bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2208: Peptide-mediated 'miniprep' isolation of extracellular vesicles is suitable for high-throughput proteomics; method evaluation and application in colon cancer
Authors: Wit Meike De , Knol Jaco , Reus Inge de , Schelfhorst Tim Tim , Bishop-Currey Logan , Grieken Nicole van , Piersma Sander , Pham Thang V. , Fijneman Remond J. , Meijer Gerrit A. , Verheul Henk , Jimenez Connie R. 
Published: July 2017

Abstract
Objective:
Extracellular vesicles (EVs) are cell-secreted membrane vesicles enclosed by a lipid bilayer derived from endosomes or from the plasma membrane. Since they are released into body fluids, and their cargo includes tissue-specific and disease-related molecules, EVs represent a rich source for disease biomarkers. However, standard ultracentrifugation methods for EV isolation (UC-EV) are laborious, time-consuming, and require high inputs.
Method:
A recently described isolation method, which can be performed at small ‘miniprep’ scale, utilizes specific Heat Shock Protein (HSP)-binding peptides to aggregate HSP-decorated EVs (Ghosh et al. (2014), PLoS ONE 9:e110443). The authors showed comparable results for their method (abbreviated HSP-EV here) and UC-EV, but a detailed proteomic comparison was lacking. Therefore, we compared both methods using label-free proteomics of replicate EV isolations from HT-29 cancer cell-conditioned medium. Subsequently we applied this technique on secretomes of fresh human colorectal cancer (CRC) (n=17) and colon adenoma (n=4) tissue as well as patient-matched normal colon tissue.
Results:
Despite a 30-fold different input scale (UC-EV: 60 ml versus HSP-EV: 2 ml), both methods yielded comparable numbers of identified proteins (3115 versus 3085), with reproducible identifications (72.5% versus 75.5%) and spectral count-based quantification (average CV 31% versus 27%). EVs obtained by either method contained established EV markers and proteins linked to vesicle-related gene ontologies. In the EV fraction of the tissue secretomes 6390 proteins were identified, of which 471 proteins were significantly 5-fold more present in CRC samples than in normal tissue EVs. Gene ontology analysis revealed enrichment of nuclear proteins involved in DNA damage response, chromosome organization and RNA processing in the CRC EVs.
Conclusions:
The HSP-EV method provides an advantageous, simple and rapid approach for EV isolation from small amounts of biological samples, enabling high-throughput analysis in a biomarker discovery setting.
Citation Format: Meike De Wit, Jaco Knol, Inge de Reus, Tim Tim Schelfhorst, Logan Bishop-Currey, Nicole van Grieken, Sander Piersma, Thang V. Pham, Remond J. Fijneman, Gerrit A. Meijer, Henk Verheul, Connie R. Jimenez. Peptide-mediated 'miniprep' isolation of extracellular vesicles is suitable for high-throughput proteomics; method evaluation and application in colon 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 2208. doi:10.1158/1538-7445.AM2017-2208
Abstract 2210: Scavenging for lethal prostate cancer biomarkers in FFPE tissue
Authors: Ramberg Håkon , Braadland Peder R. , Stensland Maria E. , Rakaee Mehrdad , Nygård Ståle , Øgren Olov , Berge Viktor , Svindland Aud , Richardsson Elin , Souza Gustavo A. de , Andersen Sigve , Taskén Kristin A. 
Published: July 2017

Abstract
The aim of this study was to identify protein biomarkers that can be used to stratify indolent and aggressive prostate cancer using formalin-fixed paraffin-embedded tissue (FFPE) from radical prostatectomy specimens. The protein expression profiles of benign and malign samples from 23 patients, who had either died of prostate cancer or were alive 10 years after diagnosis, were determined using an Easy nLC1000 Nano-LC system connected to a quadrupole-Orbitrap (Q Exactive) mass spectrometer equipped with a nanoelectrospray ion source. The generated MS raw files were submitted to MaxQuant software for protein identification. Differential expression of LFQ-normalized intensities was tested by Cox proportional hazards modeling and LASSO (least absolute shrinkage and selection operator) regression analysis within the R software environment. The label free Orbitrap analysis identified 3523 unique proteins from the FFPE tissue samples. LASSO of protein expression profiles of indolent and aggressive prostate cancer tissue samples identified ten proteins which could stratify the groups. Seven of the proteins were significantly correlated with time to prostate cancer specific mortality in Cox proportional hazards modeling (p-value &lt; 0.05). The selected proteins were found to be linked to NFkappaB signaling, cell structure, translation and metabolic pathways. Finally, proteins capable of distinguishing benign from malignant prostate tissue were identified by LASSO, and 22 of these were significantly different between the groups (Bonferroni-adjusted p-value &lt; 0.05). Selected candidate biomarkers are currently being validated by immunohistochemical analyses of 535 radical prostatectomy specimens from two independent Norwegian cohorts with long follow-up time.
Citation Format: Håkon Ramberg, Peder R. Braadland, Maria E. Stensland, Mehrdad Rakaee, Ståle Nygård, Olov Øgren, Viktor Berge, Aud Svindland, Elin Richardsson, Gustavo A. de Souza, Sigve Andersen, Kristin A. Taskén. Scavenging for lethal prostate cancer biomarkers in FFPE tissue [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 2210. doi:10.1158/1538-7445.AM2017-2210
Abstract 2214: Adaptive dynamic artificial polyligand targeting (ADAPT): a method to identify exosomal proteins from a prostate cancer cell line
Authors: Hornung Tassilo , Logie Stephen , Bondre Aniket S. , Maher Varun , Richards Melissa N. , Zarkovic Jelena , Tinder Teresa T. , O'Neill Heather A. , Miglarese Mark R. , Spetzler David B. 
Published: July 2017

Abstract
In the recent years it was demonstrated that a multitude of body fluids contains substantial amounts of exosomes, extracellular vesicles with sizes ranging between 40 and 100 nm. Those vesicles have protein profiles characteristic of their cells of origin. It was shown that exosomes play a role in cell-to-cell communication making them attractive targets to identify early disease stage biomarkers. Cancer heterogeneity is known for a long time to be an important clinical determinant of patient outcome. We developed the highly multiplexed ADAPT platform to capture systems-based biological signatures that may reflect the molecular heterogeneity of various cancer types and help to improve diagnosis of the disease. Exosomes from two prostate cancer cell lines, VCaP and LNCaP, were used to train ssDNA libraries to discriminate them. A highly diverse library of 1012 oligonucleotides (ODNs) was subjected to five rounds of positive and negative selection against exosomes from VCaP and LNCaP prostate cancer cell lines. Individual ODNs that bound preferably to exosomes from VCaP cells were identified by NGS, resynthesized and binding of co-precipitated ODNs to VCaP exosomes was verified by qPCR. LC-MS/MS was used to identify binding partners of ODNs bound to VCaP exosomes. Several of those binding partners (CHMP1b/2a/4b, VPS28, Syntenin-1) were found to be part of the ESCRT machinery, which participates in exosomes biogenesis. It was found that those proteins are overexpressed in human cancers. In addition, we identified the chemokine I-TAC, which is overexpressed in blood and tissue of men with advanced prostate adenocarcinomas. Finally, we found hnRNP-1, a cancer associated splicing factor, and the cold shock proteins RNPL and A18 hnRNP. Knock-down of these cold shock proteins has been shown to enhance chemotherapeutic cell killing of prostate cells. ADAPT is an unbiased profiling platform that identifies cancer associated proteins expressed on exosomes. This platform can be deployed against multiple cancer types and offers broad potential applications in biomarker discovery.
Citation Format: Tassilo Hornung, Stephen Logie, Aniket S. Bondre, Varun Maher, Melissa N. Richards, Jelena Zarkovic, Teresa T. Tinder, Heather A. O'Neill, Mark R. Miglarese, David B. Spetzler. Adaptive dynamic artificial polyligand targeting (ADAPT): a method to identify exosomal proteins from a prostate cancer cell line [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 2214. doi:10.1158/1538-7445.AM2017-2214
Abstract 2225: Green tea polyphenols suppress tumor growth and invasion by targeting matrix metalloproteinases, RECK and TIMP-3, in a mouse model implanted with prostate tumors
Authors: Shankar Eswar , Bhaskaran Natarajan , Kanwal Rajnee , Gupta Sanjay 
Published: July 2017

Abstract
Green tea polyphenols (GTP) and its major constituent, epigallocatechin-3-gallate (EGCG) reactivate epigenetically silenced genes in cancer cells that reduces invasiveness and migration capabilities; however, the mechanisms whereby these effects occur are not well understood. RECK, a novel tumor suppressor and the tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) genes negatively regulates matrix metalloproteinases (MMPs) and inhibits tumor invasion, angiogenesis and metastasis. In the present study, we demonstrate that GTP mediate epigenetic induction of RECK and TIMP-3 thereby playing a key role in suppressing invasiveness and gelatinolytic activity of MMP-2 and MMP-9 in vivo in an orthotopic implantation model of human prostate cancer. Athymic nude mice were implanted with human prostate cancer LNCaP tumor in the ventral prostate for 2 weeks and later treated DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (AZA), histone deacetylase inhibitor, trichostatin A (TSA) and histone methyltransferase inhibitor, 3-Deazaneplanocin A (DZNep) individually at 0.1 mg/kg body weight intraperitoneally at alternate days/week; and combination of AZA+TSA and DZNep+TSA at similar doses and times; whereas GTP was provided peroral by gavage at 7.5 and 15.0 mg/kg body weight freshly prepared in 100µl PBS. Treatment of mice with GTP resulted in marked decrease in tumor growth and its local invasion in dose dependent manner, compared to treatment with epigenetic inhibitors and their combination after 8 weeks of intervention. GTP treatment significantly reduced serum levels of MMP-2, MMP-9 and VEGF, compared to treatment with epigenetic inhibitors alone. Combination of AZA+TSA exhibited similar effect which was equivalent to the lower dose of GTP treatment. Furthermore, GTP treatment significantly reduced EZH2 and H3K27me3 and class I HDAC protein levels in tumors. GTP partially reversed the hypermethylation status of RECK and TIMP3 gene and significantly enhanced their protein expression in the tumor tissue. Inhibition of VEGF, MMP-2 and MMP-9 levels were also noted after GTP treatment in tumor tissue in dose-dependent manner. Our findings suggest that induction of RECK and TIMP-3 are key epigenetic events modulated by GTP that results in suppression of matrix degradation and angiogenesis to delay prostate cancer invasion and its subsequent progression.
Citation Format: Eswar Shankar, Natarajan Bhaskaran, Rajnee Kanwal, Sanjay Gupta. Green tea polyphenols suppress tumor growth and invasion by targeting matrix metalloproteinases, RECK and TIMP-3, in a mouse model implanted with prostate tumors [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 2225. doi:10.1158/1538-7445.AM2017-2225
Abstract 2227: Fgf2 from visceral adipose tissue stimulates neoplastic transformation of nonmalignant epithelial cells
Authors: Chakraborty Debrup , Benham Vanessa , Demireva Elena Y. , Bullard Blair , Bernard Jamie J. 
Published: July 2017

Abstract
Background: Adiposity plays a crucial role in the pathogenesis and prognosis of different types of cancers. Epidemiological evidence suggests visceral adipose tissue (VAT) and high-fat diets (HFD) are associated with increased cancer risk however the mechanism is not understood. The aim of this study was to explore the factors in VAT that stimulate neoplastic transformation.
Methods: We modeled visceral adiposity-stimulated neoplastic transformation using our novel ex vivo system of VAT-condition medium stimulated epithelial cell transformation (measured by growth in soft agar) and our in vivo murine lipectomy model of ultraviolet light B (UVB)-induced, VAT promoted skin tumor formation. FgfR1(-/-) stable cells were generated by using CRISPR-Cas9 technology and were used to investigate the role of fibroblast growth factor-2 (FGF2) and FGFR1 dependent signaling in neoplastic transformation, both in vitro and in vivo.
Results: Only the VAT of obese mice fed a HFD [not VAT from low-fat diet (LFD) fed mice] stimulated neoplastic transformation of skin epithelial cells. Furthermore, human VAT stimulated both skin and mammary epithelial cell transformation. The differences in VAT activity between LFD and HFD fed mice and human donors were associated with the levels of FGF2. Circulating levels of FGF2 were associated with non-melanoma tumor formation in vivo. Human and mouse VAT failed to stimulate transformation in FgfR1(-/-) cells and do not form tumors when injected in Nude mice in vivo.
Conclusion: Collectively, our data show FGF2 released from VAT and its interaction with FGFR1 is a novel and potential direct path of VAT-enhanced tumorigenesis. Blocking the FGFR1 induced signaling in VAT of abdominally obese individuals may be an important cancer prevention strategy as well as an adjuvant therapy for improving outcomes following cancer diagnosis.
Citation Format: Debrup Chakraborty, Vanessa Benham, Elena Y. Demireva, Blair Bullard, Jamie J. Bernard. Fgf2 from visceral adipose tissue stimulates neoplastic transformation of nonmalignant epithelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2227. doi:10.1158/1538-7445.AM2017-2227
Abstract 2232: Dietary flavonoids, CYP1A1 genetic variants, and the risk of colorectal cancer
Authors: Kim Jeongseon , Cho Young Ae , Lee Jeonghee , Oh Jae Hwan , Sohn Dae Kyung , Chang Hee Jin 
Published: July 2017

Abstract
The role of dietary flavonoid intake in colorectal carcinogenesis might differ according to flavonoid subclasses and individual genetic variants related to carcinogen metabolism. Therefore, we examined whether greater habitual dietary intake of flavonoid subclasses was associated with a lower risk of colorectal cancer and whether CYP1A1 genetic variants altered this association. A semi-quantitative food frequency questionnaire was used to assess the dietary intake of six flavonoid subclasses (flavonols, flavonones, flavanones, flavan-3-ols, anthocyanidins, and isoflavones) in 923 patients with colorectal cancer and 1,846 controls; furthermore, CYP1A1 genetic variants (rs4646903 and rs1048943) were genotyped. A logistic regression was used to investigate dietary flavonoid intake, CYP1A1 genetic variants, and their interaction in relation to colorectal cancer risk. Among the subclasses of flavonoids, flavonols (OR [95% CI] = 0.10 [0.06, 0.16], highest vs. lowest quartile) and flavan-3-ols (OR [95% CI] = 0.54 [0.41, 0.71], highest vs. lowest quartile) were significantly associated with colorectal cancer risk after adjusting for confounders. Homozygous variants of CYP1A1 rs4646903 showed a protective effect on the risk of rectal cancer (OR [95% CI] = 0.60 [0.37, 0.96], CC vs. TT/TC). Carriers of the CC homozygous variant with high flavonol intake showed a greater decrease in colorectal cancer risk compared with T allele carriers with low flavonol intake (P for interaction = 0.02), particularly regarding rectal cancer (P for interaction = 0.005). In conclusion, the effect of dietary flavonoid intake on colorectal cancer risk differs by flavonoid subclasses and CYP1A1 genetic variants.
Citation Format: Jeongseon Kim, Young Ae Cho, Jeonghee Lee, Jae Hwan Oh, Dae Kyung Sohn, Hee Jin Chang. Dietary flavonoids, CYP1A1 genetic variants, and the risk of colorectal 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 2232. doi:10.1158/1538-7445.AM2017-2232
Abstract 2233: The fatty acid amide hydrolase inhibitor URB937 ameliorates radiation-induced lung injury in a mouse model
Authors: Li Rui , Xue Jianxin , Lu You 
Published: July 2017

Abstract
Radiation-induced lung injury (RILI) is a common and potentially serious complication from radiotherapy to the thoracic region. In the current study, we examined the potential protective effects of URB937, an inhibitor of fatty acid amide hydrolase in a mouse model of RILI. Here we challenged male C57BL/6 mice with irradiation (16Gy to the thoracic region), and then injected intraperitonealy either URB937 (1 mg/kg) or vehicle three times a week for 30 days. Mice were sacrificed at either the end of treatment or120 days from irradiation to evaluate the extent of RILI and mice survival. Potential effects of URB937 on irradiation induced tumor growth inhibition were also evaluated. We found URB937 increased endocannabinoids in the lungs, attenuated the extent of RILI, prolonged the survival,inhibited mice developed pathological alterations of pneumonitis and lung fibrosis. Irradiation-induced increases of proinflammatory and profibrotic cytokines were decreased, including interleukin-1β, interleukin-6, tumor necrosis factor-α, transforming growth factor-β1 in plasma and lung tissue. Moreover, malondialdehyde in plasma and inflammation cells in bronchoalveolar lavage fluid were lower in mice treated with irradiation plus URB937. URB937 did not affect tumor growth inhibition caused by irradiation. These results suggested that inhibiting fatty acid amide hydrolase through URB937 could ameliorate the RILI, without affecting the efficacy of irradiation on tumor control.
Note: This abstract was not presented at the meeting.
Citation Format: Rui Li, Jianxin Xue, You Lu. The fatty acid amide hydrolase inhibitor URB937 ameliorates radiation-induced lung injury in a mouse 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 2233. doi:10.1158/1538-7445.AM2017-2233
Abstract 2236: High-throughput screen for chemopreventative agents that influence adipose-tissue driven epithelial neoplastic transformation
Authors: Benham Vanessa , Chakraborty Debrup , Bullard Blair , Dexheimer Thomas S. , Bernard Jamie J. 
Published: July 2017

Abstract
Epidemiological studies have established a link between excess adiposity and cancer risk. Our previous data demonstrate that adipose tissue stimulates the transformation (steps a cell undergoes to become malignant) of both skin and mammary epithelial cells as measured by anchorage-independent growth in soft agar. Therefore, agents that influence growth in soft agar may be used therapeutically for skin and breast cancer prevention. However, soft agar in a 24-well format is unsuitable for high-throughput screens. Our objective was to develop a high-throughput assay to evaluate chemopreventative agents that influence adipose-tissue driven epithelial neoplastic transformation. We demonstrate that growth in low attachment conditions in a 384-well plate format is strongly correlated with growth in soft agar. We utilized this methodology and discovered the ability of several compounds to inhibit adipose tissue-stimulated transformation. These compounds will be used in vivo in our models of adipose tissue- or HFD-promoted tumor formation.
Citation Format: Vanessa Benham, Debrup Chakraborty, Blair Bullard, Thomas S. Dexheimer, Jamie J. Bernard. High-throughput screen for chemopreventative agents that influence adipose-tissue driven epithelial neoplastic transformation [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 2236. doi:10.1158/1538-7445.AM2017-2236
Abstract 2239: Establishment of lung cancer CTCs competent for lung metastasis
Authors: Que Zujun , Luo Bin , Shi Qihui , Tian Jianhui 
Published: July 2017

Abstract
Circulating tumor cells (CTCs) have been implicated as a population of cells that may seed metastasis, which led to the 90% of dead in cancer patients. Though the detection technologies has been developing swiftly, how to culture the cells successfully and established a permanent cell line are the bottleneck in the research of metastasis mechanism. Here, we report, for the first time, the establishment of permanent cell line from CTCs of one Non-small cell lung cancer patient and identifying the biological characterization. The cell line designated CTC-0430 has been cultured for more than one and half years, and the cells have been characterized at the genome, proliferation, drug resistance, the expression of surface marker, cytokine secretion level and the characterization of tumor formation in vivo by immunohistochemisty and immunofluorescence. This thorough analysis showed that CTC-0430 cells display a stable phenotype characterized by an intermediate epithelial/mesenchymal phenotype, stem-cell like properties and the drug sensitivity test indicating stronger drug resistance compared to A549 and 95-D. When compare with A549 and 95-D cells, CTC-0430 cells high expression of ICAM-1 and ICAM-3, low expression of CX3CL1 and CXCL1. Functional studies showed that CTC-0430 cells induced rapidly in vivo tumors after subcutaneous transplantation in immunodeficient mice, and the lung metastasis were found after six months later. Besides, three months after we injected the cells into the caudal vein of NOD/SCID mice, the lung metastasis were formed. And then the metastasis lung tissue were used to make IHC and IF, which indicated that the biological characterizations of this cell line are different from A549. The establishment stable cell line of lung cancer CTCs can provide in vivo research platform for the intervention study of postoperative metastasis of lung cancer.
Citation Format: Zujun Que, Bin Luo, Qihui Shi, Jianhui Tian. Establishment of lung cancer CTCs competent for lung metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2239. doi:10.1158/1538-7445.AM2017-2239