bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumor-Initiated Inflammation Overrides Protective Adaptive Immunity in an Induced Melanoma Model in Mice
Authors: Soudja Saïdi M. , Wehbe Maria , Mas Amandine , Chasson Lionel , de Tenbossche Céline Powis , Huijbers Ivo , Van den Eynde Benoît , Schmitt-Verhulst Anne-Marie
Published: April 2010

We studied the effect of the immune system on two differentially aggressive melanomas developing in mice on conditional deletion of the INK4A/ARF tumor suppressor gene, with concomitant expression of oncogene H-RasG12V and a natural cancer-germline tumor antigen (TA). “Slow progressor” melanomas contained no activated T lymphocytes (TL). In contrast, “aggressive” melanomas were infiltrated by activated TLs lacking effector molecules and expressing high levels of PD-1, indicating an exhausted phenotype. Aggressive melanomas were also infiltrated by immature myeloid cells (IMC). Infiltration was associated with local inflammation and systemic Th2/Th17-oriented chronic inflammation that seemed to impair further activation of TLs, as tumor-specific T cells adoptively transferred into mice bearing aggressive melanomas were poorly activated and failed to infiltrate the melanoma. This immunosuppression also led to the incapacity of these mice to reject inoculated TA-positive tumors, in contrast to slow-progressing melanoma-bearing mice, which were responsive. To test the role of adaptive immunity in tumor progression, we induced melanomas in immunodeficient RagKO compound mice. These mice developed aggressive but not slow-progressing melanomas at a higher frequency and with a shorter latency than immunocompetent mice. Immunodeficient mice also developed abnormal inflammation and infiltration of IMCs in a manner similar to immunocompetent mice, indicating that this phenotype was not dependent on adaptive immunity. Therefore, tumor-intrinsic factors distinguishing the two melanoma types control the initiation of inflammation, which was independent of adaptive immunity. The latter delayed development of aggressive melanomas but was overridden by inflammation. Cancer Res; 70(9); 3515–25. ©2010 AACR.
More Modifiers Move on DNA Damage
Authors: Morris Joanna R.
Published: April 2010

In mammalian cells the accumulation of repair proteins to double-strand breaks is a phosphorylation- and ubiquitylation-regulated process. Some of the genes that encode the kinases and ubiquitin ligases in this pathway are cancer predisposition genes, most prominently the breast cancer predisposition gene BRCA1, which encodes a ubiquitin ligase. How BRCA1 ligase activity was regulated following DNA damage was poorly understood. In this review I summarize new data that show a third post-translational modification, by the small ubiquitin like modifier SUMO, is part of the same cascade, enabling and activating DNA damage-regulated processes, including the BRCA1 ligase activity. Cancer Res; 70(10); 3861–3. ©2010 AACR.
A Phase Ib Study of AMG 102 in Combination with Bevacizumab or Motesanib in Patients with Advanced Solid Tumors
Authors: Rosen Peter J. , Sweeney Christopher J. , Park Dorothy J. , Beaupre Darrin M. , Deng Hongjie , Leitch Ian M. , Shubhakar Poornima , Zhu Min , Oliner Kelly S. , Anderson Abraham , Yee Lorrin K.
Published: April 2010

Purpose: This phase Ib study evaluated the safety, pharmacokinetics, pharmacodynamics, and antitumor activity of AMG 102, a fully human monoclonal antibody against hepatocyte growth factor/scatter factor (HGF/SF), in combination with bevacizumab or motesanib in patients with advanced solid tumors.

Experimental Design: Patients with treatment-refractory advanced solid tumors were sequentially enrolled into four cohorts (3, 10, or 20 mg/kg AMG 102 plus 10 mg/kg bevacizumab i.v. every 2 weeks, or 3 mg/kg AMG 102 i.v. every 2 weeks plus 75 mg motesanib orally once daily).

Results: Fourteen patients were enrolled and received AMG 102. The combination of AMG 102 with bevacizumab (n = 12) seemed to have acceptable toxicity. The number of patients (n = 2) who received AMG 102 plus motesanib was insufficient to adequately assess safety. No dose-limiting toxicities were reported. Enrollment in the motesanib cohort was suspended because of reports of cholecystitis in other motesanib studies. Treatment-emergent adverse events among patients receiving AMG 102 plus bevacizumab were generally mild and included fatigue (75%), nausea (58%), constipation (42%), and peripheral edema (42%). No anti-AMG 102 antibodies were detected. Bevacizumab did not seem to affect AMG 102 pharmacokinetics. Circulating total HGF/SF increased from baseline throughout the study. Eight of 10 evaluable patients had reductions in tumor dimensions, and stable disease at ≥8, ≥16, and ≥24 weeks occurred in 9, 7, and 4 patients, respectively. Progression-free survival ranged from 7.9 to 121.9 weeks.

Conclusions: AMG 102 in combination with bevacizumab was well tolerated. Further evaluation of AMG 102 in combination with antiangiogenic agents is warranted. Clin Cancer Res; 16(9); 2677–87. ©2010 AACR.
New Molecular and Biological Mechanism of Antitumor Activities of KW-2478, a Novel Nonansamycin Heat Shock Protein 90 Inhibitor, in Multiple Myeloma Cells
Authors: Nakashima Takayuki , Ishii Toshihiko , Tagaya Hisashi , Seike Toshihiro , Nakagawa Hiroshi , Kanda Yutaka , Akinaga Shiro , Soga Shiro , Shiotsu Yukimasa
Published: April 2010

Purpose: The heat shock protein 90 (Hsp90) plays an important role in chaperoning oncogenic client proteins in multiple myeloma (MM) cells, and several Hsp90 inhibitors have shown antitumor activities both in vitro and in vivo. However the precise mechanism of action of Hsp90 inhibitor in MM has not been fully elucidated.

Experimental Design: We evaluated the antitumor activities of KW-2478, a nonansamycin Hsp90 inhibitor, in MM cells with various chromosomal translocations of immunoglobulin heavy chain (IgH) loci both in vitro and in vivo.

Results: Our studies revealed that exposure of KW-2478 to MM cells resulted in growth inhibition and apoptosis, which were associated with degradation of well-known client proteins as well as a decrease in IgH translocation products (FGFR3, c-Maf, and cyclin D1), and FGFR3 was shown to be a new client protein of Hsp90 chaperon complex. In addition, KW-2478 depleted the Hsp90 client Cdk9, a transcriptional kinase, and the phosphorylated 4E-BP1, a translational inhibitor. Both inhibitory effects of KW-2478 on such transcriptional and translational pathways were shown to reduce c-Maf and cyclin D1 expression. In NCI-H929 s.c. inoculated model, KW-2478 showed a significant suppression of tumor growth and induced the degradation of client proteins in tumors. Furthermore, in a novel orthotopic MM model of i.v. inoculated OPM-2/green fluorescent protein, KW-2478 showed a significant reduction of both serum M protein and MM tumor burden in the bone marrow.

Conclusions: These results suggest that targeting such diverse pathways by KW-2478 could be a promising strategy for the treatment of MM with various cytogenetic abnormalities. Clin Cancer Res; 16(10); 2792–802. ©2010 AACR.
Gene Expression Signature in Urine for Diagnosing and Assessing Aggressiveness of Bladder Urothelial Carcinoma
Authors: Mengual Lourdes , Burset Moisès , Ribal María José , Ars Elisabet , Marín-Aguilera Mercedes , Fernández Manuel , Ingelmo-Torres Mercedes , Villavicencio Humberto , Alcaraz Antonio
Published: April 2010

Purpose: To develop an accurate and noninvasive method for bladder cancer diagnosis and prediction of disease aggressiveness based on the gene expression patterns of urine samples.

Experimental Design: Gene expression patterns of 341 urine samples from bladder urothelial cell carcinoma (UCC) patients and 235 controls were analyzed via TaqMan Arrays. In a first phase of the study, three consecutive gene selection steps were done to identify a gene set expression signature to detect and stratify UCC in urine. Subsequently, those genes more informative for UCC diagnosis and prediction of tumor aggressiveness were combined to obtain a classification system of bladder cancer samples. In a second phase, the obtained gene set signature was evaluated in a routine clinical scenario analyzing only voided urine samples.

Results: We have identified a 12+2 gene expression signature for UCC diagnosis and prediction of tumor aggressiveness on urine samples. Overall, this gene set panel had 98% sensitivity (SN) and 99% specificity (SP) in discriminating between UCC and control samples and 79% SN and 92% SP in predicting tumor aggressiveness. The translation of the model to the clinically applicable format corroborates that the 12+2 gene set panel described maintains a high accuracy for UCC diagnosis (SN = 89% and SP = 95%) and tumor aggressiveness prediction (SN = 79% and SP = 91%) in voided urine samples.

Conclusions: The 12+2 gene expression signature described in urine is able to identify patients suffering from UCC and predict tumor aggressiveness. We show that a panel of molecular markers may improve the schedule for diagnosis and follow-up in UCC patients. Clin Cancer Res; 16(9); 2624–33. ©2010 AACR.
Clinical Benefit of Lapatinib-Based Therapy in Patients with Human Epidermal Growth Factor Receptor 2–Positive Breast Tumors Coexpressing the Truncated p95HER2 Receptor
Authors: Scaltriti Maurizio , Chandarlapaty Sarat , Prudkin Ludmila , Aura Claudia , Jimenez José , Angelini Pier Davide , Sánchez Gertrudis , Guzman Marta , Parra Josep Lluis , Ellis Catherine , Gagnon Robert , Koehler Maria , Gomez Henry , Geyer Charles , Cameron David , Arribas Joaquin , Rosen Neal , Baselga José
Published: April 2010

Purpose: A subgroup of human epidermal growth factor receptor 2 (HER2)–overexpressing breast tumors coexpresses p95HER2, a truncated HER2 receptor that retains a highly functional HER2 kinase domain but lacks the extracellular domain and results in intrinsic trastuzumab resistance. We hypothesized that lapatinib, a HER2 tyrosine kinase inhibitor, would be active in these tumors. We have studied the correlation between p95HER2 expression and response to lapatinib, both in preclinical models and in the clinical setting.

Experimental Design: Two different p95HER2 animal models were used for preclinical studies. Expression of p95HER2 was analyzed in HER2-overexpressing breast primary tumors from a first-line lapatinib monotherapy study (EGF20009) and a second-line lapatinib in combination with capecitabine study (EGF100151). p95HER2 expression was correlated with overall response rate (complete + partial response), clinical benefit rate (complete response + partial response + stable disease ≥24 wk), and progression-free survival using logistic regression and Cox proportional hazard models.

Results: Lapatinib inhibited tumor growth and the HER2 downstream signaling of p95HER2-expressing tumors. A total of 68 and 156 tumors from studies EGF20009 and EGF100151 were evaluable, respectively, for p95HER2 detection. The percentage of p95HER2-positive patients was 20.5% in the EGF20009 study and 28.5% in the EGF100151 study. In both studies, there was no statistically significant difference in progression-free survival, clinical benefit rate, and overall response rate between p95HER2-positive and p95HER2-negative tumors.

Conclusions: Lapatinib as a monotherapy or in combination with capecitabine seems to be equally effective in patients with p95HER2-positive and p95HER2-negative HER2-positive breast tumors. Clin Cancer Res; 16(9); 2688–95. ©2010 AACR.
Clinical Responses in a Phase II Study Using Adoptive Transfer of Short-term Cultured Tumor Infiltration Lymphocytes in Metastatic Melanoma Patients
Authors: Besser Michal J. , Shapira-Frommer Ronnie , Treves Avraham J. , Zippel Dov , Itzhaki Orit , Hershkovitz Liat , Levy Daphna , Kubi Adva , Hovav Einat , Chermoshniuk Natalia , Shalmon Bruria , Hardan Izhar , Catane Raphael , Markel Gal , Apter Sara , Ben-Nun Alon , Kuchuk Iryna , Shimoni Avichai , Nagler Arnon , Schachter Jacob
Published: April 2010

Purpose: Adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TIL) has shown promising results in metastatic melanoma patients. Although objective response rates of over 50% have been reported, disadvantages of this approach are the labor-intensive TIL production and a very high drop-out rate of enrolled patients, limiting its widespread applicability.

Previous studies showed a clear correlation between short TIL culture periods and clinical response. Therefore, we used a new TIL production technique using unselected, minimally cultured, bulk TIL (Young-TIL). The use of Young-TIL is not restricted to human leukocyte antigen (HLA)-A2 patients.

The purpose of this study is to explore the efficacy and toxicity of adoptively transferred Young-TIL following lympho-depleting chemotherapy in metastatic melanoma patients, refractory to interleukin-2 and chemotherapy.

Experimental Design: Young-TIL cultures for 90% of the patients were successfully generated, enabling the treatment of most enrolled patients. We report here the results of 20 evaluated patients.

Results: Fifty percent of the patients achieved an objective clinical response according to the Response Evaluation Criteria in Solid Tumors, including two ongoing complete remissions (20+, 4+ months) and eight partial responses (progression-free survival: 18+, 13+, 10+, 9, 6+, 4, 3+, and 3 months). All responders are currently alive. Four additional patients showed disease stabilization. Side effects were transient and manageable.

Conclusion: We showed that lympho-depleting chemotherapy followed by transfer of short-term cultured TIL can mediate tumor regression in 50% of metastatic melanoma with manageable toxicity. The convincing clinical results combined with the simplification of the process may thus have a major effect on cell therapy of cancer. Clin Cancer Res; 16(9); 2646–55. ©2010 AACR.
Phase I and Pharmacokinetic Study of Sequential Paclitaxel and Trabectedin Every 2 Weeks in Patients with Advanced Solid Tumors
Authors: Chu Quincy , Mita Alain , Forouzesh Bahram , Tolcher Anthony W. , Schwartz Gary , Nieto Antonio , Soto-Matos Arturo , Alfaro Vicente , Lebedinsky Claudia , Rowinsky Eric K.
Published: April 2010

Purpose: This phase I study evaluated the feasibility, safety, pharmacokinetics (PK), and preliminary evidence of anticancer activity of the sequential administration of paclitaxel and trabectedin on an every-2-week schedule in patients with refractory solid malignancies. The study also sought to determine the maximum tolerated dose (MTD) level on this schedule, as well as to recommend doses for disease-directed studies.

Experimental Design: Twenty-seven patients were treated with paclitaxel (80-120 mg/m2; 1-hour i.v. infusion, day 1) and trabectedin (0.525-0.775 mg/m2; 3-hour i.v. infusion, day 2) with doses increased in successive cohorts. Blood sampling for PK and drug-drug interaction studies was done.

Results: Neutropenia, which resulted in treatment delay exceeding 1 week, was the principal dose-limiting toxicity for this paclitaxel-trabectedin regimen and precluded dose escalation above 120 mg/m2 paclitaxel and 0.650 mg/m2 trabectedin. At the MTD (120 mg/m2 paclitaxel and 0.650 mg/m2 trabectedin), the safety profile was favorable in patients receiving cumulative treatment. Relevant drug-drug PK interactions between paclitaxel and trabectedin were not identified. A patient with soft tissue sarcoma had a complete response and several patients with various refractory solid malignancies showed protracted stable disease as their best response.

Conclusions: The MTD level of sequential paclitaxel 1-hour infusion (day 1) and trabectedin 3-hour infusion (day 2) administered every 2 weeks is 120 and 0.650 mg/m2, respectively. The manageable toxicities at the MTD, preliminary evidence of antitumor activity, and lack of notable PK drug-drug interactions warrant further disease-directed studies of this regimen in relevant tumor types and settings. Clin Cancer Res; 16(9); 2656–65. ©2010 AACR.
Assessment of the In vivo Antitumor Effects of ENMD-2076, a Novel Multitargeted Kinase Inhibitor, against Primary and Cell Line–Derived Human Colorectal Cancer Xenograft Models
Authors: Tentler John J. , Bradshaw-Pierce Erica L. , Serkova Natalie J. , Hasebroock Kendra M. , Pitts Todd M. , Diamond Jennifer R. , Fletcher Graham C. , Bray Mark R. , Eckhardt S. Gail
Published: April 2010

Purpose: This in vivo study was designed to investigate the efficacy of ENMD-2076, a small-molecule kinase inhibitor with activity against the Aurora kinases A and B, and several other tyrosine kinases linked to cancer, including vascular endothelial growth factor receptor 2, cKit, and fibroblast growth factor receptor 1, against murine xenograft models of human colorectal cancer (CRC).

Experimental Design: HT-29 CRC cell line xenografts were treated with either vehicle or ENMD-2076 (100 or 200 mg/kg) orally daily for 28 days. Tumor growth inhibition, dynamic contrast-enhanced magnetic resonance imaging, and 18FDG-positron emission tomography were conducted to assess the antiproliferative, antiangiogenic, and antimetabolic responses, respectively. Effects on proliferation were also analyzed by immunohistochemical methods. Additionally, three patient-derived xenografts from primary and metastatic sites were treated with ENMD-2076 (100 mg/kg) and assessed for tumor growth inhibition.

Results: In the HT-29 xenograft model, ENMD-2076 induced initial tumor growth inhibition followed by regression. Treatment was associated with significant tumor blanching, indicating a loss of vascularity and substantial reductions in tumor vascular permeability and perfusion as measured by dynamic contrast-enhanced magnetic resonance imaging. Positron emission tomography scanning showed significant decreases in 18FDG uptake at days 3 and 21 of treatment, which was associated with a marked reduction in proliferation as assessed by Ki-67. All three of the patient-derived xenografts tested were sensitive to treatment with ENMD 2076 as measured by tumor growth inhibition.

Conclusions: ENMD-2076 showed robust antitumor activity against cell line and patient-derived xenograft models of CRC that is detectable by functional imaging, supporting clinical investigation of this agent in CRC. Clin Cancer Res; 16(11); 2989–98. ©2010 AACR.
Imaging Breast Cancer Chemotherapy Response with Light
Authors: Tromberg Bruce J. , Cerussi Albert E.
Published: April 2010

Diffuse optical spectroscopy (DOS), which is used to image tumor metabolic response to neoadjuvant chemotherapy (NAC), shows large changes in tumor functional parameters with significant reductions in oxy- and deoxyhemoglobin for responders versus nonresponders. Although investigational, DOS may provide a cost-effective, risk-free method for optimizing NAC drug and dosing strategies for individual patients. Clin Cancer Res; 16(9); 2486–8. ©2010 AACR.
Driven to Death: Inhibition of Farnesylation Increases Ras Activity and Promotes Growth Arrest and Cell Death
Authors: Geryk-Hall Mandy , Yang Yanwen , Hughes Dennis P.M.
Published: April 2010

To improve cancer outcomes, investigators are turning increasingly to small molecule medicines that disrupt vital signaling cascades, inhibit malignant growth, or induce apoptosis. One vital signaling molecule is Ras, and a key step in Ras activation is membrane anchoring of Ras through prenylation, the C-terminal addition of a lipid anchor. Small molecule inhibitors of farnesyltransferase (FTI), the enzyme most often responsible for prenylating Ras, showed clinical promise, but development of FTIs such as tipifarnib has been stalled by uncertainty about their mechanism of action, because Ras seemed unimpeded in tipifarnib-treated samples. Interpretation was further complicated by the numerous proteins that may be farnesylated, as well as availability of an alternate prenylation pathway, geranylgeranylation. Our initial observations of varied response by cancer cell lines to tipifarnib led us to evaluate the role of FTI in Ras signal alteration using various tumor models. We describe our novel counterintuitive finding that endogenous Ras activity increases in cancer cell lines with low endogenous Ras activity when farnesyltransferase is inhibited by either tipifarnib or short hairpin RNA. In response to tipifarnib, variable growth arrest and/or cell death correlated with levels of activated extracellular signal–regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK). Sensitivity to tipifarnib treatment was shown by growth inhibition and by an increase in subdiploid cell numbers; cells with such sensitivity had increased activation of ERK and p38 MAPK. Because Ras must be prenylated to be active, our findings suggest that geranylgeranylated N-Ras or K-Ras B interacts differently with downstream effector proteins in sensitive cancer cells responding to tipifarnib, switching the balance from cell proliferation to growth inhibition. Mol Cancer Ther; 9(5); 1111–9. ©2010 AACR.
1,25-Dihydroxyvitamin D3 Enhances the Apoptotic Activity of MDM2 Antagonist Nutlin-3a in Acute Myeloid Leukemia Cells Expressing Wild-type p53
Authors: Thompson Thelma , Andreeff Michael , Studzinski George P. , Vassilev Lyubomir T.
Published: April 2010

The tumor suppressor p53 is often referred to as “the guardian of the genome” because of its central role in the cellular response to oncogenic stress and prevention of tumor development. Mutations of p53 in acute myeloid leukemia (AML) are rare but resistance to chemotherapy has been reported because of the deregulation of the p53 signaling and differentiation pathways. It is known that the interaction of the vitamin D metabolite 1,25-dihydroxyvitamin D3 (1,25D) with its functional vitamin D receptor leads to differentiation, G1 arrest, and increased cell survival in p53-null AML cells. However, there are no reports on the effect of 1,25D in leukemia cells expressing wild-type p53. Here, we examine vitamin D signaling in AML cells MOLM-13 and OCI-AML3 expressing wild-type p53 in the presence and absence of the MDM2 antagonist nutlin-3. We find that 1,25D alone induces monocytic differentiation in these cell lines similar to that seen in p53-null AML cells, suggesting that the presence of wild-type p53 is compatible with activation of vitamin D signaling. Combination of nutlin-3a with 1,25D accelerated programmed cell death, likely because of enhanced nutlin-induced upregulation of the proapoptotic PIG-6 protein and downregulation of antiapoptotic BCL-2, MDMX, human kinase suppressor of Ras 2, and phosphorylated extracellular signal-regulated kinase 2. Mol Cancer Ther; 9(5); 1158–68. ©2010 AACR.
Identification of c-Src as a Potential Therapeutic Target for Gastric Cancer and of MET Activation as a Cause of Resistance to c-Src Inhibition
Authors: Okamoto Wataru , Okamoto Isamu , Yoshida Takeshi , Okamoto Kunio , Takezawa Ken , Hatashita Erina , Yamada Yuki , Kuwata Kiyoko , Arao Tokuzo , Yanagihara Kazuyoshi , Fukuoka Masahiro , Nishio Kazuto , Nakagawa Kazuhiko
Published: April 2010

Therapeutic strategies that target c-Src hold promise for a wide variety of cancers. We have now investigated both the effects of dasatinib, which inhibits the activity of c-Src and several other kinases, on cell growth as well as the mechanism of dasatinib resistance in human gastric cancer cell lines. Immunoblot analysis revealed the activation of c-Src at various levels in most gastric cancer cell lines examined. Dasatinib inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) and induced G1 arrest, as revealed by flow cytometry, in a subset of responsive cell lines. In other responsive cell lines, dasatinib inhibited both ERK and AKT phosphorylation and induced apoptosis, as revealed by an increase in caspase-3 activity and cleavage of poly(ADP-ribose) polymerase. Depletion of c-Src by RNA interference also induced G1 arrest or apoptosis in dasatinib-responsive cell lines, indicating that the antiproliferative effect of dasatinib is attributable to c-Src inhibition. Gastric cancer cell lines positive for the activation of MET were resistant to dasatinib. Dasatinib had no effect on ERK or AKT signaling, whereas the MET inhibitor PHA-665752 induced apoptosis in these cells. The subsets of gastric cancer cells defined by a response to c-Src or MET inhibitors were distinct and nonoverlapping. Our results suggest that c-Src is a promising target for the treatment of gastric cancer and that analysis of MET amplification might optimize patient selection for treatment with c-Src inhibitors. Mol Cancer Ther; 9(5); 1188–97. ©2010 AACR.
Activation by Tyrosine Phosphorylation as a Prerequisite for Protein Kinase Cζ to Mediate Epidermal Growth Factor Receptor Signaling to ERK
Authors: Valkova Christina , Mertens Carmen , Weisheit Simona , Imhof Diana , Liebmann Claus
Published: April 2010

The atypical protein kinase Cζ (PKCζ) was recently shown to mediate epidermal growth factor (EGF)–induced activation of extracellular signal–regulated kinase (ERK) in head and neck squamous carcinoma (HNSCC) cells. Here, it is shown that EGF may induce tyrosine phosphorylation of PKCζ in several HNSCC cells, breast carcinoma cells, as well as mouse embryonic fibroblasts. In COS-7 cells overexpressing EGF receptor (EGFR) and PKCζ as a tumor cell model, we show that PKCζ tyrosine phosphorylation by EGF is induced by catalytic activation. Using a loss-of-function mutant of PKCζ, we can show that the tyrosine residue 417 in PKCζ plays an important role in both PKCζ activation and the ability of PKCζ to mediate activation of ERK. The importance of PKCζ in EGF-induced ERK activation can also be shown in several HNSCC and breast carcinoma cell lines as well as in PKCζ-deficient mouse embryonic fibroblasts. In addition, we present several lines of evidence suggesting the physical association of PKCζ with EGFR and the importance of the EGFR tyrosine kinase c-Src and the Src-specific phosphorylation site pY845-EGFR in the tyrosine phosphorylation as well as catalytic activation of PKCζ. This study characterizes PKCζ as a novel mitogenic downstream mediator of EGFR and indicates PKCζ as a therapeutic target in some carcinomas. Mol Cancer Res; 8(5); 783–97. ©2010 AACR.
The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis
Authors: Miki Takao , Shamma Awad , Kitajima Shunsuke , Takegami Yujiro , Noda Makoto , Nakashima Yasuaki , Watanabe Ken-ichiro , Takahashi Chiaki
Published: April 2010

Vascular endothelial cells produce considerable amounts of matrix metalloproteinases (MMP), including MMP-2, MMP-9, and membrane type 1 (MT1)–MMP. However, little is known about the regulatory mechanisms of these protease activities exhibited during vascular development. A glycosylphosphatidylinositol-anchored glycoprotein, reversion-inducing cysteine-rich protein with Kazal motifs (RECK), has been shown to attenuate MMP-2 maturation by directly interacting with MT1-MMP. Here, we show that an angiogenic factor angiopoietin-1 induces RECK expression in human umbilical vein endothelial cells (HUVEC), and RECK depletion in these cells results in defective vascular tube formation and cellular senescence. We further observed that RECK depletion downregulates β1-integrin activation, which was associated with decreased autophosphorylation of focal adhesion kinase and increased expression of a cyclin-dependent kinase inhibitor p21CIP1. In agreement, significant downregulation of β1-integrin activity was observed in vascular endothelial cells in Reck−/− mouse embryos. In HUVECs, specific inhibition of MMP-2 significantly antagonized the effect of RECK depletion on β1-integrin signaling, cell proliferation, and tube elongation. Furthermore, we observed that hypervascular tumor-derived cell lines can induce high RECK expression in convoluted vascular endothelial cells, and this in turn supports tumor growth. Targeting RECK specifically in tumor-associated vascular endothelial cells resulted in tumor regression. Therefore, we propose that RECK in tumor vascular endothelial cells can be an interesting target of cancer treatment via abortion of tumor angiogenesis. Mol Cancer Res; 8(5); 665–76. ©2010 AACR.
Altered-Function p53 Missense Mutations Identified in Breast Cancers Can Have Subtle Effects on Transactivation
Authors: Jordan Jennifer J. , Inga Alberto , Conway Kathleen , Edmiston Sharon , Carey Lisa A. , Wu Lin , Resnick Michael A.
Published: April 2010

Mutations of the sequence-specific master regulator p53 that alter transactivation function from promoter response elements (RE) could result in changes in the strength of gene activation or spectra of genes regulated. Such mutations in this tumor suppressor might lead to dramatic phenotypic changes and diversification of cell responses to stress. We have determined “functional fingerprints” of sporadic breast cancer–related p53 mutants, many of which are also associated with familial cancer proneness such as the Li-Fraumeni syndrome and germline BRCA1/2 mutant-associated cancers. The ability of p53, wild-type and mutants, to transactivate from 11 human target REs has been assessed at variable expression levels using a cellular, isogenomic yeast model system that allows for the rapid analysis of p53 function using a qualitative and a quantitative reporter. Among 50 missense mutants, 29 were classified as loss of function. The remaining 21 retained transactivation toward at least one RE. At high levels of galactose-induced p53 expression, 12 of 21 mutants that retain transactivation seemed similar to wild-type. When the level of galactose was reduced, transactivation defects could be revealed, suggesting that some breast cancer–related mutants can have subtle changes in transcription. These findings have been compared with clinical data from an ongoing neoadjuvant chemotherapy treatment trial for locally advanced breast tumors. The functional and nonfunctional missense mutations may distinguish tumors in terms of demographics, appearance, and relapse, implying that heterogeneity in the functionality of specific p53 mutations could affect clinical behavior and outcome. Mol Cancer Res; 8(5); 701–16. ©2010 AACR.
Drugs that Target Specificity Proteins Downregulate Epidermal Growth Factor Receptor in Bladder Cancer Cells
Authors: Chadalapaka Gayathri , Jutooru Indira , Burghardt Robert , Safe Stephen
Published: April 2010

The epidermal growth factor receptor (EGFR) is an important chemotherapeutic target for tyrosine kinase inhibitors and antibodies that block the extracellular domain of EGFR. Betulinic acid (BA) and curcumin inhibited bladder cancer cell growth and downregulated specificity protein (Sp) transcription factors, and this was accompanied by decreased expression of EGFR mRNA and protein levels. EGFR, a putative Sp-regulated gene, was also decreased in cells transfected with a cocktail (iSp) containing small inhibitory RNAs for Sp1, Sp3, and Sp4, and RNA interference with individual Sp knockdown indicated that EGFR expression was primarily regulated by Sp1 and Sp3. BA, curcumin, and iSp also decreased phosphorylation of Akt in these cells, and downregulation of EGFR by BA, curcumin, and iSp was accompanied by induction of LC3 and autophagy, which is consistent with recent studies showing that EGFR suppresses autophagic cell death. The results show that EGFR is an Sp-regulated gene in bladder cancer, and drugs such as BA and curcumin that repress Sp proteins also ablate EGFR expression. Thus, compounds such as curcumin and BA that downregulate Sp transcription factors represent a novel class of anticancer drugs that target EGFR in bladder cancer cells and tumors by inhibiting receptor expression. Mol Cancer Res; 8(5); 739–50. ©2010 AACR.
Focal Adhesion Kinase, a Downstream Mediator of Raf-1 Signaling, Suppresses Cellular Adhesion, Migration, and Neuroendocrine Markers in BON Carcinoid Cells
Authors: Ning Li , Chen Herbert , Kunnimalaiyaan Muthusamy
Published: April 2010

We have recently reported that activation of the Raf-1/mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase 1/2 (MEK1/2)/ERK1/2 signaling cascade in gastrointestinal carcinoid cell line (BON) alters cellular morphology and neuroendocrine phenotype. The mechanisms by which Raf-1 mediates these changes in carcinoid cells are unclear. Here, we report that activation of the Raf-1 signaling cascade in BON cells induced the expression of focal adhesion kinase (FAK) protein, suppressed the production of neuroendocrine markers, and resulted in significant decreases in cellular adhesion and migration. Importantly, inactivation of MEK1/2 by 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene or abolition of FAK induction in Raf-1–activated BON cells by targeted siRNA led to reversal of the Raf-1–mediated reduction in neuroendocrine markers and cellular adhesion and migration. Phosphorylation site-specific antibodies detected the phosphorylated FAKTyr407, but not FAKTyr397, in these Raf-1–activated cells, indicating that FAKTyr407 may be associated with changes in the neuroendocrine phenotype. Overexpression of constitutively active FAK plasmids (wild-type FAK or FAKTyr397 mutant) into BON cells reduced neuroendocrine markers, whereas the FAKTyr407 mutant plasmid did not show any decrease in the levels of neuroendocrine markers, indicating that phosphorylation of FAK at the Tyr407 residue may be important for these effects. Our results showed for the first time that FAK is an essential downstream effector of the Raf-1/MEK1/2/ERK1/2 signaling cascade and negatively regulated the neuroendocrine and metastatic phenotype in BON cells. Mol Cancer Res; 8(5); 775–82. ©2010 AACR.
STAT1 Represses Skp2 Gene Transcription to Promote p27Kip1 Stabilization in Ras-Transformed Cells
Authors: Wang Shuo , Raven Jennifer F. , Koromilas Antonis E.
Published: April 2010

The S-phase kinase-associated protein 2 (Skp2) is an F-box protein that serves as a subunit of the Skp1-Cullin-F-box ubiquitin protein ligase complex. Skp2 is overexpressed in many tumors and promotes tumor formation through its ability to induce the degradation of proteins with antiproliferative and tumor-suppressor functions, such as p27Kip1. The signal transducer and activator of transcription 1 (STAT1) is a key regulator of the immune system through its capacity to act downstream of interferons. STAT1 exhibits tumor-suppressor properties by inhibiting oncogenic pathways and promoting tumor immunosurveillance. Previous work established the antitumor function of STAT1 in Ras-transformed cells through the induction of p27Kip1 at the transcriptional level. Herein, we unveil a novel pathway used by STAT1 to upregulate p27Kip1. Specifically, we show that STAT1 impedes Skp2 gene transcription by binding to Skp2 promoter DNA in vitro and in vivo. Decreased Skp2 expression by STAT1 is accompanied by the increased stability of p27Kip1 in Ras-transformed cells. We further show that impaired expression of STAT1 in human colon cancer cells containing an activated form of K-Ras is associated with the upregulation of Skp2 and downregulation of p27Kip1. Our study identifies Skp2 as a new target gene of STAT1 in Ras-transformed cells with profound implications in cell transformation and tumorigenesis. Mol Cancer Res; 8(5); 798–805. ©2010 AACR.
Pharmacology of Sedation Agents and Reversal Agents
Authors: Fassoulaki Argyro , Theodoraki Kassiani , Melemeni Aikaterini
Published: April 2010

Sedation for gastrointestinal endoscopies is obtained by opioids, benzodiazepines, propofol, ketamine and/or droperidol. The pharmacokinetic profile of some sedatives/anesthetics renders them advantageous over others. Opioids, mainly pethidine and fentanyl, are the most popular. Though newer opioids provide a faster recovery, fentanyl is safe and advantageous due to its lower cost. Remifentanil, due to its pharmacokinetic profile (elimination half-life: 9 min), is advantageous for ambulatory patients, though it is not known whether the high cost compensates the benefits. Midazolam is the benzodiazepine of choice as it has a shorter duration of action and a better pharmacokinetic profile than diazepam. Propofol, an intravenous anesthetic, has become very popular for gastrointestinal endoscopies in sedative doses. The opioid and benzodiazepine antagonists, naloxone and flumazenil, are indicated only in particular circumstances, like deep sedation with threatening respiratory depression. Ketamine and droperidol are not popular agents for sedation in the modern endoscopic practice.
Basic Requirements for Monitoring Sedated Patients: Blood Pressure, Pulse Oximetry, and EKG
Authors: Maurer Walter G. , Walsh Michael , Viazis Nikos
Published: April 2010

The American Society of Anesthesiologists published monitoring requirements for non-anesthesia providers performing sedation and analgesia in 2002. This manuscript covered not only the monitoring of patients under sedation and analgesia but pre-procedure evaluation and preparation, personnel availability and training, emergency equipment availability, use of supplemental oxygen, sedative-analgesic agents, and discharge criteria. Current recommendations for monitoring include patient responsiveness, blood pressure, respiratory rate, heart rate, and oxygen saturation. Oxygen saturation is a critical vital sign, but there can be a significant delay between inadequate ventilation and desaturation. Supplemental oxygen can dangerously increase this disconnect. Thus, one must monitor adequacy of ventilation by direct observation, auscultation, and/or end-tidal CO2 monitoring. The most important admonition is that ‘ventilation’ and ‘oxygenation’ are not the same.
The Use of Propofol by Gastroenterologists: Medico-Legal Issues
Authors: Axon Andrew E.
Published: April 2010

The increasing use of non-anesthesiologist-administered propofol for sedation during gastrointestinal endoscopy has both clinical and legal consequences. As medical practices develop, the law of clinical negligence will be applied in the context of such developments. While the law will recognize the desirability of advanced techniques and methods, in circumstances where an injury or adverse outcome occurs, the facts of a particular case will be scrutinized to determine whether or not the duty of care between patient and clinician has been breached. This paper considers a number of specific matters likely to arise in the context of a clinical negligence/malpractice claim resulting from the use of non-anesthesiologist-administered propofol, in particular the role and standard of care expected of gastroenterologist and/or anesthetic provider, the relevance of FDA labeling, the implementation and use of protocols, the importance of patient selection and informed consent.
Advanced Cardiac Life Support Training
Authors: Despott Edward J. , Schreiber Florian
Published: April 2010

The OMED/ESGE consensus statements of the International Symposium on Sedation in Endoscopy, Athens, September 2009, in keeping with guidelines and position statements published by other societies, underline the need for sedation providers to be adequately trained in dealing with scenarios involving patients in respiratory and/or cardiovascular distress. This training should prepare the sedation provider with the necessary acumen to prevent, recognize and remedy sedation-related emergencies. Life support training that adheres to the International Liaison Committee on Resuscitation (ILCOR) guidelines should be a mandatory component of this instruction that should be governed by formal assessment and quality assurance reappraisal.
Elements of Airway Training
Authors: Arnaoutoglou Eleni
Published: April 2010

Little research has been conducted into the topic of training in sedation for endoscopic procedures. In addition, a review of the literature did not reveal any published studies regarding the elements of airway training for endoscopists. From the anesthesiology published data, it seems that common methods for airway management training include theoretical instructions and hands-on sessions on anesthetized patients, manikins or simulators. Retraining is necessary in order to maintain knowledge and to refresh and update airway skills and should be conducted on a regular basis with simulation-based practice, practice with validated airway training courses, or even with hands-on training in the anesthetic room. The need for the development of formalized airway training courses carrying the joint approval of the gastrointestinal endoscopy and anesthesiology societies seems to be essential.
Documentation of Competency on Sedation for Gastrointestinal Endoscopy
Authors: Bjorkman David J.
Published: April 2010

There is general consensus in national and societal guidelines that training for sedation should be part of basic training for endoscopy. There is no clear consensus, however, on the structure of that training. More importantly, these same guidelines are often silent on the specific requirements to demonstrate competency for administration of sedation, ways to document that competency and measures to assure that competency is maintained. In the absence of data demonstrating improved outcomes with specific approaches, the process and principles in this paper are proposed as a starting point to be modified by future research and data.
Propofol Infusion Platforms: Opportunities and Challenges
Authors: Maurer Walter G. , Philip Beverly K.
Published: April 2010

Several propofol infusion platforms have been devised to address the inherent safety problems associated with propofol administration. Patient-controlled and target-controlled infusion systems have been described in the literature, and the Sedasys™ closed-loop feedback system is in FDA review. No matter what the infusion platform, there are important safety concerns that should be addressed whenever propofol is administered by non-anesthesiologists. There must be safeguards in the facility’s sedation protocol for non-anesthesiologists that involve training, documentation, auditing/oversight, quality improvement and involvement of the facility’s pharmacy and therapeutics committee. Definitions of levels of sedation and recommendations from the American Society of Anesthesiologists on rescue, identification and management of potential complications, as well as monitoring and training of personnel are also presented.
Sedation Training Using a Human Patient Simulator
Authors: Hofmann N. , Datz C. , Schöchl H.
Published: April 2010

High-fidelity simulation uses simulators that combine all of the physiological and pharmacological responses of a human in a manikin. These simulators change and respond to the users and trainees. Using simulators for teaching sedation in the field of gastroenterology unifies all the advantages of manikins. To understand the pharmacological and pharmacodynamical principles of drugs used for sedation in different clinical scenarios, such as cardiopulmonary diseases, a high-fidelity simulator is extremely useful. Respiratory complications and airway problems are the main side effects when using sedatives. To overcome these problems, exercise of precautionary measures are highly demanded to avoid hazards. High-fidelity simulation is increasingly being used for the development of crisis resource management. There are still limitations in using simulators for education and training. At present, applying simulation for teaching is expensive. The start-up costs and the expenses for instructors and technicians are high. Moreover, no direct evidence has demonstrated that simulation training improves actual patient safety outcome and, therefore, a lot of research in this field remains to be done. Even so, confidence is growing in the validity of medical simulation as the training tool of the future.
Occupational Trichloroethylene Hypersensitivity Syndrome with Human Herpesvirus-6 and Cytomegalovirus Reactivation
Authors: Watanabe Hideaki , Tohyama Mikiko , Kamijima Michihiro , Nakajima Tamie , Yoshida Takemi , Hashimoto Koji , Iijima Masafumi
Published: April 2010

Patients having a generalised rash with severe liver dysfunction associated with exposure to trichloroethylene (TCE) have been reported mainly in Asian countries. However, no case has been reported in Japan since the 1990s. Here, we describe a case of hypersensitivity syndrome (HS) caused by TCE in a 30-year-old Japanese man. The patient developed a rash, fever and liver dysfunction 21 days after he had been exposed to TCE at his workplace. Serum human herpesvirus (HHV)-6 and cytomegalovirus (CMV) DNA were detected 4 and 7 weeks, respectively, after the onset; the IgG antibody titres to HHV-6 and CMV were significantly elevated 6 and 9 weeks, respectively, after the onset. Patch testing was positive for the metabolites of TCE (i.e. trichloroethanol, trichloroacetic acid and chloral hydrate) but not for TCE itself; these results suggest that the TCE metabolites induced this disease. Human leucocyte antigen-B*1301, which has been reported to be strongly associated with TCE-induced HS, was identified in this patient. In addition, the clinical findings, laboratory data and period of virus reactivation after onset were quite similar to those of drug-induced hypersensitivity syndrome (DIHS). We also review TCE-induced HS from the viewpoint of the similarity to DIHS in this article.
Cerebrovascular Risk Factors and Triggers in Transient Global Amnesia Patients with and without Jugular Valve Incompetence: Results from a Sample of 243 Patients
Authors: Agosti C. , Borroni B. , Akkawi N.M. , Padovani A.
Published: April 2010

Background/Aims: Different etiologies for transient global amnesia (TGA) have been proposed, such as venous flow abnormalities, spreading depression and migraine-related mechanisms like focal ischemia. Jugular vein valve incompetence (JVI) is commonly described in TGA patients, but its contributing role in TGA pathogenesis is still unknown. We investigated the possible relationship between JVI and pathogenetic mechanisms in TGA. Methods: 243 TGA patients underwent clinical and neurological assessment. Demographic characteristics, TGA-precipitating factors and vascular comorbidities were carefully recorded. In each patient, JVI was assessed by Doppler ultrasonography. Results: TGA patients were grouped according to the presence (n = 171, 70.4%) or absence (n = 72, 29.6%) of JVI. TGA patients with JVI showed a higher presence of precipitating factors, namely a Valsalva-like maneuver (JVI-positive vs. JVI-negative, 35.8 vs. 17.1%, p = 0.004) and emotional stress (36.6 vs. 21.4%, p = 0.023), but had fewer vascular comorbidities, namely hypertension (37.2 vs. 51.4%, p = 0.047) and carotid arteriosclerosis (20.5 vs. 31.9%, p = 0.050). Conclusions: These findings support the argument for the existence of at least 2 different TGA-associated mechanisms defined by JVI. On the one hand, JVI associated with a Valsalva-like maneuver and emotional stress supported the venous blood congestion hypothesis, and on the other, the JVI-negative group may have another vascular basis, unrelated to venous congestion.
Olopatadine Hydrochloride Improves Dermatitis Score and Inhibits Scratch Behavior in NC/Nga Mice
Authors: Murota Hiroyuki , El-latif Mostafa Abd , Tamura Tadafumi , Amano Toru , Katayama Ichiro
Published: April 2010

Background: Control of itch is an important issue in the treatment of atopic dermatitis (AD). Itch is mediated by a variety of pruritogens, including histamine, and promoted by neurite outgrowth in the epidermis of AD patients, probably due to the release of nerve growth factor. Objectives: We investigated the effects of orally administered olopatadine hydrochloride (olopatadine) on itching, itching mediators, and neuritogenic action in a mouse model. Materials and Methods: NC/Nga mice were treated topically with Dermatophagoides farinae body (Dfb) extract twice weekly for 4 weeks to induce AD-like lesions. They were concomitantly given oral olopatadine, distilled deionized water, or topical tacrolimus during the last 2 weeks. Results: Olopatadine significantly suppressed scratching, improved the dermatitis score, inhibited neurite outgrowth, and decreased the elevated inflammatory markers, growth factors and histamine content in the lesional skin, and serum concentration of Dfb-specific IgE. Notably, olopatadine treatment increased semaphorin 3A expression in the epidermis. Conclusions: Our study confirms the pleiotropic effects of olopatadine, i.e. inhibition of inflammation and neurite extension into the epidermis.