bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1769: The clinical outcome of stage I lung adenocarcinoma patients with or without activating EGFR mutation
Authors: Fang Yueh-Fu
Published: July 2017

Purpose: Advanced stage lung adenocarcinoma patients with activating EGFR mutation have better survival result after EGFR tyrosine kinase inhibitor developed. The prevalence of EGFR mutation is high in Asian lung adenocarcinoma patients. The difference of clinical outcome of stage I lung adenocarcinoma patients with or without activating EGFR mutation was unclear. Our aim of study is to evaluate the clinical outcome between wild and mutated stage I lung adenocarcinoma.

Patients and Methods: We had 343 Stage I lung adenocarcinoma patients who received surgical resection from January 2010 to April 2014. The preliminary data of EGFR mutation in 167 patients was done. Sanger method of direct PCR, scorpion & ARMS method or ABI allele specific TagMan PCR were used to check the EGFR mutation status of these patients’ samples. The selection of these methods was depended on purity of tumor samples.

Results: There were 93 stage IA patients and 74 stage IB patients. Sixty-four (69%) patients had activating EGFR mutations in stage IA patients, and fifty (68%) patients had activating EGFR mutations in stage IB patients. The survival of stage IA and IB were similar in patients with or without EGFR mutations. The patients without mutated EGFR had better progression free survival in IA and IB patients, especially in the IB patients, although the significance did not be reached in small preliminary sample size.

Conclusion: The survival was similar in stage IA or IB lung adenocarcinoma with or without EGFR mutations. Patient without EGFR mutation had better progression free survival although there was no statistical significance between patients with or without EGFR mutations. We will collect more patients’ data for analysis.

Citation Format: Yueh-Fu Fang. The clinical outcome of stage I lung adenocarcinoma patients with or without activating EGFR mutation [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 1769. doi:10.1158/1538-7445.AM2017-1769
Abstract 1779: Chronic NSAID use increases survival in PIK3CA-altered head and neck squamous cell carcinoma
Authors: Hedberg Matthew Louis , Peyser Noah , Gooding William , Li Hua , Brand Toni , Olivas Victor , Bivona Trever , Choisea Simion , Wang Lin , Johnson Jonas , Duvvuri Uma , Ferris Robert , Johnson Daniel , Ha Patrick , Bauman Julie , Grandis Jennifer
Published: July 2017

Background: PIK3CA is the most commonly altered oncogene in head and neck squamous cell carcinoma (HNSCC). Chronic non-steroidal anti-inflammatory (NSAID) use has been shown to reduce mortality in PIK3CA-mutated colorectal cancer through an unknown mechanism. In this retrospective cohort analysis, we evaluated the impact of NSAIDs on HNSCC survival, and sought a biologically plausible explanation for the observed effect.

Methods: The PIK3CA status of tumor tissue was determined by FISH and next generation sequencing for 266 HNSCC patients treated at the University of Pittsburgh Cancer Institute. Cox joint proportional hazards models were used to generate multivariate projections of disease-specific (DSS) and overall survival (OS). Prostaglandin E2 (PGE2) secretion and NSAID sensitivity were then assessed in HNSCC cell lines with or without PIK3CA mutation.

Results: Chronic NSAID use (≥2 days/week for ≥6 months) was associated with greater disease-specific (HR 0.24; 95% CI, 0.09 - 0.62; p = 0.0032) and overall survival (HR 0.31; 95% CI, 0.14 - 0.69; p = 0.0043) in patients whose tumors harbor PIK3CA mutation and/or amplification. Survival was unchanged by chronic NSAID use in patients with wild-type, unamplified PIK3CA. Chronic NSAID users with PIK3CA-altered tumors had an increase in estimated 5-year DSS from 25% to 72% and in OS from 45% to 78%. Benefit was independent of age, stage, and human papillomavirus (HPV) status; all of which were controlled for in the model. HNSCC cells with endogenous or engineered PIK3CA mutation secreted higher levels of PGE2 and were more sensitive to growth inhibition by NSAID treatment.

Conclusions: Chronic NSAID use was associated with markedly improved survival in patients with PIK3CA-altered HNSCC, but not in those with wild-type, unamplified PIK3CA. Preclinical studies implicate the PI3K-COX-PGE2 signaling axis as a potential mediator of NSAID response. A prospective, randomized controlled trial of NSAIDs in patients with PIK3CA-altered HNSCC is warranted.

Citation Format: Matthew Louis Hedberg, Noah Peyser, William Gooding, Hua Li, Toni Brand, Victor Olivas, Trever Bivona, Simion Choisea, Lin Wang, Jonas Johnson, Uma Duvvuri, Robert Ferris, Daniel Johnson, Patrick Ha, Julie Bauman, Jennifer Grandis. Chronic NSAID use increases survival in PIK3CA-altered head and neck squamous cell carcinoma [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 1779. doi:10.1158/1538-7445.AM2017-1779
Abstract 1781: EGFR amplification-A candidate predictive biomarker of PARP inhibitor Talazoparib sensitivity in gliomas
Authors: Wu Shaofang , Gao Feng , Li Xiaolong , Ding Jie , Zheng Siyuan , Martinez-Ledesma Emmanuel , Feng Ningping , Sulman Erik , Verhaak Roel , Groot John de , Koul Dimpy , Yung W.K.Alfred
Published: July 2017

Poly (ADP-ribose) polymerases (PARP) are enzymes involved in DNA-damage repair. Inhibition of PARP is a promising strategy for targeting cancers with defective DNA-damage repair. Several PARP inhibitors are currently in trials in the adjuvant, neoadjuvant, and metastatic settings for the treatment of ovarian, BRCA-mutated breast, and other cancers. Glioma Stem Cells (GSCs) exhibit higher oxidative base damage, single-strand DNA breaks and genomic instability and is reliant on single-strand break repair/base excision repair to tolerate additional stress. PARP critical for maintaining genomic stability by regulating a variety of DNA damage repair mechanisms is constitutively activated in GSCs. In this study, we show that talazoparib – a unique PARP inhibitor exhibits a strong single agent activity to inhibit proliferation of GSCs in vitro and suppress tumor progression in GSCs xenografts. Talazoparib cytotoxic activity was through inhibiting PARylation and by trapping PARP-DNA complexes resulting in accumulation of DNA damage during replication and ultimately leading to apoptosis. More importantly, we found EFGR amplification, which occurs in about 45% of GBM as a biomarker of response to PARPi. Gene expression and RPPA data showed that GSCs with EGFR amplification were associated with increased ROS, basal expression of DNA repair proteins and more DNA damage following talazoparib treatment, which may partially explain the sensitivity of this group of GSCs to talazoparib. EGFR enzyme activity was important for PARPi sensitivity since kinase dead mutant of EGFR or EGFR knock down cell lines were resistant to PARPi as was also shown by decreased PARP/DNA complex formation. Talazoparib showed strong inhibition of tumor growth in subcutaneous glioma model and 20% increase in median survival in an intracranial model with a 10% blood brain penetration of talazoparib. These data provide mechanistic insights into the anti-cancer activity of PARP inhibitors with mechanistic rationale of PARP inhibition and potential EGFR amplification as biomarker in the development for precision cancer.

Citation Format: Shaofang Wu, Feng Gao, Xiaolong Li, Jie Ding, Siyuan Zheng, Emmanuel Martinez-Ledesma, Ningping Feng, Erik Sulman, Roel Verhaak, John de Groot, Dimpy Koul, W.K.Alfred Yung. EGFR amplification-A candidate predictive biomarker of PARP inhibitor Talazoparib sensitivity in gliomas [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 1781. doi:10.1158/1538-7445.AM2017-1781
Abstract 1791: Spatially positioned tumor infiltrating lymphocytes predict survival in metastatic HER2 positive breast cancer treated with trastuzumab
Authors: Koivunen Jussi Pekka , Honkanen Tiia , Moilanen Tiina , Karihtala Peeter , Väyrynen Juha , Mäkinen Markus
Published: July 2017

Background: Disease outcomes of HER2+ breast cancers have dramatically changed after targeted therapies, such as trastuzumab, came to clinical practice but predictive factors for trastuzumab sensitivity and resistance are frequently unknown.

Methods: Metastatic breast cancer patients (n=48), who were treated with trastuzumab and had pre-treatment tumor samples available, were studied. The tumors were immunostained for T-cell (CD3, CD8) and NK-cell (CD56) markers and quantitative analysis of the immune cells was carried out using a computer-assisted image analysis in different tumor locations.

Results: High number of CD3 and CD8 positive T-cells was associated with significant survival benefit in the center of the tumor (CT) (p=0.007, p=0.001), but not in the invasive region. NK-cell tumor infiltration was infrequent and they could not be reliably analyzed. In a subgroup analyses, high density of CD8 CT cells was associated with significant survival benefit in non-bone only disease, in TX-3, and in ER+ tumors (p=0.006, p=0.003, p=0.001). Moreover, high CD8 CT cell density was associated with good trastuzumab responses (p=0.042).

Conclusion: High number of CD3 and CD8 positive tumor infiltrating lymphocytes in the CT area is associated with survival benefit in some patient groups with HER2+ breast cancer treated with trastuzumab. Furthermore, high number of CD8 CT cells predicts benefit from trastuzumab.

Citation Format: Jussi Pekka Koivunen, Tiia Honkanen, Tiina Moilanen, Peeter Karihtala, Juha Väyrynen, Markus Mäkinen. Spatially positioned tumor infiltrating lymphocytes predict survival in metastatic HER2 positive breast cancer treated with trastuzumab [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 1791. doi:10.1158/1538-7445.AM2017-1791
Abstract 1804: Identification of novel NM23 inhibitors as potential anti-angiogenic agents
Authors: Nordmeier Senny , Evasovic Jon , Duan Suzann , Wuebbles Ryan , Burkin Dean J. , Buxton Iain L.
Published: July 2017

Exogenous nucleoside diphosphate kinase (eNDPK or NM23) has been shown to promote endothelial cell proliferation and migration and tumor-mediated angiogenesis. This is facilitated by its transphosphorylase activity, in which a gamma terminal phosphate group from a triphosphate nucleoside is transferred to a diphosphate nucleoside, resulting in elevated ATP levels. Increased levels of ATP in the tumor microenvironment can activate purinergic receptors (P2Y1) on adjacent endothelial cells to promote angiogenesis, independent of VEGF stimulation. Triple negative human breast cancer (MDA-MB-231) cells have been shown to elaborate exosomes that contain NM23. These exosomes have implications in targeting specific tissues/cells to promote angiogenesis and tumorigenesis. Our lab has shown that inhibition of eNDPK and the P2Y1 receptor reduces endothelial cell tubulogenesis (Rumjahn, et al., 2007) and breast cancer metastasis (Yokdang, et al., 2015). Therefore, targeted inhibition of eNDPK may have implications as a treatment for early tumor-mediated angiogenesis. For this study, we utilized a transphosphorylation activity assay to perform a drug screen on compounds from the Prestwick chemical library. Recombinant NDPK-B was added to FDA-approved drug compounds along with ADP and UTP substrates. Subsequent ATP generation was detected with a luciferase luciferin reaction. Decreased luminescence signal/inhibition of NDPK was compared to ellagic acid control, a known inhibitor of NDPK. From our initial screen, we identified eight potential compounds that inhibited NDPK at comparable levels to ellagic acid. These eight compounds were further examined to determine the dose response curve. We found that Chicago Sky Blue 6B inhibited NDPK at an IC50 value of 1.88µM. The results from this study demonstrate a drug screening method to discover alternative small molecule inhibitors of NDPK as a potential treatment of angiogenesis and breast cancer metastasis.

Citation Format: Senny Nordmeier, Jon Evasovic, Suzann Duan, Ryan Wuebbles, Dean J. Burkin, Iain L. Buxton. Identification of novel NM23 inhibitors as potential anti-angiogenic agents [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 1804. doi:10.1158/1538-7445.AM2017-1804
Abstract 1807: Exosomal NM23 facilitates pro-angiogenic communication between triple-negative breast cancer cells and their vascular targets
Authors: Duan Suzann , Nordmeier Senny , Buxton Iain L.
Published: July 2017

Functional interactions between primary tumor cells and their vascular targets orchestrate the formation of a metastatic milieu enriched with oxygen and metabolic substrates. These interactions have been largely attributed to the activation of VEGF receptor pathways through localized tumor-mediated release of VEGF, in addition to indirect receptor phosphorylation. However, pro-angiogenic events are known to occur in the absence of VEGF and alternative pathways resulting in dual activation of neovascularization and enhanced vascular permeability are of growing interest. Our lab has previously shown that triple-negative human breast cancer (MDA-MB-231) cells elaborate exosomes carrying NM23, a nucleoside diphosphate kinase (eNDPK) that is suggested to act as an upstream mediator of vasodilation and pro-angiogenic activity through its action of elevating extracellular ADP/ATP levels. To further elucidate the role of eNDPK in breast cancer signaling and metastasis, we show that 231-exosomes induce a pro-angiogenic phenotype in both human umbilical vein and murine pulmonary endothelial cells. Treatment with an NDPK-specific inhibitor and an antagonist to the ADP/ATP-activated P2Y1 receptor results in amelioration of pro-angiogenic characteristics. Further, we report the development and application of a novel murine implant model to demonstrate increased specificity of 231 exosome targeting to pulmonary endothelial cells over unrelated cell lines. These results suggest the involvement of exosomes and eNDPK in pro-angiogenic communication between metastatic breast cancer cells and their vascular targets. Exosomes elaborated by these cells specifically target pulmonary endothelium in vivo, mirroring the organotropic nature of breast cancer metastasis. Implication of eNDPK in angiogenesis and metastasis may challenge current and invasive treatment methods in favor of developing novel small molecule inhibitors of eNDPK signaling. Lastly, eNDPK can be explored as a potential biomarker for early detection of metastatic breast cancer as it uniquely appears in the serum of patients with breast cancer and not in the serum of individuals with unrelated pathologies.

Citation Format: Suzann Duan, Senny Nordmeier, Iain L. Buxton. Exosomal NM23 facilitates pro-angiogenic communication between triple-negative breast cancer cells and their vascular targets [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 1807. doi:10.1158/1538-7445.AM2017-1807
Abstract 1808: Vascular endothelial growth factor receptor-2 (VEGFR-2) N-glycosylation modulates angiogenic signaling
Authors: Chandler Kevin B. , Leon Deborah R. , Meyer Rosana D. , Rahimi Nader , Costello Catherine E.
Published: July 2017

Angiogenesis, the formation of new blood vessels from pre-existing vessels, is required for tumor growth and metastasis. Vascular endothelial growth factor receptor-2 (VEGFR-2) is one of the most important receptor tyrosine kinases (RTKs) among the VEGF receptor subfamily, and activation of VEGFR-2 is essential for tumor angiogenesis. The extracellular domain of VEGFR-2 contains seven immunoglobulin-like (Ig) domains, each with multiple potential N-glycosylation sites. N-glycosylation is thought to play a central role in receptor stability, ligand binding and trafficking. However, to date the occupancy and glycoform distributions at each of the potential N-glycosylation sites and their putative role(s) in VEGFR-2 function remain largely unknown. The objective of this study is to investigate the functional importance of VEGFR-2 N-glycosylation in VEGFR-2 angiogenic signaling.

Porcine aortic endothelial (PAE) cells with ectopic expression of VEGFR-2 were treated with PNGase F to remove N-linked glycans or heat-denatured PNGase F as a control. Following PNGase F treatment for 4 hours, cells were treated with VEGF-A ligand for 0, 5, 10 or 30 minutes. VEGFR-2 phosphorylation (activation) was measured via Western blot with an anti-pTyr-1054-VEGFR-2 antibody. In addition, a polyclonal anti-VEGFR2 antibody was used to immunoprecipitate untreated and PNGase F-treated VEGFR-2 from PAE cell lysates. Evaluation of the N-glycosylation sites targeted by PNGase F was carried out by gel electrophoresis, followed by protease digestion and MS/MS analysis. MS/MS data were processed using Proteome Discoverer 1.4. To obtain site-specific glycosylation information we performed proteolysis of VEGFR-2, glycopeptide enrichment via hydrophilic interaction liquid chromatography (HILIC) and subsequent analysis of glycopeptides with an Agilent 6550 Quadrupole Time-of-Flight (Q-TOF) MS using collision-induced dissociation.

We detected a dramatic increase in ligand-mediated activation of VEGFR-2 after treatment with PNGase F, suggesting that certain N-linked glycans may hinder ligand access to the VEGF binding site, or that removal of N-linked glycans results in conformational changes that lead to increased activation of the receptor by VEGF-A. To explore this observation in greater detail, we have created a series of VEGFR-2 N-glycosylation site mutants, and we are now using the mutants to determine which glycosylation sites are involved in the observed modulation of VEGFR-2 signaling.

Citation Format: Kevin B. Chandler, Deborah R. Leon, Rosana D. Meyer, Nader Rahimi, Catherine E. Costello. Vascular endothelial growth factor receptor-2 (VEGFR-2) N-glycosylation modulates angiogenic signaling [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 1808. doi:10.1158/1538-7445.AM2017-1808
Abstract 1809: PKM2 promotes tumor angiogenesis by regulating HIF-1alpha through NF-kappaB activation
Authors: Azoitei Ninel , Becher Alexander , Steinestel Konrad , Rouhi Arefeh , Diepold Kristina , Bobrovich Susanne , Seufferlein Thomas
Published: July 2017

Initially identified as a molecule that catalyzes the phosphorylation from phosphoenolpyruvate to pyruvate in the final step of glycolysis, the M2 isoform of pyruvate kinase (PKM2) was recently reported to have a central role in the metabolic reprogramming of cancer cells as well as participating in the regulation of apoptosis and proliferation. In this study, we have identified PKM2 as a crucial molecule for progression of pancreatic cancer in which tumor microenvironment has been reported to be highly hypoxic. Our data show moderate to strong PKM2 expression in all examined human pancreatic adenocarcinoma samples. Depletion of PKM2 was associated with impaired proliferation and augmented tumor cell death in vitro, while the in vivo tumor xenograft experiments revealed a close association between impaired tumor growth and decreased blood vessel formation. Furthermore, abrogation of PKM2 prevented hypoxia-mediated HIF-1alpha accumulation and HIF-1alpha promoter activity, which negatively impacted VEGF secretion by pancreatic cancer cells deprived of oxygen. Interestingly, PKM2 expression arrest was also mirrored by impaired hypoxia-driven promoter activity of NF-kappaB which also drives the transcription of VEGF. Ectopic expression of p65 restored VEGF transcription after PKM2 ablation inferring that the kinase regulates VEGF via NF-kappaB/p65 subunit. Altogether, our study suggests that in hypoxic pancreatic tumors PKM2 interferes both with NF-kappaB/p65 and HIF-1alpha activation that ultimately triggers VEGF-A secretion and subsequent blood vessel formation.

Citation Format: Ninel Azoitei, Alexander Becher, Konrad Steinestel, Arefeh Rouhi, Kristina Diepold, Susanne Bobrovich, Thomas Seufferlein. PKM2 promotes tumor angiogenesis by regulating HIF-1alpha through NF-kappaB activation [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 1809. doi:10.1158/1538-7445.AM2017-1809
Abstract 1814: IL11 mediates tumor resistance to anti-VEGF therapy in lung cancer
Authors: Li Jie , Xie Kun , Mei Shenglin , Xu Yuanjian , Gu Hua , Li Dong , Chen Kaiming , Tao Fei , Eerdeng Jiya , Fang Jianmin
Published: July 2017

Anti-angiogenic therapy has been used for the treatment of a number of solid tumors in clinic, however, survival benefit is often limited due to development of tumor resistance to the therapy. In previous studies, we developed a mouse Lewis lung carcinoma (LLC) tumor model that is resistant to anti-VEGF therapy. In this model, we identified that Interleukin 11 (IL11) was among the most significantly upregulated genes in the resistant tumors. In current study, we demonstrated that IL11 was a pro-angiogenic factor and played a key role in the development of resistance to anti-VEGF therapy in LLC tumor. IL11 could effectively induce HUVEC proliferation, migration and capillary tube formation in vitro. It enhanced expression of MMP9 in endothelial cells. The conditioned medium from the resistant LLC cell culture could induce HUVEC migration, even VEGF was blocked, and this effect was attenuated by recombinant IL11 decoy receptor, an IL-11 inhibitor. Furthermore, combination therapy of anti-VEGF agent and IL11 blockade exhibited enhanced anti-tumor activity in lung cancer model. In addition, based on bioinformatics analysis on over 500 RNA-seq data deposited to The Cancer Genome Atlas (TCGA), we found that IL11 was highly expressed in various tumor types in patients. Data showed that high IL11 can predict a poor outcome in the lung cancer. The patients with the lower IL11 expression have significantly longer median survival time than those with higher expression. These findings support a role of IL11 signal as a compensatory angiogenic pathway that might trigger tumor resistance to anti-VEGF treatment. Therapeutically targeting IL11 could have a value in overcoming tumor resistance to anti-VEGF therapy.

Citation Format: Jie Li, Kun Xie, Shenglin Mei, Yuanjian Xu, Hua Gu, Dong Li, Kaiming Chen, Fei Tao, Jiya Eerdeng, Jianmin Fang. IL11 mediates tumor resistance to anti-VEGF therapy in lung 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 1814. doi:10.1158/1538-7445.AM2017-1814
Abstract 1820: Synergistic lethality of mAbs with an EMT reversal agent, Nintedanib, in epithelial ovarian cancer
Authors: Teo Jocelyn , Tan Heng Liang , Huang Ruby Yun-Ju , Choo Andre
Published: July 2017

Epithelial-mesenchymal transition (EMT) has long been associated with cancer progression and metastasis. As a reversible process, it has the potential to be a target for cancer therapy. However, this concept of reversing EMT has not yet been widely explored in current treatment strategies. With the identification of Nintedanib as an EMT reversal agent, we aim to broaden the utility of this triple angiokinase inhibitor by identifying agents that show synergistic lethality with it, using monoclonal antibodies (mAbs) as a combinatory therapeutic. Nintedanib alone is not significantly cytotoxic to cells, but it causes cell cycle arrest and reverses EMT. By selecting for this sub-population of cells that are sensitive to Nintedanib, their subsequent eradication can then be more intricately directed. Since the anti-cancer application of EMT reversal agents are not fully developed, it is vital to investigate mechanisms that sensitize cancer cells to cytotoxic agents following EMT reversal.Our group has generated panels of mAbs that are able to bind differentially to various breast and ovarian cancer cell lines along the EMT spectrum. Some mAbs preferentially bind to Epithelial lines, while some have stronger affinity to Mesenchymal lines. This implies that mAbs have the ability to differentiate between epithelial and mesenchymal phenotypes, uncovering a new dimension to the capability of mAbs which have yet to be explored.Using a mesenchymal ovarian cancer cell line as a starting model, Nintedanib-treated cells were screened with the mAbs and those showing a >2-fold increase in binding were shortlisted for further validation. Based on this criteria, 26 mAbs were identified and further characterized in functional Antibody Drug Conjugate (ADC) assays with mAbs conjugated to a toxic drug, Saporin (or Zap). Mabs successfully internalizing and killing the cancer cells will then be further chosen for antigen characterization and in vivo functional studies. Ultimately, there is huge potential for mAbs to be discovered that shows synergy with Nintedanib to enhance lethality to various types of cancer cells. A novel clinical strategy, the application of EMT reversal in terms of utilising a synthetic lethality-like approach allows better design of combinatory therapeutics, increasing treatment efficacy that might revolutionalise cancer treatment.

Citation Format: Jocelyn Teo, Heng Liang Tan, Ruby Yun-Ju Huang, Andre Choo. Synergistic lethality of mAbs with an EMT reversal agent, Nintedanib, in epithelial ovarian 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 1820. doi:10.1158/1538-7445.AM2017-1820
Abstract 1825: Angiopoietin-Tie-2 functional axis in colorectal cancer liver metastasis (CRCLM) provides a new marker for stratification and evaluation of tumor progression
Authors: Ibrahim Nisreen S.
Published: July 2017

Colorectal cancer (CRC) is the third leading cause of cancer in Canadians, with liver metastases being the major cause of death from this disease. Tumors induce angiogenesis, a phenomenon known as the ‘angiogenic switch’, which is an essential step in tumor progression whereby the balance of pro- and anti-angiogenic factors are important for active angiogenesis. Clinical efficacy of targeted VEGF (anti-angiogenic) treatment has been validated as a cancer therapy. Our group, together with others, has identified unique histological growth patterns HGPs (desmoplastic, replacement and pushing) within liver metastases that have different responses to anti-angiogenic therapy. The patients with Desmoplastic HGP (DHGP) that received anti-angiogenic plus chemotherapy prior to resection had a significantly better pathologic response and survival than patients with Replacement HGP (RHGP). The aim of this study was to explore the role of Ang-1, Ang-2, Tie-2 and VEGF in the development and progression of CRCLM tumors with distinct HGPs. Here, human CRCLM tumor samples were analyzed by quantitative real-time PCR (Q-PCR) and immunohistochemistry (IHC) staining. The Q-PCR results demonstrated that the expression of Ang-2 was lower in RHGP tumor samples compared with DHGP tumor samples. This data was validated by IHC, were IHC scoring results showed that the ratio of Ang-2: Ang-1 expression in DHGP tumors was higher compared to RHGP tumors. VEGF and Tie2 proteins were expressed in both tumor patterns. Thus vascular quiescence maintained by constitutive Ang-1/Tie-2 signaling, found in RHGP tumors, prevails over destabilization and pro-inflammatory Ang-2/Tie-2 signaling, which is higher in the DHGP tumor samples. Since vascular remodelling is driven by Ang-2/Tie-2 in DHGP, which is dependent on VEGF we would expect anti-angiogenic therapy to be effective on DHGP. Furthermore, the RHGP had low levels of Ang-2 and high levels of Ang-1, together with the presence of Tie-2, and then one would predict that VEGF is not required for the growth of these tumors and thus would not respond to anti-angiogenic therapy, as has been shown in our patients’ cohort. Taken together, these data suggest that the Angiopoietin/Tie-2 functional axis is an important player in CRCLM tumor progression and can be a potential target for CRCLM cancer therapy with stratification of patients by HGPs.

Citation Format: Nisreen S. Ibrahim. Angiopoietin-Tie-2 functional axis in colorectal cancer liver metastasis (CRCLM) provides a new marker for stratification and evaluation of tumor 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 1825. doi:10.1158/1538-7445.AM2017-1825
Abstract 1833: Imaging-guided brain tumor surgery strategy based on a triple-modality MRI-PET-fluorescence imaging probe and imaging analysis
Authors: Meng Hui , Jin Yushen , Zhang Shaowei , Zhang Shuai , Ma Xibo , Tian Jie
Published: July 2017

Precise brain tumor resection is valuable for increasing the 5-year survival rate. However, the delineation of tumor margin is a major challenge. Transferrin modified nanoprobes had the property of passing the blood-brain barrier (BBB). Here, we endeavored to use transferrin to fabricate a triple-modality magnetic resonance imaging-PET-fluorescence imaging molecule agent (MPF) which could improve diagnostic sensitivity and specificity of the brain tumors in the imaging acquisition and analysis. The localization of the tumor were determined by the MRI before surgery. Glucose metabolism was derived from PET imaging which would be an important influence factor for surgery strategy. Tumor margin was delineated using the quick segmentation methods according to the fluorescence imaging after craniotomy which could reduce residual tumor after resection. Finally, tissues were sliced and stained with hematoxylin and eosin every 5 mm. The histology results were processed into a whole image which can be used to evaluate the accuracy of intraoperative tumor margin. This three modality imaging-guided surgery strategy is a promising method which will probably achieve its application in clinical trials in near future.

Citation Format: Hui Meng, Yushen Jin, Shaowei Zhang, Shuai Zhang, Xibo Ma, Jie Tian. Imaging-guided brain tumor surgery strategy based on a triple-modality MRI-PET-fluorescence imaging probe and imaging analysis [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 1833. doi:10.1158/1538-7445.AM2017-1833
Abstract 1839: TGFBR1*6A mouse model mimics human breast cancer development and progression
Authors: Pennison Michael James , Wang Minghui , Jimenez Hugo , Pasche Boris
Published: July 2017

Transforming Growth Factor-ß (TGF-β) is one of the most commonly altered cellular signaling pathways in human cancer. Our group was the first to discover TGFBR1*6A (*6A), a common hypomorphic variant of the type I TGF-ß receptor (TGFBR1), which has been associated with increased risk for breast cancer in multiple epidemiological studies. Functional analysis also suggests that *6A is associated with decreased tumor suppressive TGF-β signaling in early tumor development and is involved with promoting tumorigenesis in advanced carcinomas. To better understand the functional impact of *6A and constitutively reduced TGF-ß signaling on breast cancer development and progression, we have recently developed a series of transgenic knock-in mouse model variants by replacing exon 1 of the mouse Tgfbr1 sequence with human exon 1 sequences harboring both *6A and wild-type (*9A) TGFBR1 variants and crossed them with MMTVneu mice.

We generated a total of 5 FVB double and/or triple transgenic mouse strains: MMTVneu;9A/9A, MMTVneu;6A/6A, MMTVneu;9A/6A, MMTVneu;*9A/-, and MMTVneu;6A/-. Following tumor development, measurements were performed three times per week to determine tumor volume. All mice were allowed to live up to 18 months (548 days) or until they died of natural causes. Specimens were collected for later analysis of blood, tumors, normal mammary tissue, and were evaluated for lung metastases. Statistical comparisons performed using Student’s t-test.

Only 34.4% of MMTVneu;9A/9A mice (11/32) developed mammary tumors, while 80% of MMTVneu;6A/6A mice (24/30; p=0.0002) and 62.5% of MMTVneu;6A/- mice (19/32, p=0.046) developed tumors. The percentage of MMTVneu;9A/6A and MMTVneu;*9A/- mice that developed tumors was higher than MMTVneu;9A/9A mice (48.3% and 50%, respectively) but was not statistically significant. Additionally, 66.7% of MMTVneu;6A/6A mice (20/30) developed 2 or more tumors, while only 25% of MMTVneu;9A/9A mice (8/32) developed multiple tumors (p=0.0007).

Our data suggests that we have developed transgenic mouse strains that accurately mimic human breast cancer susceptibility associated with *6A and constitutively decreased TGFBR1 expression in humans, as evidenced by other studies. These mouse models may be a tool to provide invaluable insight to better understand the role of TGFBR1 variants in the development and progression of breast cancer.

Citation Format: Michael James Pennison, Minghui Wang, Hugo Jimenez, Boris Pasche. TGFBR1*6A mouse model mimics human breast cancer development and 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 1839. doi:10.1158/1538-7445.AM2017-1839
Abstract 1844: Investigating the role of amphiregulin in breast cancer
Authors: Chiang Serena P. , Karagiannis George S. , Condeelis John S. , Segall Jeffrey E.
Published: July 2017

Breast cancer is the most prevalent cancer in women worldwide and afflicts more than 3.1 million women in the US alone. The epidermal growth factor receptor (EGFR) is frequently overexpressed in breast cancer and is associated with poor clinical outcome. Several therapies involving anti-EGFR agents have been developed to target breast cancer. However, clinical trials have shown mixed outcomes with these therapies. Amphiregulin (AREG) is a ligand for EGFR that mediates estrogen-induced mammary ductal morphogenesis during development. Expression of AREG is enriched in invasive breast carcinomas, particularly ERα-positive breast tumors. Studies have shown that AREG increases the in vitro invasiveness of breast cancer cells. Furthermore, suppression of AREG expression in transformed human breast epithelial cells reduces tumor formation when injected in nude mice. These observations suggest that AREG plays an important role in breast cancer development and progression. To investigate the role of AREG in breast cancer development and progression, we utilized a spontaneous mammary tumor mouse model where oncogenesis is driven by expression of the polyoma middle T oncoprotein (PyMT) through the mouse mammary tumor virus (MMTV) promoter. We have crossed AREG knockout mice with the MMTV-PyMT mice to generate AREG KO mice that spontaneously form mammary tumors. Tumors in AREG KO PyMT mice initially grow more quickly but eventually reach 2 cm in diameter at a relatively time as compared to tumors formed in AREG WT PyMT mice. Histologically, AREG KO tumors appear more differentiated and display a greater proportion of tumor stroma. This prompted us to further hypothesize that AREG may have diverse effects in the tumoral or stromal compartment of developing tumors. To individually assess the effects of AREG knockout in the tumor cells and the stroma, we utilized a transplantation model in which we prepared tumor chunks from AREG WT and KO PyMT tumors and transplanted them into AREG WT and KO mice. Through this model, we have found that AREG expression in the tumor has a greater impact on tumor growth and progression than the tumor stroma. Regardless of the AREG status of the recipient mice, AREG KO tumors display a significant growth impairment and reduced transplant efficiency. In summary, these results demonstrate that AREG is important for mammary tumor progression.

Citation Format: Serena P. Chiang, George S. Karagiannis, John S. Condeelis, Jeffrey E. Segall. Investigating the role of amphiregulin in 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 1844. doi:10.1158/1538-7445.AM2017-1844
Abstract 1855: Developed operation method for patient-drived orthotopic xenograft model of hepatocellular carcinoma
Authors: Byeon Hye Rim , Seo YunSung , Kong Sun-Young , Lee Seung Duk
Published: July 2017

Patient-drived xenograft(PDX) model is a biological organized platform and a patient substitute. Recent studies use the ectopic model for hepatocellular carcinoma(HCC) but this model don’t reflect likewise the tumor state of patient. For exact prognosis, it is need to establish patient-derived orthotopic xenograft(PDOX) mouse model of hepatocellular carcinoma using direct injection technique. Additionally, secretion of growth factor from bone marrow stem cells is promoted after partial hepatectomy. This mechanism is helpful to not only general liver regeneration but also tumorigenesis. Based on former PDX research, we develop the new protocol using direct injection technique. From January 2015 to June 2016, consent about getting tumor pieces to the patient of hepatocellular carcinoma. The pieces is stored in RPMI1640 and moved from operation room to research room immediately. For PDOX and PDX to subcutaneous model, 8~10weeks old NOD SCID and NSG male mouse is used. The tumor pieces cutted to the cube size 1mm. For stop the bleeding, inferior vena cava is blocked using Bulldog Clamp and the cube is directly injected median lobe to 3 places randomly. After clear the Bulldog Clamp, a left lobe is removed using tie and weighted. After 3 months, the PDOX mouse is checked for cancer growth using MRI or PET-CT. Successful PDOX or PDX to subcutaneous model is sacrificed and succeeded a generation to another Balb/C-nude male mouse 8~10weeks old. The rate of success of PDOX model and PDX to subcutaneous model is about 10% respectively. Formed tumor is verified by H&E staining. Based on protocol established, further study plan is required for improving on the rate of success to establish PDOX model and clinical application.

Note: This abstract was not presented at the meeting.

Citation Format: Hye Rim Byeon, YunSung Seo, Sun-Young Kong, Seung Duk Lee. Developed operation method for patient-drived orthotopic xenograft model of hepatocellular carcinoma [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 1855. doi:10.1158/1538-7445.AM2017-1855
Abstract 1858: Theranostic nanoparticles for detection and treatment of pancreatic cancer
Authors: Chuong Phillip , Bhutiani Neal , McNally Lacey R.
Published: July 2017

Background: Despite continued efforts to improve early detection and treatment of pancreatic adenocarcinoma, the disease still carries a poor prognosis. The combination of multispectral optoacoustic tomography (MSOT) and theranostic nanoparticles offers a possible solution by providing tumor targeting using fluorescent dyes and treatment through drug delivery. We describe a 40 nm colloidal mesoporous silica nanoparticle (CMS) with vermiform pores and a chitosan coating and tagged with the V7 peptide to provide pH-specific dual targeting to deliver a contrast agent in a tumor-specific fashion.

Methods: CMSs were synthesized using a modified Stober method, coated with chitosan, and conjugated with a V7 peptide, resulting in a dual acidic pH targeting system. Particle characterization was performed using transmission electron microscopy (TEM), UV-vis spectrophotometry, Zeta potential, and dynamic light scattering . To determine acidic pH specificity of V7-CMS, Panc1 and S2VP10 cell lines were incubated in cell culture medium at either pH 7.4 or 6.6 followed by treatment with V7-CMS. Particle uptake was determined using near infrared (NIR) fluorescence and tissue phantoms. Finally, for in vivo testing, the same CMSNs were injected into mice bearing S2VP10 pancreatic tumors. MSOT imaging was performed 8 hours after CMS injection.

Results: TEM images demonstrated successful synthesis of approximately 40 nm V7-CMSs with vermiform pores. On NIR imaging, V7-CMS demonstrated acidic pH specificity in both S2VP10 and Panc 1 cells at pH 6.6, with particle localization and signal intensity 8-fold and 5-fold higher, respectively, than that observed at pH 7.4. In tissue phantoms, increased pH specificity was also observed in both S2VP10 and Panc 1 cells at pH 6.6. Particle location and signal intensity on MSOT was 20-fold and 4-fold higher, respectively, than that observed at pH 7.4. In the murine orthotopic S2VP10 pancreatic tumor model, MSOT imaging 8 hours after IV CMS injection demonstrated V7-CMS accumulation specifically within pancreatic tumors.

Conclusion: The acidic pH specific dual targeting system using chitosan-coated and pHLIP V7-tagged CMSs results in tumor specificity. Preferential binding and dye release was increased as much as 20-fold at pH 6.6 as compared to pH 7.4 in tissue phantoms. MSOT was able to detect tumor specific accumulation of V7-CMS in vivo. These small, pH specific particles hold significant promise for effectively identifying pancreatic tumors with high sensitivity and specificity and for targeted drug delivery.

Note: This abstract was not presented at the meeting.

Citation Format: Phillip Chuong, Neal Bhutiani, Lacey R. McNally. Theranostic nanoparticles for detection and treatment 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 1858. doi:10.1158/1538-7445.AM2017-1858
Abstract 1872: Fluorescence-guided surgery in the tumor microenvironment in a syngeneic mouse model of EL-4 lymphoma
Authors: Hasegawa Kosuke , Suetsugu Atsushi , Nakamura Miki , Matsumoto Takuro , Kunisada Takahiro , Shimizu Masahito , Saji Shigetoyo , Moriwaki Hisataka , Bouvet Michael , Hoffman Robert M.
Published: July 2017

We report fluorescence-guided surgery (FGS) of the tumor microenvironment of the EL-4-RFP lymphoma cells growing subcutaneously in C57/BL6 GFP transgenic mice. Using a portable hand-held Dino-Lite digital imaging system, subcutaneous tumors were resected by FGS. The tumor surface contained both RFP cancer cells and GFP stromal cells. The tumor microenvironment was clearly visualized and resected with the Dino-Lite. Host stromal cells, including adipocyte-like cells and blood vessels with lymphocytes, were observed along the tumor by confocal microscopy in addition to cancer cells by color-coded confocal imaging. Stromal cells were within the tumor and that cancer cells also invaded the TME. Color-coded FGS, which distinguished cancer and stroma, significantly prevented local recurrence, which bright-light surgery or single-color FGS could not.

Citation Format: Kosuke Hasegawa, Atsushi Suetsugu, Miki Nakamura, Takuro Matsumoto, Takahiro Kunisada, Masahito Shimizu, Shigetoyo Saji, Hisataka Moriwaki, Michael Bouvet, Robert M. Hoffman. Fluorescence-guided surgery in the tumor microenvironment in a syngeneic mouse model of EL-4 lymphoma [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 1872. doi:10.1158/1538-7445.AM2017-1872
Abstract 1874: First demonstration that hypoxia targeting with radiation improves tumor control in a mammal tumor model
Authors: Halpern Howard J. , Epe; Boris , Maggio Matthew C. , Krzykawska-Serda Martyna , Redler Gage H. , Miller Richard C. , Barth Eugene D. , Weichselbaum Ralph R.
Published: July 2017

For over a century, resistance of radiation to living tissues has been associated with hypoxia, a local lack of molecular oxygen, or low pO2. The focus of this research has been to validate the hypothesis that has been assumed, but not proven over the past century, that radiation treatment focused specifically on hypoxic regions of tumors would improve tumor curability. This research used Electron Paramagnetic Resonance (EPR) imaging to provide absolute pO2 images in volume elements or voxels of murine tumors with 1 torr pO2 resolution and 0.7 mm spatial resolution in FSa fibrosarcomas in the legs of C3H mice. We define hypoxic regions of tumors as those whose image subvolumes or voxels have low pO2, voxels less than 10 torr. We have established EPR pO2 imaging as a reliable locator of relevant radiobiologically relevant hypoxia. To determine if pO2 based dose painting improves tumor cure, we implemented the XRAD225Cx system to deliver gantry based x-ray treatments to mouse tumors accurately registered with EPR pO2 images. All tumors were pretreated with a dose of radiation that had been determined in separate experiments to cure 30% of tumors, a TCD30. Conclusive failure to demonstrate a difference between treating crude spherical volumes including ~ 85% of hypoxic voxels with an additional curative dose compared with treatment with shells of radiation avoiding hypoxia (anti-boosts) proved tenets of radiation biology: a few surviving hypoxic clonogens cause radiation treatment failure. In response, we implemented rapid 3D printing Tungsten loaded, highly conformal plastic blocks to compare treating ~100% of hypoxic tumor voxels with hypoxia avoidance, treating similar volumes of well oxygenated tumor. Only then did we observe significant (p<0.01) tumor control differences between hypoxic boosts and anti-boosts. This is the first validation of the curative effectiveness of focusing hypoxia based dose painting in mammalian tumors.

Citation Format: Howard J. Halpern, Boris Epe;, Matthew C. Maggio, Martyna Krzykawska-Serda, Gage H. Redler, Richard C. Miller, Eugene D. Barth, Ralph R. Weichselbaum. First demonstration that hypoxia targeting with radiation improves tumor control in a mammal tumor 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 1874. doi:10.1158/1538-7445.AM2017-1874