bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3295: Microbiome profiling and immunological characterization on gastrointestinal tract tumors
Authors: Zhang Chao , Cleveland Kyle , Schnoll-Sussman Felice , McClure Bridget , Bigg Michelle , Schultz Nikolaus , Shah Manish A. , Betel Doron 
Published: July 2016

Abstract
Motivation: Conservative estimates are that around 20% of all malignancies may be attributed to microbiota. Especially in the gastrointestinal (GI) tract, infection can cause chronic inflammation and further influence immune response to create a proinflammatory microenvironment for cancer development. This can account in part for the reason that the immunotherapy in colon cancer is less efficacious than in non-GI tract cancers. We proposed a systematic computational study to profile the microbiome from next generation sequencing of gastric biopsy samples, and evaluate the relationships among microbiome composition, host immune response and genomic characterization.
Method: Patients undergoing upper endoscopy without chronic inflammatory disease or chronic NSAID use were eligible for participation, including cancer/non-cancer, Helicobacter pylori positive and negative patients. In total, 62 endoscopic biopsies from 3 locations in the gastric antrum/body were freshly frozen for whole genome sequencing and transcriptome analysis. In addition, TCGA whole genome sequencing (WGS) data from 37 gastric cancer, 102 colon cancer, and 46 rectal cancer samples were analyzed for microbiome composition.
Results: H. pylori was detected in all positive infection gastric biopsy samples by our pipeline, and the results were successfully validated by qPCR. Comparing to other traditional bacterial identification methods, our pipeline shows a strong capacity for profiling microbiome from low bacterial content biopsy samples. We observed a high positive correlation between the H. pylori content and total non-H. pylori bacteria content. And we also identified significantly differentially expressed immune-related genes between active H. pylori infection and negative samples. Finally, we analyzed collected TCGA WGS data, and identified the dominant species from the different cancer studies, such as Bacteroides fragilis in rectum adenocarcinoma, and Fusobacterium nucleatum in colon adenocarcinoma. The correlation among microbiome composition, the expression of immune-related genes and cancer genomic characterization were also demonstrated in this study.
Citation Format: Chao Zhang, Kyle Cleveland, Felice Schnoll-Sussman, Bridget McClure, Michelle Bigg, Nikolaus Schultz, Manish A. Shah, Doron Betel. Microbiome profiling and immunological characterization on gastrointestinal tract tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3295.
Abstract 33: NSCLC cells internalize ATP in vitro and in vivo using multiple endocytotic pathways
Authors: Qian Yanrong , Wang Xuan , Li Yunsheng , Cao Yanyang , Chen Xiaozhuo 
Published: July 2016

Abstract
Upregulated glycolysis in cancer, the Warburg effect, has been studied for almost one century. However, the biological reasons for the upregulation are still debated and the effect is far from fully understood. One of the major controversies related to the effect is the role of ATP synthesis. Intratumoral ATP concentrations are found to be 10⁁3-10⁁4 times higher than those in normal tissues of the same cell origin. However, whether and how cancer cells use the abundant extracellular ATP was unknown until we recently reported that cancer cells internalize ATP by macropinocytosis (1). Extracellular ATP was found to increase intracellular ATP concentration and promote cell proliferation and drug resistance in cancer cells (1). Here we report that, using a nonhydrolyzable fluorescent ATP (NHF-ATP) as an ATP surrogate, high and low molecular weight dextrans as endocytosis tracers and fluorescence microscopy detection, cultured human NSCLC A549 and H1299 cells as well as xenografted A549 tumors were found to internalize ATP at concentrations within the reported intratumoral ATP concentrations. Endocytosis inhibitor studies show that in addition to macropinocytosis, clathrin- and caveolae-mediated endocytoses contribute to the ATP internalization, which led to a ∼30% increase in intracellular ATP level after 45 min of ATP incubation. This increase cannot be accounted for by purinergic receptor signaling or increased intracellular ATP synthesis rates. Internalization of NHF-ATP and dextran exhibits time-dependent profiles parallel to the extracellular ATP-induced intracellular ATP level elevation. All these demonstrate the observed ATP increase is a result of internalization of extracellular ATP and NHF-ATP is a powerful tool for studying ATP internalization. These findings significantly deepen our understanding of the Warburg effect by showing how cancer cells in tumors take up intratumoral ATP and perform previously unrecognized biological functions. It strongly suggests ATP-sharing among cancer and stromal cells and identifies multiple novel anticancer targets (2).
1. Qian Y, Wang X, et al., Cancer Lett. 351(2014): 242-5.
2. Chen X, Qian Y, Wu S. Free. Radic. Biol. Med. 79(2015): 253-263.
Citation Format: Yanrong Qian, Xuan Wang, Yunsheng Li, Yanyang Cao, Xiaozhuo Chen. NSCLC cells internalize ATP in vitro and in vivo using multiple endocytotic pathways. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 33.
Abstract 3312: Transcriptional regulation of chemokine receptor 4 (CXCR4) by nuclear respiratory factor 1 (NRF1) controls estrogen-induced malignant transformation of breast epithelial cells to breast cancer stem cells
Authors: Das Jayanta Kumar , Roy Deodutta 
Published: July 2016

Abstract
Chemokine receptor 4 (CXCR4) is involved in the maintenance normal stem cells and metastasis of cancer stem cells. NRF1 is one of the major transcription factor that controls transcription of CXCR4. The functional regulation of transcription of NRF1 target genes has not been explored in breast cancer. In this study, we have investigated whether transcriptional regulation of CXCR4 by NRF1 regulates estrogen-induced malignant transformation of breast epithelial cells to breast cancer stem cells (CSCs). We have previously shown that NRF1 may be involved in 17 β-estradiol (E2) induced malignant transformation of breast epithelial cells, however, the mechanism of transcriptional regulation of the NRF1 target genes remains unknown. In this study we showed that NRF1 and E2 jointly contributed in reprogramming of breast epithelial cells to CSCs. Levels of breast cancer stem cell markers (CD44+CD24+ALDH1+CD133+) were significantly increased by E2 treatment in NRF1 overexpressing cells compared to cells transfected with vector receiving E2. E2-induced increases of spheroid formation, cell survival and growth of cancer stem cells were modulated by functional gain or loss of NRF1. For morphological details we used a holographic microscope (HoloMonitor M4), we observed that E2 generated breast cancer stem cells with CD44+CD24+ALDH1+CD133+ markers produced large live spheroids in NRF1 overexpressing cells compared to cells transfected with vector receiving E2. Overexpression of NRF1 promoted the transition of E2-treated breast epithelial MCF-10A cells to mesenchymal stem cell-like phenotype. The ChIP qPCR, RT-qPCR, Western blotting and immunofluorescence microscopic assays showed that NRF1 mediated transcriptional changes of its target genes CXCR4, BNIP3, and DJ-1 correlated with malignant phenotypic changes. The suppression of NRF-1 activity induced growth arrest and apoptosis, and reduced stemness, growth and tumorigenic property of cancer stem cells. In summary, our findings for the first time showed that transcriptional regulation of CXCR4 by NRF1 may contribute in the induction of a pre-malignant phenotype by estrogen presumably by promoting generation of breast cancer stem cells. These data suggest NRF1 as an emerging potential target for therapeutic intervention against breast cancer.
Citation Format: Jayanta Kumar Das, Deodutta Roy. Transcriptional regulation of chemokine receptor 4 (CXCR4) by nuclear respiratory factor 1 (NRF1) controls estrogen-induced malignant transformation of breast epithelial cells to breast cancer stem cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3312.
Abstract 3332: Targeting cancer stem cells in ovarian cancer
Authors: Kakar Sham S. , Ratajczak Mariusz Z. 
Published: July 2016

Abstract
Ovarian cancer is a disease commonly complicated by the presence of ascites in the abdominal cavity which represents a major clinical problem. Currently, the treatment for ovarian cancer entails cytoreductive surgery followed by chemotherapy, mainly cisplatin or carboplatin combined with paclitaxel. Although this regimen is initially effective in a high percentage of cases, unfortunately within few months of initial treatment, tumor relapse occurs because of platinum-resistance. This is attributed mainly due to the presence of cancer stem cells (CSCs) present in ascites which are chemo-resistant and responsible for recurrence of cancer. In our preliminary studies, we show for the first time that withaferin A (WFA), a bioactive compound isolated from the plant Withania somnifera targets putative CSCs. Treatment of nude mice bearing orthotopic ovarian tumors with WFA resulted in a 70% to 80% reduction in tumor growth, complete inhibition of metastasis, and a significant elimination of the cells expressing CSCs markers - CD44, CD24, CD34, CD117, ALDH1 and Oct4 and down regulation of Notch1 and its downstream signaling genes (Hes1 and Hey1) reported to play crucial role for self-renewal and maintenance of CSCs. In addition, WFA also resulted in a significant inhibition of securin expression an “oncogene” important in ovarian tumorigenesis. In contrast, treatment of mice with CIS alone had the opposite effects, causing an increase in cells expressing CSC markers and Notch1 signaling pathway. However, combining of WFA with CIS showed enhanced effects on suppression of securin as well as elimination of CSCs. This may explain the development of platinum-resistance and recurrence of cancer in patients treated with first line chemotherapy. Based on this information, we hypothesize that a combination of WFA with CIS should target ovarian cancer cells as well as cancer stem cells by reducing securin and CSCs thereby reducing CIS resistance and recurrence of ovarian cancer. In our study, we used cells from ascites from patients with recurrent ovarian cancer and determine the effect of WFA alone and in combination with CIS on tumorigenic functions of ascites cells and CSCs from ascites in vitro and in vivo, and defined the molecular mechanisms associated with self-renewal, maintenance and recurrence of cancer in relation to expression of securin. Because the two agents act synergistically, this combination may minimize side effects as well as induction of cisplatin resistance and recurrence of cancer. In addition, this study will justify the initiation of phase I/phase II clinical trials to assess potential toxicity and efficacy of WFA/CIS combination in advanced/recurrent ovarian cancer patients.
Citation Format: Sham S. Kakar, Mariusz Z. Ratajczak. Targeting cancer stem cells in ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3332.
Abstract 3375: Activation of the acid sphingomyelinase pathway as a potential mechanism of chemotherapy-induced normal tissue vasculature damage
Authors: Mizrachi Aviram 
Published: July 2016

Abstract
Background: Several classes of chemotherapy confer substantial risk for late-term vascular morbidity. We aimed to investigate the mechanisms of chemotherapy-induced vascular toxicity in normal tissues.
Methods: The effect of Doxorubicin and Cisplatin has been studied in-vitro using bovine aortic endothelial cells (BAEC) as a model of normal tissue vascular endothelium. Specifically, we investigated activation of the acid sphingomyelinase (ASMase) pathway, which may lead to endothelial dysfunction (vasoconstriction) and apoptosis following incubation with different concentrations of Doxorubicin and Cisplatin. ASMase activity was measured using radioenzymatic assay with [N-methyl-14C] sphingomyelin. Cellular Reactive Oxygen Species (ROS) production was assessed using the 2’,7’-dichlorofluorescin diacetate (DCFDA) detection assay kit (Abcam®). Endothelial apoptosis was quantified using the 3,5-bis-benzamide assay (Sigma-Aldrich®). For the in-vivo studies, mouse femoral arterial blood flow was measured in real-time during and after Doxorubicin (8 mg/kg intravenously) or Cisplatin (8 mg/kg) administration. Visualization of ovarian and femoral microvasculature was obtained by fluorescence optical imaging system, equipped with a confocal fiber microscope (Cell-viZio).
Results: Incubation of BAEC with Doxorubicin (0.25 μM or 0.50 μM) resulted in increased ASMase activity, production of ROS and induction of apoptosis in a dose-dependent fashion. Cisplatin (25 μM or 50 μM) generated significantly reduced effects in BAEC yet apoptosis and ROS were notable. In-vivo endoscopic fibered confocal microscopy following chemotherapy administration revealed a differential pattern between the two agents. While post Doxorubicin injection, there was a marked reduction in fluorescent signal in small vessels (&lt;15 μM) indicating vessel constriction and disintegration of vessel's wall in larger vessels, Cisplatin treatment resulted in very minor vascular changes.
Conclusions: These findings suggest that chemotherapy-induced vascular damage in normal tissues may represent different modes of toxicity in vivo and in vitro, while is mediated via activation of the ASMase pathway in response to Doxorubicin and to a lesser extent in response to Cisplatin.
Citation Format: Aviram Mizrachi. Activation of the acid sphingomyelinase pathway as a potential mechanism of chemotherapy-induced normal tissue vasculature damage. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3375.
Abstract 3378: Using multiplex immunoassays for profiling the circulating and tissue angiogenic biomarkers in mouse and human cancer
Authors: Lie Wen-Rong , Mistry Jehangir 
Published: July 2016

Abstract
Monitoring the expression levels of multiple pro- and anti-angiogenic factors in circulation and tumor microenvironment is crucial in examining the role of angiogenesis in cancer pathological processes or in identifying potential biomarkers for oncology drug development. The Luminex xMAP technology allows simultaneous detection of multiple proteins in a single sample and has been used widely in cancer biomarker discovery. Using the Luminex xMAP technology, we developed two Human Angiogenesis Panels (17-plex and 20-plex) and a Mouse Angiogenesis Panel (27-plex) for the simultaneous detection of multiple angiogenic biomarkers in less than 50 μL of human or mouse samples. We report here the protein biomarker profiles of these 37 human angiogenic factors in 1) serum samples, collected from breast cancer and colorectal cancer patients and normal donors, 2) tumor extracts, collected from breast tumor and colorectal tumor extracts and their matched adjacent normal tissue, 3) conditioned media in cultured tumor cells and stromal cells, and in addition, the protein biomarker profiles of 27 mouse angiogenic factors in the conditioned media of various mouse tumor cells and stromal cells. For example, of the 37 human angiogenesis serum protein markers tested, 16 biomarkers [angiopoietin-2 (ANGPT-2), EGF, endoglin, follistatin, G-CSF, HGF, PLGF, sAXL, PDGF-AB/BB, osteopontin (OPN), sHGFR/c-Met, suPAR, sTie-2, sHer2, soluble neuropilin-1 (sNRP-1), and tenascin-C] showed significant difference in expression levels between normal and breast cancer serum samples (p&lt;0.05) and 13 biomarkers [ANGPT-2, EGF, PLGF, sAXL, sHer3, OPN, thrombospondin-2 (TSP-2), sHGFR/c-Met, suPAR, sTie-2, sNRP-1, sIL-6R, and tenascin-C] showed differential expression levels between normal and colorectal cancer serum samples (p&lt;0.05). Altogether, the results of this study demonstrate the utility of these human and mouse angiogenesis multiplex panels as a useful tool in studying circulating and tissue protein biomarker profiles in cancer research.
Citation Format: Wen-Rong Lie, Jehangir Mistry. Using multiplex immunoassays for profiling the circulating and tissue angiogenic biomarkers in mouse and human cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3378.
Abstract 3380: Direct contact with perivascular tumor cells enhances integrin αvβ3 signaling and migration of endothelial cells
Authors: Burgett Monica , Lathia Justin , Roth Patrick , Nowacki Amy , Pugacheva Elena , Huang Ping , Vasanji Amit , Meizhang Li , Byzova Tatiana , Mikkelsen Tom , Bao Shideng , Rich Jeremy , Weller Michael , Gladson Candece 
Published: July 2016

Abstract
The secretion of soluble pro-angiogenic factors by tumor cells and stromal cells in the perivascular niche promotes the aggressive angiogenesis that is typical of glioblastoma (GBM), which requires endothelial cell (EC) migration. Here, we show that angiogenesis can also be promoted by a direct interaction between brain tumor cells, including tumor cells with cancer stem-like properties (CSCs), and ECs. The direct interaction between CSCs and ECs was mediated by binding of integrin αvβ3 expressed on ECs to the RGD-peptide in L1CAM expressed on CSCs. Using in vitro assays, we found that this interaction increased network formation and migration-associated signaling events in ECs, including activation of integrin αvβ3, FAK, bone marrow tyrosine kinase on chromosome X (BMX), p130CAS, ERK and JNK. Comparison of the effects of co-culturing CSCs with ECs versus the effects of conditioned media from CSCs co-cultured with ECs indicated that upstream and downstream effector activation was not attributed to a secreted factor(s). Activation of αvβ3 and BMX was required for migration stimulated by direct binding but not for migration stimulated by soluble factors. RGD-peptide treatment of mice with established intracerebral GBM xenografts significantly reduced the percentage of Sox2-positive tumor cells (consistent with CSCs) in close proximity to ECs, decreased integrin αvβ3 and BMX activation and p130CAS phosphorylation in the ECs, and reduced the vessel surface area. These results show that direct interactions between CSCs and ECs have potent effects on EC migration and reveal a previously unrecognized aspect of the regulation of angiogenesis in GBM that can impact therapeutic anti-angiogenic targeting.
Citation Format: Monica Burgett, Justin Lathia, Patrick Roth, Amy Nowacki, Elena Pugacheva, Ping Huang, Amit Vasanji, Li Meizhang, Tatiana Byzova, Tom Mikkelsen, Shideng Bao, Jeremy Rich, Michael Weller, Candece Gladson. Direct contact with perivascular tumor cells enhances integrin αvβ3 signaling and migration of endothelial cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3380.
Abstract 3383: Tumor vasculature normalization by recoupling nitric oxide synthase with a tetrahydrobiopterin precursor
Authors: Rabender Christopher , Bruno Ninu , Mikkelsen Ross B. 
Published: July 2016

Abstract
The abnormal, irregular nature of tumor vasculature has been well documented. The aberrant nature of the vasculature results in uneven, heterogeneous blood flow and leaky, hemorrhagic blood vessels. Due to the unusual nature of tumor vessels, areas of hypoxia tend to develop that contribute to radioresistance and inefficiency of therapeutic drug delivery. Our current studies examine the role of nitric oxide synthase (NOS) in tumor vasculature. NOS has been demonstrated to be “uncoupled” in tumor cells due to reduced levels of tetrahydrobiopterin (BH4), a necessary cofactor, resulting in peroxynitrite (ONOO-) formation in lieu of nitric oxide (NO). NO signaling is critical for vascular function and thus uncoupling of eNOS in the endothelial cells may partly explain the poor vasculature structure found within solid tumors. Having previously demonstrated that NOS can be “recoupled” and NO production restored through treatment of tumor cells with Sepiapterin (SP), a BH4 precursor, we examined whether SP could normalize tumor vasculature, promoting radiosensitivity and improving drug uptake. Multispectral optoacoustic tomography analysis of both flank tumor xenografts and a spontaneous breast tumor model (MMTV) demonstrate that SP given orally significantly enhances the percent of oxyhemoglobin in the tumor. Immunohistochemical analysis of tumors treated with SP showed a significant reduction in CD31 staining and a significant increase in smooth muscle actin (SMA), both hallmarks of vascular normalization. Ex vivo analysis of tumors revealed that the enhanced tumor oxygenation resulted in over a two-fold increase in radiation induced cell killing. These preliminary data demonstrate great potential for SP as an adjuvant in the treatment of cancer, especially when we take into consideration that SP has been demonstrated to be cytotoxic to both breast and colon tumors. Future studies will examine drug uptake in tumors as well as the mechanism behind the vascular normalization.
Citation Format: Christopher Rabender, Ninu Bruno, Ross B. Mikkelsen. Tumor vasculature normalization by recoupling nitric oxide synthase with a tetrahydrobiopterin precursor. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3383.