bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2695: Role of tumor generated acidity in immune stromal interactions during prostate carcinogenesis
Authors: El-Kenawi Asmaa , Dhillon Jasreman , Ibrahim-Hashim Arig , Abrahams Dominique , Pilon-Thomas Shari , Ruffell Brian , Gatenby Robert , Gillies Robert
Published: July 2017

Insufficiency in tumor perfusion and high rate glycolysis combine to reduce the pH of tumor microenvironment. In a TRAMP model of prostate cancer, we had shown that carcinogenesis is associated with increasing acidification of the microenvironment and that neutralization of this acidity can prevent cancer emergence or metastases. Carcinogenesis in the TRAMP model is also associated with increased fibrosis and immune cells infiltration. We thus sought to determine if fibrosis drives immune infiltration in early tumorigenesis or vice-versa; and whether this dynamics is affected by tumor acidity. To investigate this, we harvested prostates from TRAMP mice or their matching non-transgenic controls at different time points and stained serial prostate tissue sections with F4/80 (macrophages), SMA (cancer-associated fibroblasts, CAFs), and Masson’s Trichome (collagen). Quantitative image analysis reveals that increase in fibrosis occur prior to macrophage infiltration and that both events preceded tumor development. However, the relative amount of collagen fibers was unchanged across all time points. Notably, neither fibrosis nor macrophage infiltration occurred in mice treated with buffer, suggesting an involvement of acidity in this immune stromal interactions. Interestingly, macrophages isolated from latter time points in the untreated group as well as macrophages co-cultured with prostate tumor cells at acidic pH, possessed an M2-like phenotype by expressing immunosuppressive genes (e.g. Arginase 1, Arg1) and a range of scavenging receptors (e.g. mannose receptor, Cd206), as well as releasing more angiogenic factors (e.g. VEGF and MMPs). Similar results were recapitulated when M2 macrophages were stimulated at acidic pH by showing enhanced Cd206 and Arg1 expression. On the functional level, macrophages activated at acidic pH had a higher ability to uptake fluorescently labelled ovalbumin and collagen, as examples of mannosylated ligands that prevail the fibrotic microenvironment. In summary, these results suggest that tumor acidity may promote fibrosis, with subsequent macrophage infiltration and phenotypic switching, leading to increased collagen turnover. It is suspected that this extracellular matrix remodeling may be permissive for tumor progression.

Citation Format: Asmaa El-Kenawi, Jasreman Dhillon, Arig Ibrahim-Hashim, Dominique Abrahams, Shari Pilon-Thomas, Brian Ruffell, Robert Gatenby, Robert Gillies. Role of tumor generated acidity in immune stromal interactions during prostate carcinogenesis [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 2695. doi:10.1158/1538-7445.AM2017-2695
Abstract 2699: NFATc2 in non-small cell lung cancer mediates inhibition of peripheral blood mononuclear cell proliferation
Authors: Lam Ian K. , Xiao Zhi-Jie , Tin Vicky P. , Huang Fang Ping , Wong Maria P.
Published: July 2017

Lung cancer is the leading cause of cancer death worldwide. While genetic mutations that provide growth advantages to cancer cells are fundamental to malignant transformation, ability to evade the immune system is also crucial in tumorigenesis. We have previously shown that human lung cancers with high expression of nuclear factor of activated T cells, cytoplasmic 2, (NFATc2) are associated with reduced recurrence-free and overall survival. NFATc2 is expressed in most immune cells and has been extensively studied in immune cells. However, its function in cancer development is not completely understood. In this study, the role of NFATc2 in lung cancer on immune escape is explored. Lung cancer cell lines with high NFATc2 expression were co-cultured with peripheral blood mononuclear cells (PBMC) and a significant inhibition of PBMC proliferation was observed compared to PBMC only control. However, the inhibitory effect of PBMC proliferation was reversed in co-culture with NFATc2 knockdown cancer cells. When cancer cells and PBMC were separated by a membrane using Transwell inserts, the anti-proliferative effect on PBMC was not observed suggesting that cell-cell contact was required for the NFATc2-mediated inhibition. Moreover, interleukin-2 (IL-2) was produced by PBMC irrespective of the presence or absence of lung cancer cells. These results showed that the inhibition of proliferation was independent of the activation of PBMC. Taken together, we have shown NFATc2 in lung cancer plays a role in immune modulation by inhibiting PBMC proliferation and this could be a novel mechanism of immune evasion in lung cancer.

Citation Format: Ian K. Lam, Zhi-Jie Xiao, Vicky P. Tin, Fang Ping Huang, Maria P. Wong. NFATc2 in non-small cell lung cancer mediates inhibition of peripheral blood mononuclear cell proliferation [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 2699. doi:10.1158/1538-7445.AM2017-2699
Abstract 2711: Whole exome sequencing analyses of colorectal cancers associated with liver metastasis
Authors: Ma Yanlei , Weng Wenhao , Yang Yongzhi , Wei Qing , Cai Sanjun
Published: July 2017

Purpose: Liver metastasis is one of the major causes of death in colorectal cancer (CRC) patients. But the process is still unknown. Here, we analyzed genetic differences between primary tumors associated with liver metastasis and those without liver metastasis and predicted clinical significance.

Experimental design: Eight primary tumors associated with liver metastasis and twelve without liver metastasis of CRC were analyzed by whole-exome sequencing. The functional characterisation for the role of metastatic-related genes was investigated through in vitro migration and invasion assays. For clinical relevance, TMAs were used for immunohistochemistry assay.

Results: The mutation spectrum was similar to that of previous studies. TP53, APC and KRAS were the three most frequently mutated genes, with no significant difference of mutation frequency between primary tumors from patients with liver metastasis and those from other patients. Notably, four recurrently mutated genes (FBXW7, EDIL3, KCNK9 and PCDHGA7) were identified exclusively in the tumors associated with liver metastasis. Furthermore, in vitro analyses showed that EDIL3 and KCNK9 overexpression resulted in enhanced cell migration and invasion, while CRISPR-mediated deletion of FBXW7 and PCDHGA7 exhibited enhanced migratory and invasive properties as well. For clinical relevance, highly expressed EDIL3 and KCNK9 was significantly associated with liver metastasis, whereas the expression of FBXW7 and PCDHGA7 negatively correlated with it in our cohorts. Pathway enrichment analysis also indicated a role for cell adhesion signaling in liver metastasis of CRC patients.

Conclusion: Our study identified candidate genes, which might response to the initiation of metastasis, exclusively mutated in tumors associated with liver metastasis. These findings also provided potential therapeutic targets for personalized treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Yanlei Ma, Wenhao Weng, Yongzhi Yang, Qing Wei, Sanjun Cai. Whole exome sequencing analyses of colorectal cancers associated with liver metastasis [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 2711. doi:10.1158/1538-7445.AM2017-2711
Abstract 2722: Next generation targeted sequencing reveals four unknown mutations in the Mexican population with Gorlin syndrome
Authors: Veronica Fragoso , Espinosa Natalia , Dominguez Julieta , Vaca-Paniagua Felipe , Herrera Luis A. , Alvarez Rosa M.
Published: July 2017

Aim: Nevoid Basal Cell Carcinoma Syndrome (NBCCS), an autosomal dominant disorder, is characterized by the presence of a variety of developmental anomalies, and predisposition to multiple basal cell carcinomas (BCCs), and other neoplasias. NBCCS mainly results from germinal mutations in the PTCH1 gene, that encodes a transmembrane glycoprotein, component of the hedgehog signalling pathway. The incidence of NBCCS in general population is estimated at 1 in 150000. Data related to NBCCS in Mexican patients is reduced to clinical case reports, without molecular testing. Therefore, we conducted next-generation sequencing (NGS) of PTCH1, in 9 Mexican patients with diagnosis of NBCCS.

Material and methods: NGS of PTCH1 was performed, in the context of a hereditary cancer gene panel. The PTCH1 regions with insufficient coverage (<30× read depth), were complemented by Sanger sequencing, to ensure 100% coverage of the gene. In-silico analysis was perfomed for missenses mutations. The identified pathogenic mutations were verified by capillary sequencing.

Results: Targeted analysis of PTCH1 gene, looking for deletions, insertions, splice-site alterations, nonsense and missense mutations, was done. Four PTCH1 mutations (4/9) were identified: a deletion (c.1044delG); one insertion (c.631dupA); a non-sense mutation (c.3406G>T), and a missense mutation (c.1488C>A). None of the mutations had been previously reported.

Conclusions : NGS of PTCH1 gene in 9 Mexican patients with clinical criteria for NBCCS allowed the identification of four novel pathogenic mutations (44% frequency). This is the first molecular overview of PTCH1 in NBCCS patients of our population. The perspectives of the current study lie in the molecular testing of other genes involved in NBCCS (PTCH2, SUFU), and other mechanisms (whole gene deletion), as well as the review of possible phenotype-genotype correlations.

Citation Format: Fragoso Veronica, Natalia Espinosa, Julieta Dominguez, Felipe Vaca-Paniagua, Luis A. Herrera, Rosa M. Alvarez. Next generation targeted sequencing reveals four unknown mutations in the Mexican population with Gorlin 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 2722. doi:10.1158/1538-7445.AM2017-2722
Abstract 2741: Assessing the utility of circulating tumor DNA as a surveillance tool for sarcomas and Li-Fraumeni syndrome using a pre-clinical model
Authors: Paramathas Sangeetha , Lewis Nathan , Guha Tanya , Motala Zainab , Malkin David
Published: July 2017

Li-Fraumeni Syndrome (LFS) is a hereditary cancer predisposition syndrome commonly characterized by the presence of inherited mutations in the tumor suppressor gene TP53. This leads to an early onset of a wide spectrum of tumors in multiple organ systems. Current surveillance protocols for early tumor detection include biochemical screening, MRI and ultrasound scans, colonoscopy and mammography (for adults). While early detection is associated with improved survival, the complexity of testing, potential to ‘misdiagnose’ tumors (false positive/negative) and requirement for multiple imaging modalities makes clinical surveillance challenging to implement and interpret. Liquid biopsies are a recently described diagnostic and prognostic tool that takes advantage of analyzing circulating tumor DNA - fragmented genomic material that are released into the blood from dying tumor cells.

In this proof-of-principle study, we use mouse xenograft tumor models to assess the dynamic relationship between tumor burden and ctDNA concentrations, as well as resolve the sensitivity of capturing the presence of lesions in their earliest stages of growth. Longitudinal analyses of ctDNA from serially collected blood were performed on mice with xenografts of rhabdomyosarcoma (Rh4 and Rh30), osteosarcoma (HOS) and non-small cell lung carcinoma (H1975) of both localized and simulated metastatic tumors. We characterized ctDNA using droplet digital PCR (ddPCR) for known gene mutations that were specific for the cancer cell lines used. We observe obvious increases in ctDNA with increased tumor burden and a complete clearance of ctDNA after tumor resection. In addition, using the metastatic model, we have conducted synchronized imaging and blood-based biopsies to determine the smallest lesion able to be detected in the blood by ctDNA.

These studies provide the foundation for early tumor detection with ctDNA in Trp53 mutant mice that develop spontaneous tumors analogous to LFS. This model will help further our understanding on the utility of ctDNA as a surveillance and diagnostic tool for LFS as well as assess its potential for clinical use.

Citation Format: Sangeetha Paramathas, Nathan Lewis, Tanya Guha, Zainab Motala, David Malkin. Assessing the utility of circulating tumor DNA as a surveillance tool for sarcomas and Li-Fraumeni syndrome using a pre-clinical model [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 2741. doi:10.1158/1538-7445.AM2017-2741
Abstract 2751: Automatic DNA extraction system can improve the EGFR point mutation detection rate of liquid biopsy
Authors: Nakashima Chiho , Sato Akemi , Abe Tomonori , Nakamura Tomomi , Komiya Kazutoshi , Sueoka Eisaburo , Kimura Shinya , Sueoka-Aragane Naoko , Kato Junichi , Hirai Mitsuharu
Published: July 2017

The usefulness of liquid biopsy to detect mutations from cancer patients has been well recognized today. However, because the mutation detection rates from plasma DNA were relatively lower than those of tissue re-biopsy, its clinical utility has not been confirmed yet. As previously we reported, we have developed fully automatic high-sensitive point mutation detecting system named mutation-biased PCR and quenched probe (MBP-QP) system for liquid biopsy. Recently, the importance of pre-analytical procedures for plasma DNA anazysis has been highlighted. In this study, we examined whether the automatic DNA extraction system can improve the mutation detection rate in our MBP-QP system. Sixty-one plasma samples were obtained from advanced non-small cell lung cancer patients, and plasma DNA extraction was performed from 200μl plasma by manually (200-M), and 200μl (200-A), 1000μl (1000-A) plasma by automatically. We used silica membrane spin column system for manual DNA extraction, and magnet beads system for automatic DNA extraction procedure. The median DNA concentrations quantified by quantitative real-time PCR of 200-M, 200-A, 1000-A were 4.92, 6.00, 20.1 ng/mL plasma, respectively. In terms of the epidermal growth factor receptor (EGFR) L858R point mutation detection, the sensitivity of 200-M, 200-A, 1000-A were 36.6%, 58.5%, 77.5%, that of the specificity were 93.3%, 100%, 96.7%, and the concordance rates were 60.6%, 76.1%, 85.7%, respectively. The size distribution of automatically extracted plasma DNA represented two peaks characteristics at 170 bp and 5 kb. In this study, we indicate the automatic DNA extraction can improve mutation detection rates in plasma DNA.

Citation Format: Chiho Nakashima, Akemi Sato, Tomonori Abe, Tomomi Nakamura, Kazutoshi Komiya, Eisaburo Sueoka, Shinya Kimura, Naoko Sueoka-Aragane, Junichi Kato, Mitsuharu Hirai. Automatic DNA extraction system can improve the EGFR point mutation detection rate of liquid biopsy [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 2751. doi:10.1158/1538-7445.AM2017-2751
Abstract 2752: Liquid biopsy analysis of FGFR3 and PIK3CA hotspot mutations for disease surveillance in bladder cancer
Authors: Christensen Emil , Birkenkamp-Demtröder Karin , Nordentoft Iver , Høyer Søren , Keur Kirstin Van Der , Kessel Kim Van , Zwarthoff Ellen , Agerbæk Mads , Ørntoft Torben Falck , Jensen Jørgen Bjerggaard , Dyrskjøt Lars
Published: July 2017

Background: Disease surveillance in patients with bladder cancer is important for early diagnosis of progression and metastasis and for optimal therapeutic treatment.

Experimental procedures: Droplet digital PCR (ddPCR) assays were developed and tumour DNA from two patient cohorts was screened for FGFR3 and PIK3CA hotspot mutations. One cohort (NMIBC cohort) included 363 patients with non-muscle invasive bladder cancer (NMIBC). Another cohort (Cx cohort) included 468 patients with bladder cancer undergoing radical cystectomy. Urine supernatants (NMIBC cohort: n=216, Cx cohort: n=27) and plasma samples (NMIBC cohort: n=39, Cx cohort: n=27) from patients harbouring mutations were subsequently screened using ddPCR assays. Liquid biopsies were collected from 2003-2015 with up to 11.3 years of follow-up for the NMIBC cohort and from 1995-2009 with up to 13.5 years of follow-up for the Cx cohort.

Results and limitations: In total, 36% of the patients in the NMIBC cohort (129/363) and 11% of the patients in the Cx cohort (44/403) harboured at least one hotspot mutation in FGFR3 or PIK3CA. Screening of DNA from serial urine supernatants from the NMIBC cohort showed high levels of tumour DNA (tDNA) to be significantly associated with later disease progression (p=0.003). Furthermore, high levels of tDNA in preoperative urine supernatants and plasma samples were associated with recurrence in the Cx cohort (p=0.068 and p=0.016 respectively). A positive correlation between the levels of tDNA in urine and plasma was observed (corr.=0.6). The retrospective study design and the low volumes of plasma available for analysis were limitations of the study.

Conclusions: Increased levels of FGFR3 and PIK3CA mutated DNA in urine and plasma is indicative of later progression and metastasis in bladder cancer.

Citation Format: Emil Christensen, Karin Birkenkamp-Demtröder, Iver Nordentoft, Søren Høyer, Kirstin Van Der Keur, Kim Van Kessel, Ellen Zwarthoff, Mads Agerbæk, Torben Falck Ørntoft, Jørgen Bjerggaard Jensen, Lars Dyrskjøt. Liquid biopsy analysis of FGFR3 and PIK3CA hotspot mutations for disease surveillance in bladder 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 2752. doi:10.1158/1538-7445.AM2017-2752
Abstract 2754: A novel liquid biopsy method for development of aptamer libraries that bind blood plasma exosomes from breast cancer patients
Authors: Domenyuk Valeriy , Levenberg Symon , Stark Adam , Miglarese Mark , Spetzler David
Published: July 2017

Improved technologies capable of characterizing system-wide changes associated with complex diseases will be required to be able to detect millions of proteins and their isoforms as well as multi-molecular complexes. We present a method for developing aptamer libraries using blood plasma exosomes that provides unprecedented system-wide coverage of native exosomal complexes. To train a naïve aptamer library toward cancer samples (positive selection), the library (~1013 biotinylated ssODN species) was incubated with plasma from individual cancer patients and aptamer-bound exosomes were isolated using polymer-based precipitation. Negative selection was performed by contacting the aptamer library with exosomes from donors without breast cancer and recovering unbound aptamers from the supernatant. In all, 12 libraries trained toward 12 individual breast cancer patients were used to probe additional samples. Exosome-bound aptamers were identified and quantified by Next Generation Sequencing (NGS) to build highly accurate signatures for cancer/healthy donor classification. Using these signatures, cancer patients’ binding profiles were easily distinguishable from controls. Interestingly, cancer-trained libraries did not distinguish any of the negative control samples from each other, indicating that the selection pressure for cancer was high and noise due to inherent inter-healthy donor heterogeneity was minimal. Full validation studies are ongoing. Aptamer libraries may ultimately be deployed as a minimally-invasive diagnostic adjunct in breast and other cancers.

Citation Format: Valeriy Domenyuk, Symon Levenberg, Adam Stark, Mark Miglarese, David Spetzler. A novel liquid biopsy method for development of aptamer libraries that bind blood plasma exosomes from breast cancer patients [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 2754. doi:10.1158/1538-7445.AM2017-2754
Abstract 2765: Psychosocial predictors of life satisfaction among married women with breast cancer
Authors: Asuzu Chioma , Efosa-Urhoghide Ngozi
Published: July 2017

Introduction: Breast cancer presents a challenge to the totality of a woman’s life. Correlates of life satisfaction among married women with breast cancer have not been very well explored. Purpose: This study seeks to determine the relationship that exists between life satisfaction with depression, cognitive emotional regulation, sexuality, self-concept, dejection, interpersonal conflict and social support among married women with breast cancer in Southwest Nigeria. It also seeks to ascertain which of these variables predict life satisfaction among the respondents.

Method: The variables in the study were assessed using modified standardized instruments which were revalidated for cultural suitability after ethical approvals were obtained in each of the 5 centres involved in the study. The instruments were given to breast cancer patients after informed consent to participate in the study have been obtained.

Result: The larger proportion {148 or 40.5%)} of the participants were in the age range of 46-55 years and 112 (30.7%) had stage III disease. Analysis of data showed that significant linear relationship exists between life satisfaction of married women with breast cancer and depression (r = .200**; p < .001), self-concept (r = .306**; p < .001), cognitive emotional regulation (r = .398**; p < .001), dejection (r = .226**; p < .001), interpersonal conflict (r = .352**; p < .001), anxiety (r =.316**; p < .001), sexuality (r =.305*; p < .001) and social support (r = .534; p < .001). There were also significant correlations among the independent variables.

Conclusion: Psychosocial factors are potent predictors of life satisfaction among married women with breast cancer. Based on the findings, it was recommended that psycho-social intervention based on the factors identified as causative agents of life satisfaction should be developed and implemented for married individuals.

Note: This abstract was not presented at the meeting.

Citation Format: Chioma Asuzu, Ngozi Efosa-Urhoghide. Psychosocial predictors of life satisfaction among married women with breast 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 2765. doi:10.1158/1538-7445.AM2017-2765
Abstract 2770: Bcl-2 overexpression is a predictive biomarker for the combination of ABT-199 with chemotherapy in diffuse large B-cell lymphoma cell lines
Authors: McConnell Ian , Anderson Elyse , Yazbeck Victor
Published: July 2017

Diffuse Large B-Cell Lymphoma (DLBCL) is the most common subtype of Non-Hodgkin’s Lymphoma. The anti-apoptotic Bcl-2 protein is expressed in certain DLBCL subtypes such as “Double-Hit” where it mediates resistance to chemotherapy. ABT-199 is a BH3-mimetic that specifically targets Bcl-2, and has shown excellent clinical activity in several hematological malignancies. The anti-apoptotic protein Mcl-1 mediates resistance to ABT-199, and can be inhibited with chemotherapy. However, despite variability in the Bcl-2 expression levels, current clinical trials involving combinations of ABT-199 and chemotherapy for the treatment of patients with DLBCL use a one-size-fits-all approach regardless of Bcl-2 expression levels. We hypothesized that high Bcl-2 expression levels correlates with sensitivity to the combination of ABT-199 and chemotherapy in DLBCL. We examined the activity of ABT-199 in four DLBCL cell lines with varying Bcl-2 levels (CARNAVAL, OCI-ly18, SU-DHL-4 and SU-DHL-8) as determined by cell viability (Cell Titer Glo Assay) and apoptosis (Annexin-V/PI staining and FACS) analyses. CARNAVAL and SU-DHL-8 have the highest and lowest Bcl-2 expression levels, respectively, as revealed by immunoblotting. Of the four cell lines, CARNAVAL displayed the highest sensitivity (IC50 = ~7 nM) while SU-DHL-8 was resistant (IC50 = >10 µM) with the other 2 cell lines exhibiting intermediate sensitivity (45.9 nM for OCI-ly18 and 2.6 µM for SU-DHL-4). We then selected CARNAVAL and SU-DHL-8 in order to further examine the effect of the combination of ABT-199 with classic chemotherapy agents (etoposide and doxorubicin) used in DLBCL treatment. In CARNAVAL cells, the combination of either chemotherapy with ABT-199 led to a significant decrease in cell viability at 48 hrs compared to each single agent alone. This was associated with apoptosis induction as measured by Annexin-V/PI, caspase-3 and PARP cleavage, an increase in γ-H2AX (associated with DNA damage), and decrease in Mcl-1. Unlike CARNAVAL cells, ABT-199 did not enhance the activity of chemotherapy in SU-DHL-8 cells that lack Bcl-2 expression. These preclinical results support the use of Bcl-2 overexpression as a predictive biomarker for selecting patients that are likely to benefit from the combination of ABT-199 with chemotherapy in DLBCL. Current efforts are investigating the mechanism of resistance to the combination.

Citation Format: Ian McConnell, Elyse Anderson, Victor Yazbeck. Bcl-2 overexpression is a predictive biomarker for the combination of ABT-199 with chemotherapy in diffuse large B-cell lymphoma cell lines [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 2770. doi:10.1158/1538-7445.AM2017-2770
Abstract 2781: Exosomal Del-1 as a potent diagnostic marker for breast cancer: A prospective cohort study
Authors: Lee Soo Jung , Lee Jeeyeon , Chae Yee Soo , Jung Jin Hyang , park Ho Yong , Baek Moon-Chang , Moon Pyong-Gon
Published: July 2017

Background: We previously demonstrated a diagnostic role of exosomal del-1 with two separated groups of breast cancer patients. In the current study, therefore, we aimed to confirm the diagnostic role in a prospective study with breast cancer by measuring plasma exosomal del-1 before and after surgery.

Patients and methods: To identify the optimal time of sampling after surgery, serial blood at day 1, 3, 5, and 7 after surgery was collected from 22 patients with early breast cancer. Thereafter, One hundred fifteen patients with breast cancer who underwent curative surgery were enrolled in the prospective cohort study to compare difference in plasma exosomal del-1 measured by ELISA at the time of diagnosis and post-surgery.

Results: Among all 22 patients for optimal sampling time after surgery, exosomal del-1 was higher than 0.5 at the time of diagnosis and then normalized at POD1. Among 115 patients for the confirmatory set, 109 (94.8%) patients showed a normalization of del-1 lower than 0.5 after surgery and 10 patients showed del-1> 0.4 . Median f/u duration of 22 months, 9 patients experienced relapse (4 locoregional and 5 distant), where 3 out of 6 in high group (>0.5), and 2 out of 4 in borderline group (0.4-0.5), and 4 out of 105 in normalized group (<0.4). In particular, patients who relapsed in higher del-1 group showed relatively earlier relapse compared to lower del-1 group.

Conclusion: In a prospective cohort study, we confirmed that exosomal del-1 has a potent diagnostic role in breast cancer. Furthermore, del-1 was also identified to dramatically decrease after curative surgery. Our current findings suggest its potential prognostic role as well as diagnostic role in breast cancer patients.

Changes of Exosomal Del-1 level between baseline and after-surgery (n=115)sampling timebaselineafter surgeryp-value(paired t-test)Del-1 (mean±SD)1.237±0.4530.188±0.124<0.001

Citation Format: Soo Jung Lee, Jeeyeon Lee, Yee Soo Chae, Jin Hyang Jung, Ho Yong park, Moon-Chang Baek, Pyong-Gon Moon. Exosomal Del-1 as a potent diagnostic marker for breast cancer: A prospective cohort study [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 2781. doi:10.1158/1538-7445.AM2017-2781
Abstract 2797: MiR-137 and MiR-496 target Del-1 and affect triple negative breast cancer progression
Authors: Lee Jeeyeon , Chae Yee Soo , Lee Soo Jung , Jung Jin Hyang , Park Ho Yong , Baek Moon-Chang
Published: July 2017

Background: Although Del-1 was recently proposed as a new biomarker for early breast cancer in our previous studies, the mechanisms of Del-1 expression are barely understood. In the current study, we selected two microRNAs (miR-137 and - 496), potentially affecting Del-1 expression in breast cancer and examined their impact on Del-1 expression in a variety of breast cancer cell lines to identify their potential role in Del-1 expression and thereby breast cancer development or progression.

Methods: Del-1 mRNA and miR-137/- 496 levels were measured by qRT-PCR among breast epithelial (MCF10A) and cancer cells (MDA-MB-231, MCF7, SK-BR3 and T-47D). The effects of miR-137/- 496 on cell proliferation and invasion were detected using MTT, wound healing and Transwell assays. Furthermore, luciferase reporter assay was used to identify the direct regulation of Del-1 by miR-137 or - 496 in MDA-MB-231 cells. Plus, we analyzed the expressions of miR-137 or - 496 and Del-1 mRNA from 20 patients with triple negative early breast cancer.

Results: miR-137 and - 496 levels were low in all breast cancer cell lines. As Del-1 mRNA expression was remarkably higher in MDA-MB-231 compared to the other breast cancer cell lines, further functional analyses were done with MDA-MB-231 representing triple negative breast cancer subtype. Both miR-137 and miR-496 were revealed to directly bind at the 3’-UTR of Del-1. Del-1 by Luciferase reporter assay and Del-1 expression was upregulated by inhibitors and reversed by both mimics of both miR-137 and miR-496. Furthermore, both miR-137 and miR-496 were also demonstrated to inhibit cell proliferation, migration and invasion of MDA-MB-231, suggesting that these miRNAs affect cancer progression via Del-1. MiR-137 and miR-496 were remarkably down-regulated in 7 out of 12 triple negative breast cancer tissues, in particular with high Ki67 and high histologic grade.

Conclusion: Although Del-1 was recently introduced as a new biomarker for triple negative breast cancer, the mechanisms of Del-1 expression were barely identified. The current study firstly demonstrated that microRNA 137 and 496 are involved in Del-1 regulation by binding at Del-1 gene, affecting cancer progression by altering Del-1 expression.

Citation Format: Jeeyeon Lee, Yee Soo Chae, Soo Jung Lee, Jin Hyang Jung, Ho Yong Park, Moon-Chang Baek. MiR-137 and MiR-496 target Del-1 and affect triple negative breast 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 2797. doi:10.1158/1538-7445.AM2017-2797
Abstract 280: The maximum tumor length and lymph node metastasis in esophageal squamous cell carcinomas
Authors: Lv Shuang , Lv Shuang , Zhou Fu You , Yang Hai Jun , Li Bei , Liu Li Na , Xia Yan Jie , Qin Yan Ru , Fan Rui Tai , Xiao Fan Kai , Wang Li Dong
Published: July 2017

The lymph node metastasis (LNM) is one of the crucial risk factors for esophageal squamous cell carcinomas (ESCC) prognosis. However, it is difficult to precisely determine the LNM status before surgery. The maximum tumor length (MTL) may reflect the ESCC growth speed and could be calculated accurately either with computed tomography or ultrasound endoscopy before esophagectomy. Therefore the present study was designed to determine if the MTL could reflect the LNM status for ESCC. A total of 22,660 ESCC patients was enrolled in this study from the ESCC database (1973-2015) in Henan Key Laboratory for Esophageal Cancer Research of the First Affiliated Hospital, Zhengzhou University. Of these patients, there were 13,793 males with an mean age of 60±8, and 8,867 females with an mean age of 61±8. All the cases had been performed radical esophagectomy. The MTL and LNM records were retrieved from the medical records in hospitals. Based on the 6th version of UICC criteria, the LNM was classified as N0 and N1. The MTL was classified as 5 groups: <2cm, ≥2 and <4cm, ≥4 and <6cm , ≥6 and <8cm , ≥8cm. The SPSS21.0 software and Spearman rank correlation were used to determine the correlation of MTL and LNM. The results demonstrated that the average and median of MTL were 4.1±1.8 cm and 4.0 cm (range: 0.1-18 cm) in these ESCC patients. The positive lymph node metastasis rates in each MTL groups were 19.1%, 34.1%, 45.8%, 51.5%, and 54.1%, respectively. Spearman rank correlation analysis indicated that MTL was strongly correlated with LNM (r=0.997, P<0.001), suggesting that MTL could reflect the LNM status in ESCC and that MTL could be used as one of clinical staging parameters for ESCC. [Supported by Major Science and Technology Projects of Henan Province (161100311300). National Key Research and Development Program: Precision Medicine (2016).]

Note: This abstract was not presented at the meeting.

Citation Format: Shuang Lv, Shuang Lv, Fu You Zhou, Hai Jun Yang, Bei Li, Li Na Liu, Yan Jie Xia, Yan Ru Qin, Rui Tai Fan, Fan Kai Xiao, Li Dong Wang. The maximum tumor length and lymph node metastasis in esophageal squamous cell carcinomas [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 280. doi:10.1158/1538-7445.AM2017-280
Abstract 2814: Effect of kif14 overexpression on tumor formation in mice in a lifespan study
Authors: Pitt Natalie , Sishtla Kamakshi , Shadmand Mehdi , Sulaiman Rania , Sinn Anthony L. , Condon Keith , Sandusky George E. , Corson Timothy W.
Published: July 2017

KIF14 is a mitotic kinesin and microtubule-associated molecular motor that plays an essential role in the last stages of cytokinesis. In multiple cancer types, KIF14, overexpression in tumors correlates with stage, aggressiveness, and poor patient outcomes. There is considerable interest in KIF14 as a possible oncogene, since the KIF14 locus is in a common region of genomic gain in multiple cancers.

In this study, wild type BDF-1 mice and a strain constitutively overexpressing Kif14, known as Kif14-Tg mice, were evaluated at the end of their natural lifespan, specifically for the occurrence of tumors. There were approximately fifty mice per genotype, and eighty of these mice have died. These mice were necropsied and the following tissues were collected: kidney, liver, lung, spleen, seminal vesicles, ovary, mammary gland, bone marrow, pituitary gland, eye, and brain. Any gross nodules were also collected. These tissues were then fixed in 10% neutral buffered formalin, processed, sectioned, and stained with hematoxylin and eosin.

Although there was no significant difference in mouse weights through the lifespan or in the survival curve between the Kif14-Tg and their wild type littermates, a significant increase in follicular lymphoma and diffuse large B cell lymphoma was seen in Kif14-Tg mice compared to the wild type mice. Thirty-three mice developed either a follicular lymphoma or a diffuse large B cell lymphoma, and twenty-four of those mice were Kif14-Tg mice. These were the two most common tumors in Kif14-Tg mice. Other lesions and tumors that were seen in both strains included thymic lymphoma, sarcomas, myeloid dysplasia, and other carcinomas. Nontumorous lesions were seen in both strains of mice including hydronephrosis and telangiectasia in multiple organs. Ballooning degeneration of the lens of the eyes was observed in five mice from the strain Kif14-Tg.

Our finding of increased follicular lymphoma and diffuse large B cell lymphoma proves the first evidence that Kif14 can promote tumor formation in a wild-type background. This is the first evidence that Kif14 may have a role in lymphoma, but complements our earlier finding that Kif14 overexpression can accelerate tumor formation in a mouse model of retinoblastoma. Together, these outcomes further support Kif14’s potential as an oncogene.

Citation Format: Natalie Pitt, Kamakshi Sishtla, Mehdi Shadmand, Rania Sulaiman, Anthony L. Sinn, Keith Condon, George E. Sandusky, Timothy W. Corson. Effect of kif14 overexpression on tumor formation in mice in a lifespan study [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 2814. doi:10.1158/1538-7445.AM2017-2814
Abstract 2818: PDX model of the enriched pleural effusion from advanced lung cancer patient for testing on targeted drug sensitivity and resistance
Authors: Zhang Feifei , Wang Ke , Xu Wenhua , Yang Yang , Ouyang Kedong , Lu Shun , Wen Danyi , Chen Taiping
Published: July 2017

Advanced lung cancer of late stage patients is selectively sensitive to available targeted therapeutics and often developing resistance to the early sensitive drug treatment. Surgery tumor tissues are usually not available to establish patient derived xenograft (PDX) models which is critical to evaluate drug sensitivity and resistance. We applied CD45 magnet beads to enrich the tumor cell population from pleural effusion specimens of advanced lung cancer. Seven PDX models were successfully established in this way. Based on genetic profiling of these models, TKI drug related EGFR mutations and ALK fusion were confirmed (EGFR: L858R, T790M, Exon 19 insertion; ALK fusion). Further in vivo efficacy studies were designed to test on multiple drug sensitivity and resistance including Gefitinib, Erlotinib, Ceritinib, Alectinib, AZD9291. Tumor growth inhibition (TGI) were well matched with corresponding gene mutations among the models. In particularly, T790M model was resistant to Erlotinib and quite sensitive for AZD9291. In conclusion, the models established here are very useful to test on potency of many current investigational TKI drugs and provide base for drug resistant model establishment.

Citation Format: Feifei Zhang, Ke Wang, Wenhua Xu, Yang Yang, Kedong Ouyang, Shun Lu, Danyi Wen, Taiping Chen. PDX model of the enriched pleural effusion from advanced lung cancer patient for testing on targeted drug sensitivity and resistance [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 2818. doi:10.1158/1538-7445.AM2017-2818
Abstract 2819: Intestinal estrogen receptor beta attenuates colon cancer signaling pathways in vivo
Authors: Pettersson Linnea , Saxena Ashish , Vu Trang , Gustafsson Jan-Åke , Archer Amena , Williams Cecilia
Published: July 2017

Colorectal cancer (CRC) is the third most common form of cancer and the second leading cause for cancer deaths. Several epidemiological, in vitro and in vivo studies suggest that Estrogen receptor β (ERβ) could be a possible target for CRC prevention and treatment, but the potential mechanism is not demonstrated. ERβ is expressed in intestinal epithelial cells, as well as in some intestinal immune cells and other tissues. Previous studies have seen increased CRC after full ERβ knockout, but it is not clear if this is mediated by intestinal ERβ. The specific aim of our study is to determine the role of intestinal ERβ during colon cancer development in vivo. We deleted ERβ specifically in the intestine epithelia of mice (iERβKO), and induced CRC using Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS) treatment. After 9 weeks treatment, all mice presented colitis and two iERβKO mice but none of the wild-type (WT) mice had developed tumors. qPCR showed that the expression of pro-inflammatory markers was higher in iERβKO treated mice than in untreated or WT mice, Immunohistochemistry (IHC) of proliferative markers showed that iERβKO exhibited higher cell proliferation in the top of the crypts compared to controls. After 16 weeks treatment, all mice developed tumors. qPCR showed that the expression of mesenchymal markers was higher in treated iERβKO mice compared to treated WT mice. Our results demonstrate that intestinal ERβ attenuates colon cancer-related pathways.

Citation Format: Linnea Pettersson, Ashish Saxena, Trang Vu, Jan-Åke Gustafsson, Amena Archer, Cecilia Williams. Intestinal estrogen receptor beta attenuates colon cancer signaling pathways in vivo [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 2819. doi:10.1158/1538-7445.AM2017-2819
Abstract 2827: Establishing a mouse model to investigate GRB7-mediated signaling in development and triple-negative breast cancer
Authors: Lofgren Kristopher A. , Meier David R. , Cash Steven E. , Kenny Paraic A.
Published: July 2017

Triple-negative breast cancer (TNBC) lacks expression of the estrogen and progesterone steroid receptors and HER2; thus TNBC patients cannot benefit from the recent successes of targeted therapies, and instead rely on cytotoxic chemotherapeutic combinations. As such, tumor recurrence is high and patient prognosis is poor. GRB7 is a cytoplasmic adaptor protein required for cell migration, invasion, and proliferation in TNBC. We have previously reported that a high GRB7 mRNA level was the strongest predictor of tumor recurrence in a cohort of 246 TNBC patients treated with doxorubicin. GRB7 was confirmed in a smaller, independent cohort as an indicator of poor prognosis. After the observation of both decreased doxorubicin sensitivity and elevated GRB7 expression levels in TNBC tumors, we hypothesized that GRB7 was mediating a signaling pathway critical for drug resistance and cell survival.

To determine the role of GRB7 in mouse development and tumorigenesis, the current study focuses on the Grb7tm1a(EUCOMM)Wtsi knockout-first transgenic mouse, in which the insertion of the 1L2_Bact_P cassette upstream of the critical exons of GRB7 renders an effective knockout of GRB7, and the inclusion of loxP sites flanking critical exons of GRB7 facilitates the creation of tissue-specific, conditional GRB7 deletions. Herein, we investigate GRB7 expression in the homozygous Grb7tm1a mouse and derivative tissue-specific deletions, and evaluate their suitability for modeling GRB7-mediated events in mammary gland development and tumorigenesis in a TNBC model. Primary cell lines from the Grb7tm1a mouse and breast cancer cell lines will then be used to delineate the effects of altered GRB7 levels on the mechanism of developmental and TNBC cell signaling, respectively.

Citation Format: Kristopher A. Lofgren, David R. Meier, Steven E. Cash, Paraic A. Kenny. Establishing a mouse model to investigate GRB7-mediated signaling in development and triple-negative breast 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 2827. doi:10.1158/1538-7445.AM2017-2827
Abstract 2829: Identification of transcriptomic signatures of organotropism in pancreatic cancer metastasis
Authors: Zhong Yi , Fan Jun , Huang Jinlong , Iacobuzio-Donahue Christine A.
Published: July 2017

Metastatic disease is the most common cause of death in pancreatic ductal adenocarcinoma (PDA) patients. The most common sites of metastatic spread of PDA are to the liver, lung, and peritoneum. The acquisition of advanced stage PDA tissue samples has greatly improved our descriptive understanding of pancreatic cancer metastasis; however, important aspects of PDA metastatic cell biology remain poorly understood. We have previously shown that distant metastases arise from genetically evolved subclones within the primary site of PDA. To understand the biologic basis of PDA metastatic subclone formation we established a transgenic mouse model of PDA based on heterozygous inactivation of Tgfbr2 in association with p48-Cre mediated activation of mutant Kras and Trp53 alleles (KPTC mice). This model revealed that metastatic efficiency and organotropism are related but independently controlled phenomena by mutant Trp53 gain of function in association with TGFb signaling. Pertubation of TGFb signaling reduced liver but not lung metastasis due to a profound extravasation deficiency characterized by sinusoidal growth and lack of desmoplastic stroma compared to KPC mice. Analogous findings were confirmed in liver samples from patients indicating their clinical relevance. Portal vein colonization as a direct mode of access to the liver was observed in both mice and humans. To better understand the cell- autonomous features that promote organ specific metastasis, we have established cell lines from primary and metastatic PDAs of KPC mice representing five different target organ sites (liver, lung, peritoneum, omentum, lymph node). RNAseq and WGCNA performed on extracted RNA from these established cell lines indicate gene expression modules that are unique to metastases. Among metastases, gene expression modules have also been identified that are unique to local versus distant metastases. Analyses are ongoing to functionally validate these modules in additional samples of mouse and human PDA. These findings establish a novel paradigm for understanding pancreatic cancer metastasis and the observed clinical latencies of liver versus lung metastases specifically. Identification of cell-autonomous and micro-environmental mechanisms that support PDA organotropism have the potential to reveal novel targets for therapeutic intervention and modulation of the lethal metastatic phenotype.

Citation Format: Yi Zhong, Jun Fan, Jinlong Huang, Christine A. Iacobuzio-Donahue. Identification of transcriptomic signatures of organotropism in pancreatic cancer metastasis [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 2829. doi:10.1158/1538-7445.AM2017-2829