bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5501: Antagonism of sorafenib and regorafenib actions by epidermal growth factor in hepatoma cells
Authors: Carr Brian I. , D'Alessandro Rosalba , Refolo Maria Grazia , Lippolis Catia , Messa Caterina , Cavallini Aldo
Published: August 2015

Background. Platelet numbers are related to growth and aggressiveness of several tumor types, including hepatoma (hepatocellular carcinoma). We previously found that platelet lysates also stimulated growth and migration of hepatoma lines in vitro and antagonized the growth-inhibitory effects on hepatoma cells of both multikinase inhibitors sorafenib and regorafenib (BMC Cancer 2014; 14:351).

Aims. To investigate the mechanisms for the platelet-mediated multikinase drug resistance, we examined the actions of several mitogens that are known to be contained by platelets (PDGF, VEGF, TGF beta, FGF, IGF-1, EGF).

Results. We found that epidermal growth factor (EGF) alone antagonized the growth inhibitory actions of Sorafenib or Regorafenib on HepG2, PLC/PRF/5 and Hep3B human hepatoma cells, using an MTT assay. Detailed studies revealed that EGF also antagonized Regorafenib-mediated inhibition of cell migration and invasion. Furthermore, EGF also blocked regorafenib-induced apoptosis, measured by Annexin V using a MUSE cell analyzer. These effects of EGF were antagonized by concomitant addition to the cultures of EGF receptor antagonist Erlotinib, showing that the EGF receptor was involved in the mechanisms of EGF-mediated blocking of Regorafenib effects. In addition, Erlotinib also partially blocked the effects of platelet lysates in antagonizing regorafenib-mediated growth inhibition. Similar antagonism was found using IGF-1, and those effects were blocked by GSK IGF-1 receptor antagonist. Other platelet factors had only weak actions in these assays.

Conclusions. EGF antagonized regorafenib-mediated growth and migration inhibition and apoptosis induction in hepatoma cells. These EGF actions were mediated via the EGF receptor, as were the similar platelet lysate actions.

Citation Format: Brian I. Carr, Rosalba D'Alessandro, Maria Grazia Refolo, Catia Lippolis, Caterina Messa, Aldo Cavallini. Antagonism of sorafenib and regorafenib actions by epidermal growth factor in hepatoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5501. doi:10.1158/1538-7445.AM2015-5501
Abstract 5507: Characterization of ADC bystander killing in admixed tumor model
Authors: Li Fu , Emmerton Kim Kissler , Jonas Mechthild , Zhang Xinqun , Law Che-Leung
Published: August 2015

We have previously described an admixed tumor model, consisting of the CD30+ ALCL cell line Karpas 299 and CD30- Karpas-35R (SGN-35 resistant) cells, to study the phenomenon of ‘bystander killing’ of antigen negative tumor cells by antibody-drug conjugates (ADCs) in tumors with a heterogeneous target expression profile. Here we study the contribution of payload membrane permeability and target expression in bystander killing.

We hypothesize that membrane permeability of payloads may impact bystander killing in addition to their intrinsic potency. Monomethyl auristatin E (MMAE)-based ADCs utilizing the cleavable linkers valine-citrulline (vc) or glucuronide (gluc) release the membrane permeable MMAE. As a result, antiCD30-vcMMAE and antiCD30-glucMMAE showed similar bystander killing in vivo. We then compared the more membrane permeable payloads, MMAE and pyrrolobenzodiazepine dimer (PBD), to those that are less permeable, monomethyl auristatin F (MMAF) and auristatin T (AT), as antiCD30 conjugates in this model. We found that antiCD30-AT and antiCD30-vcMMAF killed only CD30+ tumor cells in admixed tumors, while antiCD30-MMAE, antiCD30-GlucMMAE and antiCD30-PBD killed both CD30+ and CD30- cells. These results suggest that membrane permeability is one of the key factors determining in vivo bystander killing.

To elucidate the role of antigen positive cells in bystander killing in this model, we varied the implant ratio of CD30+ cells and evaluated ADC activity in vivo. Flow cytometric analysis and CD30 immunohistochemistry confirmed a corresponding change of CD30+ cell percentage in the resulting admixed tumors (∼40%, 70%, or 80%). Treatment with ADCs in these tumors demonstrated that antiCD30-vcMMAE induced remissions in tumors with 70 or 80% CD30+ cells, whereas antiCD30-PBD produced remissions in all three admixed tumor ratios.

Collectively, these data confirm that membrane permeable payloads can kill antigen negative cells in vivo. We hypothesize that the enhanced bystander killing by PBD-based ADCs may result from the intrinsic potency of the small molecule and/or membrane permeability. This information will be useful to consider when designing future ADCs.

Citation Format: Fu Li, Kim Kissler Emmerton, Mechthild Jonas, Xinqun Zhang, Che-Leung Law. Characterization of ADC bystander killing in admixed tumor model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5507. doi:10.1158/1538-7445.AM2015-5507
Abstract 551: Identification of gene-expression biomarkers in urine pathology specimens for the detection of bladder cancer
Authors: Chapman Karen B. , Qiu Liqun , Kidd Jennifer , Baxi Aparna , Lachter Markus D. , Wagner Joseph , Rosenthal Dorothy L. , Olson Matthew T.
Published: August 2015

Bladder cancer recurrence screening is typically managed with a combination of urine cytology and cystoscopy, each with its inherent caveats. Cystoscopy is generally considered the gold standard for diagnosing bladder cancer and can yield a valuable biopsy, but is too invasive and expensive for routine screening and surveillance. Voided urine cytology is inexpensive and readily available, but lacks the desired level of sensitivity. Furthermore, up to 20% of urine cytology specimens fall into one of two indeterminate categories that require follow-up testing, typically in the form of an invasive cystoscopy procedure. With the goal of developing a urine-based molecular test for bladder cancer recurrence surveillance, we examined gene expression biomarkers in urine cytopathology specimens. 90 patient urine samples were analyzed: 45 high-grade urothelial carcinoma (HG-UC) and 45 benign samples. All patient samples were submitted to the Johns Hopkins Cytopathology Department for evaluation for bladder cancer via urine cytology. The samples represent patients undergoing bladder cancer recurrence screening and patients without a prior history of bladder cancer presenting with hematuria. The urine sediments were stored in liquid based cytology solution at 4°C for a period of 7 - 10 days, during which time the cytopathology diagnosis was determined from a portion of the sample. Post-diagnosis, the pathology sample “leftover” was analyzed for gene expression. RNA was extracted, amplified and microarray analysis was performed and a panel of molecular biomarkers was identified. Individual genes in this panel discriminate between HG-UC and benign in this training set with an average ROC of 0.86 (p <0.0001) while a combined panel of 10 genes yields a ROC of 0.97 (p <0.0001). Differential expression of some of the individual genes from the panel was confirmed by qPCR. Additional evaluation of this gene panel in a validation set is ongoing and results from this validation study will be discussed.

Citation Format: Karen B. Chapman, Liqun Qiu, Jennifer Kidd, Aparna Baxi, Markus D. Lachter, Joseph Wagner, Dorothy L. Rosenthal, Matthew T. Olson. Identification of gene-expression biomarkers in urine pathology specimens for the detection of bladder cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 551. doi:10.1158/1538-7445.AM2015-551
Abstract 5517: Polymeric nanogel-based treatment regimen for enhanced efficacy and sequential administration of synergistic drug combination in pancreatic cancer
Authors: Soni Kruti S. , Hollingsworth Michael A. , Radhakrishnan Prakash , Bronich Tatiana K.
Published: August 2015

Pancreatic cancer (PC) is one of the most lethal malignancies, due to aggressive tumorigenicity, early metastasis and development of drug resistance to standard care chemotherapy. Since its approval in 1997, Gemcitabine has been the first-line treatment for advanced disease. However, there is no standard second-line therapy after Gemcitabine failure. The combination of Gemcitabine with Cisplatin (CDDP) has been explored in clinical trials for metastatic disease and our preliminary in vitro studies with the free drugs on T3M4 Simple Cells (COSMC deleted cells) showed that synergy of the combination is schedule-dependent, and Gemcitabine administration followed by CDDP showed the most potent cytotoxic activity. However, this combination proved to be only marginally effective in actual practice due to combined increased toxicity of both the agents. We have shown that encapsulation of CDDP in polymeric nanogels with cross-linked ionic cores enhanced its tumor accumulation and improved its safety profile. Additionally, sustained release profile of CDDP from nanogels allows for the administration of free Gemcitabine and CDDP loaded nanogels in a single injection while still retaining schedule-dependent synergy of the combination. Pancreatic ductal adenocarcinoma cells are known to express truncated O-glycans (Tn and STn antigens) and it is anticipated that decorating the nanogels with an antibody directed against this antigen will further enhance their uptake by tumor cells while reducing off-target accumulation.

Citation Format: Kruti S. Soni, Michael A. Hollingsworth, Prakash Radhakrishnan, Tatiana K. Bronich. Polymeric nanogel-based treatment regimen for enhanced efficacy and sequential administration of synergistic drug combination in pancreatic cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5517. doi:10.1158/1538-7445.AM2015-5517
Abstract 5536: KB-164, a novel and highly potent Src family kinase inhibitor, reduces cancer cell invasion and abrogates downstream oncogenic pathways in triple negative breast cancer
Authors: Gilani Rabia A. , Brandvold Kristoffer R. , Bao Li Wei , Phadke Sameer , Steffey Michael E. , Soellner Matthew B. , Merajver Sofia D.
Published: August 2015

Src is a non-receptor tyrosine kinase, which acts as an integrator of diverse signaling pathways used by cancer cells for cell proliferation, migration and metastasis. Recent interest in developing target specific therapy for improved benefits with manageable toxic effects has sharply increased the interest in testing new Src family kinase inhibitors for their potential anticancer therapeutic properties.

In this study, we sought to investigate the chemical characteristics and the biological effects of the novel Src family kinase inhibitor KB-164, in Src expressing triple negative breast cancer cells. We show that KB-164 acts as a potent Src kinase inhibitor (IC50 1.3 nmol/L) in MDA-MB 231 cells, 100-fold more potent than the most widely used Src family kinase inhibitor dasatinib. At concentrations of 50nmol/L, KB-164 led to complete inhibition of Src activation for 48 hours. Analysis of downstream effectors of Src revealed that treatment with KB-164 inhibits the phosphorylation of several oncogenic pathways i.e. EGFR, ERK1/2, P38MAPK and AKT. A dramatic decrease in cancer cell motility and invasion was observed on treatment with KB-164 in both SUM 149 and MDA-MB 231 triple negative breast cancer cell lines. We also show that triple negative breast cancer cells derived from the patients are equally sensitive to growth inhibition by KB-164 within an IC50 of 1.8-nmol range. No cytotoxicity was observed both in vitro (18 micro mol/L in normal human mammary epithelial cells) and in vivo (70mg/kg in mice). Our findings indicate that KB-164 inhibits signaling pathways involved in controlling tumor cell proliferation and survival, suggesting that KB-164 might prove to be a promising therapeutic agent for breast cancer.

Citation Format: Rabia A. Gilani, Kristoffer R. Brandvold, Li Wei Bao, Sameer Phadke, Michael E. Steffey, Matthew B. Soellner, Sofia D. Merajver. KB-164, a novel and highly potent Src family kinase inhibitor, reduces cancer cell invasion and abrogates downstream oncogenic pathways in triple negative breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5536. doi:10.1158/1538-7445.AM2015-5536
Abstract 5542: Flaxseed and its lignan component protect from asbestos-induced inflammation in mice
Authors: Albelda Steven M. , Pietrofesa Ralph A. , Menges Craig W. , Testa Joseph R. , Christofidou-Solomidou Melpo
Published: August 2015

Introduction/Background: Malignant mesothelioma (MM) is a highly lethal form of thoracic cancer with high mortality and poor treatment options. Development of MM has been linked directly to exposure to asbestos fibers, but with a long latency period. Recent studies have indicated that the pathogenesis of asbestos-induced cancers is due, in part, to chronic inflammation and oxidative tissue damage caused by persistent asbestos fibers. Whole grain flaxseed (FS) has known antioxidant, anti-inflammatory and cancer chemopreventive properties. Rationale: As a prelude to future chemoprevention studies, we tested the ability of oral FS and its lignan component, FLC which is enriched in the lignan secoisolariciresinol diglucoside (SDG), to prevent acute asbestos-induced inflammation and inflammatory cytokine release in MM-prone Nf2+/mut;Cdkn2a+/mut mice. Methods: Mice were given a single ip bolus of 400 mM of crocidolite asbestos. They were then placed on 10% FS or 10% FLC supplemented diets 1 day prior (-1) to or after (+1) asbestos instillation. All mice were evaluated 3 days after injection of asbestos for abdominal inflammation and proinflammatory cytokine release. The Nf2+/mut;Cdkn2a+/mut model was selected as it develops an accelerated form of MM when exposed to asbestos. Results: Using liquid chromatography and tandem mass spectrometry (LC/MS/MS), we showed that systemic levels (plasma) of flaxseed lignan metabolites (such as the mammalian lignans Enterolactone (EL) and Enterodiol (ED)) were comparable to those in other mouse models where FS was shown to be an effective chemopreventive agent. The numbers of macrophages (MF) and neutrophils (PMN) in peritoneal lavage fluid indicated that both FS and FLC blunted acute abdominal inflammation induced by asbestos. In addition, the levels of pro-inflammatory cytokines TNF alpha and IL-1 beta in lavage fluid were also decreased by the dietary agents. Conclusions: Our findings suggest that the known chemopreventive properties of FS and its lignan component appear to reduce short-term asbestos-induced inflammation and may thus prove to be a promising dietary agent in the chemoprevention of MM.

Funding: This study was supported in part by Penn's SRP Center Grant (P42 ES027320), “Asbestos fate, exposure, remediation and adverse health effects” from the National Institute of Environmental Health (NIEHS) and 1R03CA180548-02 (MCS).

Citation Format: Steven M. Albelda, Ralph A. Pietrofesa, Craig W. Menges, Joseph R. Testa, Melpo Christofidou-Solomidou. Flaxseed and its lignan component protect from asbestos-induced inflammation in mice. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5542. doi:10.1158/1538-7445.AM2015-5542
Abstract 5544: Attempts to synthesize 5,7-dihydroxy-3H-1,2-dithiolo[4,3-d]- pyrimidine, M-47, a potential new treatment for drug- resistant breast cancer
Authors: Marasco Canio J. , Dunn Joseph A. , Pera Paula J. , Macubbin Alexander E.
Published: August 2015

M-47, 5,7-Dihydroxy-3H-1,2-dithiolo[4,3-d]pyrimidine, represents a novel class of antineoplastic agents both in structure and apparent mechanism of action. The synthesis of M-47 has been shown to be problematic. Mass spectroscopy and HPLC suggest the presence of certain oxidative byproducts in minor quantities along with the desired analog. Repeated efforts to purify M-47 via crystallization, extraction, chemical modification, and various forms of chromatography have failed. Although enrichment of the sample in excess of 90% is possible, absolute purity has not been achieved. Despite the presence of minor impurities this analog was tested against a variety of cancer cell lines in vitro. Interestingly, M-47 displayed equipotent activity against certain breast cancer lines and their multidrug resistant progeny Furthermore, in vitro testing of various ratios of M-47 with its oxidative byproducts indicated that they possessed similar IC50's suggesting that cellular oxidation of M-47 might be a necessary step in its cytotoxic pathway. The profound activity of M-47, especially against drug-resistant tumor cells, warrants further investigation into the chemistry of these compounds, their biological activity and their possible mechanism of action.

Note: This abstract was not presented at the meeting.

Citation Format: Canio J. Marasco, Joseph A. Dunn, Paula J. Pera, Alexander E. Macubbin. Attempts to synthesize 5,7-dihydroxy-3H-1,2-dithiolo[4,3-d]- pyrimidine, M-47, a potential new treatment for drug- resistant breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5544. doi:10.1158/1538-7445.AM2015-5544
Abstract 5547: Anti-proliferative activity of hydnocarpin, a natural lignan, is associated with the inhibition of Wnt/β-catenin signaling pathway via axin turnover in colon cancer cells
Authors: KIM Won Kyung , LEE Min Ai , PARK Hyen Joo , HONG Ji-Young , KANG Sam Sik , LEE Sang Kook
Published: August 2015

Abnormal activation of the canonical Wnt/β-catenin pathway and up-regulation of the β-catenin/T-cell factor (TCF) response to transcriptional signaling play critical roles early in colorectal carcinogenesis. Therefore, the Wnt/β-catenin signaling is considered an attractive target for cancer chemotherapeutic of chemopreventive agents, natural compounds were evaluated for β-catenin-mediated transcriptional activity. Hydnocarpin (HC), a natural lignan from Lonicera japonica was identified as a promising candidate because it effectively inhibited β-catenin/TCF reporter gene (TOPflash) activity. HC also exhibited potent growth inhibition of SW480 human colon cancer cells. Further investigation of HC on the regulation of the Wnt/β-catenin signaling revealed that HC had minimal effect on GSKβ and CK1α, while the protein levels of both axin1 and axin2 were remarkably increased by HC treatment. In addition, the interaction of axin and β-catenin by HC was significantly increased as analyzed by immunoprecipitation assay. These events led to up-regulation of the β-catenin destruction complex formation and phosphorylation of β-catenin. To further explore the role of axin1 up-regulation by HC in the mechanism of action of Wnt inhibitory effects on SW480 cells, siRNA experiments were conducted. The growth inhibitory effects of HC on SW480 cells were abolished by axin1 siRNA, implying that HC inhibited the Wnt/β-catenin signaling in an axin-dependent fashion. These finding suggests that HC might be a novel Wnt inhibitor and has a potential of signaling regulator in β-catenin-mediated signaling pathways.

Citation Format: Won Kyung KIM, Min Ai LEE, Hyen Joo PARK, Ji-Young HONG, Sam Sik KANG, Sang Kook LEE. Anti-proliferative activity of hydnocarpin, a natural lignan, is associated with the inhibition of Wnt/β-catenin signaling pathway via axin turnover in colon cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5547. doi:10.1158/1538-7445.AM2015-5547
Abstract 555: A workflow for enrichment and whole genome amplification (WGA) of circulating tumor cells (CTCs) for next generation sequencing
Authors: Nisic Merisa , Hao Sijie , Harouaka Ramdane , Zheng Si-Yang
Published: August 2015

Metastasis is the leading cause of cancer deaths. Circulating tumor cells (CTCs) are thought to be responsible for the seeding of metastases. These tumor cells are heterogeneous with variation occurring at the single-cell level. Using a flexible micro spring array (FMSA) device for label-free mechanical enrichment of CTCs from whole blood samples, we established a workflow from isolation followed by whole genome amplification (WGA) and finally sequencing of circulating tumor cells. Briefly, circulating tumor cells were collected from the FMSA using the Zeiss Laser Capture Microdissection System. Isolated cells underwent whole genome amplification using a commercial WGA kit. The quality of the amplified genomic DNA was analyzed using the TaqMan® Real-Time PCR Assays. Specific loci were then amplified with the Ion AmpliSeq™ Cancer Hotspot Panel v2 and sequenced using the Ion PGM™ System. These studies were performed to understand the genomic heterogeneity observed in circulating tumor cells.

Citation Format: Merisa Nisic, Sijie Hao, Ramdane Harouaka, Si-Yang Zheng. A workflow for enrichment and whole genome amplification (WGA) of circulating tumor cells (CTCs) for next generation sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 555. doi:10.1158/1538-7445.AM2015-555
Abstract 5552: Medicinal plants show anticancer splicing activity of apoptotic genes
Authors: Dlamini Zodwa , Bates David
Published: August 2015

Oncogenesis is associated with splicing switches and a disruption of splicing regulation being specific for cancer formation and tumour progression. An association between differential expression of splicing isoforms and tumour progression has been shown for several proteins, such as MDM2 and survivin. Currently the analysis of cancer-specific alternative splicing is a promising step forward in basic and translational molecular biology. Traditional medicine has a long history and is still the major source of medicine in developing countries. Approximately 70% of the South African population consults traditional healers, perpetuating the need for scientific appraisal of traditional medicine as a means to establish its efficiency and safety Also, pharmacological and phytochemical insights into several plants have led to the discovery of novel chemicals and therefore novel drugs. Alternatively, such novel chemical structures can serve as lead compounds/templates for the design of new drugs. The aim was to ascertain if the South African medicinal plants have anticancer splicing activity. 10c cells were treated with Tulbhaghia violacea and Cotyledon orbiculata, followed by mRNA extraction and RT-PCR. The results showed that Tulbhaghia violacea and Cotyledon orbiculata extracts have anti-cancer splicing activity on the BCLX and the AXL apoptosis genes. Additionally Cotyledon orbiculata extract has an anticancer splicing activity of the angiogenesis gene VEGF165. VEGF Elisa also confirmed the VEGF165 VEGF165b splicing switch. We have shown that South African medicinal plants have anti-cancer splicing activity. The anti-cancer splicing activity of the VEGF gene by Cotyledon orbiculata could mean that this extract has both angiostatic and apoptotic anti-cancer activities. We are continuing to screen more medicinal plants and will select those extracts with anti-cancer splicing activity for further studies. These further studies should identify numerous splicing pathways and completely elucidate the splicing target compounds that may serve as novel anti-cancer drugs or lead compounds.

Note: This abstract was not presented at the meeting.

Citation Format: Zodwa Dlamini, David Bates. Medicinal plants show anticancer splicing activity of apoptotic genes. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5552. doi:10.1158/1538-7445.AM2015-5552
Abstract 5562: BreastDefend enhances the effects of tamoxifen in estrogen receptor-positive human breast cancer in vitro and animal models in vivo
Authors: Cheng Shujie , Alvarado Mark , Loganathan Jagadish , Castillo Victor , Sandusky Georg , Eliaz Isaac , Sliva Daniel
Published: August 2015

In this study we evaluated the combined effects of the natural product/dietary supplement BreastDefend (BD) and tamoxifen (TAM) on MCF-7 estrogen receptor-positive (ER+) human breast cancer cells in vitro and an orthotopic mouse model in vivo. BD has been studied previously in preclinical models, and shown to exert significant effects on MDA-MB-231 triple-negative breast cancer cells. The proliferation of MCF-7 cells was evaluated using MTT assays, whereas the expression of genes involved in the molecular mechanisms of cancer was evaluated using DNA microarrays. MCF-7 cells were stimulated with estradiol (10 nM) and treated with TAM (1 μM), BD (10 μg/ml), and the combination of TAM and BD for 3 days and 6 days. Compared with control, BD enhanced the anti-proliferative activity of TAM significantly (P < 0.05), and the combination of BD and TAM induced apoptosis. The results of DNA microarray analyses suggested that the pro-apoptotic activity of BD and TAM in MCF-7 cells was associated with the significant upregulation of BRAF (an oncogene that can block proliferation and induce apoptosis) and downregulation of BCL2 (an apoptosis inhibitor) and FN1 (fibronectin, a glycoprotein involved in tumorigenesis and metastasis) expression. In the orthotopic model MCF-7 cells were implanted into the mammary fat pads of female ovariectomized nude mice (n = 12-13). The mice received subcutaneous pellets that released estradiol and TAM, whereas BD was administered orally via intragastric gavage. All groups, including control, received estradiol. The combination of BD and TAM suppressed tumor size in vivo markedly compared with the BD and TAM alone groups. The inhibition of tumor growth was associated with the induction of the cell cycle inhibitor protein p21 in MCF-7-generated breast tumors. In summary, this study suggests that BD could be considered for use in clinical studies as an adjuvant therapy in conjunction with TAM for the treatment of ER+ human breast cancer.

Citation Format: Shujie Cheng, Mark Alvarado, Jagadish Loganathan, Victor Castillo, Georg Sandusky, Isaac Eliaz, Daniel Sliva. BreastDefend enhances the effects of tamoxifen in estrogen receptor-positive human breast cancer in vitro and animal models in vivo. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5562. doi:10.1158/1538-7445.AM2015-5562
Abstract 5565: Antitumor activities of psammaplin A analogues in human lung cancer cells
Authors: Shin Yoonho , Hong Suckchang , Shin Jongheon , Park Hyeung-geun , Lee Sang Kook
Published: August 2015

Histone deacetylase inhibitors (HDACi) are considered to be potential candicates for cancer therapeutic agents. HDACi suppresses the progression of tumorigenesis through epigenetic regulation of target protein acetylation. Psammaplin A (PsA), originated from a marine product Pseudoceratina purpurea, also showed a potential HDAC inhibition. Based on the potential anticancer activity of PsA, various PsA analogs were designed and synthesized to explore novel candidates in the development of anticancer agents. Among compounds tested, a β-napthyl-derivertive of PsA (compound 30) exhibited a strong growth-inhibitory activity against human lung cancer cells and the activity was comparable to PsA. Compound 30 also effectively inhibited HDAC and up-regulated the level of ac-Histone3, p-AKT, p-ERK in human A549 lung cancer cells. In in vivo xenograft animal model implanted A549 cells, the administration of compound 30 intraperitoneally for 35 days exhibited a potential growth inhibitory activity of the tumor (47.6% inhibition at 30 mg/kg). The immunohistochemical analysis of tumor tissues also revealed the inhibition of the cell proliferation biomarker Ki-67 expression by compound 30. Taken together, these data suggest that a β-napthyl analog of PsA has the potential antitumor activity and thus to be prioritized in the development of cancer chemotherapeutic agents.

Citation Format: Yoonho Shin, Suckchang Hong, Jongheon Shin, Hyeung-geun Park, Sang Kook Lee. Antitumor activities of psammaplin A analogues in human lung cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5565. doi:10.1158/1538-7445.AM2015-5565
Abstract 5568: Inhibitory Effect of Partial Electron Decoupling Agents as an Alternate Therapy for Cancer
Authors: Bagasra Omar , Heslop Kareem , Livingston Krystal , Corbett Keneisha , Ejiawoko Amarachi O. , Sealey Leanna
Published: August 2015

Cancer therapies are generally non-specific for cancer cells and affect all cells in the body, resulting in severe side effects. We are working on an alternate method to present and treat cancer. Hypothesis: All replicating cells require energy in the forms of ATP and row material to reproduce. Cancer in a human body generally is the most energy consuming foci; therefore, it will be logical to down regulate the highest energy output pathways that do not harm the human being treated for a cancer. Two of the most prevalent cancers in are adenocarcinomas of prostate and breast. We hypothesized that if we treat cancer cells to a partial electron transport inhibitor that results in reduction in total ATP output it may significantly reduce cancer cell proliferation without interfering with normal functioning of other physiologic and immune functions. We chose Amygdalin (D-mandelonitrile-β-gentiobioside) which is found in seeds of prunasin family plants, such as peaches, apricots, almonds, apples, and other rosaceous plants and have been used as a traditional alternate medicine for treating terminal cancers and other illnesses. We evaluated the optimal dose that can inhibit various breast and prostate cancer cell lines as well two stem cell lines derived from neuroblastomas. We determine that cancer cell line and stem cell line proliferation rates were significantly inhibited 0.9 μM concentration of Amygdalin. This level of Amygdalin can be achieved by consuming 4-6 bitter almond and 2-3 apricot seed daily. In addition, we observed that Amygdalin induced apoptosis in the prostate and breast cancer cell lines.

Citation Format: Omar Bagasra, Kareem Heslop, Krystal Livingston, Keneisha Corbett, Amarachi O. Ejiawoko, Leanna Sealey. Inhibitory Effect of Partial Electron Decoupling Agents as an Alternate Therapy for Cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5568. doi:10.1158/1538-7445.AM2015-5568
Abstract 5572: Tracking the functional analysis of cancer risk variants
Authors: Tingle Sharna , Carrick Danielle , Schully Sheri , Clyne Mindy , Nelson Stefanie A.
Published: August 2015

Genome-wide association studies (GWAS) have identified thousands of alleles associated with cancer risk, but understanding the functional effects of these variants has proceeded more slowly. In addition, although catalogs of variants exist, tracking progress to determine variant function is challenging. We conducted a literature search and review to assess the trajectory of functional characterization of GWAS results and describe the types of analysis conducted and tools and databases used. We used the PubTator Web-based tool, which uses advanced text-mining techniques to accelerate manual literature curation, to search the literature for articles describing functional annotation of cancer GWAS hits. We generated a list of 646 unique SNPs from the Cancer Genomics and Epidemiology Navigator (CGEN) and imported the list to PubTator. We identified a total of 292 articles that included both variant discovery by GWAS and functional analysis, and reviewed the articles for information including cancer type, variant functional class (e.g., intron, exon, intergenic, etc.), and type of functional analysis conducted. These articles included 40 different cancer types or subtypes. The majority of cancer risk variants were intronic or intergenic, with only a small number located in coding regions. Most of the functional analyses were focused on the potential modification/disruption of regulatory processes by the risk allele (e.g., alteration of transcription factor binding sites, putative expressed quantitative trait loci), in line with current hypotheses regarding cancer risk variant function. Most studies also described effects of variants at the molecular, or cellular, rather than organismal level. We also reviewed the literature pertaining to variants identified for three cancers with possible pleiotropy - ovarian, breast, and prostate - to understand the trajectory of functional analyses beyond the initial discovery of the variants. We describe our use of the PubTator tool and provide a literature-based overview of progress in understanding the function of cancer risk variants identified via GWAS.

Citation Format: Sharna Tingle, Danielle Carrick, Sheri Schully, Mindy Clyne, Stefanie A. Nelson. Tracking the functional analysis of cancer risk variants. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5572. doi:10.1158/1538-7445.AM2015-5572
Abstract 558: Correlations of FGFR1 with ligand signaling in small-cell lung cancer
Authors: Yu Hui , Badzio Andrzej , Boyle Theresa , Lu Xian , Rivard Christopher J. , Kowalewski Ashley , Rikke Brad , Ellison Kim , Rozeboom Leslie , Hassan Biftu , Hirsch Fred R.
Published: August 2015

Introduction: Small cell lung cancer (SCLC) accounts for 15% of all lung cancers and has been under-studied relative to novel therapies. Signaling through fibroblast growth factors (FGF2, FGF9) and their high-affinity receptor (FGFR1) has recently emerged as a contributing factor in the pathogenesis and progression of non-small cell lung cancer (NSCLC). However, knowledge of FGFR1 and ligand expression in SCLC is limited.

Methods: FGFR1 gene copy number, mRNA levels and protein expression were determined by silver in situ hybridization (SISH), mRNA ISH and immunohistochemistry (IHC) in primary tumors from 78 SCLC patients. Expression of the ligands FGF2 and FGF9 mRNA and protein were determined by mRNA ISH and IHC, respectively. Spearman's correlations were conducted using SAS 9.4.

Results: FGFR1 protein expression by IHC demonstrated a significant correlation with FGFR1 gene copy number (p = 0.03) and FGFR1 mRNA levels (p<0.0001). The positivity of FGFR1 mRNA in the SCLC cohort was 21%. FGFR1 mRNA expression correlated with both FGF2 (p = 0.0001) and FGF9 (p = 0.0018) mRNA levels, as well as with FGF2 (0.0105) and FGF9 (0.0012) protein levels. There was no significant association between FGFR1 and ligands with clinical characteristics or prognosis.

Conclusions: FGFR1 amplification and mRNA levels correlate with FGFR1 protein expression. The SCLC cohort has a substantially higher proportion of FGFR1 mRNA positive cases (21%) compared to FGFR1 gene copy number positive cases (7.8%). Combined analysis of FGFR1 and ligand expression may be valuable to determine which SCLC patients may respond to FGFR1 inhibitor therapy.

Key Words: small cell lung cancer; FGFR1; FGF2; FGF9

Citation Format: Hui Yu, Andrzej Badzio, Theresa Boyle, Xian Lu, Christopher J. Rivard, Ashley Kowalewski, Brad Rikke, Kim Ellison, Leslie Rozeboom, Biftu Hassan, Fred R. Hirsch. Correlations of FGFR1 with ligand signaling in small-cell lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 558. doi:10.1158/1538-7445.AM2015-558
Abstract 5585: Analysis of saliva samples from healthy adults in north central Florida for human papillomavirus infection
Authors: Hwang Michelle M. , Choi Hyun-Ji , Langaee Taimour Y. , Logan Henrietta L. , Raisch Kevin P.
Published: August 2015

Background: Prevalence rates of healthy adults shedding HPV in their saliva have varied depending on the socio-economical status of the subjects. We hypothesize that samples collected from high-poverty and low-education areas of North Central Florida will have a high prevalence of HPV positive subjects. From published studies, we would expect an infection rate of 2-10% with male and female rates being similar.

Methods: Through health fairs in North Central Florida, we consented and collected saliva from 148 healthy adults. Of the 148 study samples, 64% were between age 18-60 and 22% were above age 60. 32% of the saliva samples were from males and 68% were from females. Dividing the saliva samples by race, we had 41% white, 51% African American. DNA was isolated from the saliva samples and PCR was used to amplify HPV-specific DNA. Degenerate PCR primers were used for the PCR and sequencing the amplicons. All DNA samples were coded so the investigators do not know the identity of the samples until completion of all experiments.

Results: We optimized the degenerate primer sets, MY09/MY11 and GP5+/GP6+, for magnesium concentration and annealing temperature. In addition we optimized a nested PCR using the degenerate primers to increase the sensitivity of HPV detection. From our direct DNA sequence analysis, we identified 10/148 (7%) HPV-positive samples. HPV-types found include: 16, 12, 32, 38, 61, 83, 72, 90, and 110. The inclusion of the primer set GP5+/GP6+ with MY09/11 resulted in an increase of total HPV-positivity from 5% to 7% and the detection of high-risk HPV types 32 and 90. Of the HPV-positive saliva samples, 9% were from 18-60 year old healthy adults and 3% were from over 60-year old adults. Adult females had a similar infection rate as adult males, 7% vs. 6%. Upon comparing race, African Americans were slightly more likely to be infected than whites, 7% vs. 5%. Of the 10 adults with detectable virus in their saliva, 70% were of the high risk HPV type.

Conclusion: Our sample size was small but showed an overall infection rate of healthy adults to be similar to rates found in other studies. In our study, male and females had similar infection rates. The age 60 and younger group were found to be three times more likely to be infected than the older than age 60 group. We found no significant difference between race in our study.

Citation Format: Michelle M. Hwang, Hyun-Ji Choi, Taimour Y. Langaee, Henrietta L. Logan, Kevin P. Raisch. Analysis of saliva samples from healthy adults in north central Florida for human papillomavirus infection. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5585. doi:10.1158/1538-7445.AM2015-5585
Abstract 563: Evaluation of EGFR mutation analysis performed by Sanger sequencing versus real-time PCR (EGFR RGQ PCR Kit)
Authors: Dietmaier Wolfgang , Schardt Irene , Rümmele Petra , Geissinger Eva , Hartmann Arndt , Stöhr Robert
Published: August 2015

EGFR mutation analysis of exons 18, 19 and 21 is a standard analysis in routine molecular pathology for prediction of response of EGFR-tyrosine kinase inhibitors (EGFR-TKI) in non-small cell lung cancer (NSCLC). The aim of this study was to evaluate the mutation detection using two widely used mutation analysis systems. For this purpose we analyzed exons 18, 19 and 21 of EGFR in 718 NSCLC with both nested PCR followed by direct Sanger sequencing and the EGFR RGQ PCR Kit (Qiagen). 694/718 samples were amplifiable and 676 samples allowed interpretable results by both methods. We found mutations in 74/676 (10,1%) samples by Sanger sequencing and in 91/676 (13,0%) using the RGQ Kit. There were 10/676 (1,5%) cases showing mutations only by Sanger Sequencing but not by the RGQ Kit. These mutations comprised four common EGFR-TKI sensitivity mutations in Exon21 (p.L858R (n = 2) and p.L861Q (n = 2)) as well as a complex deletion-insertion in exon 19 (n = 1). The other 5 mutations detected only by Sanger sequencing were uncommon and very rare mutations located in exon 18 and 21 which cannot be detected by the RGQ Kit due to its primer design. Vice versa, we found mutations in 21/676 samples using the RGQ Kit but not with Sanger sequencing. 20 of these mutations were common EGFR-TKI sensitivity mutations in exon 19 (deletions or complex delins, n = 15) and in exon 21 (p.L858R, n = 5) as well as a “p.G719X” mutation (n = 1) in exon 18. According to these data which includes mutations only in exons 18, 19 and 21 but not in exon 20 the mutation detection rate is higher using the RGQ-Kit than using the Sanger sequencing method (13,0% vs. 10,1%) especially in detecting common EGFR mutations associated with EGFR-TKI sensitivity. However the RGQ Kit fails in detecting uncommon and rare EGFR mutations for that this Kit is not designed and may produce non-informative results in some cases when ΔCT values are out of valid ranges whereas nested PCR followed by Sanger sequencing may still work.

Citation Format: Wolfgang Dietmaier, Irene Schardt, Petra Rümmele, Eva Geissinger, Arndt Hartmann, Robert Stöhr. Evaluation of EGFR mutation analysis performed by Sanger sequencing versus real-time PCR (EGFR RGQ PCR Kit). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 563. doi:10.1158/1538-7445.AM2015-563
Abstract 566: ChemoPlex SRM assay predicts response to specific chemotherapeutic agents in NSCLC
Authors: An Eunkyung , Kim Tae-Jung , Monga Manish , Bengali Kathleen , Drilea Alexi , Reilly Joseph , Darfler Marlene , Burrows Jon , Hembrough Todd
Published: August 2015

Current cancer treatment regimens rely on the use of chemotherapy agents that inhibit DNA replication and repair machinery. Several proteins are involved in this mechanism and these proteins have been identified as predictive biomarkers of response for many chemotherapy agents. While it has been demonstrated that expression levels of these biomarkers can predict chemotherapy response, chemotherapy in both the adjuvant and metastatic settings for NSCLC are not based on biomarker analysis. We have developed a quantitative, multiplex, mass spectrometry based diagnostic assay that can identify multiple biomarkers associated with chemotherapy benefit.

At the time of abstract submission, the expression levels of each biomarker were surveyed in 23 NSCLC patients, and the biomarker level for each chemotherapy agent (ERCC1 and XRCC1 for Platinum based chemotherapy; FR alpha and GART for pemetrexed; hENT1 and RRM1 for gemcitabine; and TUBB3 for taxane) was evaluated with clinical outcome. We are extending these analyses with a population of 100 NSCLC tumors with associated outcome.

Our pilot study shows a wide range of expression for each biomarker in patient biopsies. Among these biomarkers, FR alpha in NSCLC shows over 4 orders of magnitude in expression range. Also as expected, each biomarker level was found to be correlated with clinical outcome. For example, patients with high level of FR alpha showed improved response to pemetrexed; patients with lower level of TUBB3 showed improved response to taxane and patients with lower level of hENT1 showed poorer response to gemcitabine.

Our preliminary retrospective data analysis suggests that the ChemoPlex SRM assay can help predict response to specific chemotherapeutic agents. This test can be used to characterize individual tumor biology and identify actionable targets that can better inform patient care.

Citation Format: Eunkyung An, Tae-Jung Kim, Manish Monga, Kathleen Bengali, Alexi Drilea, Joseph Reilly, Marlene Darfler, Jon Burrows, Todd Hembrough. ChemoPlex SRM assay predicts response to specific chemotherapeutic agents in NSCLC. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 566. doi:10.1158/1538-7445.AM2015-566
Abstract 570: PD-L1 expression in paired non-small cell lung cancer tumor samples
Authors: Kim Jhingook , Sorensen Steffen Filskov , Choi Yoon-La , Wang Zhen (Adelle) , Sun Jong-Mu , Choi Hyejoo , Georgsen Jeanette Baehr , Dolled-Filhart Marisa , Emancipator Kenneth , Wu Dianna Y. , Meldgaard Peter , Zhou Wei , Hager Henrik
Published: August 2015

Background There is limited information on changes over time in programmed death ligand 1 (PD-L1) expression in non-small cell lung cancer (NSCLC).

Methods PD-L1 expression was assessed in paired tumor samples collected at different time points from the same patient using a prototype immunohistochemistry (IHC) assay with the 22C3 antibody. PD-L1 strong and weak positivity were defined to be traceable to the 1% and 50% cutoffs used in the clinical trial version of the assay. Concordance of PD-L1 expression between samples was analyzed by treating the IHC proportional score as a continuous and categorical variable.

Results 90 patients were included (73 from Korea, 17 from Denmark). 75% were male, 37% had squamous cell carcinoma, and 83% had stage I-IIIA disease. 97% of first samples and 53% of second samples were from lung. 89% and 63%, respectively, were surgical samples, with a median interval of 20.9 months (range, 0.1-94.0) between collection dates; 91% of paired samples were collected >3 months apart. A significant correlation of the PD-L1 IHC proportional score was observed for the paired samples (Pearson correlation coefficient, 0.62; P < 0.001). At the second time point, scores were identical in 39% of paired samples, higher in 32%, and lower in 29%. Of note, 12% of PD-L1-negative cases became positive and 20% of PD-L1-positive cases become negative at the second time point. The concordance rate was 56% (95% CI, 46%-67%) when PD-L1 expression was categorized as strong or weak positive or negative (Table).

Conclusion Although a significant correlation of the PD-L1 IHC proportional score was observed, this preliminary analysis suggested a discordance rate of 44% between initial and subsequent NSCLC tumor samples when patients were categorized by PD-L1 expression level. Similar results were observed when analyzing paired melanoma samples. This discordance should be considered if PD-L1 expression is used for selection of enrollment in clinical trials.

Table. Concordance of PD-L1 Expression in Paired NSCLC Samples (N = 90)

Later Time PointPD-L1 ExpressionStrong positive, n (%)Weak positive, n (%)Negative, n (%)Earlier Time PointStrong positive3 (3)3 (3)1 (1)Weak positive7 (8)20 (22)17 (19)Negative011 (12)28 (31)

Citation Format: Jhingook Kim, Steffen Filskov Sorensen, Yoon-La Choi, Zhen (Adelle) Wang, Jong-Mu Sun, Hyejoo Choi, Jeanette Baehr Georgsen, Marisa Dolled-Filhart, Kenneth Emancipator, Dianna Y. Wu, Peter Meldgaard, Wei Zhou, Henrik Hager. PD-L1 expression in paired non-small cell lung cancer tumor samples. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 570. doi:10.1158/1538-7445.AM2015-570