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Supplementary Fig. 5. Kaplan-Meier analysis of the survival of patients with different mutations.

Legends for Supplementary Figures

Current standard of care for patients with pediatric acute lymphoblastic leukemia (ALL) is mainly effective, with high remission rates after treatment. However, the genetic perturbations that give rise to this disease remain largely undefined, limiting the ability to address resistant tumors or develop less toxic targeted therapies. Here, we report the use of next-generation sequencing to interrogate the genetic and pathogenic mechanisms of 240 pediatric ALL cases with their matched remission samples. Commonly mutated genes fell into several categories, including RAS/receptor tyrosine kinases, epigenetic regulators, transcription factors involved in lineage commitment, and the p53/cell-cycle pathway. Unique recurrent mutational hotspots were observed in epigenetic regulators CREBBP (R1446C/H), WHSC1 (E1099K), and the tyrosine kinase FLT3 (K663R, N676K). The mutant WHSC1 was established as a gain-of-function oncogene, while the epigenetic regulator ARID1A and transcription factor CTCF were functionally identified as potential tumor suppressors. Analysis of 28 diagnosis/relapse trio patients plus 10 relapse cases revealed four evolutionary paths and uncovered the ordering of acquisition of mutations in these patients. This study provides a detailed mutational portrait of pediatric ALL and gives insights into the molecular pathogenesis of this disease. Cancer Res; 77(2); 390–400. ©2016 AACR.

Supplementary Fig. 4. p53 pathway is dysregulated in ALL.

Supplementary Fig. 2. Mutations found in the ASXL family of genes.

Supplementary Fig. 1. Mutational spectrum of pediatric ALL.

Supplementary Table 1. Samples used for whole exome and targeted sequencing. Supplementary Table 2. Clinical information of patients. Supplementary Table 3. 560 Genes screened in the targeted sequencing. Supplementary Table 4. Sequences of shRNA or CRISPR-Cas9 sgRNA used in this study. Supplementary Table 5. Sequences of real-time PCR primers used in this study. Supplementary Table 6. Sequences of primers used for Sanger validation of CRISPR-Cas9 indel. Supplementary Table 7. Mutations of KRAS identified in this study have been found in a variety of cancers. Supplementary Table 8. Mutations of PTPN11 in other cancers occurring in the same location as in our ALL cohort. Supplementary Table 9. Recurrent mutation sites of FLT3 in different cancers. Supplementary Table 10. Cell lines or cancer samples harboring either E1099K or T1150A hotspot mutation of WHSC1. Supplementary Table 11. R1446 mutations of CREBBP recurrently occurred in a variety of cancers. Supplementary Table 12. Mutational hotspot (D1399) of EP300 occurring in other cancers occurring in the same location as in our ALL cohort.Supplementary Table 13. Rare mutations occurring in our ALL cohort (mutated in one individual each), but these mutations have been recurrently documented in the COSMIC cancer mutations database. Supplementary Table 14. Mutations of genes involved in DNA repair pathway in relapse samples.

Supplementary Figure 2. Pathways regulated by PARK2 in glioma cells.

Supplementary Information includes Supplementary Materials and Methods, Supplementary Figure Legends and References. (FOR PUBLICATION)

Supplementary Figure 3. PARK2 interacts with multiple forms of β-catenin proteins.

Supplementary Figure 4. Wnt-β-catenin and EGFR-AKT pathways in glioma.

Supplementary Table 1.Summary of somatic PARK2 mutations. Supplementary Table 2. Sequences of shRNAs and siRNAs. Supplementary Table 3. Sequences of primers used for qRT-PCR.

PARK2 (PARKIN) is an E3 ubiquitin ligase whose dysfunction has been associated with the progression of Parkinsonism and human malignancies, and its role in cancer remains to be explored. In this study, we report that PARK2 is frequently deleted and underexpressed in human glioma, and low PARK2 expression is associated with poor survival. Restoration of PARK2 significantly inhibited glioma cell growth both in vitro and in vivo, whereas depletion of PARK2 promoted cell proliferation. PARK2 attenuated both Wnt- and EGF-stimulated pathways through downregulating the intracellular level of β-catenin and EGFR. Notably, PARK2 physically interacted with both β-catenin and EGFR. We further found that PARK2 promoted the ubiquitination of these two proteins in an E3 ligase activity-dependent manner. Finally, inspired by these newly identified tumor-suppressive functions of PARK2, we tested and proved that combination of small-molecule inhibitors targeting both Wnt–β-catenin and EGFR–AKT pathways synergistically impaired glioma cell viability. Together, our findings uncover novel cancer-associated functions of PARK2 and provide a potential therapeutic approach to treat glioma. Cancer Res; 75(9); 1815–27. ©2015 AACR.

Supplementary Figure 5. Dual inhibition of glioma cells with combinational use of MK2206 and ICG001.

Supplementary Figure 1. Prognostic value of PARK2 expression in GBM and ovarian serious cystadenocarcinoma.

Esophageal squamous cell carcinoma (ESCC) is a lethal disease due to its high aggressiveness. The aim of the present study was to investigate the role of xeroderma pigmentosum complementation group D (XPD) in the growth and invasion of ESCC and to elucidate the potential underlying molecular mechanisms. Western blot analysis and RT‑qPCR were used to detect the expression level of XPD in ESCC tissue samples and adjacent normal esophageal tissue samples. The pEGFP‑N2/XPD plasmid was transfected into human ESCC cell lines (EC9706 and EC109). The proliferation, apoptosis, migration and invasion of EC9706 or EC109 cells were assessed following transfection with the XPD overexpression plasmid. The chemosensitivity of EC9706 or EC109 cells to cisplatin or fluorouracil was evaluated by CCK‑8 assay. The expression levels of phosphoinositide 3‑kinase (PI3K)/AKT, nuclear factor (NF)‑κB, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) and mitogen‑activated protein kinase (MAPK) signaling pathway‑related genes were detected by RT‑qPCR and western blot analysis. The results demonstrated that the expression level of XPD was markedly lower in ESCC tissue samples than in adjacent normal esophageal tissue samples. The pEGFP‑N2/XPD plasmid was successfully transfected into EC9706 or EC109 cells, inducing XPD overexpression. A High XPD expression markedly suppressed cell proliferation, migration and invasion, and increased the apoptotic rate of EC9706 and EC109 cells. Furthermore, the overexpression of XPD significantly increased the chemosensitivity of EC9706 and EC109 cells to cisplatin or fluorouracil. Following XPD overexpression, the expression levels of PI3K, p‑AKT, c‑Myc, Cyclin D1, Bcl‑2, vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)‑9 were markedly downregulated, while the expression level of p21 was markedly upregulated. On the whole, the findings of the present study demonstrate that XPD inhibits the growth and invasion of EC9706 and EC109 cells, whilst also enhancing the chemosensitivity of EC9706 and EC109 cells to cisplatin or fluorouracil by regulating the PI3K/AKT signaling pathway. XPD may thus be an underlying target for ESCC treatment and drug resistance.

Exploring the composition of soil organic carbon (SOC) and aggregate stability over a range of elevations is essential to evaluate the stability of SOC and the conversion of carbon source or sink of mountain ecosystems. However, information of those changes in mountain ecosystems in arid and semi-arid regions is still scarce to date. In this study, we estimated the response of SOC stability along elevation gradient (1,380–2,438 m) by combining physical protection of aggregates with chemical protection of SOC using a redundancy analysis method in the Helan Mountains, northwest China. Results showed that the dominated aggregates of Helan Mountains changed from microaggregates to macroaggregates with increasing elevation, and the mean weight diameter (MWD) in mid and high elevations (2,139–2,248 m) was significantly higher than that in low elevations (1,380–1,650 m). Solid-state 13C NMR spectra showed that O-alkyl C was the main chemical composition of SOC in the different elevations. Regression analysis indicated that with the increase of elevation, the proportion of aromatic C and aromaticity gradually increased, peaked at the mid elevation and subsequently decreased. These findings indicated that the soil structure and chemical composition of SOC are more stable in the mid elevation. Macroaggregates were positively correlated with O-alkyl C and aromatic C, while silt and clay were positively correlated with alkyl C, indicative of a lower decomposition rate of macroaggregates. The RDA analysis confirmed that aggregate stability were significant associated with SOC content and chemical composition such as O-alkyl C, indicating that the soil labile carbon pool was most related to affect aggregate stability. Our findings suggest that the Helan Mountains had a better soil structure and a more stable SOC chemical composition at mid elevation ranges. The soil structure at low elevations is sensitive to environment change; therefore, a reduction in anthropogenic disturbance is suggested for improved sustainability of the ecosystem.

Aim: To compare and evaluate the prognostic value of various pretreatment combinations of inflammatory factors in patients with lung cancer (LC). Materials & methods: This study enrolled 1005 patients with LC and categorized into a discovery cohort and a validation cohort. Results: A combination of Lymphocyte-to-C-reactive protein levels (LCR) demonstrated the highest correlation with poor first-line progression-free survival (PFS) and overall survival (OS) (p 0.05) compared with other parameters in LC patients. Decreased preoperative LCR was an independent prognostic factor for first-line PFS and OS (p 0.05) in patients. Conclusion: Pretreatment LCR is a promising biomarker for first-line PFS and OS in patients with LC.

Hepatocellular carcinoma (HCC) is one of the primary causes of cancer-associated deaths worldwide. Current treatment methods include surgical resection, chemotherapy and radiotherapy; however the curative rate remains low, thus novel treatments are required. The aim of the present study was to investigate the role of targeting protein for Xenopus kinesin-like protein 2 (TPX2) in the growth of HCC and its underlying molecular mechanism. Immunohistochemistry staining, reverse transcription-quantitative (RT-q)PCR and western blotting were used to detect the expression of TPX2 mRNA and protein in liver cancer tissue samples, adjacent normal liver tissue samples, and the HCC cell lines Huh7, Hep3B, PLC/PRF/5 and MHCC97-H. The recombinant plasmid pMagic4.1-shRNA-TPX2 was constructed and transfected into Huh7 and Hep3B HCC cells to silence TPX2 expression. The proliferation, apoptosis, migration and invasion of Huh7 cells and Hep3B cells were evaluated before and after TPX2 silencing. The mRNA and protein expression levels of multiple signaling pathway-associated genes were detected by RT-qPCR and western blotting. The expression levels of TPX2 mRNA and protein were significantly higher in HCC tissue samples compared with adjacent normal liver tissue sample. TPX2 mRNA and protein expression levels were detected in the different HCC cell lines. The recombinant plasmid pMagic4.1-shRNA-TPX2 was successfully transfected into Huh7 and Hep3B cells, resulting in TPX2 silencing. TPX2 knockdown significantly reduced cell proliferation, cell migration and cell invasion of Huh7 and Hep3B cells, whilst also increasing the rate of apoptosis in these cells. Following TPX2 silencing, the expression levels of PI3K, phospho-AKT, Bcl-2, c-Myc and Cyclin D1 were significantly decreased, whereas the expression levels of P21 and P27 were significantly increased. In conclusion, TPX2 may suppress the growth of HCC by regulating the PI3K/AKT signaling pathway and thus, TPX2 may be a potential target for the treatment of liver cancer.

Diagnosis and relapse: cytogenetically normal acute myelogenous leukemia without FLT3 -ITD or MLL -PTD

Partial tandem duplication of MLL (MLL-PTD) characterizes acute myeloid leukemia (AML) patients often with a poor prognosis. To understand the order of occurrence of MLL-PTD in relation to other major AML mutations and to identify novel mutations that may be present in this unique AML molecular subtype, exome and targeted sequencing was performed on 85 MLL-PTD AML samples using HiSeq-2000. Genes involved in the cohesin complex (STAG2), a splicing factor (U2AF1) and a poorly studied gene, MGA were recurrently mutated, whereas NPM1, one of the most frequently mutated AML gene, was not mutated in MLL-PTD patients. Interestingly, clonality analysis suggests that IDH2/1, DNMT3A, U2AF1 and TET2 mutations are clonal and occur early, and MLL-PTD likely arises after these initial mutations. Conversely, proliferative mutations (FLT3, RAS), typically appear later, are largely subclonal and tend to be unstable. This study provides important insights for understanding the relative importance of different mutations for defining a targeted therapeutic strategy for MLL-PTD AML patients.

Acute myeloid leukemia (AML) with an FLT3 internal tandem duplication (FLT3-ITD) mutation is an aggressive hematologic malignancy with a grave prognosis. To identify the mutational spectrum associated with relapse, whole-exome sequencing was performed on 13 matched diagnosis, relapse, and remission trios followed by targeted sequencing of 299 genes in 67 FLT3-ITD patients. The FLT3-ITD genome has an average of 13 mutations per sample, similar to other AML subtypes, which is a low mutation rate compared with that in solid tumors. Recurrent mutations occur in genes related to DNA methylation, chromatin, histone methylation, myeloid transcription factors, signaling, adhesion, cohesin complex, and the spliceosome. Their pattern of mutual exclusivity and cooperation among mutated genes suggests that these genes have a strong biological relationship. In addition, we identified mutations in previously unappreciated genes such as MLL3, NSD1, FAT1, FAT4, and IDH3B. Mutations in 9 genes were observed in the relapse-specific phase. DNMT3A mutations are the most stable mutations, and this DNMT3A-transformed clone can be present even in morphologic complete remissions. Of note, all AML matched trio samples shared at least 1 genomic alteration at diagnosis and relapse, suggesting common ancestral clones. Two types of clonal evolution occur at relapse: either the founder clone recurs or a subclone of the founder clone escapes from induction chemotherapy and expands at relapse by acquiring new mutations. Relapse-specific mutations displayed an increase in transversions. Functional assays demonstrated that both MLL3 and FAT1 exert tumor-suppressor activity in the FLT3-ITD subtype. An inhibitor of XPO1 synergized with standard AML induction chemotherapy to inhibit FLT3-ITD growth. This study clearly shows that FLT3-ITD AML requires additional driver genetic alterations in addition to FLT3-ITD alone.

PARK2 (PARKIN) is an E3 ubiquitin ligase whose dysfunction has been associated with the progression of Parkinsonism and human malignancies, and its role in cancer remains to be explored. In this study, we report that PARK2 is frequently deleted and underexpressed in human glioma, and low PARK2 expression is associated with poor survival. Restoration of PARK2 significantly inhibited glioma cell growth both in vitro and in vivo, whereas depletion of PARK2 promoted cell proliferation. PARK2 attenuated both Wnt- and EGF-stimulated pathways through downregulating the intracellular level of β-catenin and EGFR. Notably, PARK2 physically interacted with both β-catenin and EGFR. We further found that PARK2 promoted the ubiquitination of these two proteins in an E3 ligase activity-dependent manner. Finally, inspired by these newly identified tumor-suppressive functions of PARK2, we tested and proved that combination of small-molecule inhibitors targeting both Wnt–β-catenin and EGFR–AKT pathways synergistically impaired glioma cell viability. Together, our findings uncover novel cancer-associated functions of PARK2 and provide a potential therapeutic approach to treat glioma. Cancer Res; 75(9); 1815–27. ©2015 AACR.

We investigated the oncogenic role of SETDB1, focusing on non-small cell lung cancer (NSCLC), which has high expression of this protein. A total of 387 lung cancer cases were examined by immunohistochemistry; 72% of NSCLC samples were positive for SETDB1 staining, compared to 46% samples of normal bronchial epithelium (106 cases) (p

Protein structure prediction is a main task in the field of bioinformatics, and the prediction of protein secondary structure is the key point of this task. Extracting representative features and efficient classification methods are the basis of the prediction process. In this paper, a prediction method based on BP neural network is proposed. We use valid protein features extracted by variable-sized sliding window and two different encoding modes (5-bit encoding and Profile encoding) as input data, to make predictions for the secondary structure of proteins. The prediction accuracies are calculated by Jackknife test on three commonly used low-similarity protein datasets: 25PDB, 1189 and 640, and this method achieves a high overall accuracy upon these three datasets.

In order to filter out the PPI (protein-protein interaction) data noise and improve the accuracy of protein complexes prediction, this paper proposed a method to integrate biological information and topology information as the weight of the PPI network. There are also some traditional methods to deal with the data noise by topology information of biological information, but fewer of them integrate those two aspect. In this paper, we introduced a method to extract the feature vectors for protein couples or the interactions instead of single proteins based on both topology and biology information, then the cosine theory is used to measure the weight of PPI network. At last, it used four classical predicting protein complexes algorithms to detect protein complexes from different weighted PPI network. The result shows that our method performs better both on filtering out data noise and predicting protein complexes.

Current standard of care for patients with pediatric acute lymphoblastic leukemia (ALL) is mainly effective, with high remission rates after treatment. However, the genetic perturbations that give rise to this disease remain largely undefined, limiting the ability to address resistant tumors or develop less toxic targeted therapies. Here, we report the use of next-generation sequencing to interrogate the genetic and pathogenic mechanisms of 240 pediatric ALL cases with their matched remission samples. Commonly mutated genes fell into several categories, including RAS/receptor tyrosine kinases, epigenetic regulators, transcription factors involved in lineage commitment, and the p53/cell-cycle pathway. Unique recurrent mutational hotspots were observed in epigenetic regulators CREBBP (R1446C/H), WHSC1 (E1099K), and the tyrosine kinase FLT3 (K663R, N676K). The mutant WHSC1 was established as a gain-of-function oncogene, while the epigenetic regulator ARID1A and transcription factor CTCF were functionally identified as potential tumor suppressors. Analysis of 28 diagnosis/relapse trio patients plus 10 relapse cases revealed four evolutionary paths and uncovered the ordering of acquisition of mutations in these patients. This study provides a detailed mutational portrait of pediatric ALL and gives insights into the molecular pathogenesis of this disease. Cancer Res; 77(2); 390–400. ©2016 AACR.

Anaplastic thyroid carcinoma (ATC) is an aggressive malignancy having no effective treatment. Laminin subunit-γ-2 (LAMC2) is an epithelial basement membrane protein involved in cell migration and tumor invasion and might represent an ideal target for the development of novel therapeutic approaches for ATC.The objective of the investigation was to study the role of LAMC2 in ATC tumorigenesis.LAMC2 expression was evaluated by RT-PCR, Western blotting, and immunohistochemistry in tumor specimens, adjacent noncancerous tissues, and cell lines. The short hairpin RNA (shRNA) approach was used to investigate the effect of LAMC2 knockdown on the tumorigenesis of ATC.LAMC2 was highly expressed in ATC samples and cell lines compared with normal thyroid tissues. Silencing LAMC2 by shRNA in ATC cells moderately inhibited cell growth in liquid culture and dramatically decreased growth in soft agar and in xenografts growing in immunodeficient mice. Silencing LAMC2 caused cell cycle arrest and significantly suppressed the migration, invasion, and wound healing of ATC cells. Rescue experiments by overexpressing LAMC2 in LAMC2 knockdown cells reversed the inhibitory effects as shown by increased cell proliferation and colony formation. Microarray data demonstrated that LAMC2 shRNA significantly altered the expression of genes associated with migration, invasion, proliferation, and survival. Immunoprecipitation studies showed that LAMC2 bound to epidermal growth factor receptor (EGFR) in the ATC cells. Silencing LAMC2 partially blocked epidermal growth factor-mediated activation of EGFR and its downstream pathway. Interestingly, cetuximab (an EGFR blocking antibody) or EGFR small interfering RNA additively enhanced the antiproliferative activity of the LAMC2 knockdown ATC cells compared with the control cells.To our knowledge, this is the first report investigating the effect of LAMC2 on cell growth, cell cycle, migration, invasion, and EGFR signaling in ATC cells, suggesting that LAMC2 may be a potential therapeutic target for the treatment of ATC.

Discovery of cancer genes through interrogation of genomic dosage is one of the major approaches in cancer research. In this study, we report that phosphodiesterase subtype 4D (PDE4D) gene was homozygously deleted in 198 cases of 5,569 primary solid tumors (3.56%), with most being internal microdeletions. Unexpectedly, the microdeletions did not result in loss of their gene products. Screening PDE4D expression in 11 different types of primary tumor samples ( n = 165) with immunohistochemistry staining revealed that its protein levels were up-regulated compared with corresponding nontransformed tissues. Importantly, depletion of endogenous PDE4D with three independent shRNAs caused apoptosis and growth inhibition in multiple types of cancer cells, including breast, lung, ovary, endometrium, gastric, and melanoma, which could be rescued by reexpression of PDE4D. We further showed that antitumor events triggered by PDE4D suppression were lineage-dependently associated with Bcl-2 interacting mediator of cell death (BIM) induction and microphthalmia-associated transcription factor (MITF) down-regulation. Furthermore, ectopic expression of the PDE4D short isoform, PDE4D2, enhanced the proliferation of cancer cells both in vitro and in vivo. Moreover, treatment of cancer cells with a unique specific PDE4D inhibitor, 26B, triggered massive cell death and growth retardation. Notably, these antineoplastic effects induced by either shRNAs or small molecule occurred preferentially in cancer cells but not in nonmalignant epithelial cells. These results suggest that although targeted by genomic homozygous microdeletions, PDE4D functions as a tumor-promoting factor and represents a unique targetable enzyme of cancer cells.

The present study was designed to investigate the phenotypic transformation and migration of adventitial fibroblasts using 5-bromo-2′-deoxyuridine (BrdU) labeling following angioplasty and to explore the correlation between adventitial cells and post-angioplasty restenosis. A vascular restenosis model was established in 23 rats by injuring the common carotid artery with a wire. BrDU was used to label the fibroblasts followed by immunohistochemistry for α-actin. Blood vessels were observed under light microscopy and scanning electron microscopy followed by image analysis. The number of BrDU-positive fibroblasts in the intima, media and adventitia of the blood vessels was determined 3, 7, 41 and 28 days after injury. The results demonstrated that at different time points, the number of BrDU-positive cells was significantly different in the intima, media and adventia (P

With expanding admissions in every college in our country this recent years,future education is changing forn elite to popular. Students and parents pay more and more attention in school’location.environment and other conditions, but some colleges are too far away form home for field observation, so they had to get some unintuition information about school form traditional 2D web pages ,now Virtual Campus if used with corresponding explain can work out this problem very well, and it can observe school in 360 degree comprehensivity. Virtual Campus’s design applied computer’s3D modeling method and internet informations, through computer shows the campus humanities and campus style and features, 3D Rhino is a classical “NURBS” software. It makes up surfaces rely on points and lines, and then makes up the basic grapes we need, it can establishes complex model conveniently and fastly the author select 3D Rhino as develop software make innovation study and realize in Virtual Campus. This passage introduces how to use 3D Rhino and Coogle SketchUP to make virtual school scenes, school’s 3D scene model’s making and narratives how to visit virtual campus through Google Earth ,the major content can divided into three parts. First: Study how to establish college surface 3D building’s outside scenes model ingreat detail through remote sensing satellites pictures and inside seenes shooting means get scenes datas, use 3DRhino modeling skill establish scene models.3DRhino can build complex surface effects, such as school’s sculptures\roofs\complex fences and walls’ scenes models. Second: Take Google SketchUP map measure make materials map into models to get lifelike 3D sculpture, use Google SketchUP’s truly grain or get form take photos add to model surface, Delete useless buildings details to optimization dealing with models, to prominent the features about buildings. With the advent of the Internet age , today, the computer has been deeply every family.Many famous universities such as Zhejiang University , Shanghai Jiaotong University , etc., adopting virtual reality technology to build a virtual campus .Virtual campus provides a platform ,which to help achieve the University 's resource management, environmental planning and school development , remote access, etc.In today's changing the technology will be applied to more schools with eelectronic information technology(Electronics IT) improve. In most of the campus network has been widely used, with the development of colleges and universities in our country, the improvement of the education system, more and more students have the opportunity to enter colleges and universities for further study. Many colleges and universities have to start expansion.Based on a series of teaching , accommodation and other facilities continue to increase, the effective management of the campus began to be taken seriously.Therefore, campus information management has become the most effective management.Most schools began to use the network to establish their own teaching , living, entertainment and other systems.And compared to traditional campus , virtual campus can give students a more intuitive, more effective understanding, so this technology has also vigorously developed in China. During the globalization of information, the virtual campus created the conditions for the country 's schools to the world.Online virtual campus provides many opportunities not only for Chinese students but also for foreign students to to understand each other's university , college life, and even the campus culture .Through virtual campus , schools can have a more in-depth exchanges and contacts , which not only promotes academic exchanges,but also provides favorable conditions for enhancing the competitiveness of schools and optimizing Campus. International Industrial Informatics and Computer Engineering Conference (IIICEC 2015) © 2015. The authors Published by Atlantis Press 943 Image-based modeling techniques to draw the photo images of buildings , based on a large number of environmental stresses multi-angle photos into the computer for analysis, using computer vision technology to help analyze and establish the geometry of buildings and the environment.This method does not have to establish a separate complex model to analyze automatically by a computer model , if the technology is mature , will greatly enhance the efficiency of the building modeling .In recent years, this modeling approach concern of many scholars , and is committed to improving the technology and the popularity of talk .But now computer vision technology is not mature enough , modeled by this method still requires producers to enter a lot of matching data.How to achieve between the face image matching , how to eliminate or reduce the image noise , also remains to be further studied.Image-based modeling method flow chart shown in Figure 1 . Figure 1 image modeling method 1.Virtual Campus Problems With the popularization of network technology and google , more and more universities have begun to build their own virtual campus .From a functional point of view , the virtual campus is divided into two categories: One virtual campus is mainly used to serve the campus teaching and management,and it is through the establishment of digital campus to expand the campus network functions;The other is mainly to show the campus scenery and school style, while making an inquiry and guided tours . Technically,no matter what type of virtual campuses are using virtual reality technology to show more intuitive campus buildings and scenery.But as we can see ,there are still many shortcomings in the virtual campus technology ,mainly in the interaction between the viewer and the navigation is not good enough .That there are many problems to solve and improve the process of building a virtual campus , mainly in the following aspects: 1.1 At present, virtual campus mainly adopts three-dimensional(3D) panoramic way fixed-point observation , it can not be close observation , unable to see the scenes details.In order to maximize the display of three-dimensional(3D) scenes , its preimage acquisition makes high demands . 1.2After a three-dimensional(3D) construction, virtual campus scenes are rendered into an image to be displayed. When a user browsing ,it only for fixed-point observation and not at liberty to browse ,, and it is more passive in terms of information inquiry , interaction is also poor. 1.3Realistic virtual campus scenes need to build a large geometric model, which is bound to take up a lot of storage space, the attendant problems is the browser opens the page in the network will take a long time .Therefore , a need to study the optimization method , making the model more realistic and less storage space on a balance can be achieved in improving the network transmission speed and fidelity scene coordination in hand, which in the future development of the virtual campus has very far-reaching significance. Virtual campus landscape model according to the theory of cognitive research , geographical scope of the performance of the spatial objects within the scope encompasses part of the campus .In

Nowdays, Chinese business pay more attention to build their commercial Stores Identity System. In recent years, during the process of the “ Second upgrade of corporate visual image”, academic and business circles focus on how to express the inherent brand ‘s cultural concepts and national characteristic effectively, and how to reflects psychological need of the crowd accurately. use different levels of design symbols coding rules, and get rid of some unnecessary information. Finally they will find the distinctive characteristics of the corporate brand image to enhance the brand culture and competitiveness. 1. Explore how to spread the space image with brand differences in the design methodology Based on years of practical experience, the author thinks that, in the process of national or brand characteristic symbol design, which means the encoding process of the symbol spreading, has a certain scientific laws and specific method. First in terms of methodology, we should realize clearly, in both the expression and the choice of construction materials, should be subject to the basic requirement of modern design theory and information communication, only in this way can get the accurate good symbol recognition and information arrival rate. Not split using some traditional Chinese symbols, but conformity the symbol by contemporary audience allow forms, instead of a variety of symbol of national culture, mining the most representative code combination according to scientific research, And spread these symbols according to the function different communication theory in a planned way hierarchical, is not only use ancestor or an area commonly used construction materials and technology as the carrier of the dissemination of information, but as far as possible use the most common material on the market at present, or the most easy to implement process the information. So no matter how the nationalization of the characteristic design, to meet the basic requirements of commercial space SI systemIdentify effective, moderate volume, no noise interference, convenient construction simple, easy to copy. As Mr LiZhong hair says in the book : A qualified modern designers should have three kinds of abilities-1 information understanding and application 2 constitute ability of design modelling and color 3 convey information and the ability to communicate through design. The three abilities actually represents the information of import and integration ability, design codes and performance ability, symbol of output control and message control ability. 2. Information coding level and rule of the difference brand space system According to the basic model of information communication of Shannon and Weaver., the communication process of a complete design information should abide by the following rules: From the information source(designer) get the information encoded(The design process) and then transformed into symbols or symbols groups(information)the symbol or information through certain information spreading media(channel) Interpretation by the sink(the information receiver), this process of interpretation symbols called decoding. In the links spreading channel, there is a noise interference problem, there are also symbol similar with spreading target symbolor confusing significance make the audience cannot accurately identify the symbol. So the accurate and effective coding link、control channel noise has become the key problem of this communication link which designers need to focus on. Ensure that the two links, the decoding process will fall into place, To International Industrial Informatics and Computer Engineering Conference (IIICEC 2015) © 2015. The authors Published by Atlantis Press 951 ensure the communication intention understands by the recipient. In short, the encoding process need to be aware of three major problems: The first is disseminate information for different purposes – analysis what types of information propagation, and then to determine the type of information design and code in what order which is design program, the last is the type of information needed to follow those rules or methods in the process of coding which is the design method as figure1shows. Fig 1 the symbol image Figure 2 communication link As the main brand’s space recognition, brand channel store exists in various forms、 business models and morphology,whether the brand interior stores in large shopping malls or similar kind of McDonald's outdoor shops on both sides of the street, whether it is as independent as Sinopec gas station chain form or large stores as Dazhong, Gome, as long as enterprises or brand after system of SI regulated, there are following five symbol characteristics in space system of communication channels in its business1 :1、logo and the word standard brands such as VI symbols2 performance style and modeling 3、the standard store color 4、system4 uniform material 5、The brand unique spatial aided shape。 In the multi view interpretation Symbols the designer,Symbolic interpretation’s horizons fusion shows a multiple loop structure, that is take the interpreter and the designer as the main body, the former’s attention is the interpreter perspective effect on the symbolic interpretation of behavior, the latter concern about the impact on the interpretation of the behavior of the designer perspective. As designers, we cannot control the decoding process, but can accurately interpretation symbols by careful arrangement of information and extensive research influence the information receiver, standing in the designer angle, the author believes that the above 5 big symbol feature groups can be divided into three sequentially coded information hierarchy according to their different purpose of propagation. The first is the necessary information hierarchy, including the logo and the standard word etc. the core brand visual symbols. The so-called necessary information refers to is the information need to spread, like a personal understanding another person must first know its name. Company logo and the standard word are the external performance of the enterprise internal culture, industry, and other aspects of the intrinsic characteristics of the concept, the core of the visual image or text point directly to the connotation of enterprise. No matter what stores need to first communicated to consumers is name or logo. It is known as the necessary information. Secondly the need to disseminate information hierarchy is homogeneous information hierarchy, the three main aspects of the hierarchical information including style and shape, store standard color system, unified material. The so-called homogeneous meaning is to code a dissemination of information and the same industry or the same population subdivision location nearest rival in all aspects of the requirements. This encoding is to ensure that the specification characteristics possible propagation of this industry in the practical application of SI space. After the study, different industries have different performance characteristics of the symbol, according to the features and the industry or the location of population subdivision in many industries tend to show a homogeneous rules. Take the color symbol as an example,the fast food industry such as McDonald's, KFC, pizza hut, real time, Yoshinoya etc. Volkswagen brand, its Wailimian main tone often focus on

This paper briefly introduces the clothing poster, explains the process of making this poster and simply talk about the inspiration origin of clothing designer. It points out that the corresponding poster background is produced according to related topics and images. It focuses on the poster image processing, font design, color design as well as layout design, and then orchestrate them reasonably. 1. Preparation for Poster Production 1.1 Materials collection Before producing this poster, we understand relevant contents first, and then collect relevant information and images, check out some books, search for some reference materials as comparison on the Internet and revise constantly. Designers from department of design ask about the category and inspiration of clothing and know that in most instances, clothing in each season can be divided into four series: woman series, office series, modern series and high-end series. While theme of this season is “Ocean Impression” which belongs to modern and leisure series. 1.2 Theme source Theme inspiration of this season comes from the movie “Avatar”. It draws on the ocean color system as well as the epidermal mimicry pattern of original ecological animals, and combines the heart geometric art in 1960s. In addition, it uses bionic, variation and reconstruction design techniques which makes the design modeling present a kind of relaxed and dynamic atmosphere, and highlights the modern, mysterious and cool ocean feeling of this season. 2. Poster Design Process 2.

This paper takes soybean milk machine as an example. Nowadays, soybean milk machines on the market all adapt rotational flow to cut the beans and electric heating tube to heat. Whether they have filter nets or not or the beans are dry or wet is not the main factor to classify soybean milk machines. Instead, design is the most competitive factor. In other words, the overall design and user experience are important factors to subdivide soybean milk machines. Figure 1 shows the family income structure of people who drink soybean milk.According to the consumer survey in 2014, different family incomes have little effect on preference of people who drink soybean milk. Structure charts of urban market levels show that in 2014 the sales volume of soybean milk machine in markets of mega cities takes 40% of the total, and mega cities are the largest markets of soybean milk machine; markets in megapolis and other markets both take half of the remaining 60% share. (Mega cities refer to cities with a population over two million; megapolis means cities with a population between 100 to 200 million; The large cities and under refer to cities with 1 million people or less). Figure 1 Yield analysis Figure 2 Classification Figure 2 shows owning rate of small electrical appliances in urban families. Based on a consumer survey in 2014, 48% of urban families have soybean milk machine, which is much lower than the owning rate of electric cooker and induction cooker. In summary, it can be seen that soybean milk machine will be a necessity of our life just like TV and computer. As more and more people concern about fitness and health maintenance, there will be an increasing number of Chinese families using soybean milk machines. Less affected by age and family income, people will not pay too much attention to its price comparing to food safety and delicacy. Besides, soybean milk machine has stronger price advantage than others. Market is an important factor affecting the design, and in some time, it is the market that plays a vital role in design. 1. Analysis on the existing soybean milk machines From the aspect of soybean milk machine market, Joyoung, Midea, Donlim, Philips and other brands take more than 80% of total market share. Next, soybean milk machine of these three brands-Joyoung, Philips and Midea analyzed below. 3rd International Conference on Mechanical Engineering and Intelligent Systems (ICMEIS 2015) © 2015. The authors Published by Atlantis Press 500 1.1Analysis on structures As is shown in Figure 3, soybean milk machines on the market basically use this structure.In this separation structure, technical and functional components are gathered in the upper part of the soybean milk machine, and the lower part made of stainless steel is only used as a container. Control room is responsible for the entire computer-controlled process of making soybean milk. Blades are responsible for the high-speed rotation for breaking the soybeans. Level electrodes are responsible for detecting the water level to prevent dry heating. Anti-overflow electrode is responsible for detecting the water level to prevent overflow. Electrical heated tube is responsible for heating the entire soybean milk. Figure 3 Analysis on structures 1.2 Analysis on appearance After analysis on structure, appearance of soybean milk machines will be analyzed in the following part. Because of the separation structure, the upper part of the machine has a handle to control panel while the lower part has a side handle. Although the locations of socket holes are different, they are basically in the upper part. As a whole,the appearance of the machine honestly reflects the function and use, but the overall image of the product need to be improved because it lacks integrity and doesn’t fully consider about the using process. For example, after the making of soybean milk is finished, the whole upper part has a certain temperature, but there is only one handle in the upper part, so it is clear that there is no suitable placing angle. The only solution is to put it in a bowl or on a board to wait for cleaning. All the functional opponents are gathered in the upper part, so the machine has a certain weight and is too heavy to move, so it is both inconvenient and insecure.Instead, the lower part just plays the role as a “reaction chamber”and is not made full use of. Comparisons among soybean milk machines of brands Yoyoung, Philips and Midea indicates that although they have various styles and qualities, the basic pattern is the same, and has never been greatly improved. So there is a wide space as well as a big challenge for further efforts. 1.3 Analysis on materials In aspects of the material of soybean milk machine, the material selection of the inner side is very strict because it has to withstand high temperature. The common materials of the inner side are mainly stainless steel and ABS with high-temperature resistance. As to the outside of soybean milk machine, there is a vast space for development. The appearance of the soybean milk machine is more beautiful and of finer quality. The outside materials are stainless steel, ABS and so on. Surface treatment techniques include brushed metal, electroplating, LED decoration and so forth. There is a wide-open space on exterior design. 1.4Analysis of Operating Mode The existing soybean milk machines almost adapt keys operation. Under the guidance of a given computer program, press the key to start and it needs no more time to manage. The operation mode is a direct response to functions, which need to be improved with people’s higher standard for flavor. The design of operation interface is an important aspect. The operation interface is used to communicate and exchange information, so the quality of the operation interface directly determines the humanization of the product. The design could also refer to other technologies that other products

This paper based on study of humanized packaging design factors and specific implementation, it should include more humanity, emotion, aesthetic connotation, reflects the human nature concern, establish a kind of harmonious and unified relationship between people and products, human and environment. With the establishment and perfecting of market economy in our country, the competition of product market increasingly fierce, survival and development of an enterprise depends mainly on two aspects: One is that if the product is used for the society, the second is whether it can profit from it. The key to these two aspects is the product itself, whether that is really can buy and loved by consumers. Elegant and delicate product packaging can add the brightness grace for the product, increase the product sales, profit for the enterprise. Therefore also put forward higher requirements for product packaging design, the product packaging design on the premise of meet its basic function, should give full consideration to consumers' physiological and psychological feeling, that means it should consider people's aesthetic demand from form, color, structure, material and visual interface design, as well as the safety and comfort with the nature of such claim. 1. Human packaging design concept Humanized package design is the packing design of the "people-oriented", is the essence of human requirement, Through product outer packing technology, art and other aspects of integrated design to meet the needs of use function and aesthetic function and show the spirit of a kind of humanistic care. This requires the designer should be in line with people-oriented thoughts to feel, mining and summarise the mass consumer's psychological feeling and desire during the packaging design process, Grope for the regularity of the activities in the packaging design and consumer psychology to improve the effect of packaging design of human nature, prompt consumers to purchase products, profits for the enterprise. 2. The connotation of humanized packaging design Emotional design:The packing of the product adapt to the consumer's subjective experience and the design of the inner feelings, Make people satisfy their emotional needs in life with the help of product packaging color of text, graphics, structure, shape, improving consumer psychological aesthetic. The character design: Product packaging on the basis of meeting the demand of universal human physiology, psychology and intelligence, meet consumers' different age, sex, status the design of the packaging requirements.In the products modern society, humanized product packaging demand has become increasingly strong. The cost design: Design thinking of paying attention to the cost of the product packaging, reduce consumer burden. Since society appear "too far packaging" often make the businessman to packag ordinary product too fine at all costs, lead to unnecessary waste of economic, without considering the additional financial burden to consumers. Ecological design: Means to save packing material resources, and encourage recycling product packaging design can be used to reduce packaging waste, also known as sustainable packaging International Industrial Informatics and Computer Engineering Conference (IIICEC 2015) © 2015. The authors Published by Atlantis Press 947 design. Such as the application of different kinds of bottles, cans can be recycled packaging materials and containers, and even on the destruction of convenience, not damage the environment, such as ecological environment requirements. 3. The implementation of humanized packaging design 3.1 color and words Packaging color and text are important factors to convey the product information and beautify the product, it not only can reflect the characteristics of decorative products, also has an important influence on consumers' psychology. Different product has its unique visual color feeling, such as food commonly used bright rich colors, through such as red, yellow, orange to emphasis the sense of taste, highlight food’s fresh, delicious and nutritious. Medicine commonly use pure, warm and cool colors, use grey cool color reflect anti-inflammatory, analgesic, warm color reflect tonic, health care and nutrition among commonly used color, make the patient not stimulate too much or feeling boredom. And packaging design must be concise, clear, fully embodies the product attributes, difficult to use too intricate fonts and words shouldn’t be used, we should use easy to understand, easy to read, easy to identify font, and must take into account the consumer's discrimination, to make the person be clear at a glance. As shown in figure 1. Figure 1 color integral collocation 3.2 Structure and shape Packaging structure modelling is a closely integrated and industrial modernization, science and technology and the unity of artistic form, aesthetic and associated modelling design. Modelling and structure of the product packaging design should satisfy all the basic protection and the transportation, on the basis of product function, put themselves in the angle of consumer to consider, mainly by combining ergonomics principle, pay attention to consumer behavior, the aesthetics and the combination of mechanical design, to meet consumers' aesthetic and reach the purpose of easy to use. Such as tea, pigments, pharmaceuticals (boxes into barrels from dozens of grams of a small bag, pieces of a small bag of small doses of simple packaging), convenient for consumer to use and carry, liquid and particle state into the measuring tool for consumers to measuring, and design the packaging structure to comply with the principle of the man-machine engineering form and style, make it can be opened, can carry, can hang, etc. Using aspects Packing should be easy to open, easy to close. No matter what the packing of the product, should be easy to open and close, easy to fall out of use, as shown in figure 2. At the time of packaging should put yourself in the Angle of consumers to consider using details, such as packaging, how to fast and convenient to package the product, how not to use other tools easily open, now a lot of packing design into a swan. Figure 2 convenient mouth Figure 3 split line Convenient to carry. So in the time of product design should consider the product using and carrying, can be implemented by packaging, can also be implemented by the product itself

In this study, we performed whole-exome and targeted sequencing on 85 MLL-PTD AML patients. These AMLs have oncogenic tandem duplication of the MLL gene. At least one well-known oncogenic driver mutation was identified in over 90% of the MLL-PTD patients. In line with earlier sequencing studies of other AML subtypes and the TCGA-AML-sequencing project, DNMT3A was the most often mutated epigenetic regulator (25%); IDH1/2 hotspot mutations were identified in 31% of patients. TET family was the third most prominently mutated epigenetic regulator (TET1 (5%), TET2 (16.3%). Mutations of epigenetic regulators also occurred in polycomb-associated proteins (EZH2, ASXL family members), chromatin remodelers (ARID2, ARID1A), genes associated with histone acetylation (CREBBP, EP300, KAT6A, KAT6B) and histone methylation (MLL2, MLL3). Proliferation-related pathway was extensively mutated, with 54 of 80 MLL-PTD patients (67.5%) carrying at least one mutation of proliferative genes. Specifically, FLT3 mutations were found in 46% of patient samples. Notably, some FLT3-ITD patients had more than one type of internal tandem duplication (ITD) insertion, probably reflecting existence of multiple subclones in these leukemias. We found highly prevalent mutations of cohesin genes: STAG2 (16%), SMC1A (6%), SMC3 (1%), RAD21 (1%) and CTCF (6%). Cohesin pathway is more frequently mutated in MLL-PTD patients (26%) than the AML samples from either TCGA (13%) or a meta-analysis of 1000 AML (9.1%). Remarkably, an extremely high proportion of the mutations had a strong tendency to disrupt the coding sequence in STAG2, emphasizing their crucial tumor-suppressor role in this AML subtype (16% in MLL-PTD vs 3% in TCGA-AML. RNA processing pathway was also strikingly altered in MLL-PTD patients. The most prominently mutated genes within this category were the splicing factors. They included U2AF1 (13%, S34F/Y), SRSF2 (3%), SF3A1 (5%), ZRSR2 (3%), DHX15 (1%) and CWC22 (1%). Multiple mutations co-occur with MLL-PTD which are usually acquired in a sequential manner. A potential ordering for acquisition of many mutations include IDH2/DNMT3A/U2AF1/TET2→MLL-PTD→RAS-receptor tyrosine kinase based on the following reasons: #1, real-time-PCR showed that MLL-PTD was absent in remission while mutations of IDH2, DNMT3A, TET2 and U2AF1 were still retained with a high VAF. This suggests that MLL-PTD was acquired after mutations of IDH2, DNMT3A, TET2 and U2AF1; #2, MLL-PTD is highly stable during disease progression as compared with mutations of the RAS-RTK. On the other hand, RAS-RTK mutations frequently exist as subclonal mutations and tend to be unstable during disease progression. These observations support a notion that MLL-PTD was acquired prior to RAS-RTK. Taken together, MLL-PTD is acquired after those remission-persisting, initiating mutations (IDH2, DNMT3A, TET2 and U2AF1), but prior to lesions of the proliferation-related drivers. Citation Format: Lingwen Ding, Qiaoyang Sun, Kar-Tong Tan, Wenwen Chien, Anand Mayakonda, Dechen Lin, Xinyi Loh, Jinfen Xiao, Manja Meggendorfer, Tamara Alpermann, Manoj Garg, Su-Lin Lim, Vikas Madan, Norimichi Hattori, Yasunobu Nagata, Satoru Miyano, Allen Yeoh Eng Juh, Hsin-An Hou, Yan-Yi Jiang, Yan-Yi Jiang, Sumiko Takao, Li-Zhen Liu, Siew-Zhuan Tan, Siew-Zhuan Tan, Michael Lill, Mutsumi Hayashi, Akitoshi Kinoshita, Hagop M. Kantarjian, Steven M. Kornblau, Seishi Ogawa, Torsten Haferlach, Henry Yang, H. Phillip Koeffler. Mutational profiling of MLL-PTD acute myeloid leukemia [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 2450. doi:10.1158/1538-7445.AM2017-2450

With the increasing number of online comments, it was hard for buyers to find useful information in a short time so it made sense to do research on automatic summarization which fundamental work was focused on product reviews mining. Previous studies mainly focused on explicit features extraction whereas often ignored implicit features which hadn't been stated clearly but containing necessary information for analyzing comments. So how to quickly and accurately mine features from web reviews had important significance for summarization technology. In this paper, explicit features and “feature-opinion” pairs in the explicit sentences were extracted by Conditional Random Field and implicit product features were recognized by a bipartite graph model based on random walk algorithm. Then incorporating features and corresponding opinions into a structured text and the abstract was generated based on the extraction results. The experiment results demonstrated the proposed methods outpreferred baselines.

Anaplastic thyroid cancer (ATC) has no effective treatment, resulting in a high rate of mortality. We established cell lines from a primary ATC and its lymph node metastasis, and investigated the molecular factors and genomic changes associated with tumor growth.The aim of the study was to understand the molecular and genomic changes of highly aggressive ATC and its clonal evolution to develop rational therapies.We established unique cell lines from primary (OGK-P) and metastatic (OGK-M) ATC specimen, as well as primagraft from the metastatic ATC, which was serially xeno-transplanted for more than 1 year in NOD scid gamma mice were established. These cell lines and primagraft were used as tools to examine gene expression, copy number changes, and somatic mutations using RNA array, SNP Chip, and whole exome sequencing.Mice carrying sc (OGK-P and OGK-M) tumors developed splenomegaly and neutrophilia with high expression of cytokines including CSF1, CSF2, CSF3, IL-1β, and IL-6. Levels of HIF-1α and its targeted genes were also elevated in these tumors. The treatment of tumor carrying mice with Bevacizumab effectively decreased tumor growth, macrophage infiltration, and peripheral WBCs. SNP chip analysis showed homozygous deletion of exons 3-22 of the PARD3 gene in the cells. Forced expression of PARD3 decreased cell proliferation, motility, and invasiveness, restores cell-cell contacts and enhanced cell adhesion. Next generation exome sequencing identified the somatic changes present in the primary, metastatic, and primagraft tumors demonstrating evolution of the mutational signature over the year of passage in vivo.To our knowledge, we established the first paired human primary and metastatic ATC cell lines offering unique possibilities for comparative functional investigations in vitro and in vivo. Our exome sequencing also identified novel mutations, as well as clonal evolution in both the metastasis and primagraft.

Lateral access lumbar interbody fusion (LLIF) is a minimally invasive technique that has an increasing popularity. It offers unique advantages and circumvents risk of certain serious complications encountered in other conventional spinal approaches. This study provides a statistical analysis defining the lateral access learning curve in the Asian population. This prospective study included 32 consecutive patients who underwent LLIF from April 2012 to August 2014. The surgeries were performed by two senior spine surgeons and follow-up was conducted at 6 weeks, 3, 6, 9 months and 1 year post-operation. The breakpoint in operating time occurred at the 22nd level operated, from a mean of 71 min in the early phase group to a mean of 42 min in the steady state group. LLIF at L4/5 level is technically more demanding but technically feasible as competency is achieved. During the learning process, there was no compromise of perioperative or clinical outcomes. It should be feasibly incorporated into a spine surgeon’s repertoire of procedures for the lumbar spine.

Esophageal squamous cell carcinoma (ESCC) is a world-wide prevalent cancer, which is particularly common in certain regions of Asia. Here we report the whole-exome or targeted deep sequencing of 139 paired ESCC cases, and analysis of somatic copy number variations (SCNV) of over 180 ESCCs. We identified novel significantly mutated genes such as FAT1, FAT2, ZNF750 and KMT2D, in addition to previously discovered ones (TP53, PIK3CA and NOTCH1). Further SCNV evaluation, immunohistochemistry and biological analysis suggested their functional relevance in ESCC. Notably, RTK-MAPK-PI3K pathways, cell cycle and epigenetic regulation are frequently dysregulated by multiple molecular mechanisms in this cancer. Moreover, our approaches uncovered many novel druggable candidates, and XPO1 was further explored as a therapeutic target because of its mutation and protein overexpression. Together, our integrated study unmasks a number of novel genetic lesions in ESCC and provides an important molecular foundation for understanding esophageal tumors and developing therapeutic targets.

Pediatric ALL is the most common childhood tumor and the leading cause of childhood cancer deaths. To gain a better understanding of the landscape of somatic mutations in ALL, we performed whole exome and targeted sequencing of 240 pediatric B-ALL patients with their matched remission samples. The significantly mutated genes fall into several common categories: RAS/receptor tyrosine kinases, epigenetic regulators, transcription factors involved in lineage commitment and p53/cell cycle pathway. RAS/receptor tyrosine kinases: the most frequently mutated genes were members of RAS signaling (NRAS, KRAS, FLT3, PTPN11). Besides the well know hotspot mutations [G12D/V/C (NRAS 13 cases, KRAS 13 cases), G13D (NRAS 14 cases, KRAS 11 cases) and Q61H/L/R/K (NRAS 15 cases, KRAS 1 case)], novel mutational sites were also identified for KRAS: A146T/P (3 cases), K117N/T (4 cases) and V14I (1 case). High frequency missense mutations of PTPN11 clustered in SH2 domain (included the canonical hotspot A72T (5 cases) and E76K/V (4 cases)) and tyrosine-phosphatase catalytic domain (G503R/V). For FLT3, well-appreciated activating hotspot mutations in the kinase domain (D835Y/Y842C) and several novel recurrent mutationswere identified. Epigenetic regulators: hotspot mutations were identified in histone H3K36 methyltransferase WHSC1. Mutation E1099K located in the SET domain, was identified in 10 patients as well as two of the 5 ALL cell lines that we sequenced (RS4;11, SEM). Stable silencing of E1099K mutant WHSC1 in RS4;11 cells by either lentiviral shRNA or CRISPR guide RNA (sgRNA) markedly reduced clonogenic growth both in vitro and in vivo, underscoring the critical role of WHSC1 in lymphoid malignancies. Two highly-related histone/non-histone acetyltransferases, CREBBP and EP300, were also prominently mutated in our cohort. Mutations of CREBBP predominantly occurred in the acetyltransferase domain, particularly in the hotspot R1446C/H. Mutations of chromatin remodeling genes (ARID1A and ARID2) have been identified in a number of cases. Silencing of ARID1A in ALL cell lines by lentiviral shRNA resulted in upregulation of the pro-growth regulator c-MYC, while forced expression of ARID1A reduced c-MYC luciferase reporter activity. In addition, silencing of ARID1A by either shRNA or CRISPR-sgRNA resulted in enhanced clonogenic growth, suggesting that ARID1A may be involved in the c-MYC pathway and modulates the ALL cell proliferation. Mutations of epigenetic regulators were also found in the polycomb complex (EZH2, EED, SUZ12), chromatin/nucleosome structure modifying proteins (CHD2, CHD3, CHD4), TET family proteins [TET1 (2 cases), TET2 (5 cases)] and histone modification proteins (HDAC1, SIRT1, BCOR, BRD8, lysine demethylase PHF2/KDM6A, histone acetyltransferase KAT6B). Transcription factors and p53/cell cycle pathway: a number of alterations of transcription factors essential for hematopoietic and lymphoid differentiation were noted including the lineage regulator PAX5 (5 missense, 3 indels) and ETV6 (6 cases, 3 were frameshift indel and 1 was a splice-site mutations). In addition, mutations were also found in other lineage transcription factors (IKZF2, IKZF3, EBF1), WT1 (6 cases, including 3 indels and 1 stop-gain mutations), RUNX family member [RUNX2 (7 cases), RUNX1 (1 case)], ERG1 (3 cases), GATA1/3 (1 case each) and CTCF. Somatic mutations of genes involved in the p53 pathway occurred in 18 patients, including TP53, ATM and the kinases that regulate p53 activities (HIPK1, HIPK2). Germline TP53 pathogenic variants were found in these 2 patients. Taken together, we extensively interrogated the mutational landscape of a large cohort of pediatric ALL samples by exome and targeted resequencing. This study provides a detailed mutational portrait of pediatric ALL and gives new insights into the molecular pathogenesis of this disease. Disclosures Kantarjian: Amgen: Research Funding; ARIAD: Research Funding; Bristol-Myers Squibb: Research Funding; Pfizer Inc: Research Funding; Delta-Fly Pharma: Research Funding; Novartis: Research Funding. Ogawa:Sumitomo Dainippon Pharma: Research Funding; Kan research institute: Consultancy, Research Funding; Takeda Pharmaceuticals: Consultancy, Research Funding.

Chromosomal translocation t(8;21) (q22;q22) leading to generation of oncogenic RUNX1-RUNX1T1 fusion is a cytogenetic abnormality observed in about 10% of acute myelogenous leukemia (AML). Studies in animal models and recent next generation sequencing approaches have suggested cooperativity of secondary genetic lesions with t(8;21) in inducing leukemogenesis. In this study, we used targeted and whole exome sequencing of 93 cases (including 30 with matched relapse samples) to profile the mutational landscape of t(8;21) AML at initial diagnosis and post-therapy relapse. We identified recurrent mutations of KIT, TET2, MGA, FLT3, NRAS, DHX15, ASXL1 and KMT2Dgenes in this subtype of AML. In addition, high frequency of truncating alterations in ASXL2 gene (19%) also occurred in our cohort. ASXL2 is a member of mammalian ASXL family involved in epigenetic regulation through recruitment of polycomb or trithorax complexes. Unlike its closely related homolog ASXL1, which is mutated in several hematological malignancies including AML, MDS, MPN and others; mutations of ASXL2 occur specifically in t(8;21) AML. We observed that lentiviral shRNA-mediated silencing of ASXL2 impaired in vitro differentiation of t(8;21) AML cell line, Kasumi-1, and enhanced its colony forming ability. Gene expression analysis uncovered dysregulated expression of several key hematopoiesis genes such as IKZF2, JAG1, TAL1 and ARID5B in ASXL2 knockdown Kasumi-1 cells. Further, to investigate implications of loss of ASXL2 in vivo, we examined hematopoiesis in Asxl2 deficient mice. We observed an age-dependent increase in white blood cell count in the peripheral blood of Asxl2 KO mice. Myeloid progenitors from Asxl2 deficient mice possessed higher re-plating ability and displayed altered differentiation potential in vitro. Flow cytometric analysis of >1 year old mice revealed increased proportion of Lin-Sca1+Kit+ (LSK) cells in the bone marrow of Asxl2 deficient mice, while the overall bone marrow cellularity was significantly reduced. In vivo 5-bromo-2'-deoxyuridine incorporation assay showed increased cycling of LSK cells in mice lacking Asxl2. Asxl2 deficiency also led to perturbed maturation of myeloid and erythroid precursors in the bone marrow, which resulted in altered proportions of mature myeloid populations in spleen and peripheral blood. Further, splenomegaly was observed in old ASXL2 KO mice and histological and flow cytometric examination of ASXL2 deficient spleens demonstrated increased extramedullary hematopoiesis and myeloproliferation compared with the wild-type controls. Surprisingly, loss of ASXL2 also led to impaired T cell development as indicated by severe block in maturation of CD4-CD8- double negative (DN) population in mice >1 year old. These findings established a critical role of Asxl2 in maintaining steady state hematopoiesis. To gain mechanistic insights into its role during hematopoietic differentiation, we investigated changes in histone marks and gene expression affected by loss of Asxl2. Whole transcriptome sequencing of LSK population revealed dysregulated expression of key myeloid-specific genes including Mpo, Ltf, Ngp Ctsg, Camp and Csf1rin cells lacking Asxl2 compared to wild-type control. Asxl2 deficiency also caused changes in histone modifications, specifically H3K27 trimethylation levels were decreased and H2AK119 ubiquitination levels were increased in Asxl2 KO bone marrow cells. Global changes in histone marks in control and Asxl2 deficient mice are being investigated using ChIP-Sequencing. Finally, to examine cooperativity between the loss of Asxl2 and RUNX1-RUNX1T1 in leukemogenesis, KO and wild-type fetal liver cells were transduced with retrovirus expressing AML1-ETO 9a oncogene and transplanted into irradiated recipient mice, the results of this ongoing study will be discussed. Overall, our sequencing studies have identified ASXL2 as a gene frequently altered in t(8;21) AML. Functional studies in mouse model reveal that loss of ASXL2 causes defects in hematopoietic differentiation and leads to myeloproliferation, suggesting an essential role of ASXL2 in normal and malignant hematopoiesis. *LH and NH contributed equally Disclosures Ogawa: Takeda Pharmaceuticals: Consultancy, Research Funding; Sumitomo Dainippon Pharma: Research Funding; Kan research institute: Consultancy, Research Funding.

PARK2 is an E3 ubiquitin ligase whose dysfunction has been associated with the progression of autosomal recessive juvenile Parkinson's disease and human malignancies. However, its role in cancer remains to be explored. In this study, we report that PARK2 is frequently deleted and underexpressed in human glioma, and low PARK2 expression is associated with poor survival in cohorts of both low-grade glioma and glioblastoma multiforme (GBM). Functional studies revealed a tumor-suppressive role of PARK2 in GBM cells. Restoration of PARK2 significantly inhibited glioma cell growth both in vitro and in vivo, while depletion of endogenous PARK2 promoted cell proliferation. cDNA microarray analysis showed that PARK2 expression strongly altered the activity of both the Wnt and EGFR pathways. Immunoblot analysis confirmed that ectopic expression of PARK2 suppressed the intracellular levels of β-catenin, EGFR, as well as their down-stream targets including Cyclin D1, TCF4, c-Myc, p-AKT and p-S6K, etc. Notably, PARK2 physically interacted with both β-catenin and EGFR, and promoted their ubiquitination in an E3-ligase dependent manner. Similar to the ligase-dead PARK2 mutant (C431S), three PARK2 mutants harboring cancer-derived somatic mutations (T173A, T240M and P294S) showed decreased ability to ubiquitinate either β-catenin or EGFR proteins. We further found that PARK2 attenuated the cellular response to both Wnt3a and EGF stimulation, suggesting PARK2 is a negative regulator of both the Wnt and EGFR pathways. Lastly, inspired by these newly identified functions of PARK2, we tested and proved that the combination of small-molecule inhibitors targeting both Wnt-β-catenin and EGFR-AKT pathways synergistically impaired glioma cell viability. In aggregate, our findings uncover novel, cancer-associated functions of PARK2 and provide a potential therapeutic approach to treat glioma. Citation Format: Liang Xu, De-Chen Lin, Ye Chen, Haiyan Yan, Masaharu Hazawa, Ngan Doan, Jonathan W. Said, Ling-Wen Ding, Li-Zhen Liu, Henry Yang, Shi-zhu Yu, Michael Kahn, Dong Yin, Phillip Koeffler. PARK2 is a negative regulator of Wnt and EGFR pathways in glioma. [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 122. doi:10.1158/1538-7445.AM2015-122

Background Nasopharyngeal carcinoma (NPC) arises from the epithelial tissue of the nasopharynx. This cancer has remarkable ethnic and geographic distributions, with a particularly high prevalence in Southern China and Southeast Asia. Genetic susceptibility and Epstein-Barr virus infections have been implicated in the pathogenesis of NPC. However, genomic abnormalities of this neoplasm remain largely obscure, and no effective targeted therapy has been established. Therefore, a strong need exists to characterize the genetic alterations of this type of tumour for guiding the development of more effective and innovative therapeutic regimens. Methods Whole exomes sequencing (WES) was performed on 56 NPC germline/tumor pairs and 5 NPC cell lines. Transcriptome sequencing (RNA-seq) was conducted on 4 tumors from this cohort. An additional 61 NPC germline/tumor pairs and 5 non-paired primary tumors were further subjected to targeted regional deep-sequencing (TS). The somatic copy number variations (SCNV) of 52 primary tumors were profiled by SNP-array hybridization (50 of them also had WES data). Finally, both in vitro and in vivo biological and biochemical experiments were performed to evaluate the newly-identified mutations in NPC cell lines. Results We identified a total of 1,577 non-silent somatic mutations affecting 1,413 genes from WES, revealing a relatively low mutational rate and wide mutational diversity. Integration of the results from WES, TS and RNA-seq revealed a distinct mutational signature and a number of significantly mutated genes (such as TP53, ARID1A, MLL2, BAP1, PIK3CA, etc.) in NPC. Pathway enrichment analysis of genetic lesions identified several important cellular processes and pathways in NPC including chromatin modification, ERBB-PI3K signaling and autophagy machinery. We further characterized the biological functions of both ARID1A and BAP1 proteins in NPC cell lines. Depletion of wild-type endogenous ARID1A expression with shRNAs resulted in increased anchorage-independent colony formation, cell migration and xenograft growth in vivo, whereas ectopically expressed ARID1A suppressed both cell proliferation and migration. Similarly, ectopic expression of wild-type BAP1 suppressed anchorage-independent colony formation of NPC cells. Conclusion We characterized the genomic landscape of 128 NPC cases, and identified a number of novel driver genes with statistical and biological evidence. Integrated analysis showed enrichment of genetic lesions affecting chromatin modification, ERBB-PI3K signaling and autophagy machinery, offering opportunities for developing novel treatments. These results in aggregate provide an important genetic foundation for further study of the molecular pathology and etiology of this fatal disease. Citation Format: DECHEN LIN, Xuan Meng, Masaharu Hazawa, Yasunobu Nagata, Ana Maria Varela, Liang Xu, Yusuke Sato, Li-Zhen Liu, Ling-Wen Ding, Arjun Sharma, Boon Cher Goh, Soo Chin Lee, Bengt Fredrik Petersson, Feng Gang Yu, Paul Macary, Min Zin Oo, Soh Ha Chan, Henry Yang, Seishi Ogawa, H. Phillip Koeffler. The genomic landscape of nasopharyngeal carcinoma. [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 3939. doi:10.1158/1538-7445.AM2015-3939

Context: Anaplastic thyroid carcinoma (ATC) is an aggressive malignancy having no effective treatment. Laminin subunit gamma-2 (LAMC2) is an epithelial basement membrane protein involved in cell migration and tumour invasion and might represent an ideal target for the development of novel therapeutic approaches for ATC. Objective: Study the role of LAMC2 in ATC tumorigenesis. Design: LAMC2 expression was evaluated by RT-PCR, western blotting and immunohistochemistry in tumor specimens, adjacent non-cancerous tissues and cell lines. shRNA approach was used to investigate the effect of LAMC2 knockdown on tumorigenesis of ATC. Results: LAMC2 was highly expressed in ATC samples and cell lines compared to normal thyroid tissues. Silencing LAMC2 by shRNA in ATC cells moderately inhibited cell growth in liquid culture and dramatically decreased growth in soft agar and in xenografts growing in immunodeficient mice. Silencing LAMC2 caused cell cycle arrest and significantly suppressed migration, invasion and wound healing of ATC cells. Rescue experiments by overexpressing LAMC2 in LAMC2 knockdown cells, reversed the inhibitory effects as shown by increased cell proliferation and colony formation. Microarray data demonstrated that LAMC2 shRNA significantly altered expression of genes associated with migration, invasion, proliferation and survival. Immunoprecipitation and co-localization experiments showed that LAMC2 bound to EGFR in ATC cells. Silencing LAMC2 partially blocked EGF-mediated activation of EGFR and its downstream pathway. Interestingly, cetuximab (EGFR blocking antibody) or EGFR siRNA additively enhanced the anti-proliferative activity of the LAMC2 knockdown ATC cells compared to control cells. Conclusions: To our knowledge, this is the first report investigating the effect of LAMC2 on cell growth, cell cycle, migration, invasion and EGFR signaling in ATC cells, suggesting that LAMC2 may be a potential therapeutic target for treatment of ATC. Citation Format: Manoj Garg, Deepika Kanojia, Ryoko Okamoto, Vikas Madan, Wenwen Chien, Abhishek Sampath, Ling-Wen Ding, Meng Xuan, Jonathan W Said, Ngan Doan, Li-Zhen Liu, Henry Yang, Sigal Gery, Gleen D Braunstein, H.Phillip Koeffler. Laminin-5 gamma-2 (LAMC2) is highly expressed in anaplastic thyroid carcinoma and is associated with tumor progression, migration and invasion by modulating signaling of EGFR. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5570. doi:10.1158/1538-7445.AM2014-5570

Background Esophageal squamous cell carcinoma (ESCC) is a world-wide prevalent cancer, which is particularly common in certain regions of Asia. However, the genetic alterations of this disease have not been fully described. Methods We determined the mutational landscape of ESCC through whole-exome or targeted deep sequencing of 139 tumor/germline pairs. The somatic copy number variations (SCNV) of over 180 ESCC tumors were analyzed by SNP-array or array-CGH. Candidate driver mutations were mathematically calculated by the most updated algorithm MutSigCV, and the protein expression of which were thoroughly evaluated by immunohistochemistry. Finally, biological effects of candidate drivers were examined through functional studies in vitro and in vivo. Results Deep sequencing and computational analysis identified frequent and novel driver mutations affecting genes such as FAT1 (11.5%), ZNF750 (7.2%), EP300 (7.9%) and MLL2 (18.0%). FAT1 and ZNF750 genes were homozygouly deleted in 3.5% and 3.4% ESCCs, respectively, and their RNA transcripts and proteins were expressed at much lower levels in primary tumors compared to adjacent normal esophageal epithelial. Importantly, silencing of wild-type FAT1 or ZNF750 expression with siRNA/shRNAs significantly promoted cell proliferation and decreased cell differentiation, whereas ectopic expression of these genes resulted in opposite biological consequences. These genetic alterations and biological evidence strongly suggest that FAT1 and ZNF750 likely encode tumor suppressors which are frequently disrupted in ESCC. Furthermore, by integrating the landscape of somatic mutation, SCNV and protein dysregulation, we proposed that RTK-PI3K-MAPK pathways, cell cycle and epigenetic regulation are frequently altered by multiple molecular mechanisms in esophageal tumors. Finally, our systematic approaches uncovered many novel druggable candidates in this disease, such as FGFR1, ALK and XPO1. Conclusions By demonstrating the genomic landscape of over 180 ESCC cases, our integrated study unmasks a number of novel genetic lesions in ESCC and provides an important molecular foundation for understanding esophageal tumors and developing therapeutic targets. Citation Format: Dechen Lin, Xuan Meng, Liang Xu, Lingwen Ding, Manoj Garg, Henry Yang, Lizhen Liu, Jiajie Hao, Mingrong Wang, Yasunobu Nagata, Yusuke Sato, Yusuke Okuno, Seishi Ogawa, Phillip Koeffler. Comprehensive molecular characterization of esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2225. doi:10.1158/1538-7445.AM2014-2225