Hwan Kim, Hoon Gu Kim, Bong Seog Kim, Young Kee Shin. Quantification of EGFR allele frequency predicts tumor response to EGFR tyrosine kinase inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-241. doi:10.1158/1538-7445.AM2017-LB-241
Abstract LB-244: Implication of secretory mucin MUC5AC and its glycan modification for detection of pancreatic cancer
Authors: Hayashi Chihiro , Carmicheal Joseph , Krasnoslobodtsev Alexey , Haung Ying , Brand Randall E. , Batra Surinder K. , Kaur Sukhwinder
Published: July 2017

Background and Aims

Pancreatic Cancer (PC) is often discovered in late stages, which contributes to its extreme

lethality. Early detection is key to improved curative response and increased survival. This study

explored the potential of secretory mucin MUC5AC and its glycosylation (Sialyl-Lewis A and

Disialyl-Lewis A) states in combination with carbohydrate antigen CA19.9 for early diagnosis of

PC.

Methods

Sandwich ELISA and radioimmunoassay were utilized to measure patient serum for levels of

MUC5AC and CA19.9, respectively. A blinded reference set was obtained from the NCI’s Early

Detection Research Network Program (n=242): healthy control (HC, N=50), chronic pancreatitis

(CP, N=59), acute biliary obstruction (ABO, N= 30), stage IA/IB/IIA (N=52), and stage IIB PC

(N=41), to assess the diagnostic performance of MUC5AC. Further, glycans on MUC5AC

(Disialyl Lewis A, Lewis A, Blood group H (BGH) antigen) were measured via Surface

Enhanced Raman Spectroscopy (SERS).

Results

MUC5AC levels were significantly increased in patients with stage IA/IB/IIA/IIB PC in

comparison to HC, ABO and CP. MUC5AC efficiently differentiated stage IA/IB/IIA PC from

healthy controls [AUC- 0.707], ABO [AUC- 0.662], and CP [AUC- 0.745]. CA19.9

differentiated stage IA/IB/IIA PC from healthy controls [AUC- 0.727], ABO [AUC-0.537] and

CP [AUC- 0.675]. The MUC5AC combination with CA19.9 significantly improved the

performance of CA19.9 alone: healthy controls [AUC- 0.761], ABO [AUC-0 .679], and CP

[AUC- 0.753]. Further, PC serum was shown to express MUC5AC with greater Sialyl-Lewis A

glycosylation in comparison to Disialyl-Lewis A glycosylation. Further, the reverse trend for

these glycosylation was observed in CP and BC group.

Conclusions

MUC5AC is an early-stage PC biomarker that offers improved performance with CA19.9 for the

detection of PC. Also, measurement of MUC5AC glycan modification will aid in the better

differentiation of PC from healthy controls and high risk groups (CP, ABO). Further, the ratio of

disialyl to CA19.9 glycosylation of MUC5AC is a novel way to differentiate between PC and

benign conditions.

Citation Format: Chihiro Hayashi, Joseph Carmicheal, Alexey Krasnoslobodtsev, Ying Haung, Randall E. Brand,, Surinder K. Batra, Sukhwinder Kaur. Implication of secretory mucin MUC5AC and its glycan modification for detection of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-244. doi:10.1158/1538-7445.AM2017-LB-244
Abstract LB-252: Single-cell mRNA profiling reveal transcriptional heterogeneity among pancreatic circulating tumor cells
Authors: Lapin Morten , Tjensvoll Kjersti , Oltedal Satu , Javle Milind , Smaaland Rune , Gilje Bjørnar , Nordgård Oddmund
Published: July 2017

Background: Single-cell analysis of the transcriptional heterogeneity of circulating tumor cells have the prospect of better understanding the biology of these cells and their involvement in the metastatic process. In addition, such analyses may reveal new knowledge about the mechanisms leading to chemotherapy resistance and tumor progression in pancreatic cancer patients.

Methods: Single circulating tumor cells were enriched from blood samples (n = 50) from patients with advanced pancreatic cancer using our previously established negative depletion strategy MINDEC, and detected by immuno-fluorescence microscopy. The single cells were isolated by micromanipulation, reverse transcribed, pre-amplified, and analyzed using single cell multiplex mRNA profiling to reveal transcriptional heterogeneity.

Results: Circulating tumor cells were detected in 29 (58%) of the examined blood samples. A total of 48 potential circulating tumor cells were isolated, in which 17 cells had detectable mRNA levels and were identified as circulating tumor cells by expression of

epithelial (KRT8, KRT19, EPCAM, E-Cadherin) or mesenchymal (Vimentin, N-Cadherin, ZEB1) markers. These pancreatic circulating tumor cells were revealed to heterogeneously express the examined transcripts, and also showed aberrant expression of cancer stem cell (CD24, CD44, ALDH1A1) markers and the extracellular matrix marker SPARC. Hierarchical clustering and principal component analyses revealed epithelial-like and mesenchymal-like circulating tumor cell subpopulations, which were distinct from white blood cells and cancer cell line cells. Further analyses of the transcript SPARC suggested a relation to epithelial-mesenchymal transition in pancreatic circulating tumor cells.

Conclusion: The analysis of single pancreatic circulating tumor cells defines distinct subpopulations and reveals aberrant expression of transcripts relevant for the dissemination of circulating tumor cells to distant sites.

Citation Format: Morten Lapin, Kjersti Tjensvoll, Satu Oltedal, Milind Javle, Rune Smaaland, Bjørnar Gilje, Oddmund Nordgård. Single-cell mRNA profiling reveal transcriptional heterogeneity among pancreatic circulating tumor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-252. doi:10.1158/1538-7445.AM2017-LB-252
Abstract LB-254: Thymidylate synthase gene polymorphisms is negatively associated with gastric cancer survival
Authors: Chen Jinfei , Wang Maggie Haitian , Zee Benny Chung-Ying
Published: July 2017

Background: Gastric cancer (GC) is a major public health problem in China and its carcinogenesis has not been fully understood. It was found that genetic variation played an important role in the etiology. Thymidylate synthase (TYMS) is a critical target for cancer chemo-therapy, which functions in DNA replication, synthesis and repair. However, the results of the relationship between TYMS and cancer is unclear. To find the genomic susceptibility in TYMS, bio-markers are necessary for preventing and understanding the etiology of gastric cancer.

Objectives: The research explores the relationship between TYMS and survival time of 876 Chinese gastric cancer patients.

Methods: A retrospective cohort of 876 surgically resected gastric cancer patients with clinicopathologic and follow up information were collected. The genotypes of SNPs were examined by SNaPshot method. The median survival time (MST) for each categorical variable and survival curve were estimated by Kaplan-Meier method. The overall survival was analyzed by univariate and multivariate Cox proportional hazard model.

Results: TYMS was significantly associated with survival time. The study found eight samples (1.0%) with the novel allele (4R). In univariate analysis, the group of including 4R allele had lowest survival condition (P value = 0.006). The hazard ratio of individual with 4R allele was 2.48 (95% CI: 1.10-5.58) after controlling age, gender, smoking, drinking and TNM stage.

Conclusion: Our study found a novel allele (4R) in TYMS in gastric cancer patients.

Key words: Gastric cancer; TYMS; overall survival

Citation Format: Jinfei Chen, Maggie Haitian Wang, Benny Chung-Ying Zee. Thymidylate synthase gene polymorphisms is negatively associated with gastric cancer survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-254. doi:10.1158/1538-7445.AM2017-LB-254
Abstract LB-258: Orphan nuclear hormone receptor, DAX-1, expression during progressive stages of invasive ductal carcimona
Authors: Dishington Erin
Published: July 2017

The orphan nuclear hormone receptor DAX-1 (Dosage Sensitive Sex Reversal Adrenal Hypoplasia Congenita on the X Chromosome, gene 1) plays an important role in the development of adrenal and gonadal tissues and functions as a global negative-regulator of steroidogenesis. In addition, it is known to play a role in several cancers including breast. DAX-1 immunoreactivity had been found to directly correlate with androgen receptor (AR), estrogen receptor (ER) expression and lymph node metastasis. Beyond this, DAX-1’s role in breast cancer is relatively poorly understood. In this study we investigated, using fluorescent immunohistochemistry, the expression of DAX-1 and AR during progressive stages of invasive ductal carcinoma with known estrogen receptor ER status. Additionally correlations with patient age, tumor grade, TNM score, progesterone receptor (PR), antigen Ki67 expression and human epidermal growth factor receptor 2 (Her2), were analysed. Comparing both bioinformatic and protein staining methodology we attempted to correlate DAX-1, AR and ER to cancer stage. In order to understand the mechanisms involved in cancer progression we employed epigenetic studies to determine if changes in DAX-1 expression correlated to changes in methylation status. Our data suggests DAX-1 is expressed more frequently in early stage breast cancer. Understanding the time points and mechanisms by which DAX-1 acts in breast cancer could potentially provide a novel target for therapeutic treatments.

Citation Format: Erin Dishington. Orphan nuclear hormone receptor, DAX-1, expression during progressive stages of invasive ductal carcimona [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-258. doi:10.1158/1538-7445.AM2017-LB-258
Abstract LB-263: P53 promotes replication genome stability by epigenetics-enabled MRE11 restart
Authors: Schlacher Katharina , Roy Sunetra , Tomaszowski Karl-Heinz
Published: July 2017

DNA damage activates p53, the most frequently mutated tumor suppressor, to guard the genome and control gene transcription, epigenetics and apoptosis. Yet, how p53 suppresses genome instability remains ill-defined and is clouded by its genome-wide alterations. Here, enabled by our kPOND technology for single-cell nascent DNA/protein interactions and single-molecule DNA spreading, we define p53’s role in promoting genomic stability by enabling replication restart of stalled replication forks. Mechanistically, p53 enables MRE11 nuclease recruitment for stalled replication fork restart via control of epigenetic factor. We find an apoptosis-proficient and tumor promoting p53 mutant allele to be replication restart defective, while an apoptosis defective and poorly tumor-promoting murine p53 allele remains mostly restart proficient, revealing an improved correlation of restart functions with tumorigenesis compared to apoptosis functions. Importantly, both null and p53 gain-of-function (GOF) mutant cells show restart defects resulting in replication dependent genomic instability, uncovering a common function defect aside gene transcription. Both mutant and p53 null cells show increased recruitment of error-prone repair factors and DNA polymerases, providing a mechanism for genomic instability observed in these cells by pathway homeostasis imbalance. Collective results unveil a common transcription-independent pathway defect for both p53 null and GOF mutants laying groundwork for new targeting strategies of p53 defects in cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Katharina Schlacher, Sunetra Roy, Karl-Heinz Tomaszowski. P53 promotes replication genome stability by epigenetics-enabled MRE11 restart [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-263. doi:10.1158/1538-7445.AM2017-LB-263