bluuebunny/crossref_metadata_2025 huggingface dataset:
In-Depth Global Analysis of Transcript Abundance Levels in Porcine Alveolar Macrophages Following Infection with Porcine Reproductive and Respiratory Syndrome Virus
Authors: Miller Laura C. , Neill John D. , Harhay Gregory P. , Lager Kelly M. , Laegreid William W. , Kehrli Marcus E.
Published: January 2011

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. Identifying specific cell signaling or activation pathways that associate with variation in PRRSV replication and macrophage function may lead to identification of novel gene targets for the control of PRRSV infection. Serial Analysis of Gene Expression (SAGE) was used to create and survey the transcriptome ofin vitromock-infected and PRRSV strain VR-2332-infected porcine alveolar macrophages (PAM) at 0, 6, 12, 16, and 24 hours after infection. The transcriptome data indicated changes in transcript abundance occurring in PRRSV-infected PAMs over time after infection with more than 590 unique tags with significantly altered transcript abundance levels identified (P<.01). Strikingly, innate immune genes (whose transcript abundances are typically altered in response to other pathogens or insults including IL-8, CCL4, and IL-1β) showed no or very little change at any time point following infection.
Time and Space in Digital Game Storytelling
Authors: Wei Huaxin , Bizzocchi Jim , Calvert Tom
Published: January 2011

The design and representation of time and space are important in any narrative form. Not surprisingly there is an extensive literature on specific considerations of space or time in game design. However, there is less attention to more systematic analyses that examine both of these key factors—including their dynamic interrelationship within game storytelling. This paper adapts critical frameworks of narrative space and narrative time drawn from other media and demonstrates their application in the understanding of game narratives. In order to do this we incorporate fundamental concepts from the field of game studies to build a game-specific framework for analyzing the design of narrative time and narrative space. The paper applies this framework against a case analysis in order to demonstrate its operation and utility. This process grounds the understanding of game narrative space and narrative time in broader traditions of narrative discourse and analysis.
Comparative Analysis on Applicability of Satellite and Meteorological Data for Prediction of Malaria in Endemic Area in Bangladesh
Authors: Rahman Atiqur , Roytman Leonid , Goldberg Mitch , Kogan Felix
Published: January 2011

Relationships between yearly malaria incidence and (1) climate data from weather station and (2) satellite-based vegetation health (VH) indices were investigated for prediction of malaria vector activities in Bangladesh. Correlation analysis of percent of malaria cases with Advanced Very High Resolution Radiometer- (AVHRR-) based VH indices represented by the vegetation condition index (VCI—moisture condition) and the temperature condition index (TCI—estimates thermal condition) and with rainfall, relative humidity, and temperature from ground-based meteorological stations. Results show that climate data from weather stations are poorly correlated and are not applicable to estimate prevalence in Bangladesh. The study also has shown that AVHRR-based vegetation health (VH) indices are highly applicable for malaria trend assessment and also for the estimation of the total number of malaria cases in Bangladesh for the period of 1992–2001.
Lipid Rafts: Keys to Sperm Maturation, Fertilization, and Early Embryogenesis
Authors: Kawano Natsuko , Yoshida Kaoru , Miyado Kenji , Yoshida Manabu
Published: January 2011

Cell membranes are composed of many different lipids and protein receptors, which are important for regulating intracellular functions and cell signaling. To orchestrate these activities, the cell membrane is compartmentalized into microdomains that are stably or transiently formed. These compartments are called “lipid rafts”. In gamete cells that lack gene transcription, distribution of lipids and proteins on these lipid rafts is focused during changes in their structure and functions such as starting flagella movement and membrane fusion. In this paper, we describe the role of lipid rafts in gamete maturation, fertilization, and early embryogenesis.
Animal Models of Colitis‐Associated Carcinogenesis
Authors: Kanneganti Manasa , Mino-Kenudson Mari , Mizoguchi Emiko
Published: January 2011

Inflammatory bowel disease (IBD) is a group of chronic inflammatory disorders that affect individuals throughout life. Although the etiology and pathogenesis of IBD are largely unknown, studies with animal models of colitis indicate that dysregulation of host/microbial interactions are requisite for the development of IBD. Patients with long‐standing IBD have an increased risk for developing colitis‐associated cancer (CAC), especially 10 years after the initial diagnosis of colitis, although the absolute number of CAC cases is relatively small. The cancer risk seems to be not directly related to disease activity, but is related to disease duration/extent, complication of primary sclerosing cholangitis, and family history of colon cancer. In particular, high levels and continuous production of inflammatory mediators, including cytokines and chemokines, by colonic epithelial cells (CECs) and immune cells in lamina propria may be strongly associated with the pathogenesis of CAC. In this article, we have summarized animal models of CAC and have reviewed the cellular and molecular mechanisms underlining the development of carcinogenic changes in CECs secondary to the chronic inflammatory conditions in the intestine. It may provide us some clues in developing a new class of therapeutic agents for the treatment of IBD and CAC in the near future.
Innate Immune Effectors in Mycobacterial Infection
Authors: Saiga Hiroyuki , Shimada Yosuke , Takeda Kiyoshi
Published: January 2011

Tuberculosis, which is caused by infection withMycobacterium tuberculosis(Mtb), remains one of the major bacterial infections worldwide. Host defense against Mtb is mediated by a combination of innate and adaptive immune responses. In the last 15 years, the mechanisms for activation of innate immunity have been elucidated. Toll-like receptors (TLRs) have been revealed to be critical for the recognition of pathogenic microorganisms including mycobacteria. Subsequent studies further revealed that NOD-like receptors and C-type lectin receptors are responsible for the TLR-independent recognition of mycobacteria. Several molecules, such as active vitamin D3, secretary leukocyte protease inhibitor, and lipocalin 2, all of which are induced by TLR stimulation, have been shown to direct innate immune responses to mycobacteria. In addition, Irgm1-dependent autophagy has recently been demonstrated to eliminate intracellular mycobacteria. Thus, our understanding of the mechanisms for the innate immune response to mycobacteria is developing.
High-Level Synthesis of In-Circuit Assertions for Verification, Debugging, and Timing Analysis
Authors: Curreri John , Stitt Greg , George Alan D.
Published: January 2011

Despite significant performance and power advantages compared to microprocessors, widespread usage of FPGAs has been limited by increased design complexity. High-level synthesis (HLS) tools have reduced design complexity but provide limited support for verification, debugging, and timing analysis. Such tools generally rely on inaccurate software simulation or lengthy register-transfer-level simulations, which are unattractive to software developers. In this paper, we introduce HLS techniques that allow application designers to efficiently synthesize commonly used ANSI-C assertions into FPGA circuits, enabling verification and debugging of circuits generated from HLS tools, while executing in the actual FPGA environment. To verify that HLS-generated circuits meet execution timing constraints, we extend the in-circuit assertion support for testing of elapsed time for arbitrary regions of code. Furthermore, we generalize timing assertions to transparently provide hang detection that back annotates hang occurrences to source code. The presented techniques enable software developers to rapidly verify, debug, and analyze timing for FPGA applications, while reducing frequency by less than 3% and increasing FPGA resource utilization by 0.7% or less for several application case studies on the Altera Stratix-II EP2S180 and Stratix-III EP3SE260 using Impulse-C. The presented techniques reduced area overhead by as much as 3x and improved assertion performance by as much as 100% compared to unoptimized in-circuit assertions.
AlN/GaN-Based MOS-HEMT Technology: Processing and Device Results
Authors: Taking S. , MacFarlane D. , Wasige E.
Published: January 2011

Process development of AlN/GaN MOS-HEMTs is presented, along with issues and problems concerning the fabrication processes. The developed technology uses thermally grown Al2O3as a gate dielectric and surface passivation for devices. Significant improvement in device performance was observed using the following techniques: (1) Ohmic contact optimisation using Al wet etch prior to Ohmic metal deposition and (2) mesa sidewall passivation. DC and RF performance of the fabricated devices will be presented and discussed in this paper.
Elongator: An Ancestral Complex Driving Transcription and Migration through Protein Acetylation
Authors: Creppe Catherine , Buschbeck Marcus
Published: January 2011

Elongator is an evolutionary highly conserved complex. At least two of its cellular functions rely on the intrinsic lysine acetyl‐transferase activity of the Elongator complex. Its two known substrates—Histone H3 and α‐Tubulin—reflect the different roles of Elongator in the cytosol and the nucleus. A picture seems to emerge in which nuclear Elongator could regulate the transcriptional elongation of a subset of stress‐inducible genes through acetylation of Histone H3 in the promoter‐distal gene body. In the cytosol, Elongator‐mediated acetylation of α‐Tubulin contributes to intracellular trafficking and cell migration. Defects in both functions of Elongator have been implicated in neurodegenerative disorders.
Serum Neopterin Is Not Increased in Obese Juveniles
Authors: Mangge Harald , Freytag Florian , Almer Gunter , Weghuber Daniel , Bauer-Denk Carmen , Fuchs Dietmar
Published: January 2011

Objective. Cardiovascular disease is associated with inflammation and immune activation, concentrations of immune activation markers like neopterin predict outcome in adults.Methods. Serum neopterin concentrations and early metabolic and pre-atherosclerotic symptoms were analyzed in 295 obese juveniles and 101 normal weight controls of similar age. Additionally, the influence of a 12 months weight reduction program on neopterin levels was investigated in 31 obese juveniles.Results. Intima-media thickness of common carotid arteries (IMT) and the concentrations of C-reactive protein (CRP) were increased in the obese juveniles (P<.001). Also triglycerides, oxidized LDL, fasted insulin levels, HOMA-index, leptin, liver transaminases and uric acid were increased compared to the controls. However, serum neopterin was decreased in the obese versus non-obese juveniles (P<.03). The intervention consisting of regular sports, nutritional devices, and a psychologic attendance led after 12 months to an increase of neopterin concentration (P<.05; paired test).Conclusions. Neopterin concentrations in juvenile obesity behaved considerably different from what was demonstrated in adults, levels did not correlate with metabolic and pre-atherosclerotic symptoms found in early phases although early vascular burden and chronic low grade inflammation was indicated by increased IMT and CRP. Neopterin concentrations increased after a 12 months intervention program.
Breast Cancer Stem Cells Are Regulated by Mesenchymal Stem Cells through Cytokine Networks
Authors: Liu Suling , Ginestier Christophe , Ou Sing J. , Clouthier Shawn G. , Patel Shivani H. , Monville Florence , Korkaya Hasan , Heath Amber , Dutcher Julie , Kleer Celina G. , Jung Younghun , Dontu Gabriela , Taichman Russell , Wicha Max S.
Published: January 2011

We have used in vitro and mouse xenograft models to examine the interaction between breast cancer stem cells (CSC) and bone marrow–derived mesenchymal stem cells (MSC). We show that both of these cell populations are organized in a cellular hierarchy in which primitive aldehyde dehydrogenase expressing mesenchymal cells regulate breast CSCs through cytokine loops involving IL6 and CXCL7. In NOD/SCID mice, labeled MSCs introduced into the tibia traffic to sites of growing breast tumor xenografts where they accelerated tumor growth by increasing the breast CSC population. With immunochemistry, we identified MSC–CSC niches in these tumor xenografts as well as in frozen sections from primary human breast cancers. Bone marrow–derived MSCs may accelerate human breast tumor growth by generating cytokine networks that regulate the CSC population. Cancer Res; 71(2); 614–24. ©2011 AACR.
A Novel Transgenic Mouse Model of the Human Multiple Myeloma Chromosomal Translocation t(14;16)(q32;q23)
Authors: Morito Naoki , Yoh Keigyou , Maeda Atsuko , Nakano Takako , Fujita Akiko , Kusakabe Manabu , Hamada Michito , Kudo Takashi , Yamagata Kunihiro , Takahashi Satoru
Published: January 2011

Multiple myeloma (MM) is a currently incurable neoplasm of terminally differentiated B cells. The translocation and/or overexpression of c-MAF have been observed in human MM. Although c-MAF might function as an oncogene in human MM, there has been no report thus far describing the direct induction of MM by c-MAF overexpression in vivo. In this study, we have generated transgenic (TG) mice that express c-Maf specifically in the B-cell compartment. Aged c-Maf TG mice developed B-cell lymphomas with some clinical features that resembled those of MM, namely, plasma cell expansion and hyperglobulinemia. Quantitative RT-PCR analysis demonstrated that Ccnd2 and Itgb7, which are known target genes of c-Maf, were highly expressed in the lymphoma cells. This novel TG mouse model of the human MM t(14;16)(q32;q23) chromosomal translocation should serve to provide new insight into the role of c-MAF in tumorigenesis. Cancer Res; 71(2); 339–48. ©2011 AACR.
A Novel Sialyltransferase Inhibitor Suppresses FAK/Paxillin Signaling and Cancer Angiogenesis and Metastasis Pathways
Authors: Chen Jia-Yang , Tang Yen-An , Huang Sin-Ming , Juan Hsueh-Fen , Wu Li-Wha , Sun Ying-Chieh , Wang Szu-Chi , Wu Kuan-Wei , Balraj Gopula , Chang Tzu-Ting , Li Wen-Shan , Cheng Hung-Chi , Wang Yi-Ching
Published: January 2011

Increased sialyltransferase (ST) activity promotes cancer cell metastasis, and overexpression of cell surface sialic acid correlates with poor prognosis in cancer patients. To seek therapies targeting metastasis for cancer treatment, we developed a novel ST inhibitor, Lith-O-Asp, and investigated its antimetastatic and antiangiogenic effects and mechanisms. We found that cells treated with Lith-O-Asp showed a reduction of activity on various ST enzymes by in vitro and cell-based activity analyses. Lith-O-Asp inhibited migration and invasion abilities in various cancer cell lines and showed inhibitory effect on the angiogenic activity of human umbilical vein endothelial cells. Indeed, Lith-O-Asp treatment consequently delayed cancer cell metastasis in experimental and spontaneous metastasis assays in animal models. Importantly, Lith-O-Asp decreased the sialic acid modification of integrin-β1 and inhibited the expression of phospho-FAK, phospho-paxillin, and the matrix metalloprotease (MMP) 2 and MMP9. Lith-O-Asp attenuated the Rho GTPase activity leading to actin dynamic impairment. In addition, 2DE-MS/MS and immunoblotting analyses showed that Lith-O-Asp altered the protein expression level and phosphorylation status of various proteins involved in crucial metastasis and angiogenesis pathways such as vimentin and ribonuclease/angiogenin inhibitor RNH1. Furthermore, Lith-O-Asp treatment significantly inhibited the invasive ability exerted by ectopic overexpression of various ST enzymes catalyzing α-2,6- or α-2,3-sialylation. Our results provide compelling evidence that the potential pan-ST inhibitor, Lith-O-Asp, suppressed cancer cell metastasis likely by inhibiting FAK/paxillin signaling and expressing antiangiogenesis factors. Lith-O-Asp is worthy for further testing as a novel antimetastasis drug for cancer treatment. Cancer Res; 71(2); 473–83. ©2011 AACR.
Peptides and Aptamers Targeting HSP70: A Novel Approach for Anticancer Chemotherapy
Authors: Rérole Anne-Laure , Gobbo Jessica , De Thonel Aurelie , Schmitt Elise , Pais de Barros Jean Paul , Hammann Arlette , Lanneau David , Fourmaux Eric , Deminov Oleg , Micheau Olivier , Lagrost Laurent , Colas Pierre , Kroemer Guido , Garrido Carmen
Published: January 2011

The inhibition of heat shock protein 70 (HSP70) is an emerging strategy in cancer therapy. Unfortunately, no specific inhibitors are clinically available. By yeast two-hybrid screening, we have identified multiple peptide aptamers that bind HSP70. When expressed in human tumor cells, two among these peptide aptamers—A8 and A17—which bind to the peptide-binding and the ATP-binding domains of HSP70, respectively, specifically inhibited the chaperone activity, thereby increasing the cells' sensitivity to apoptosis induced by anticancer drugs. The 13-amino acid peptide from the variable region of A17 (called P17) retained the ability to specifically inhibit HSP70 and induced the regression of subcutaneous tumors in vivo after local or systemic injection. This antitumor effect was associated with an important recruitment of macrophages and T lymphocytes into the tumor bed. Altogether, these data indicate that peptide aptamers or peptides that target HSP70 may be considered as novel lead compounds for cancer therapy. Cancer Res; 71(2); 484–95. ©2011 AACR.
Heat Shock Protein 27 Differentiates Tolerogenic Macrophages That May Support Human Breast Cancer Progression
Authors: Banerjee Sanjib , Lin Chuen-Fu L. , Skinner Kristin A. , Schiffhauer Linda M. , Peacock James , Hicks David G. , Redmond Eileen M. , Morrow David , Huston Alissa , Shayne Michelle , Langstein Howard N. , Miller-Graziano Carol L. , Strickland Jennifer , O'Donoghue Lauren , De Asit K.
Published: January 2011

Tumor cells release several factors that can help the progression of the tumor by directly supporting tumor growth and/or suppressing host antitumor immunity. Here, we report that human primary breast tumor cells not only express elevated levels of heat shock protein 27 (Hsp27) at the intracellular level but also release extremely high levels of Hsp27 compared with the same patients' serum Hsp27 levels, predicting an acutely increased concentration of soluble Hsp27 in the human breast tumor microenvironment (HBTM). We demonstrate that Hsp27 levels in the HBTM can be extremely elevated as evidenced by high soluble Hsp27 levels in patients' tumor interstitial fluid. Because increasing numbers of tumor-associated macrophages (TAM) in the HBTM negatively correlate to patients' clinical outcomes and we have previously reported the immunoregulatory activity of soluble Hsp27, here, we tested for any specific effects of soluble Hsp27 on human monocyte to macrophage differentiation. We demonstrate that soluble Hsp27 causes the differentiation of monocytes to macrophages with immuno-tolerizing phenotypes (HLA-DRlow, CD86low, PD-L1high, ILT2high, and ILT4high). We detected the presence of TAMs with similar phenotypes in breast cancer patients. Hsp27-differentiated macrophages induce severe unresponsiveness/anergy in T cells. Moreover, these macrophages lose tumoricidal activity but become extremely proangiogenic, inducing significant neovascularization, a process that is critically important for tumor growth. Thus, our data demonstrate a novel immune escape and tumor growth–supporting mechanism mediated by soluble Hsp27 that may be operative in human breast cancer. Cancer Res; 71(2); 318–27. ©2011 AACR.
Phospholipase D Meets Wnt Signaling: A New Target for Cancer Therapy
Authors: Kang Dong Woo , Choi Kang-Yell , Min Do Sik
Published: January 2011

Phospholipase D (PLD) has been increasingly recognized as a critical regulator of cell proliferation and tumorigenesis. PLD regulates downstream effectors by generating phosphatidic acid (PA), and the expression and activity of PLD are elevated in many different types of human cancer. Aberrant activation of Wnt/β-catenin signaling, followed by hyper-activation of target genes, is linked to a wide range of cancers. New studies reveal a direct connection between the PLD and the Wnt signaling pathways; PLD is a transcriptional target of β-catenin/T-cell factor (TCF) and reinforces Wnt/β-catenin signaling related with cellular transformation. In this review, we discuss the emerging importance of PLD and PA in the Wnt/β-catenin signaling network, which is associated with tumorigenesis, and suggest that the PLD/PA signaling pathway is a potential therapeutic target for the treatment of cancer. Cancer Res; 71(2); 293–7. ©2011 AACR.
HMGA2 Overexpression-Induced Ovarian Surface Epithelial Transformation Is Mediated Through Regulation of EMT Genes
Authors: Wu Jingjing , Liu Zhaojian , Shao Changshun , Gong Yaoqin , Hernando Eva , Lee Peng , Narita Masashi , Muller William , Liu Jinsong , Wei Jian-Jun
Published: January 2011

The AT-hook transcription factor HMGA2 is an oncogene involved in the tumorigenesis of many malignant neoplasms. HMGA2 overexpression is common in both early and late-stage high-grade ovarian serous papillary carcinoma. To test whether HMGA2 participates in the initiation of ovarian cancer and promotion of aggressive tumor growth, we examined the oncogenic properties of HMGA2 in ovarian surface epithelial (OSE) cell lines. We found that introduction of HMGA2 overexpression was sufficient to induce OSE transformation in vitro. HMGA2-mediated OSE transformation resulted in tumor formation in the xenografts of nude mice. By silencing HMGA2 in HMGA2-overexpressing OSE and ovarian cancer cell lines, the aggressiveness of tumor cell growth behaviors was partially suppressed. Global gene profiling analyses revealed that HMGA2-mediated tumorigenesis was associated with expression changes of target genes and microRNAs that are involved in epithelial-to-mesenchymal transition (EMT). Lumican, a tumor suppressor that inhibits EMT, was found to be transcriptionally repressed by HMGA2 and was frequently lost in human high-grade serous papillary carcinoma. Our findings show that HMGA2 overexpression confers a powerful oncogenic signal in ovarian cancers through the modulation of EMT genes. Cancer Res; 71(2); 349–59. ©2011 AACR.
NCI Image-Guided Drug Delivery Summit
Authors: Tandon Pushpa , Farahani Keyvan
Published: January 2011

On April 17, 2010, scientists from academia, the National Cancer Institute (NCI), and the Food and Drug Administration (FDA) assembled at “The NCI Image Guided Drug Delivery Summit,” in Washington D.C., to discuss recent advances, barriers, opportunities, and regulatory issues related to the field. The meeting included a scientific session and an NCI/FDA session, followed by a panel discussion of speakers from both sessions. Image-guided drug delivery (IGDD) in cancer is a form of individualized therapy where imaging methods are used in guidance and monitoring of localized and targeted delivery of therapeutics to the tumor. So, a systematic approach to IGDD requires mechanisms for targeting, delivery, activation, and monitoring of the process. Although the goal in IGDD is to optimize the therapeutic ratio through personalized image-guided treatments, a major challenge is in overcoming the biological barriers to the delivery of therapeutics into tumors and cells. Speakers discussed potential challenges to clinical translation of nano-based drug delivery systems including in vivo characterization of nanocarriers, preclinical validation of targeting and delivery, studies of biodistribution, pharmacokinetics, pharmacodynamics, and toxicity as well as scale-up manufacturing of delivery systems. Physiologic and quantitative imaging techniques may serve as enabling tools that could potentially transform many existing challenges into opportunities for advancement of the field. Cancer Res; 71(2); 314–7. ©2011 AACR.
Physical Oncology: A Bench-to-Bedside Quantitative and Predictive Approach
Authors: Frieboes Hermann B. , Chaplain Mark A.J. , Thompson Alastair M. , Bearer Elaine L. , Lowengrub John S. , Cristini Vittorio
Published: January 2011

Cancer models relating basic science to clinical care in oncology may fail to address the nuances of tumor behavior and therapy, as in the case, discussed herein, of the complex multiscale dynamics leading to the often-observed enhanced invasiveness, paradoxically induced by the very antiangiogenic therapy designed to destroy the tumor. Studies would benefit from approaches that quantitatively link the multiple physical and temporal scales from molecule to tissue in order to offer outcome predictions for individual patients. Physical oncology is an approach that applies fundamental principles from the physical and biological sciences to explain certain cancer behaviors as observable characteristics arising from the underlying physical and biochemical events. For example, the transport of oxygen molecules through tissue affects phenotypic characteristics such as cell proliferation, apoptosis, and adhesion, which in turn underlie the patient-scale tumor growth and invasiveness. Our review of physical oncology illustrates how tumor behavior and treatment response may be a quantifiable function of marginally stable molecular and/or cellular conditions modulated by inhomogeneity. By incorporating patient-specific genomic, proteomic, metabolomic, and cellular data into multiscale physical models, physical oncology could complement current clinical practice through enhanced understanding of cancer behavior, thus potentially improving patient survival. Cancer Res; 71(2); 298–302. ©2011 AACR.
Suppression of Colonic Polyposis by Homeoprotein CDX2 through its Nontranscriptional Function that Stabilizes p27Kip1
Authors: Aoki Koji , Kakizaki Fumihiko , Sakashita Hiromi , Manabe Toshiaki , Aoki Masahiro , Taketo Makoto M.
Published: January 2011

Caudal-related homeoprotein CDX2 is expressed in intestinal epithelial cells, in which it is essential for their development and differentiation. A tumor suppressor function is suggested by evidence that CDX2 levels are decreased in human colon cancer specimens and that an inactivating mutation of Cdx2 in ApcΔ716 mice markedly increases the incidence of colonic polyps. In this study, we investigated roles for transcriptional and nontranscriptional functions of CDX2 in suppression of colonic tumorigenesis. Mutagenic analysis of CDX2 revealed that loss of function stabilizes CDK inhibitor p27Kip1 by a nontranscriptional but homeodomain-dependent mechanism that inhibits cyclin E-CDK2 activity and blocks G0/G1-S progression in colon cancer cells. p27Kip1 stabilization was mediated by an inhibition of ubiquitylation-dependent proteolysis associated with decreased phosphorylation of Thr187 in p27Kip1. siRNA-mediated knockdown of p27Kip1 relieved the decrease in cyclin E-CDK2 activity and S-phase cell fraction elicited by CDX2 expression. Together, these results implicate a nontranscriptional function of CDX2 in tumor suppression mediated by p27Kip1 stabilization. Up to approximately 75% of low-CDX2 human colon cancer lesions show reduced levels of p27Kip1, whereas approximately 68% of high-CDX2 lesions retain expression of p27Kip1. These results show that low levels of CDX2 accelerate colon tumorigenesis by reducing p27Kip1 levels. Cancer Res; 71(2); 593–602. ©2011 AACR.
SDHAF2 (PGL2-SDH5) and Hereditary Head and Neck Paraganglioma
Authors: Kunst Henricus P.M. , Rutten Martijn H. , de Mönnink Jan-Pieter , Hoefsloot Lies H. , Timmers Henri J.L.M. , Marres Henri A.M. , Jansen Jeroen C. , Kremer Hannie , Bayley Jean-Pierre , Cremers Cor W.R.J.
Published: January 2011

Purpose: Hereditary head and neck paraganglioma (HNPGL) syndromes are associated with mutations in the SDHD(PGL1), SDHC(PGL3), and SDHB(PGL4) genes encoding succinate dehydrogenase subunits. We recently described mutations in a previously uncharacterized human gene, now called SDHAF2, and showed that this was the long-sought “imprinted” PGL2 gene. Here, we present a new branch of the Dutch SDHAF2 (PLG2-SDH5) family.

Experimental Design: The SDHAF2 family has been collected over a 30-year period. The family described here was linked to PGL2 and at-risk family members were invited to participate in this study. Patients were investigated and treated dependent on tumor size and localization. All family members have now been analyzed for the SDHAF2 mutation status.

Results: Among the 57 family members, 23 were linkage positive including 7 risk-free carriers (maternal imprinting). Of the 16 at-risk individuals, 11 had a total of 24 tumors with primarily carotid (71%) and vagal locations (17%). Multifocality of tumors was prominent (91%). Malignancy was not detected. The average age at onset was 33 years, and many patients (42%) were asymptomatic prior to screening. SDHAF2 mutation analysis confirmed the findings of the previously performed linkage analysis without detection of discrepancies.

Conclusions: We established the SDHAF2 mutation status of PGL2 family members. Phenotypic characterization of this family confirms the currently exclusive association of SDHAF2 mutations with HNPGL. This SDHAF2 family branch shows a young age at onset and very high levels of multifocality. A high percentage of patients were asymptomatic at time of detection. Clin Cancer Res; 17(2); 247–54. ©2011 AACR.
Primary CNS Lymphoma in Children and Adolescents: A Descriptive Analysis from the International Primary CNS Lymphoma Collaborative Group (IPCG)
Authors: Abla Oussama , Weitzman Sheila , Blay Jean-Yves , O'Neill Brian Patrick , Abrey Lauren E. , Neuwelt Edward , Doolittle Nancy D. , Baehring Joachim , Pradhan Kamnesh , Martin S. Eric , Guerrera Michael , Shah Shafqat , Ghesquieres Hervé , Silver Michael , Betensky Rebecca A. , Batchelor Tracy
Published: January 2011

Purpose: To describe the demographic and clinical features and outcomes for children and adolescents with primary CNS lymphoma (PCNSL).

Experimental Design: A retrospective series of children and adolescents with PCNSL was assembled from 10 cancer centers in 3 countries.

Results: Twenty-nine patients with a median age of 14 years were identified. Sixteen (55%) had Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 1 or greater. Frontline therapy consisted of chemotherapy only in 20 patients (69%), while 9 (31%) had chemotherapy plus cranial radiotherapy. Most patients received methotrexate (MTX)-based regimens. Overall response rate was 86% (complete remission 69%, partial remission 17%). The 2-year progression-free survival (PFS) and overall survival (OS) rates were 61% and 86%, respectively; the 3-year OS was 82%. Univariate analyses were conducted for age (≤14 vs. >14 years), PS (0 or 1 vs. >1), deep brain lesions, MTX dose, primary treatment with chemotherapy alone, intrathecal chemotherapy, and high-dose therapy. Primary treatment with chemotherapy alone was associated with better overall response rates with an odds ratio (OR) of 0.125 (P = 0.02). There was a marginally significant relationship between higher doses of MTX and response (OR = 1.5, P = 0.06). ECOG-PS of 0 to 1 was the only factor associated with better outcome with hazard ratios of 0.136 (P = 0.017) and 0.073 (P = 0.033) for PFS and OS, respectively.

Conclusion: This is the largest series collected of pediatric PCNSL. The outcome of children and adolescents seems to be better than in adults. PS of 0 to 1 is associated with better survival. Clin Cancer Res; 17(2); 346–52. ©2011 AACR.
Phase I and Pharmacokinetic Study of CT-322 (BMS-844203), a Targeted Adnectin Inhibitor of VEGFR-2 Based on a Domain of Human Fibronectin
Authors: Tolcher Anthony W. , Sweeney Christopher J. , Papadopoulos Kyri , Patnaik Amita , Chiorean Elena G. , Mita Alain C. , Sankhala Kamalesh , Furfine Eric , Gokemeijer Jochem , Iacono Lisa , Eaton Cheryl , Silver Bruce A. , Mita Monica
Published: January 2011

Purpose: To determine the maximum tolerated dose (MTD), safety, pharmacokinetics, pharmacodynamics, immunogenicity, and preliminary antitumor activity of CT-322 (BMS-844203), a VEGFR-2 inhibitor and the first human fibronectin domain–based targeted biologic (Adnectin) to enter clinical studies.

Experimental Design: Patients with advanced solid malignancies were treated with escalating doses of CT-322 intravenously (i.v.) weekly (qw), or biweekly (q2w). Plasma samples were assayed for CT-322 concentrations, plasma VEGF-A concentrations, and antidrug antibodies.

Results: Thirty-nine patients completed 105 cycles of 0.1 to 3.0 mg/kg CT-322 i.v. either qw or q2w. The most common treatment-emergent grade 1/2 toxicities were fatigue, nausea, proteinuria, vomiting, anorexia, and hypertension. Grade 3/4 toxicities were rare. Reversible proteinuria, retinal artery, and vein thrombosis, left ventricular dysfunction, and reversible posterior leukoencephalopathy syndrome were dose limiting at 3.0 mg/kg. The MTD was 2 mg/kg qw or q2w. CT-322 plasma concentrations increased dose proportionally. Plasma VEGF-A levels increased with dose and plateaued at 2 mg/kg qw. Anti–CT-322 antibodies developed without effects on pharmacokinetics, VEGF-A levels, or safety. Minor decreases in tumor measurements occurred in 4 of 34 evaluable patients and 24 patients had stable disease.

Conclusions: CT-322 can be safely administered at 2 mg/kg i.v. qw or q2w and exhibits promising antitumor activity in patients with advanced solid tumors. The absence of severe toxicities at the MTD, demonstration of plasma drug concentrations active in preclinical models, and clinical pharmacodynamic evidence of VEGFR-2 inhibition warrant further development of CT-322 and suggest strong potential for Adnectin-based targeted biologics. Cancer Res; 17(2); 363–71. ©2011 AACR. Clin Cancer Res; 17(2); 363–71. ©2011 AACR.
Exosomes and Cancer: A Newly Described Pathway of Immune Suppression
Authors: Zhang Huang-Ge , Grizzle William E.
Published: January 2011

Exosomes are small (30 to 100 nm) membrane-bound particles that are released from normal, diseased, and neoplastic cells and are present in blood and other bodily fluids. Exosomes contain a variety of molecules including signal peptides, mRNA, microRNA, and lipids. Exosomes can function to export from cells unneeded endogenous molecules and therapeutic drugs. When exosomes are taken up by specific cells, they may act locally to provide autocrine or paracrine signals or, at a distance, as a newly described nanoparticle-based endocrine system. Specifically, mRNA transferred to cells by exosomes can result in the production of new proteins. In cancer, signals via exosomes affect the immune system by inhibiting the functions of T cells and normal killer (NK) cells and by inhibiting the differentiation of precursors to mature antigen-presenting cells. Also, exosomes increase the number and/or activity of immune suppressor cells, including myeloid-derived suppressor cells, T-regulatory cells, and CD14+, HLA-DR−/low cells. The effects of exosomes on the development and progression of cancers, with an emphasis on suppression of immune surveillance, is described. Also discussed are potential uses of exosomes clinically, in the development of vaccines, in targeting tumors, and in diagnosis and/or early detection. Clin Cancer Res; 17(5); 959–64. ©2011 AACR.
Therapeutic Drug Monitoring for the Individualization of Docetaxel Dosing: A Randomized Pharmacokinetic Study
Authors: Engels Frederike K. , Loos Walter J. , van der Bol Jessica M. , de Bruijn Peter , Mathijssen Ron H.J. , Verweij Jaap , Mathot Ron A.A.
Published: January 2011

Purpose: Docetaxel pharmacokinetic (PK) parameters, notably clearance and exposure (AUC), are characterized by large interindividual variability. The purpose of this study was to evaluate the effect of PK-guided [area under the plasma concentration versus time curve (AUC) targeted], individualized docetaxel dosing on interindividual variability in exposure.

Experimental Design: A limited sampling strategy in combination with a validated population PK model, Bayesian analysis, and a predefined target AUC was used. Fifteen patients were treated for at least 2 courses with body surface area–based docetaxel and 15 with at least 1 course of PK-guided docetaxel dosing.

Results: Interindividual variability (SD of ln AUC) was decreased by 35% (N = 15) after 1 PK-guided course; when all courses were evaluated, variability was decreased by 39% (P = 0.055). PK-guided dosing also decreased the interindividual variability of percentage decrease in white blood cell and absolute neutrophil counts by approximately 50%.

Conclusions: Further research is required to determine whether the decrease in PK variability can contribute to a reduction in interindividual variability in efficacy and toxicity. Clin Cancer Res; 17(2); 353–62. ©2011 AACR.
Circulating Tumor Cells and EpCAM Expression in Neuroendocrine Tumors
Authors: Khan Mohid S. , Tsigani Theodora , Rashid Mohammed , Rabouhans Jeremy S. , Yu Dominic , Luong Tu Vinh , Caplin Martyn , Meyer Tim
Published: January 2011

Purpose: Neuroendocrine tumors (NET) are heterogeneous tumors with widely variable survival. It is unknown whether they express EpCAM (epithelial cell adhesion molecule) and thus whether NET circulating tumor cells (CTC) are detectable. We systematically investigated EpCAM expression and CTC detection in patients with metastatic NETs and evaluated the potential of CTCs to predict radiological progression.

Experimental Design: EpCAM protein expression was evaluated in 74 samples of formalin-fixed, paraffin-embedded NET tissue by immunohistochemistry. Seventy-nine patients with metastatic NETs (42 midgut, 5 unknown primary, 19 pancreatic, 13 bronchopulmonary) had blood samples drawn for CTC isolation and enumeration utilizing the CellSearch platform. Patients were classified as having progressive or nonprogressive disease on the basis of serial imaging.

Results: Strong homogeneous, membranous EpCAM expression was observed in all ileal (n = 26) and pancreatic NETs (n = 16), whereas variable EpCAM expression was observed in bronchopulmonary NETs (n = 13). Forty-three percent of midgut and 21% of pancreatic NETs had CTCs detected with a range of 0–62 and 0–11, respectively. The absence of CTCs was strongly associated with stable disease (P < 0.001). There was a moderate correlation between CTC levels and urinary 5-hydroxyindole acetic acid (r = 0.5, P = 0.007) and between CTC levels and burden of liver metastases (B = 8.91, P < 0.001). There was no or low correlation between CTC levels and Ki-67 (r = 0.08, P = 0.59) and serum chromogranin A (r = 0.246, P = 0.03).

Conclusions: This is the first systematic analysis showing EpCAM expression and CTC detection in NETs. CTCs seem to be associated with progressive disease and may provide useful prognostic information given the variable survival rates in these tumors. Clin Cancer Res; 17(2); 337–45. ©2011 AACR.
Impact of Exploratory Biomarkers on the Treatment Effect of Bevacizumab in Metastatic Breast Cancer
Authors: Jubb Adrian M. , Miller Kathy D. , Rugo Hope S. , Harris Adrian L. , Chen Dafeng , Reimann James D. , Cobleigh Melody A. , Schmidt Maike , Langmuir Virginia K. , Hillan Kenneth J. , Chen Daniel S. , Koeppen Hartmut
Published: January 2011

Purpose: The addition of bevacizumab to cytotoxic chemotherapy has demonstrated a progression-free survival (PFS) benefit in the first-line and second-line treatment of advanced or metastatic breast cancer (MBC). However, the addition of bevacizumab to capecitabine in heavily pretreated MBC patients did not show a PFS benefit (AVF2119g phase III trial). The aim of this study was to evaluate the expression of novel putative biomarkers as predictors of benefit from bevacizumab in retrospective subset analyses of the AVF2119g trial.

Experimental Design: In the AVF2119g trial, 462 patients with MBC were randomly assigned to receive capecitabine or capecitabine plus bevacizumab. Primary tumor tissue and outcome data were available for 223 patients. Biomarker expression was assessed by in situ hybridization (VEGF-A, VEGF-B, thrombospondin-2 and Flt4) or immunohistochemistry (VEGF-C, PDGF-C, neuropilin-1, delta-like ligand (Dll) 4, Bv8, p53 and thymidine phosphorylase) on formalin-fixed, paraffin-embedded tissue. PFS was associated with these variables in retrospective subset analyses.

Results: Patients with low scores for Dll4, VEGF-C, and neuropilin-1 showed trends toward improvement in PFS associated with the addition of bevacizumab to capecitabine (P values = 0.01, 0.05, and 0.07, respectively). These observations were not statistically significant following correction for multiple hypothesis testing.

Conclusion: These retrospective subset analyses suggest that expression of Dll4, VEGF-C, and neuropilin-1 may predict benefit from bevacizumab. Such observations are not conclusive but warrant additional testing. Clin Cancer Res; 17(2); 372–81. ©2011 AACR.
Modulating Endogenous NQO1 Levels Identifies Key Regulatory Mechanisms of Action of β-Lapachone for Pancreatic Cancer Therapy
Authors: Li Long Shan , Bey Erik A. , Dong Ying , Meng Jieru , Patra Biswanath , Yan Jingsheng , Xie Xian-Jin , Brekken Rolf A. , Barnett Carlton C. , Bornmann William G. , Gao Jinming , Boothman David A.
Published: January 2011

Purpose: Pancreatic cancer is the fourth leading cause of cancer-related deaths, in which the 5-year survival rate is less than 5%. Current standard of care therapies offer little selectivity and high toxicity. Novel, tumor-selective approaches are desperately needed. Although prior work suggested that β-lapachone (β-lap) could be used for the treatment of pancreatic cancers, the lack of knowledge of the compound's mechanism of action prevented optimal use of this agent.

Experimental Design: We examined the role of NAD(P)H:quinone oxidoreductase-1 (NQO1) in β-lap–mediated antitumor activity, using a series of MIA PaCa-2 pancreatic cancer clones varying in NQO1 levels by stable shRNA knockdown. The antitumor efficacy of β-lap was determined using an optimal hydroxypropyl-β-cyclodextran (HPβ-CD) vehicle formulation in metastatic pancreatic cancer models.

Results: β-Lap–mediated cell death required ∼90 enzymatic units of NQO1. Essential downstream mediators of lethality were as follows: (i) reactive oxygen species (ROS); (ii) single-strand DNA breaks induced by ROS; (iii) poly(ADP-ribose)polymerase-1 (PARP1) hyperactivation; (iv) dramatic NAD+/ATP depletion; and (v) programmed necrosis. We showed that 1 regimen of β-lap therapy (5 treatments every other day) efficaciously regressed and reduced human pancreatic tumor burden and dramatically extended the survival of athymic mice, using metastatic pancreatic cancer models.

Conclusions: Because NQO1 enzyme activities are easily measured and commonly overexpressed (i.e., >70%) in pancreatic cancers 5- to 10-fold above normal tissue, strategies using β-lap to efficaciously treat pancreatic cancers are indicated. On the basis of optimal drug formulation and efficacious antitumor efficacy, such a therapy should be extremely safe and not accompanied with normal tissue toxicity or hemolytic anemia. Clin Cancer Res; 17(2); 275–85. ©2011 AACR.
Unique Functional Status of Natural Killer Cells in Metastatic Stage IV Melanoma Patients and Its Modulation by Chemotherapy
Authors: Fregni Giulia , Perier Aurélie , Pittari Gianfranco , Jacobelli Simon , Sastre Xavier , Gervois Nadine , Allard Mathilde , Bercovici Nadège , Avril Marie Françoise , Caignard Anne
Published: January 2011

Purpose: Immunotherapy is an alternative for metastatic melanoma patients resistant to chemotherapy. Natural killer (NK) cells are powerful antileukemia effectors and their role in solid tumors is suspected. NK cell activation is regulated by a balance between activating receptors, which detect stress molecules on tumor cells, and HLA-I specific inhibitory receptors. Here, we studied the phenotype and function of NK cells in stage IV metastatic melanoma patients.

Experimental Design: Circulating NK cells from 35 healthy donors and 51 patients were studied: 24 patients before chemotherapy (prechemotherapy), 17 patients 1 month after 1 to 4 lines of chemotherapy (postchemotherapy), and 10 patients analyzed pre- and postchemotherapy. NK functionality was carried out toward 2 primary metastatic melanoma cell lines, analyzed for the expression of NK receptor ligands.

Results: NK cells from prechemotherapy patients exhibit an NKp46dim/NKG2Adim phenotype. In contrast, NK cells from postchemotherapy patients display high expression of NKp46 and NKG2A receptors. Purified NK cells from patients are efficiently activated in response to melanoma cells. Melanoma cells express different level of NKG2D ligands and HLA-I molecules. In agreements with their phenotype, NK cells from pre- and postchemotherapy patients present distinct functional status toward these primary melanoma cells. A dynamic label free assay was used to determine the pathways involved in the lysis of melanoma cells by IL-2–activated NK cells. NKG2D, NCR (natural cytotoxicity receptor), and DNAM-1 are involved in the NK-mediated lysis of melanoma cells.

Conclusions: These results provide new arguments and clues to design NK cell–based immunotherapeutic strategies for melanoma patients. Clin Cancer Res; 17(9); 2628–37. ©2011 AACR.
Pretreatment Transcriptional Profiling for Predicting Response to Neoadjuvant Chemoradiotherapy in Rectal Adenocarcinoma
Authors: Brettingham-Moore Kate H. , Duong Cuong P. , Greenawalt Danielle M. , Heriot Alexander G. , Ellul Jason , Dow Christopher A. , Murray William K. , Hicks Rodney J. , Tjandra Joe , Chao Michael , Bui Andrew , Joon Daryl Lim , Thomas Robert J. S. , Phillips Wayne A.
Published: January 2011

Purpose: Patients presenting with locally advanced rectal cancer currently receive preoperative radiotherapy with or without chemotherapy. Although pathologic complete response is achieved for approximately 10% to 30% of patients, a proportion of patients derive no benefit from this therapy while being exposed to toxic side effects of treatment. Therefore, there is a strong need to identify patients who are unlikely to benefit from neoadjuvant therapy to help direct them toward alternate and ultimately more successful treatment options.

Experimental Design: In this study, we obtained expression profiles from pretreatment biopsies for 51 rectal cancer patients. All patients underwent preoperative chemoradiotherapy, followed by resection of the tumor 6 to 8 weeks posttreatment. Gene expression and response to treatment were correlated, and a supervised learning algorithm was used to generate an original predictive classifier and validate previously published classifiers.

Results: Novel predictive classifiers based on Mandard's tumor regression grade, metabolic response, TNM (tumor node metastasis) downstaging, and normal tissue expression profiles were generated. Because there were only 7 patients who had minimal treatment response (>80% residual tumor), expression profiles were used to predict good tumor response and outcome. These classifiers peaked at 82% sensitivity and 89% specificity; however, classifiers with the highest sensitivity had poor specificity, and vice versa. Validation of predictive classifiers from previously published reports was attempted using this cohort; however, sensitivity and specificity ranged from 21% to 70%.

Conclusions: These results show that the clinical utility of microarrays in predictive medicine is not yet within reach for rectal cancer and alternatives to microarrays should be considered for predictive studies in rectal adenocarcinoma. Clin Cancer Res; 17(9); 3039–47. ©2011 AACR.