bluuebunny/crossref_metadata_2025 huggingface dataset:
The Challenge of Prenatal Diagnostic Work-Up of Maternally Inherited X-Linked Opitz G/BBB: Case Report and Literature Review
Authors: Spinelli Marialuigia , Sica Carmine , Dallapiccola Bruno , Novelli Antonio , Di Meglio Letizia , Martinelli Pasquale
Published: May 2015

Background. Prenatal diagnosis of Optiz G/BBB syndrome (OS) is challenging because the characteristic clinical features, such as facial and genitourinary anomalies, may be subtle at sonography and rather unspecific. Furthermore, molecular testing of the disease gene is not routinely performed, unless a specific diagnosis is suggested.Method. Both familial and ultrasound data were used to achieve the diagnosis of X-linked OS (XLOS), which was confirmed by molecular testing ofMID1gene (Xp22.3) at birth.Results. Sequencing ofMID1gene disclosed the nucleotide change c.1285 +1 G>T, previously associated with XLOS.Conclusions. This case illustrates current challenges of the prenatal diagnostic work-up of XLOS and exemplifies how clinical investigation, including family history, and accurate US foetal investigations can lead to the correct diagnosis.
Bone Loss Triggered by the Cytokine Network in Inflammatory Autoimmune Diseases
Authors: Amarasekara Dulshara Sachini , Yu Jiyeon , Rho Jaerang
Published: May 2015

Bone remodeling is a lifelong process in vertebrates that relies on the correct balance between bone resorption by osteoclasts and bone formation by osteoblasts. Bone loss and fracture risk are implicated in inflammatory autoimmune diseases such as rheumatoid arthritis, ankylosing spondylitis, inflammatory bowel disease, and systemic lupus erythematosus. The network of inflammatory cytokines produced during chronic inflammation induces an uncoupling of bone formation and resorption, resulting in significant bone loss in patients with inflammatory autoimmune diseases. Here, we review and discuss the involvement of the inflammatory cytokine network in the pathophysiological aspects and the therapeutic advances in inflammatory autoimmune diseases.
Left Diaphragmatic Herniation following Orthotopic Liver Transplantation in an Adult
Authors: Rosat Adriá , Alonso Ayaya , Padilla Javier , Sanz Pablo , Varona M. Aránzazu , Méndiz Javier , Moneva Enrique , Barrera Manuel
Published: May 2015

Diaphragmatic herniation is an uncommon complication in the postquirurgic follow of the liver transplant. The associated symptoms are unspecific and may not suggest the correct diagnosis. It may explain why in many patients the diagnosis remains unmade or it is made only after a long interval of time. We present the case of a fifty-seven-year-old male who required an orthotopic liver transplant in 2010 due to a trifocal hepatocarcinoma. In postoperatory follow-up the patient showed alimentary regurgitation, vomiting, and dyspepsia. The diagnosis was made by an oesophagogastroduodenal transit with barium and an abdominal CT scan that showed a left diaphragmatic herniation with the gastric fundus into the thorax. With these findings we decided to perform a programmed surgery. After takedown of adhesions and replacement of the stomach into the upper abdomen, the palm-sized diaphragmatic opening was closed with a synthetic material. The patient’s condition remained stable throughout the entire operation. The postoperative course was uneventful and he was discharged at the fifth day after surgery with a normal digestive intake. In a 12-month follow-up the patient shows no symptoms.
SnO2‐Based Nanomaterials: Synthesis and Application in Lithium‐Ion Batteries and Supercapacitors
Authors: Zhao Qinqin , Ma Lisha , Zhang Qiang , Wang Chenggang , Xu Xijin
Published: May 2015

Tin dioxide (SnO2) is an important n‐type wide‐bandgap semiconductor, and SnO2‐based nanostructures are presenting themselves as one of the most important classes due to their various tunable physicochemical properties. In this paper, we firstly outline the syntheses of phase‐pure SnO2 hierarchical structures with different morphologies such as nanorods, nanosheets, and nanospheres, as well as their modifications by doping and compositing with other materials. Then, we reviewed the design of SnO2‐based nanostructures with improved performance in the areas of lithium‐ion batteries (LIBs) and supercapacitors.
Transitioning the Adolescent with IBD from Pediatric to Adult Care: A Review of the Literature
Authors: Bollegala Natasha , Nguyen Geoffrey C.
Published: May 2015

The incidence of inflammatory bowel disease (IBD), comprising Crohn’s disease (CD) and ulcerative colitis (UC), has increased in pediatric populations over the last decade. Patients diagnosed during childhood often survive well into adulthood, and therefore their healthcare requires transfer to an adult gastroenterologist, usually at age 18 years. Transition has been defined in the literature as the “purposeful planned movement of adolescents and young adults with chronic conditions from child-centered to adult-oriented health care systems” (Blum et al., 1993). The purpose of this review is to establish the current state of knowledge regarding the transition from pediatric to adult care in IBD. This review highlights that developmentally appropriate transitional care is now recognized as a healthcare priority and thoughtful targeted intervention is needed.
Emergence of Carbapenem-ResistantKlebsiella pneumoniae: Progressive Spread and Four-Year Period of Observation in a Cardiac Surgery Division
Authors: Lombardi Fortunata , Gaia Paola , Valaperta Rea , Cornetta Maria , Tejada Milvana Rosa , Di Girolamo Luca , Moroni Alessandra , Ramundo Federica , Colombo Alessio , Valisi Massimiliano , Costa Elena
Published: May 2015

Frequent use of carbapenems has contributed to the increase toK. pneumoniaestrains resistant to this class of antibiotics (CRKP), causing a problem in the clinical treatment of patients. This investigation reports the epidemiology, genetic diversity, and clinical implication of the resistance to drugs mediated by CRKP in our hospital. A total of 280K. pneumoniaestrains were collected; in particular 98/280 (35%) were CRKP. Sequencing analysis of CRKP isolated strains showed that 9/98 of MBL-producing strains carried theblaVIM-1gene and 89/98 of the isolates were positive forblaKPC-2. Antimicrobial susceptibility tests revealed a complete resistance to third-generation cephalosporins and a moderate resistance to tigecycline, gentamicin, and fluoroquinolones with percentages of resistance of 61%, 64%, and 98%, respectively. A resistance of 31% was shown towards trimethoprim-sulfamethoxazole. Colistin was the most active agent against CRKP with 99% of susceptibility. Clonality was evaluated by PFGE and MLST: MLST showed the same clonal type, ST258, while PFGE analysis indicated the presence of a major clone, namely, pulsotype A. This finding indicates that the prevalent resistant isolates were genetically related, suggesting that the spread of these genes could be due to clonal dissemination as well as to genetic exchange between different clones.
Kinetoelastodynamics Modeling and Analysis of Spatial Parallel Mechanism
Authors: Chen Xiulong , Li Yunfeng , Deng Yu , Li Wenbin , Wu Haibin
Published: May 2015

The nonlinear elastodynamic modeling and analysis of the 4-UPS-UPU spatial 5-degree-of-freedom parallel mechanism are investigated. The kinetoelastodynamics theory is used to derive the elastic dynamic equations of 4-UPS-UPU spatial parallel mechanism. In order to grasp the effect of geometric nonlinearity on dynamic behaviors, such as displacement error output, velocity error output, acceleration error output, stress of driving limbs, and natural frequencies, the variations of dynamic behaviors considering geometric nonlinearity and without considering geometric nonlinearity are discussed, respectively. The numerical simulation results show the nonlinear elastodynamic model established can reasonably reflect the dynamic behaviors of 4-UPS-UPU spatial parallel mechanism with flexible driving limbs. And geometric nonlinearity is demonstrated to have significant impact on dynamic response and dynamic characteristics of spatial parallel mechanism. The researches can provide important theoretical base for the optimal design of spatial parallel mechanism.
Abstract 1470: The association of miR-21, HER-2/neu, and PTEN expression and clinical outcome of breast cancer
Authors: Lin Hsiao-Ching , Cheng Ya-Wen , Hsu Nan-Yung
Published: May 2015

Objective: Over the last decade, the incidence of breast cancer increased year by year, and the mortality rate is the top of female cancer in Taiwan. Previous study showed that overexpression of miR-21 was associated with recurrence in breast cancer. The aim of this study was to analyze the association of miR-21 expression and clinic pathologic data and to investigate whether the effect of miR-21 through PTEN pathway.

Materials and Methods: A total of 120 female breast cancer patients, who were enrolled at a single medical center in Taichung. MiR-21, PTEN, hormone receptor (estrogen receptor,progesterone receptor) and

HER-2 (human Epidermal Growth Factor Receptor 2) receptor were analyzed by quantitative RT-PCR. The function of invasion and migration of four breast cancer cell lines (T-47D, MDA-MB-231, MCF-7 and BT-474) were analyzed by invasion and migration assay and wound healing assay.

Results: Our data showed that no significantly difference was found between miR-21 and PTEN. MiR-21 expression was positive correlated with HER-2/neu mRNA levels (P=0.038). Patients with high miR-21 level had poor disease-free survival (p=0.047) compared with low miR-21 level patients. In addition, patients with high PTEN level had poor disease-free survival (P=0.024). We also found that patients with miR-21 high/ HER-2/neu high or miR-21 high/ PTEN high had poor disease-free survival (miR-21/HER-2/neu, p=0.009; miR-21/PTEN, p=0.003). Thus, we suggested that the expressions of miR-21, HER-2/neu and PTEN play important roles in prognosis and associated with recurrence in breast cancer. These results were also confirmed in cell model experiments.

Conclusion: In this study, we found that the expression of miR-21, HER-2/neu, and PTEN were associated with the recurrence of breast cancer in Taiwan. Thus, we suggested that the other pathways maybe involved in miR-21 mediated pathogenesis of breast cancer.

Citation Format: Hsiao-Ching Lin, Ya-Wen Cheng, Nan-Yung Hsu. The association of miR-21, HER-2/neu, and PTEN expression and clinical outcome of breast cancer. [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 1470. doi:10.1158/1538-7445.AM2014-1470
Abstract 1513: Use of complex oligonucleotide libraries for concurrent high-resolution fluorescence imaging of both DNA and RNA in various sample types
Authors: Ach Robert A. , Tsang Peter , Scheffer-Wong Alicia , Bruhn Laurakay , Powell Weston , Engreitz Jesse , LaSalle Janine , Guttman Mitchell , Yamada Alice
Published: May 2015

Fluorescence in situ Hybridization (FISH) is a powerful technique for determining the localization specific nucleic acid sequences within individual cells. Previously, the use of FISH has often been dependent upon access to cloned template DNA for the generation of probes, which can be difficult if clones for specific regions are unavailable, or if the genomic region of interest contains repetitive and/or other problematic sequences. We have developed the ability to chemically synthesize DNA in massively parallel reactions, which we have used to produce libraries of oligonucleotides up to 200 bases in length that can be utilized for the generation of FISH probes. The sequences of the oligonucleotides in these libraries are selected in silico using empirically determined criteria so as to avoid repetitive elements or regions homologous to other non-targeted loci. We have found that these oligonucleotide library-derived FISH probes can detect human genomic regions as small as 1.8 kb and as large as whole chromosomes in both metaphase and interphase cells, using the same simple assay protocol. Because of the inherent flexibility in our probe design methods, we can readily visualize regions rich in repeats and/or GC content. We have also used these oligonucleotide library-derived FISH probes to detect the localization of a variety of both coding and non-coding RNAs in fixed tissue culture cells and formalin-fixed paraffin-embedded tissue sections, using both conventional fluorescence and structured illumination microscopy. Simultaneous hybridization of FISH probes labeled with different fluorophores enables visualization of multiple sequences at once. Using probes designed specifically to transcribed vs. non-transcribed regions has enabled the simultaneously detect DNA and RNA from the same locus, or from two different loci, in the same FISH assay. The ability to generate high performance FISH probes using chemically synthesized oligo libraries that can simultaneously detect DNA and RNA yields a valuable tool for studies of how localization of specific nucleic acids impacts biological function.

Citation Format: Robert A. Ach, Peter Tsang, Alicia Scheffer-Wong, Laurakay Bruhn, Weston Powell, Jesse Engreitz, Janine LaSalle, Mitchell Guttman, Alice Yamada. Use of complex oligonucleotide libraries for concurrent high-resolution fluorescence imaging of both DNA and RNA in various sample types. [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 1513. doi:10.1158/1538-7445.AM2014-1513
Abstract 1529: Ovarian low grade serous cancer: Mutation analysis with a comprehensive 409 cancer gene panel
Authors: Tsang Yvonne T. , Izaguirre Daisy , Kwan Suet-Yan , Mok Samuel C. , Gershenson David , Wong Kwong-Kwok
Published: May 2015

Patients with ovarian serous cancer are usually younger and survive longer than patients with high-grade serous cancer. Unfortunately, most low-grade serous cancer patients eventually die of the disease because recurrent low-grade serous carcinoma is relative chemoresistant. So far, the known mutations identified in low-grade ovarian serous carcinomas and their putative precursor ovarian serous borderline tumors are BRAF and KRAS mutations. BRAF and KRAS mutations together have been identified in approximately 60% of serous borderline tumors and early stage low-grade serous carcinomas. However, BRAF mutation is rare in advanced-stage low-grade serous carcinomas. To further investigate the molecular pathogenesis of low-grade serous carcinoma, mutation analyses of 409 cancer related genes were performed in 21 low-grade ovarian serous carcinoma, 8 ovarian serous borderline tumors and one ovarian cystadenoma using next generation sequencer. A total of nineteen missense mutations were identified in 20 samples but ten samples had no detectable mutations. BRAF mutation was detected in 3 serous borderline tumors and one low-grade serous carcinoma. KRAS mutations were detected in three low-grade serous carcinomas. Other than BRAF and KRAS mutations, we also detected other prevalent mutations: ATRX mutations were detected in 3 low-grade ovarian serous carcinomas and a cystadenoma; KMT2C mutations were detected in 3 low-grade ovarian serous carcinomas; ATM mutations were detected in 2 low-grade ovarian serous carcinomas. Interestingly, both ATRX and MLL3 genes are involved in the chromatin remodeling. Moreover, both ATRX and ATM are involved in DNA damaging response. Thus, the pathogenesis of low-grade serous carcinoma may involve de-regulation of chromatin remodeling pathway and DNA damaging response. Further analysis of a large cohort of low-grade serous carcinomas for these prevalent mutations is ongoing.

Citation Format: Yvonne TM Tsang, Daisy Izaguirre, Suet-Yan Kwan, Samuel C. Mok, David Gershenson, Kwong-Kwok Wong. Ovarian low grade serous cancer: Mutation analysis with a comprehensive 409 cancer gene panel. [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 1529. doi:10.1158/1538-7445.AM2014-1529
Abstract 1535: Immunohistochemistry predicts presence and type of TP53 mutation in high-grade serous carcinoma
Authors: Köbel Martin , Piskorz Anna , Liu Shuhong , Brenton James D.
Published: May 2015

TP53 mutations are ubiquitous to ovarian high-grade serous carcinomas (HGSC) but uncommon in other types such as endometrioid carcinomas. Despite the advances in direct sequencing, this technology is not widely available for routine diagnostics. Significant improvements in standardization of immunohistochemistry (IHC) have dramatically improved sensitivity. We hypothesized that the pattern of p53 expression by IHC can predict the presence and group of TP53 mutations: complete absence indicating null mutations versus overexpression indicating missense mutation.

DNA extracted from 91 fresh frozen HGSC samples obtained from the Canadian Ovarian Experimental Unified Resource (COEUR) and from 66 formalin fixed paraffin embedded HGSC samples obtained from Calgary Laboratory Services were sequenced using TAmSeq methods for NGS. Additional Sanger sequencing was performed for confirmation or for identification of indel mutations. Accompanying tissue microarrays with 0.6 mm cores were constructed and immunohistochemistry for p53 was performed on a Leica Bond MAX platform using the DO-7 monoclonal antibody. The expression of p53 was scored in a 3-tier system: complete absence, wild type (any staining from 1-70%), overexpression (>70% strong nuclear staining). Cytoplasmic localization was noted separately.

TP53 mutations were detected in 147/157 HGSC (94%). The type of mutation in descending order were missense 104/147 (71%), frameshift 17/147 (12%), stopgain 13/147 (9%), and splice site 13/147 (13%). 101/104 cases with TP53 missense mutations showed p53 overexpression (98% concordance) while 12/17 cases with frameshift, 11/13 cases with stopgain, and 7/13 with splicing (combined 70% concordance) showed the expected complete absence of p53 expression. The overall sensitivity of p53 IHC to predict the group of mutation was 89% (131/147). Only 6/147 (4%) cases with TP53 mutations showed a p53 wild type pattern, four those were splice site mutations, hence the sensitivity of p53 IHC to predict the presence of mutation is 94%. Interestingly, 7/10 cases with TP53 wild type or synonymous silent mutations showed aberrant p53 expression as either complete absence or overexpression, suggesting alternative abnormalities in the p53 pathway. The three cases with TP53 wild type and p53 wild type expression may not be HGSC.

We conclude that contemporary p53 immunohistochemistry is a useful, widely accessible, inexpensive complimentary test for direct TP53 sequencing.

Citation Format: Martin Köbel, Anna Piskorz, Shuhong Liu, James D. Brenton. Immunohistochemistry predicts presence and type of TP53 mutation in high-grade serous 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 1535. doi:10.1158/1538-7445.AM2014-1535
Abstract 1559: TTC36, a novel chaperone of heat shock protein 70, functions as a tumor suppressor in hepatocellular carcinoma
Authors: Jiang Lingxi , Liu Ming , Li Yan , Guan Xinyuan
Published: May 2015

Hepatocellular carcinoma (HCC) is one of the most devastating human diseases characterized by high incidences and mortality rates. However, there is still limited knowledge of molecular mechanisms leading to the development of HCC. By using transcriptome sequencing profiling of non-tumor and HCC samples, a subset of differentially expressed genes have been identified. One of those genes, the TTC36 gene is down-regulated in HCC tissues compared with non-tumor tissues. Reduced expression of TTC36 is correlated with advanced tumor stages and decreased overall survival. Ectopic expression of TTC36 in HCC cells show decreased tumorigenic ability in vitro and vivo function assays. Silencing TTC36 expression with short hairpin RNA had the opposite effects. We had found that TTC36 promoted apoptosis of HCC cells by interacting with heat shock protein 70 and induced the activation of caspases-9, caspase-3 and PARP. Further characterization of TTC36 may provide a novel prognostic biomarker for HCC and shed light on better management of HCC treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Lingxi Jiang, Ming Liu, Yan Li, Xinyuan Guan. TTC36, a novel chaperone of heat shock protein 70, functions as a tumor suppressor in hepatocellular 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 1559. doi:10.1158/1538-7445.AM2014-1559
Abstract 1572: Dystrophin Is a tumor suppressor in human cancers with myogenic programs
Authors: Wang Yuexiang , Marino-Enriquez Adrian , Bennett Richard , Zhu Meijun , Eilers Grant , Antonescu Cristina , Fletche Christopher , Raut Chandrajit , Rijn Matt van de , Kunkel Louis , Demetri George , Fletcher Jonathan
Published: May 2015

Many common human mesenchymal tumors, including gastrointestinal stromal tumor (GIST), rhabdomyosarcoma (RMS), and leiomyosarcoma (LMS), feature myogenic differentiation. Although presumptive initiating mutations have been identified in these cancers, the subsequent mechanisms of malignant progression are not known. Here we report that intragenic deletion of the dystrophin-encoding and muscular dystrophy-associated DMD gene is a frequent mechanism by which myogenic tumors progress to high-grade, lethal sarcomas. Genome-wide SNP screens demonstrated DMD intragenic deletions in 23 of 38 myogenic sarcomas (61%) compared to 0 of 58 non-myogenic sarcomas (p < 0.0001). Although DMD is an X-linked gene, somatic DMD deletions affected both sexes equally and DMD deletions in female patients involved the active X chromosome, resulting in functional DMD nullisomy. Dystrophin was expressed strongly in nonneoplastic and benign counterparts for GIST, RMS and LMS, and the DMD deletions in the malignant myogenic sarcomas clustered at the 5′ end of the gene, inactivating larger dystrophin isoforms, including 427kDa dystrophin, while preserving expression of an essential 71kDa isoform (Dp71) which is encoded by the DMD 3′ end. Dystrophin restoration inhibited myogenic sarcoma cell migration, invasion, anchorage independence, and invadopodia formation, and dystrophin inactivation was found in 96%, 100%, and 62% of metastatic GIST, embryonal RMS, and LMS, respectively. The genomic, clinicopathological and functional evidence validate dystrophin as a tumor suppressor and likely anti-metastatic factor, suggesting that therapies in development for muscular dystrophies may also have relevance in treatment of cancer.

Citation Format: Yuexiang Wang, Adrian Marino-Enriquez, Richard Bennett, Meijun Zhu, Grant Eilers, Cristina Antonescu, Christopher Fletche, Chandrajit Raut, Matt van de Rijn, Louis Kunkel, George Demetri, Jonathan Fletcher. Dystrophin Is a tumor suppressor in human cancers with myogenic programs. [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 1572. doi:10.1158/1538-7445.AM2014-1572
Abstract 1591: Src is activated in cervical adenocarcinoma
Authors: Nishimura Masato , Tanimura Natsumi , Kawakita Takako , yoshidaa Kanako , Irahara Minoru
Published: May 2015

Src family tyrosine kinases are important factors for cell growth. Src is reported to be activated in ovarian mucinous adenocarcinoma, and correlates with oxaliplatin resistance. Cervical adenocarcinoma is thought to have worse prognosis compared with squamous cell carcinoma due to low response to chemotherapy and radiotherapy. So new therapy has been expected for cervical adenocarcinoma. We examined src expression by immunohistochemistry in cervical adenocarcinoma treated with surgery during 2001 to 2011 and association with clinico pathological findings. 36 invasive cancers and 1 adenocarcinoma in situ (AIS) and 12 normal cervical glands were examined. In invasive cancer, src was activated in 16/36 (44%) cases, especially in 9/15 (60%) of mucinous type. Src was also activated in AIS, but not in normal cervical glands. The frequency of parametrial invasion and lymphnode metastasis in Src positive and negative cases were 1/15 (7%) versus 1/18 (6%) and 2/15 (13%) versus 3/20 (15%) in respectively. There were no statistical differences between src positive and negative cases. 3 cases in src positive and 1 case in src negative recurred. Moreover, src was activated in I case of AIS, but not activated in adjacent normal gland. In summary, src activation is an early event in the carcinogenesis of cervical adenocarcinoma, and has possibility to be a new target in the treatment of cervical adenocarcinoma.

Citation Format: Masato Nishimura, Natsumi Tanimura, Takako Kawakita, Kanako yoshidaa, Minoru Irahara. Src is activated in cervical adenocarcinoma. [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 1591. doi:10.1158/1538-7445.AM2014-1591
Abstract 1593: GSK3β mediated arsenite induction of spindle abnormalities
Authors: Lin Tz-Chi , Chen Ren-Meei , Kuo Hsiao-Hui , Yih Ling-Huei
Published: May 2015

Arsenite induces centrosome amplification and spindle abnormalities, leading to chromosome missegregation and consequently resulting in mitotic cell apoptosis or aneuploidy. However, the underlying mechanism of how arsenite disrupts mitotic spindles is not known. In this study, we showed that CGL2 cells treated with arsenic trioxide (ATO) were arrested at mitotic stage in company with the formation of substantial spindle abnormalities and significant reduction of acetylated- and detyrosinated-tubulin, characteristics of decreased MT stability. Immunofluorescence staining of EB1, a plus-end microtubule-binding protein, revealed discrete EB1 spots distributing along microtubule (MT) nucleated from the two spindle poles in untreated mitotic cells. However, it showed a disorganized pattern in ATO-arrested mitotic cells with the spot-like EB1 clusters splashing all over the cells. In addition, immunoblotting and immunofluorescence staining revealed that the mitosis-specific inhibitory phospho-GSK3 was diminished in ATO-arrested mitotic cells. Pharmacological inhibition of GSK3 could prevent ATO induction of spindle abnormalities, mitotic arrest, and cell death. Since GSK3β has been reported to reduce MT stability by phosphorylating several microtubule-associated proteins, our current results indicated that GSK3β may mediate ATO induction of spindle abnormalities.

Citation Format: Tz-Chi Lin, Ren-Meei Chen, Hsiao-Hui Kuo, Ling-Huei Yih. GSK3β mediated arsenite induction of spindle abnormalities. [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 1593. doi:10.1158/1538-7445.AM2014-1593
Abstract 1602: Clobetasol treatment of vulvar carcinoma cell lines influences growth rates
Authors: Lewis Jani E. , Mehta Mehek , Treichel Alison , Colligan Sean , Nguyen Vinh , Foster David C.
Published: May 2015

Vulvar cancer is an aggressive cancer often masked by diseases such as the vulvar rash known as lichen sclerosis (LS). Ultrapotent corticosteroids such as clobetasol are commonly used to treat LS but there is debate over the safety of long-term clobetasol use and development of vulvar cancer. Growing cells from LS biopsies has proven difficult so our objective was to decipher if clobetasol promotes changes in vulvar cancers in vitro that parallel the formation of more aggressive cancer types. Previously we have shown that clobetasol actually promotes transformation of one vulvar cancer cell line into a more aggressive phenotype. Our present studies used MTT assays to examine the effect that short-term (72 hours) and long-term (4 weeks) clobetasol treatment had on the growth rate of vulvar cancer cell lines. Our preliminary data indicates that the growth response varies between cell lines with at least two cell lines displaying increased cell growth after extended treatment with clobetasol when compared to controls.

Citation Format: Jani E. Lewis, Mehek Mehta, Alison Treichel, Sean Colligan, Vinh Nguyen, David C. Foster. Clobetasol treatment of vulvar carcinoma cell lines influences growth rates. [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 1602. doi:10.1158/1538-7445.AM2014-1602
Abstract 1616: Phosphoprotein enrichment pathway analysis tools for studying cancer signaling
Authors: Wiredja Danica , Liu Yu , Schlatzer Daniela , Gokulrangan Giridharan , Narla Goutham , Chance Mark
Published: May 2015

Cancer development and progression is driven by dysregulated signaling transduction, which is predominantly regulated by serine, threonine, and tyrosine phosphorylation. Consequently, MS-based phosphoproteomics, which can identify and quantitate thousands of phosphopeptides, provides a powerful quantitative tool to detect and comprehensively catalog alterations in cancer signaling cascades that cannot be captured by analysis of DNA or RNA alone. Unfortunately, the complexity of the datasets generated by these advanced proteomic methodologies are still not matched with well-established or fully tested bioinformatics tools to extract critical pathways from these high-dimensional phosphoprotein data.

Here, we present two approaches for identifying pathways with altered phosphorylation patterns: Protein Set Enrichment Analysis (PSEA) and Protein Interaction Enrichment Network Analysis (PIENA). Development of these tools was inspired by two published methods utilizing gene expression datasets to identify disrupted gene sets: Gene Set Enrichment Analysis (GSEA) and Gene Interaction Enrichment Network Analysis (GIENA), respectively. PSEA, through pathway scoring, extracts pathways that show a uniform directional phosphorylation change through all protein members of that set. As in the traditional GSEA method, an enrichment score is calculated for each pathway based on intensity values from all detected phosphopeptides. The second method, PIENA, looks for notable differences in phosphorylation between phosphopeptide pairs. Ultimately, this approach identifies pathways enriched with altered phosphoprotein interactions.

Both methods have been used to identify changes in signaling states for two lung adenocarcinoma cell lines, A549 and H358, treated with either control or a novel phosphatase activator with anti-cancer properties. Both PSEA and PIENA identify pathways classically implicated in cancer, including p53, mitosis and cell cycle progression, insulin secretion, immune regulation, and RAS signaling cascades. Furthermore, the ERK pathway is consistently found to be significantly disrupted in both A549 and H358. This corroborates biological findings that this novel drug has activity against KRAS mutated cell lines. Although future cellular experiments will help to confirm the detailed mechanism of this drug's effects, our current results demonstrate the advantage of these new pathway identification tools in providing unbiased and global identification of key drivers of signaling.

Citation Format: Danica Wiredja, Yu Liu, Daniela Schlatzer, Giridharan Gokulrangan, Goutham Narla, Mark Chance. Phosphoprotein enrichment pathway analysis tools for studying cancer signaling. [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 1616. doi:10.1158/1538-7445.AM2014-1616
Abstract 1628: Direct anti-cancer effects of zoledronic acid on human cancer cell
Authors: Jiang Pengfei , Mukthavavam Rajesh , Nomura Natsuko , Pingle Sandeep C. , Kesari Santosh
Published: May 2015

Zoledronic Acid (ZA) is a potent nitrogen-containing bisphosphonates (NBP) has been extensively used to limit bone turnover in a various diseases including tumors. Recently, clinical trails of Zoledronic Acid demonstrated direct anti-cancer effects as well as the known skeletal-related events. We investigated the effect of 4 NBPs on human tumor cells proliferation and found ZA is most potent for GBM cell, breast cell and GBM patients' stem-cell-like cells. ZA also effectively inhibited GBM tumor growth in mouse model. ZA stimulated strong autophagy but not apoptotic signals in all tested cells. One intermediate product of cholesterols synthesis pathway—-geranylgeranyl diphosphate (GGPP) rescued cell from the cytotoxicity of ZA. Knock-down RABGGTA, which encoded a subunit of the Rab geranylgeranyltransferase proteins, induced a similar effect as Zoledronic Acid in cancer cell lines. These data suggested great potential for Zoledronic Acid against human cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Pengfei Jiang, Rajesh Mukthavavam, Natsuko Nomura, Sandeep C. Pingle, Santosh Kesari. Direct anti-cancer effects of zoledronic acid on human cancer cell. [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 1628. doi:10.1158/1538-7445.AM2014-1628
Abstract 163: Identification of a bone marrow-derived MDSC subset that promotes the growth of colon carcinoma liver metastases
Authors: Ham Boram
Published: May 2015

The hepatic microenvironment can play an important role in promoting liver metastasis. We and others have shown that when metastatic carcinoma cells enter the liver, a rapid inflammatory response mediated by TNF-α is triggered, enabling tumor transendothelial migration and metastasis. TNF-α binds to the cell surface through two receptors; the p55/p60 TNFR1 and the p75/p80 TNFR2. To elucidate the roles of these receptors in the inflammatory cascade and in liver colonization by tumor cells, we used mice with tnfr1, tnfr2 and combined tnfr1/tnfr2 gene deficiencies and investigated parameters of the host response and the outcome of metastasis in these mice using the highly metastatic mouse colon carcinoma MC-38 and lung carcinoma H-59 cells. We found that experimental liver metastasis following intrasplenic/portal inoculation of the tumor cells was markedly reduced in TNFR2-/- and even further in TNFR1-/-/TNFR2-/- female mice as compared to wild-type controls but no reduction was observed in TNFR1-/- mice. To identify the underlying mechanism(s), we investigated changes in the liver microenvironment following tumor inoculation using a combination of immunohistochemistry, confocal microscopy, qRT-PCR, and cytokine/chemokines profiling. We found that while in wild-type and TNFR1-/- mice, VCAM-1 expression on the sinusoidal endothelium was significantly upregulated within 16 hr post tumor injection, it was unchanged in TNFR2-/- and double negative female mice. In addition, qRT-PCR analysis revealed that the production of the neutrophil chemokine CXCL1, while upregulated in control mice, was suppressed in TNFR2-/- mice. These changes were associated with a parallel decrease in Ly6G+ neutrophil recruitment into extravascular, pro-metastatic niches 16-48 hr post tumor inoculation. Interestingly, these reductions were not observed in male TNFR2-/- mice and correspondingly, their metastatic burden was not reduced relative to control mice. Taken together, the results suggest that TNFR2 plays a critical and gender-specific role in Ly6G+ cell recruitment into the tumor-microenvironment by regulating sinusoidal endothelial VCAM-1 expression and chemokine production. In turn, the recruitment of these cells into extravascular pro-metastatic niches appears to promote liver metastasis.

Citation Format: Boram Ham. Identification of a bone marrow-derived MDSC subset that promotes the growth of colon carcinoma liver metastases. [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 163. doi:10.1158/1538-7445.AM2014-163
Abstract 1640: A novel small molecule inhibitor of lysophosphatidic acid acyltransferase-beta inhibits pancreatic cancer cell proliferation and mTOR signaling
Authors: Blaskovich Michelle A. , Luo Yunting , Ge Yiyu , Sebti Saïd M. , Lawrence Harshani R. , Lawrence Nicholas J. , Springett Gregory M.
Published: May 2015

Lysophosphatidic acid acyltransferase (LPAAT-β), a phosphatidic acid (PA)-generating enzyme, plays an essential role in triglyceride synthesis. A less familiar role for LPAAT-β is that of a regulator of cell growth and differentiation by means of the function of its product, PA, which plays a vital role in the activation of key signal transduction proteins including Raf, PKC-ζ and mTOR. It is in this role that LPAAT-β presents as a potential therapeutic target in cancer. Accordingly, we developed a library of small molecule inhibitors of LPAAT-β with nanomolar activity against LPAAT-β enzyme activity in vitro (2012 Annual Meeting, Abstract #2470). We have now performed additional SAR and lead optimization to produce a potent small molecule that inhibits proliferation, mTOR signaling pathways, and total cellular PA levels at low micromolar concentrations. The lead compound also inhibits anchorage-independent growth of pancreatic cancer cell lines at nanomolar concentrations. This compound demonstrates effects on cellular signaling that are similar to the effects of LPAAT-β siRNA.

Citation Format: Michelle A. Blaskovich, Yunting Luo, Yiyu Ge, Saïd M. Sebti, Harshani R. Lawrence, Nicholas J. Lawrence, Gregory M. Springett. A novel small molecule inhibitor of lysophosphatidic acid acyltransferase-beta inhibits pancreatic cancer cell proliferation and mTOR signaling. [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 1640. doi:10.1158/1538-7445.AM2014-1640
Abstract 1658: Enhanced shedding of extracellular vesicles from amoeboid prostate cancer cells: Potential effects on the tumor microenvironment
Authors: Kim Jayoung , Morley Samantha , Le Minh , Bedoret Denis , Umetsu Dale , Vizio Dolores D. , Freeman Michael
Published: May 2015

The gene encoding the cytoskeletal regulator DIAPH3 is lost at high frequency in metastatic prostate cancer, and silencing of DIAPH3 evokes a transition to an amoeboid phenotype in multiple tumor cell backgrounds. This amoeboid transformation is accompanied by increases in plasma membrane deformations (blebbing), cell migration, invasion, and metastasis. DIAPH3 silencing also promotes the shedding of atypically large (>1μm) extracellular vesicles (EV) containing bioactive cargo. Whether loss of DIAPH3 also stimulates the release of bioactive nano-sized EV (exosomes) is not established. Here we examined the mechanism of release and potential biological functions of EV shed from DIAPH3-silenced and other prostate cancer cells. We observed that stimulation of LNCaP cells with the prostate stroma-derived growth factor heparin-binding EGF-like growth factor (HB-EGF), combined with p38MAPK inhibition caused exosome shedding, a process mediated by ERK1/2 hyperactivation. DIAPH3 silencing in DU145 cells also increased rates of exosome production. EV isolated from DIAPH3-silenced cells activated AKT1 and androgen signaling, increased proliferation of recipient tumor cells, and suppressed proliferation of human macrophages and peripheral blood mononuclear cells (PBMC). DU145-derived EV contained miR-125a, which suppressed AKT1 expression and proliferation in recipient human peripheral blood mononuclear cells and macrophages. Our findings suggest that EV across a wide size range (∼70nm to >1um) are produced by amoeboid prostate cancer cells as a result of DIAPH3 loss and/or growth factor stimulation. These EV may condition the tumor microenvironment through multiple mechanisms, including promoting the growth of cancer cells and suppression of tumor-infiltrating immune cells.

Citation Format: Jayoung Kim, Samantha Morley, Minh Le, Denis Bedoret, Dale Umetsu, Dolores Di Vizio, Michael Freeman. Enhanced shedding of extracellular vesicles from amoeboid prostate cancer cells: Potential effects on the tumor microenvironment. [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 1658. doi:10.1158/1538-7445.AM2014-1658
Abstract 167: Roles of adipocytes and M2-macrophages in high-fat diet-stimulated lymph node metastasis of B16F10 melanoma
Authors: Cho Han Jin , Jung Jae I. , Jung Yoo Jin , Lee Ki Won , Sung Mi-Kyung , Park Jung Han Yoon
Published: May 2015

The tumor microenvironment consists of a number of distinct cell types including tumor cells, fibroblasts, endothelial cells, immune cells, and adipocytes, and produces a wide variety of factors, which contribute to tumor progression. We previously observed that high-fat diet (HFD) feeding stimulates solid tumor growth and lymph node (LN) metastasis in mice subcutaneously injected with B16F10 melanoma cells. We also observed that HFD feeding increases the number of lipid vacuoles and M2-macrophage (M2-Mϕ)s as well as the expression of various cytokines/growth factors in tumor tissues. In the present study, we noted that HFD increased the expression of CCR7 in the tumor. The concentrations of CCL19 and CCL21, the CCR7 ligands, were significantly higher in the LN than those in tumor, and HFD feeding increased the concentrations of these ligands in the LN with negligible changes in the tumor tissue creating higher concentration gradients of these two ligands between the LN and tumor. In order to explore the interactions between tumor cells, adipocytes, lymphatic endothelial cell (LEC)s, and/or M2-Mϕs, we established in vitro co-culture models involving 3 distinct cell types. The mRNA levels of cytokines and growth factors were increased in B16F10 cells and M2-Mϕs when B16F10 cells, M2-Mϕs, and/or mature adipocyte (MA)s were co-cultured in a transwell system. Among these cytokines/growth factors, the mRNA levels of MCP-1 were synergistically increased in B16F10 cells when co-cultured with M2-Mϕs and MAs. The migration of monocytes was tremendously elevated when media conditioned by B16F10 cells/M2-Mϕs/MAs co-cultures was used as a chemoattractant. Furthermore, MAs increased the expression of CCR7 mRNA in B16F10 cells and the expression of CCL19 and CCL21 mRNAs in LECs. These results indicate that in HFD-fed mice, adipocytes induce MCP-1 expression and monocyte infiltration which play important roles in solid tumor growth and LN metastasis. Additionally, adipocyte-stimulated activation of CCL19, CCL21/CCR7 axis in lymphatic endothelial cells and tumor cells helps tumor cells to enter the lymphatic vessels and increased ligand concentration gradients between the LN and tumor facilitate tumor cell migration to the LN.

Citation Format: Han Jin Cho, Jae In Jung, Yoo Jin Jung, Ki Won Lee, Mi-Kyung Sung, Jung Han Yoon Park. Roles of adipocytes and M2-macrophages in high-fat diet-stimulated lymph node metastasis of B16F10 melanoma. [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 167. doi:10.1158/1538-7445.AM2014-167
Abstract 168: Understanding the role of the carcinoid associated fibroblasts in the neuroendocrine tumor microenvironment
Authors: Bowden Michaela , Sicinska Ewa , Kulke Matthew , Loda Massimo
Published: May 2015

A lack of cell lines and animal models has been an obstacle in developing novel therapeutic agents for advanced carcinoid tumors. In the past two years we have developed a collection of short-term carcinoid cell lines. Carcinoid cells in culture grow slowly and are importantly attached to, much faster, growing stromal fibroblasts, referred to as CAF's (Carcinoid Associated Fibroblasts). This phenotype is conserved in vivo and closely mimics the status of these tumors in humans. CAF's are known to have pro-tumorigenic effects; are attractive targets for cancer therapy; as well as valuable clinical biomarkers of prognosis. With the availability of an in-house panel of primary cell lines and control cultured fibroblasts, teasing apart the tumor-fibroblast relationship is feasible through collection of cell culture supernatants and in vitro screening of secreted proteins. We present here our initial screening studies using three cell culture environments, namely normal human fibroblasts, carcinoid-associated fibroblasts (CAF's) and the tumor microenvironment co-culture of stromal cells and CAF's.

We screened 55 secreted proteins with known carcinogenic associations by multiplexed bead-based (BioPlex, BioRad) analysis. We have identified a potential 5-marker CAF signature encompassing elevated secretion of IL-6 and MCP-1; and the suppression of IL-8 and FGF basic, coupled with VEGF expression. As part of the next phase we plan to utilize our collection of banked whole blood, serum, and plasma specimens comprising over 1000 cases of patients with neuroendocrine tumors, to validate our secreted protein signature in early diagnosis and / or assessing progression and prognosis in patients with neuroendocrine tumors based on stage and tumor subtype.

Citation Format: Michaela Bowden, Ewa Sicinska, Matthew Kulke, Massimo Loda. Understanding the role of the carcinoid associated fibroblasts in the neuroendocrine tumor microenvironment. [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 168. doi:10.1158/1538-7445.AM2014-168