bluuebunny/crossref_metadata_2025 huggingface dataset:
Ugene, a Newly Identified Protein That Is Commonly Overexpressed in Cancer and Binds Uracil DNA Glycosylase
Authors: Guo Chunguang , Zhang Xiaodong , Fink Stephen P. , Platzer Petra , Wilson Keith , Willson James K.V. , Wang Zhenghe , Markowitz Sanford D.
Published: August 2008

Expression microarrays identified a novel transcript, designated as Ugene, whose expression is absent in normal colon and colon adenomas, but that is commonly induced in malignant colon cancers. These findings were validated by real-time PCR and Northern blot analysis in an independent panel of colon cancer cases. In addition, Ugene expression was found to be elevated in many other common cancer types, including breast, lung, uterus, and ovary. Immunofluorescence of V5-tagged Ugene revealed it to have a nuclear localization. In a pull-down assay, uracil DNA glycosylase 2 (UNG2), an important enzyme in the base excision repair (BER) pathway, was identified as a partner protein that binds to Ugene. Coimmunoprecipitation and Western blot analysis confirmed the binding between the endogenous Ugene and UNG2 proteins. Using deletion constructs, we find that Ugene binds to the first 25 amino acids of the UNG2 NH2 terminus. We suggest that Ugene induction in cancer may contribute to the cancer phenotype by interacting with the BER pathway. [Cancer Res 2008;68(15):6118–26]
Loss of Integrin α1β1 Ameliorates Kras-Induced Lung Cancer
Authors: Macias-Perez Ines , Borza Corina , Chen Xiwu , Yan Xuexian , Ibanez Raquel , Mernaugh Glenda , Matrisian Lynn M. , Zent Roy , Pozzi Ambra
Published: August 2008

The collagen IV binding receptor integrin α1β1 has been shown to regulate lung cancer due to its proangiogenic properties; however, it is unclear whether this receptor also plays a direct role in promoting primary lung tumors. To investigate this possibility, integrin α1-null mice were crossed with KrasLA2 mice that carry an oncogenic mutation of the Kras gene (G12D) and develop spontaneous primary tumors with features of non–small cell lung cancer. We provide evidence that KrasLA2/α1-null mice have a decreased incidence of primary lung tumors and longer survival compared with KrasLA2/α1 wild-type controls. Tumors from KrasLA2/α1-null mice were also smaller, less vascularized, and exhibited reduced cell proliferation and increased apoptosis, as determined by proliferating cell nuclear antigen and terminal deoxynucleotidyl-transferase–mediated dUTP nick-end staining, respectively. Moreover, tumors from the KrasLA2/α1-null mice showed diminished extracellular signal-regulated kinase (ERK) but enhanced p38 mitogen-activated protein kinase activation. Primary lung tumor epithelial cells isolated from KrasLA2/α1-null mice showed a significant decrease in anchorage-independent colony formation, collagen-mediated cell proliferation, ERK activation, and, most importantly, tumorigenicity when injected into nude mice compared with KrasLA2/α1 wild-type tumor cells. These results indicate that loss of the integrin α1 subunit decreases the incidence and growth of lung epithelial tumors initiated by oncogenic Kras, suggesting that both Kras and integrin α1β1 cooperate to drive the growth of non–small cell lung cancer in vivo. [Cancer Res 2008;68(15):6127–35]
Synergistic Inhibition of Lung Tumorigenesis by a Combination of Green Tea Polyphenols and Atorvastatin
Authors: Lu Gang , Xiao Hang , You Hui , Lin Yong , Jin Huanyu , Snagaski Blake , Yang Chung S.
Published: August 2008

Purpose: The present study investigated the possible synergistic inhibitory effect of a novel combination of polyphenon E (PPE, a standardized green tea polyphenol preparation) and atorvastatin (trade name Lipitor) in a mouse tumorigenesis model and in human lung cancer H1299 and H460 cell lines.

Experimental Design: Female A/J mice were given two weekly i.p. injections of 4-(methylnitrosaminao)-1-(3-pyridyl)-1-butanone (150 mg/kg total dose); 1 week later, mice were treated with PPE (0.25% or 0.5% in drinking fluid), atorvastatin (200 or 400 ppm in diet), or PPE (0.25%) plus atorvastatin (200 ppm) for 16 weeks. The interaction of these two agents was also studied in human lung cancer H1299 and H460 cells.

Results: The individual agents, PPE or atorvastatin, were not effective in inhibiting lung tumorigenesis. The low-dose combination of PPE and atorvastatin, however, significantly reduced both the tumor multiplicity and tumor burden (by 56% and 55%, respectively, P < 0.05). Isobologram analysis of the interaction of the two agents indicated that the combination synergistically decreased tumor multiplicity (P = 0.0006) and tumor burden (P = 0.0009). The inhibition was associated with enhanced apoptosis and suppressed myeloid cell leukemia 1 (Mcl-1) level in adenoma as determined by immunohistochemistry and Western blots. Treatment with combinations of PPE and atorvastatin also synergistically decreased the number of viable H1299 and H460 cells as determined by isobologram analysis. This synergistic effect was associated with increased apoptosis as determined by the terminal deoxyribonucleotide transferase–mediated nick-end labeling assay. The combination of PPE and atorvastatin was more efficient in reducing the antiapoptotic protein Mcl-1 level and increasing the cleaved caspase-3 and cleaved poly(ADP)-ribose polymerase level than the single-agent treatment.

Conclusions: The present work showed that PPE and atorvastatin synergistically inhibited 4-(methylnitrosaminao)-1-(3-pyridyl)-1-butanone–induced lung tumorigenesis in mice and the growth of lung cancer H1299 and H460 cells, possibly through enhanced apoptosis. The results provide leads for future research on the application of this combination for the prevention and treatment of lung cancer.
The Retinoblastoma Family Member pRb2/p130 Is an Independent Predictor of Survival in Human Soft Tissue Sarcomas
Authors: Masciullo Valeria , Berardengo Ester , Boglione Antonella , Sgambato Alessandro , Bernardi Amelia , Forni Marco , Linari Alessandra , Cito Letizia , Scambia Giovanni , Comandone Alessandro , Giordano Antonio
Published: August 2008

Purpose: pRb2/p130, a member of the Retinoblastoma gene family, has been shown to be a powerful prognostic factor in several malignancies. We sought to evaluate pRb2/p130 protein expression and its clinical effect in patients affected with soft tissue sarcomas (STS).

Experimental Design: Expression of pRb2/p130 was evaluated by immunohistochemistry on formalin-fixed, paraffin-embedded sections in 41 STSs. Results obtained were correlated with clinicopathologic variables and disease-free and overall survival (OS) in univariate and multivariate analysis.

Results: Expression of pRb2/p130 was diminished in 25 (61%) tumors, whereas the remaining ones (39%) were classified as high expressors. No correlation between pRb2/p130 expression and clinicopathologic variables was observed. However, a direct relationship between pRb2/p130 expression and clinical outcome of the patients was found in the subgroup of nonmetastatic tumors (n = 31). In univariate analysis, reduced pRb2/p130 expression was a negative prognostic factor and correlated with shorter disease-free survival (P = 0.021) and OS (P = 0.017) survival. In multivariate analysis, reduced pRb2/p130 expression was confirmed to be an independent predictor of shorter OS when considered together with tumor stage and grading (risk ratio, 7.893; confidence interval, 1.618-38.509; P = 0.011).

Conclusions: This study shows for the first time the potential prognostic value of pRb2/130 expression evaluated on formalin-fixed, paraffin-embedded sections in STSs patients. pRb2/p130 immunoreactivity can be used to predict OS in patients with nonmetastatic STSs and, therefore, may represent a new prognostic marker.
A Pilot Study of Consolidative Immunotherapy in Patients with High-Risk Pediatric Sarcomas
Authors: Mackall Crystal L. , Rhee Eunice H. , Read Elizabeth J. , Khuu Hanh M. , Leitman Susan F. , Bernstein Donna , Tesso Merertu , Long Lauren M. , Grindler David , Merino Margret , Kopp William , Tsokos Maria , Berzofsky Jay A. , Helman Lee J.
Published: August 2008

Purpose: Patients with metastatic or recurrent Ewing’s sarcoma family of tumors and alveolar rhabdomyosarcoma have <25% 5-year survival in most studies. This study administered a novel immunotherapy regimen aimed at consolidating remission in these patients.

Experimental Design: Fifty-two patients with translocation positive, recurrent, or metastatic Ewing’s sarcoma family of tumors or alveolar rhabdomyosarcoma underwent prechemotherapy cell harvest via apheresis for potential receipt of immunotherapy. Following completion of standard multimodal therapy, 30 patients ultimately initiated immunotherapy and were sequentially assigned to three cohorts. All cohorts received autologous T cells, influenza vaccinations, and dendritic cells pulsed with peptides derived from tumor-specific translocation breakpoints and E7, a peptide known to bind HLA-A2. Cohort 1 received moderate-dose recombinant human interleukin-2 (rhIL-2), cohort 2 received low-dose rhIL-2, and cohort 3 did not receive rhIL-2.

Results: All immunotherapy recipients generated influenza-specific immune responses, whereas immune responses to the translocation breakpoint peptides occurred in 39%, and only 25% of HLA-A2+ patients developed E7-specific responses. Toxicity was minimal. Intention-to-treat analysis revealed a 31% 5-year overall survival for all patients apheresed (median potential follow-up 7.3 years) with a 43% 5-year overall survival for patients initiating immunotherapy.

Conclusions: Consolidative immunotherapy is a scientifically based and clinically practical approach for integrating immunotherapy into a multimodal regimen for chemoresponsive cancer. Patients receiving immunotherapy experienced minimal toxicity and favorable survival. The robust influenza immune responses observed suggest that postchemotherapy immune incompetence will not fundamentally limit this approach. Future studies will seek to increase efficacy by using more immunogenic antigens and more potent dendritic cells.
A Phase I Trial of the Oral, Multikinase Inhibitor Sorafenib in Combination with Carboplatin and Paclitaxel
Authors: Flaherty Keith T. , Schiller Joan , Schuchter Lynn M. , Liu Glenn , Tuveson David A. , Redlinger Maryann , Lathia Chetan , Xia Chenghua , Petrenciuc Oana , Hingorani Sunil R. , Jacobetz Michael A. , Van Belle Patricia A. , Elder David , Brose Marcia S. , Weber Barbara L. , Albertini Mark R. , O'Dwyer Peter J.
Published: August 2008

Purpose: This study evaluated the safety, maximum tolerated dose, pharmacokinetics, and antitumor activity of sorafenib, a multikinase inhibitor, combined with paclitaxel and carboplatin in patients with solid tumors.

Patients and Methods: Thirty-nine patients with advanced cancer (24 with melanoma) received oral sorafenib 100, 200, or 400 mg twice daily on days 2 to 19 of a 21-day cycle. All patients received carboplatin corresponding to AUC6 and 225 mg/m2 paclitaxel on day 1. Pharmacokinetic analyses were done for sorafenib on days 2 and 19 of cycle 1 and for paclitaxel on day 1 of cycles 1 and 2. Pretreatment tumor samples from 17 melanoma patients were analyzed for BRAF mutations.

Results: Sorafenib was well tolerated at the doses evaluated. The most frequent severe adverse events were hematologic toxicities (grade 3 or 4 in 33 patients, 85%). Twenty-seven (69%) patients had sorafenib-related adverse events, the most frequent of which were dermatologic events (26 patients, 67%). Exposure to paclitaxel was not altered by intervening treatment with sorafenib. Treatment with sorafenib, paclitaxel, and carboplatin resulted in one complete response and nine partial responses, all among patients with melanoma. There was no correlation between BRAF mutational status and treatment responses in patients with melanoma.

Conclusions: The recommended phase II doses are oral 400 mg twice daily sorafenib, carboplatin at an AUC6 dose, and 225 mg/m2 paclitaxel. The tumor responses observed with this combined regimen in patients with melanoma warrant further investigation.
Human Papillomavirus Type 16/18 Up-Regulates the Expression of Interleukin-6 and Antiapoptotic Mcl-1 in Non–Small Cell Lung Cancer
Authors: Cheng Ya-Wen , Lee Huei , Shiau Ming-Yuh , Wu Tzu-Chin , Huang Tsung-Teng , Chang Yih-Hsin
Published: August 2008

Purpose: Human papillomavirus (HPV) 16/18 infection is reported to be associated with nonsmoking Taiwanese female lung cancer. In this study, we attempted to further reveal the association between HPV infection with Mcl-1 and interleukin (IL)-6 expressions and to elucidate the roles of HPV infection in lung tumorigenesis.

Experimental Design: IL-6 and Mcl-1 expressions were investigated in 79 tumor tissues from lung cancer patients by immunohitochemistry. Secreting IL-6 levels and Mcl-1 expressions were examined by ELISA and Western blot, respectively, in HPV 16/18 E6- and E7-transfected A549 human lung cancer cells, as well as in the HPV16-infected TL-1 lung cancer cells established from lung cancer patients.

Results: Lung tumors (70.9% and 57.0%) had positive IL-6 and Mcl-1 immunostainings, respectively. Significant correlation between IL-6 and Mcl-1 expression were observed (P < 0.0001). Both IL-6 and Mcl-1 expression were significantly associated with HPV 16/18 infection (P = 0.014 and P = 0.004, respectively). IL-6 and Mcl-1 protein levels were not only elevated in HPV 16/18 E6- and E7-transfected A549 cells but also in TL-1 cells. Phosphatidylinositol-3-OH kinase pathway was the major pathway contributing to the up-regulation of Mcl-1 by IL-6 in HPV-infected lung cancer cells.

Conclusions: The up-regulating effects of HPV 16/18 E6 and E7 to IL-6 and Mcl-1 expressions were observed in E6- and E7-transfected A549 cells and in HPV16-infected TL-1 cells, mainly through the phosphatidylinositol-3-OH kinase pathway. The involvement of HPV infection in lung tumorigenesis may be partly through a concomitant increased expression of autocrine and/or paracrine IL-6 and the downstream Mcl-1.
Mismatch Repair Deficiency Does Not Mediate Clinical Resistance to Temozolomide in Malignant Glioma
Authors: Maxwell Jill A. , Johnson Stewart P. , McLendon Roger E. , Lister David W. , Horne Krystle S. , Rasheed Ahmed , Quinn Jennifer A. , Ali-Osman Francis , Friedman Allan H. , Modrich Paul L. , Bigner Darell D. , Friedman Henry S.
Published: August 2008

Purpose: A major mechanism of resistance to methylating agents, including temozolomide, is the DNA repair protein O6-alkylguanine-DNA alkyltransferase (AGT). Preclinical data indicates that defective DNA mismatch repair (MMR) results in tolerance to temozolomide regardless of AGT activity. The purpose of this study was to determine the role of MMR deficiency in mediating resistance in samples from patients with both newly diagnosed malignant gliomas and those who have failed temozolomide therapy.

Experimental Design: The roles of AGT and MMR deficiency in mediating resistance in glioblastoma multiforme were assessed by immunohistochemistry and microsatellite instability (MSI), respectively. The mutation status of the MSH6 gene, a proposed correlate of temozolomide resistance, was determined by direct sequencing and compared with data from immunofluorescent detection of MSH6 protein and reverse transcription-PCR amplification of MSH6 RNA.

Results: Seventy percent of newly diagnosed and 78% of failed-therapy glioblastoma multiforme samples expressed nuclear AGT protein in ≥20% of cells analyzed, suggesting alternate means of resistance in 20% to 30% of cases. Single loci MSI was observed in 3% of patient samples; no sample showed the presence of high MSI. MSI was not shown to correlate with MSH6 mutation or loss of MSH6 protein expression.

Conclusions: Although high AGT levels may mediate resistance in a portion of these samples, MMR deficiency does not seem to be responsible for mediating temozolomide resistance in adult malignant glioma. Accordingly, the presence of a fraction of samples exhibiting both low AGT expression and MMR proficiency suggests that additional mechanisms of temozolomide resistance are operational in the clinic.
Radiation Therapy with Tositumomab (B1) Anti-CD20 Monoclonal Antibody Initiates Extracellular Signal-Regulated Kinase/Mitogen-Activated Protein Kinase–Dependent Cell Death that Overcomes Resistance to Apoptosis
Authors: Ivanov Andrey , Krysov Sergei , Cragg Mark S. , Illidge Tim
Published: August 2008

Purpose: The use of targeted radiation therapy (RT) in conjunction with anti-CD20 monoclonal antibodies (mAb) delivers high clinical response rates in B-cell lymphomas as part of radioimmunotherapy. The mechanisms underlying these impressive responses, particularly in patients whose lymphomas have become refractory to chemotherapy, are poorly understood.

Experimental Design: In this study, we have investigated the signaling pathways and mode of cell death induced in B-cell lymphoma cells after the combination of RT and either type I (rituximab) or type II (tositumomab/B1) anti-CD20 mAb.

Results: Increased tumor cell death was observed when RT was combined with tositumomab, but not rituximab. This additive cell death was found to be mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)–dependent and could be reversed with mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors, as well as small interfering RNA targeting MEK1/2. Furthermore, we found that this increased death was associated with ERK1/2 nuclear accumulation after tositumomab treatment, which was enhanced in combination with RT. Importantly, although Bcl-2 overexpression resulted in resistance to RT-induced apoptosis, it had no effect on the tumor cell death induced by tositumomab plus RT, indicating a nonapoptotic form of cell death.

Conclusions: These findings indicate that RT and type II anti-CD20 mAb combine to stimulate a prodeath function of the MEK-ERK1/2 pathway, which is able to overcome apoptotic resistance potentially explaining the efficacy of this modality in treating patients with chemoresistant disease.
Lung Cancer with Epidermal Growth Factor Receptor Exon 20 Mutations Is Associated with Poor Gefitinib Treatment Response
Authors: Wu Jenn-Yu , Wu Shang-Gin , Yang Chih-Hsin , Gow Chien-Hung , Chang Yih-Leong , Yu Chong-Jen , Shih Jin-Yuan , Yang Pan-Chyr
Published: August 2008

Purpose: Clinical reports about responsiveness to gefitinib treatment in patients of non-small cell lung cancer (NSCLC) with mutations in exon 20 of epidermal growth factor receptor (EGFR) are limited. To increase understanding of the influence of exon 20 mutations on NSCLC treatment with gefitinib, we investigated the clinical features of lung cancer in patients with exon 20 mutations and analyzed the gefitinib treatment response.

Experimental Design: We surveyed the clinical data and mutational studies of NSCLC patients with EGFR exon 20 mutations in the National Taiwan University Hospital and reviewed the literature reports about EGFR exon 20 mutations and the gefitinib treatment response.

Results: Twenty-three patients with mutations in exon 20 were identified. Nine (39%) had coexisting mutations in EGFR exons other than exon 20. Sixteen patients received gefitinib treatment, and a response was noted in 4 patients. The gefitinib response rate of NSCLC with exon 20 mutations was 25%, far lower than those with deletions in exon 19 and L858R mutations. Interestingly, different exon 20 mutations and coexisting mutations seemed to have a different influence on gefitinib response.

Conclusions: EGFR exon 20 mutations of NSCLC patients result in poorer responsiveness to gefitinib treatment, but variability exists between different individuals.
ASPM Is a Novel Marker for Vascular Invasion, Early Recurrence, and Poor Prognosis of Hepatocellular Carcinoma
Authors: Lin Shih-Yeh , Pan Hung-Wei , Liu Shu-Hsiang , Jeng Yung-Ming , Hu Fu-Chang , Peng Shian-Yang , Lai Po-Lin , Hsu Hey-Chi
Published: August 2008

Purpose: Abnormal spindle-like microcephaly associated (ASPM) plays an important role in neurogenesis and cell proliferation. This study is to elucidate its role in hepatocelllular carcinoma (HCC), particularly early tumor recurrence (ETR) and prognosis.

Experimental Design: We used reverse transcription-PCR assays to measure the ASPM mRNA levels in 247 HCC and correlated with clinicopathologic and molecular features.

Results: ASPM mRNA levels were high in fetal tissues but very low in most adult tissues. ASPM mRNA was overexpressed in 162 HCC (66%) but not in benign liver tumors. ASPM overexpression correlated with high α-fetoprotein (P = 1 × 10-8), high-grade (grade II-IV) HCC (P = 2 × 10-6), high-stage (stage IIIA-IV) HCC (P = 1 × 10-8), and importantly ETR (P = 1 × 10-8). ETR is the most critical unfavorable clinical prognostic factor. Among the various independent histopathologic (tumor size, tumor grade and tumor stage) and molecular factors (p53 mutation, high α-fetoprotein, and ASPM overexpression), tumor stage was the most crucial histologic factor (odds ratio, 14.7; 95% confidence interval, 6.65-33.0; P = 1 × 10-8), whereas ASPM overexpression (odds ratio, 6.49; P = 1 × 10-8) is the most important molecular factor associated with ETR. ASPM overexpression was associated with vascular invasion and ETR in both p53-mutated (all P values = 1 × 10-8) and non-p53-mutated HCC (P = 1 × 10-8 and 0.00088, respectively). Hence, patients with APSM-overexpressing HCC had lower 5-year survival (P = 0.000001) in both p53-mutated (P = 0.00008) and non-p53-mutated HCC (P = 0.0027). In low-stage (stage II) HCC, ASPM overexpression also correlated with higher ETR (P = 0.008).

Conclusion: ASPM overexpression is a molecular marker predicting enhanced invasive/metastatic potential of HCC, higher risk of ETR regardless of p53 mutation status and tumor stage, and hence poor prognosis.
Genetic Polymorphisms of Metastasis Suppressor Gene NME1 and Breast Cancer Survival
Authors: Qu Shimian , Long Jirong , Cai Qiuyin , Shu Xiao-Ou , Cai Hui , Gao Yu-Tang , Zheng Wei
Published: August 2008

Purpose: Ample evidence supports an important role of tumor metastasis suppressor genes in cancer metastatic processes. We evaluated the association of genetic polymorphisms of metastasis suppressor gene NME1 with breast cancer prognosis in a follow-up study of patients with primary breast cancer and further investigated the functions of these polymorphisms.

Experimental Design: NME1 genotypes were analyzed in a cohort of 1,134 breast cancer patients recruited as part of the Shanghai Breast Cancer Study who were followed for a median of 7.1 years. In vitro biochemical analyses were carried out to examine the function of NME1 gene polymorphisms.

Results: Single nucleotide polymorphisms (SNP) in the promoter region of the NME1 gene were found to be associated with breast cancer prognosis. Patients carrying the C allele in rs16949649 were associated with higher breast cancer–specific mortality [hazard ratio (HR), 1.4; 95% confidence interval (95% CI), 1.1-1.9] compared with those carrying the wild-type allele, and the association was more evident in patients with an early-stage cancer (HR, 1.7; 95% CI, 1.2-2.5). SNP rs2302254 was also associated with breast cancer prognosis, and the association was statistically significant for the risk of breast cancer relapse, metastasis, and death (HR, 1.3; 95% CI, 1.0-1.6). In vitro biochemical analyses showed that minor alleles in rs2302254 and rs3760468, which is in strong linkage disequilibrium with rs16949646, altered nuclear proteins binding capacity and reduced NME1 promoter activity, supporting the results from an association study of these SNPs with breast cancer survival.

Conclusion: Promoter polymorphisms in the NME1 gene may alter its expression and influence breast cancer survival.
Erythropoiesis-Stimulating Agent Use in Cancer: Preclinical and Clinical Perspectives
Authors: Arcasoy Murat O.
Published: August 2008

Erythropoiesis-stimulating agents (ESA) used for the treatment of chemotherapy-induced anemia in cancer patients have been associated with adverse outcomes of enhanced tumor progression and impaired survival in a series of recent clinical trials. As clinical practice guidelines for ESA administration in cancer patients have evolved to improve safety, the mechanisms underlying the adverse outcomes and whether ESAs exert direct and/or indirect effects in primary tumors to modulate tumor cell growth, survival, and chemoradiotherapy responses remain uncertain. Erythropoietin receptor (EpoR) expression in tumor cells has raised the simplistic possibility that Epo signaling mediated via a functional cellular receptor may contribute to tumor progression in a direct manner. However, Epo biology in cancer is likely to be complex and an interplay of multiple factors is potentially involved in the overall tumor response to exogenous Epo. Optimization of ESA use as an important supportive therapy modality in cancer patients, and further investigation of the role of Epo-EpoR in cancer biology will require a combination of carefully designed preclinical and clinical studies designed to ascertain not only the effect of ESA therapy on clinical outcomes such as tumor response, progression-free, and overall survival but also to investigate the potential effects of Epo on biomarkers of EpoR activation and factors related to tumor biology and chemoradiation responsiveness.
Combination Chemotherapy and ALVAC-CEA/B7.1 Vaccine in Patients with Metastatic Colorectal Cancer
Authors: Kaufman Howard L. , Lenz Heinz-Josef , Marshall John , Singh Deepti , Garett Chris , Cripps Christine , Moore Malcolm , von Mehren Margaret , Dalfen Richard , Heim William J. , Conry Robert M. , Urba Walter J. , Benson Al B. , Yu Maria , Caterini Judy , Kim-Schulze Seunghee , DeBenedette Mark , Salha Danielle , Vogel Thorsten , Elias Ileana , Berinstein Neil L.
Published: August 2008

Purpose: The combination of vaccines and chemotherapy holds promise for cancer therapy, but the effect of cytotoxic chemotherapy on vaccine-induced antitumor immunity is unknown. This study was conducted to assess the effects of systemic chemotherapy on ALVAC-CEA/B7.1–induced T-cell immunity in patients with metastatic colorectal cancer.

Experimental Design: Patients with metastatic colorectal cancer were treated with fluorouracil, leucovorin, and irinotecan and were also given ALVAC-CEA/B7.1 vaccine with or without tetanus toxoid adjuvant. Eligible patients were randomized to ALVAC followed by chemotherapy and booster vaccination (group 1), ALVAC and tetanus toxoid followed by chemotherapy (group 2), or chemotherapy alone followed by ALVAC in patients without disease progression (group 3). Humoral immune responses were measured by standard ELISA assay, and carcinoembryonic antigen (CEA)-specific T-cell responses were measured by IFN-γ enzyme-linked immunospot assay.

Results: One hundred eighteen patients were randomized to receive either ALVAC before and concomitantly with chemotherapy (n = 39), ALVAC with tetanus adjuvant before and concomitantly with chemotherapy (n = 40), or chemotherapy followed by ALVAC (n = 39). Serious adverse events were largely gastrointestinal (n = 30) and hematologic (n = 24). Overall, 42 patients (40.4%) showed objective clinical responses. All patients developed antibody responses against ALVAC, but increased anti-CEA antibody titers were detected in only three patients. Increases in CEA-specific T cells were detected in 50%, 37%, and 30% of patients in groups 1, 2, and 3, respectively. There were no differences in clinical or immune responses between the treatment groups.

Conclusion: The combination of ALVAC-CEA/B7.1 vaccine and systemic chemotherapy has an acceptable safety profile in patients with metastatic colorectal cancer. Systemic chemotherapy did not affect the generation of CEA-specific T-cell responses following vaccination.
Motexafin Lutetium-Photodynamic Therapy of Prostate Cancer: Short- and Long-Term Effects on Prostate-Specific Antigen
Authors: Patel Hiral , Mick Rosemarie , Finlay Jarod , Zhu Timothy C. , Rickter Elizabeth , Cengel Keith A. , Malkowicz S. Bruce , Hahn Stephen M. , Busch Theresa M.
Published: August 2008

Purpose: The time course of serum prostate-specific antigen (PSA) response to photodynamic therapy (PDT) of prostate cancer was measured.

Experimental Design: Seventeen patients were treated in a phase I trial of motexafin lutetium-PDT. PDT dose was calculated in each patient as the product of the ex vivo measured pre-PDT photosensitizer level and the in situ measured light dose. Serum PSA level was measured within 2 months before PDT (baseline), and at day 1; weeks 1 to 3; months 1, 2, and 3; months 4 to 6; and months 7 to 11 after PDT.

Results: At 24 hours after PDT, serum PSA increased by 98% ± 36% (mean ± SE) relative to baseline levels (P = 0.007). When patients were dichotomized based on median PDT dose, those who received high PDT dose showed a 119% ± 52% increase in PSA compared with a 54% ± 27% increase in patients treated at low PDT dose. Patients treated with high versus low PDT dose showed a median biochemical delay of 82 versus 43 days (P = 0.024), with biochemical delay defined as the length of time between PDT and a nonreversible increase in PSA to a value greater than or equal to baseline.

Conclusions: Results show PDT to induce large, transient increases in serum PSA levels. Patients who experienced high PDT dose showed greater short-term increase in PSA and a significantly more durable PSA response (biochemical delay). These data strongly promote the need for individualized delivery of PDT dose and assessment of treatment effect in PDT of prostate cancer. Information gained from such patient-specific measurements could facilitate the introduction of multiple PDT sessions in patients who would benefit.
Loss of Protein Inhibitors of Activated STAT-3 Expression in Glioblastoma Multiforme Tumors: Implications for STAT-3 Activation and Gene Expression
Authors: Brantley Emily C. , Nabors L. Burton , Gillespie G. Yancey , Choi Youn-Hee , Palmer Cheryl Ann , Harrison Keith , Roarty Kevin , Benveniste Etty N.
Published: August 2008

Purpose: STATs activate transcription in response to numerous cytokines, controlling proliferation, gene expression, and apoptosis. Aberrant activation of STAT proteins, particularly STAT-3, is implicated in the pathogenesis of many cancers, including GBM, by promoting cell cycle progression, stimulating angiogenesis, and impairing tumor immune surveillance. Little is known about the endogenous STAT inhibitors, the PIAS proteins, in human malignancies. The objective of this study was to examine the expression of STAT-3 and its negative regulator, PIAS3, in human tissue samples from control and GBM brains.

Experimental Design: Control and GBM human tissues were analyzed by immunoblotting and immunohistochemistry to determine the activation status of STAT-3 and expression of the PIAS3 protein. The functional consequence of PIAS3 inhibition by small interfering RNA or PIAS3 overexpression in GBM cells was determined by examining cell proliferation, STAT-3 transcriptional activity, and STAT-3 target gene expression. This was accomplished using [3H]TdR incorporation, STAT-3 dominant-negative constructs, reverse transcription-PCR, and immunoblotting.

Results and Conclusions: STAT-3 activation, as assessed by tyrosine and serine phosphorylation, was elevated in GBM tissue compared with control tissue. Interestingly, we observed expression of PIAS3 in control tissue, whereas PIAS3 protein expression in GBM tissue was greatly reduced. Inhibition of PIAS3 resulted in enhanced glioblastoma cellular proliferation. Conversely, PIAS3 overexpression inhibited STAT-3 transcriptional activity, expression of STAT-3–regulated genes, and cell proliferation. We propose that the loss of PIAS3 in GBM contributes to enhanced STAT-3 transcriptional activity and subsequent cell proliferation.
Suppression of Tumor Formation by a Cyclooxygenase-2 Inhibitor and a Peroxisome Proliferator-Activated Receptor γ Agonist in an In vivo Mouse Model of Spontaneous Breast Cancer
Authors: Mustafa Aladdin , Kruger Warren D.
Published: August 2008

Purpose: Activation of COX-2 and inhibition of PPARγ have been observed in human and animal models of breast cancer. Both inhibition of COX-2 and activation of PPARγ can inhibit proliferation of breast cancer cells in vitro. Here, we examine the effects of the COX-2 inhibitor celecoxib and the PPARγ agonist N-(9-fluorenyl-methyloxycarbonyl)-l-leucine (F-L-Leu) on mouse breast tumor cells in vitro and in vivo.

Experimental Design: We created and characterized a mouse mammary adenocarcinoma cell (MMAC-1) line from C3 (1)-SV40 tumor antigen mice to study COX-2 and PPARγ expression and response to celecoxib and F-L-Leu in vitro. To study the in vivo effects, C3 (1)-SV40 tumor antigen mice were given either control diet or diets containing three different concentrations of celecoxib and F-L-Leu as well as a combination of both agents. Mice were then followed for tumor formation up to 1 year.

Results: MMAC-1 cells express both COX-2 and PPARγ mRNA and exhibited cooperative growth inhibition with a combination of celecoxib and F-L-Leu. In mice, the median age of death due to mammary tumors was significantly delayed in celecoxib-treated animals at all three concentrations but was not significantly affected by F-L-Leu treatment alone. A combination of celecoxib and F-L-Leu was significantly more effective than celecoxib alone.

Conclusions: Our findings suggest that a combination of a COX-2 inhibitor and PPARγ agonist can delay breast cancer in a mouse model and suggest that these agents should be studied in the context of human populations with high breast cancer risk.
Moving Toward Individualized Cancer Therapies
Authors: Tonon Giovanni , Anderson Kenneth C.
Published: August 2008

In vivo analysis of the gene expression profiles of cancer cells before and after treatment in patients may define mechanisms of sensitivity and resistance to specific drugs and ultimately allow for the selection of optimal individualized therapy to improve outcome in cancer.
Death Receptors and Their Ligands in Atherosclerosis
Authors: Kavurma Mary M. , Tan Nicole Y. , Bennett Martin R.
Published: August 2008

Atherosclerosis is characterized by the accumulation of a fibro-fatty plaque consisting of immune cells, vascular smooth muscle cells (VSMCs), vascular endothelial cells (ECs), and extracellular matrix, surrounding a lipid-rich core. The complexity of atherosclerosis is highlighted by the multifaceted effects that apoptosis and proliferation of specific cell types can have on vessels at different stages of the disease. Death receptors are membrane-bound protein complexes that on binding their cognate ligand, activate an intracellular signaling cascade that results in apoptosis. More recently, signaling from these receptors has been shown to activate multiple other processes, including cell proliferation. This review summarizes our current understanding of signaling events after death receptor activation and the role of death receptors and their ligands in atherosclerosis.
Chemokines and Cardiovascular Risk
Authors: Aukrust Pål , Halvorsen Bente , Yndestad Arne , Ueland Thor , Øie Erik , Otterdal Kari , Gullestad Lars , Damås Jan K.
Published: August 2008

Based on the importance of inflammation in atherogenesis, recent work has focused on whether plasma markers of inflammation can noninvasively diagnose and prognosticate atherosclerotic disorders. Although several studies support an important pathogenic role of chemokines in atherosclerosis, potentially representing attractive therapeutic targets in atherosclerotic disorders, this does not necessarily mean that chemokines are suitable parameters for risk prediction. In fact, the ability to reflect upstream inflammatory activity, stable levels in individuals, and high stability of the actual protein (eg, long half-life and negligible circadian variation) are additional important criteria for an ideal biomarker in cardiovascular disease. Although plasma/serum levels of certain chemokines (eg, interleukin- 8/CXCL8 and monocyte chemoattractant protein-1/CCL2) have shown to be predictive for future cardiac events in some studies, their role as clinical biomarkers is unclear, and their ability to predict subclinical atherosclerosis has been disappointing. Further prospective studies, including a larger number of patients, are needed to make any firm conclusion. Based on the participation of several chemokines in atherogenesis, it is possible that in the future, combined measurements of multiple chemokines could reveal as a “signature of disease” that can serve as a highly accurate method to assess for the presence of atherosclerotic disease.
Adenovirus-Mediated VEGF Gene Therapy Enhances Venous Thrombus Recanalization and Resolution
Authors: Modarai B. , Humphries J. , Gossage J.A. , Waltham M. , Burnand K.G. , Kanaganayagam G.S. , Afuwape A. , Paleolog E. , Smith A. , Wadoodi A.
Published: August 2008

Objective—

Rapid thrombus recanalization reduces the incidence of post–thrombotic complications. This study aimed to discover whether adenovirus-mediated transfection of the vascular endothelial growth factor gene (ad.VEGF) enhanced thrombus recanalization and resolution.

Methods and Results—

In rats, thrombi were directly injected with either ad.VEGF (n=40) or ad.GFP (n=37). Thrombi in SCID mice (n=12) were injected with human macrophages transfected with ad.VEGF or ad.GFP. Thrombi were analyzed at 1 to 14 days. GFP was found mainly in the vein wall and adventitia by 3 days, but was predominantly found in cells within the body of thrombus by day 7. VEGF levels peaked at 4 days (376±299 pg/mg protein). Ad.VEGF treatment reduced thrombus size by >50% (47.7±5.1 mm
2
to 22.0±4.0 mm
2
,
P
=0.0003) and increased recanalization by >3-fold (3.9±0.69% to 13.6±4.1%,
P
=0.024) compared with controls. Ad.VEGF treatment increased macrophage recruitment into the thrombus by more than 50% (
P
=0.002). Ad.VEGF-transfected macrophages reduced thrombus size by 30% compared with controls (12.3±0.89 mm
2
to 8.7±1.4 mm
2
,
P
=0.04) and enhanced vein lumen recanalization (3.39±0.34% to 5.07±0.57%,
P
=0.02).

Conclusion—

Treatment with ad.VEGF enhanced thrombus recanalization and resolution, probably as a consequence of an increase in macrophage recruitment.
Site-Specific Effects of PECAM-1 on Atherosclerosis in LDL Receptor–Deficient Mice
Authors: Goel Reema , Schrank Benjamin R. , Arora Shikha , Boylan Brian , Fleming Barbara , Miura Hiroto , Newman Peter J. , Molthen Robert C. , Newman Debra K.
Published: August 2008

Objective—

Atherosclerosis is a vascular disease that involves lesion formation at sites of disturbed flow under the influence of genetic and environmental factors. Endothelial expression of adhesion molecules that enable infiltration of immune cells is important for lesion development. Platelet/endothelial cell adhesion molecule-1 (PECAM-1; CD31) is an adhesion and signaling receptor expressed by many cells involved in atherosclerotic lesion development. PECAM-1 transduces signals required for proinflammatory adhesion molecule expression at atherosusceptible sites; thus, it is predicted to be proatherosclerotic. PECAM-1 also inhibits inflammatory responses, on which basis it is predicted to be atheroprotective.

Methods and Results—

We evaluated herein the effect of PECAM-1 deficiency on development of atherosclerosis in LDL receptor–deficient mice. We found that PECAM-1 has both proatherosclerotic and atheroprotective effects, but that the former dominate in the inner curvature of the aortic arch whereas the latter dominate in the aortic sinus, branching arteries, and descending aorta. Endothelial cell expression of PECAM-1 was sufficient for its atheroprotective effects in the aortic sinus but not in the descending aorta, where the atheroprotective effects of PECAM-1 also required its expression on bone marrow–derived cells.

Conclusion—

We conclude that PECAM-1 influences initiation and progression of atherosclerosis both positively and negatively, and that it does so in a site-specific manner.
Hydrogen Peroxide Potentiates the EDHF Phenomenon by Promoting Endothelial Ca
2+
Mobilization
Authors: Edwards David H. , Li Yiwen , Griffith Tudor M.
Published: August 2008

Objective—

The purpose of this study was to test the hypothesis that H
2
O
2
contributes to the EDHF phenomenon by mobilizing endothelial Ca
2+
stores.

Methods and Results—

Myograph studies with rabbit iliac arteries demonstrated that EDHF-type relaxations evoked by the SERCA inhibitor cyclopiazonic acid (CPA) required activation of K
Ca
channels and were potentiated by exogenous H
2
O
2
and the thiol oxidant thimerosal. Preincubation with a submaximal concentration of CPA unmasked an ability of exogenous H
2
O
2
to stimulate an EDHF-type response that was sensitive to K
Ca
channel blockade. Imaging of cytosolic and endoplasmic reticulum [Ca
2+
] in rabbit aortic valve endothelial cells with Fura-2 and Mag-fluo-4 demonstrated that H
2
O
2
and thimerosal, which sensitizes the InsP
3
receptor, both enhanced CPA-evoked Ca
2+
release from stores, and that the potentiating effect of H
2
O
2
was suppressed by the cell-permeant thiol reductant glutathione monoethylester. CPA-evoked relaxations were attenuated by exogenous catalase and potentiated by the catalase inhibitor 3-aminotriazole, and were abolished by the connexin-mimetic peptide
43
Gap26, which interrupts intercellular communication via gap junctions constructed from connexin 43.

Conclusions—

H
2
O
2
can enhance EDHF-type relaxations by potentiating Ca
2+
release from endothelial stores, probably via redox modification of the InsP
3
receptor, leading to the opening of hyperpolarizing endothelial K
Ca
channels and an electrotonically-mediated relaxant response.
Cardiovascular Magnetic Resonance for Direct Assessment of Anatomic Regurgitant Orifice in Mitral Regurgitation
Authors: Buchner Stefan , Debl Kurt , Poschenrieder Florian , Feuerbach Stefan , Riegger Günter A.J. , Luchner Andreas , Djavidani Behrus
Published: August 2008

Background—

In patients with mitral regurgitation (MR), assessment of the severity of valvular dysfunction is crucial. Recently, regurgitant orifice area has been proposed as the most useful indicator of the severity of MR. The purpose of our study was to determine whether planimetry of the anatomic regurgitant orifice (ARO) in patients with MR is feasible by cardiovascular magnetic resonance (CMR) and correlates with invasive catheterization and echocardiography effective regurgitant orifice [ECHO-ERO] by proximal isovelocity surface area.

Methods and Results—

Planimetry of ARO was performed with a 1.5-T CMR scanner using a breath-hold balanced gradient echo sequence true fast imaging with steady state precession (TrueFISP). CMR planimetry of ARO was possible in 35 of 38 patients and was closely correlated with angiographic grading (
r
=0.84,
P
<0.0001). In patients with MR grade ≥III on catheterization, CMR-ARO (0.60�0.29 cm
2
versus 0.30�0.19 cm
2
,
P
<0.0001) as well as ECHO-ERO (0.49�0.17 cm
2
versus 0.27�0.10 cm
2
) were significantly elevated in comparison with MR grade <III. Further, CMR-ARO was closely correlated to CMR regurgitant fraction and volume (
r
=0.90 and
r
=0.91,
P
<0.0001, respectively) and catheterization regurgitant fraction and volume (
r
=0.86 and 0.83,
P
<0.0001, respectively). The correlation between CMR-ARO and ECHO-ERO was 0.81 (
P
<0.0001) and CMR slightly overestimated ECHO-ERO by 0.06 cm
2
(
P
<0.05). As assessed by receiver operating characteristic analysis, CMR-ARO at a threshold of 0.40 cm
2
detected MR grade ≥III as defined by catheterization, with a sensitivity and specificity of 94% and 94%, respectively.

Conclusion—

CMR planimetry of the anatomic mitral regurgitant lesion in patients with MR is feasible and permits quantification of MR with good agreement with the accepted invasive and noninvasive methods. Direct measurement by CMR is a promising new method for the precise assessment of ARO area and the severity of MR.
Aortic Root Measurement by Cardiovascular Magnetic Resonance
Authors: Burman Elisabeth D. , Keegan Jennifer , Kilner Philip J.
Published: August 2008

Background—

Cardiovascular magnetic resonance is widely used for aortic root visualization and measurement, but methods still need to be standardized. Our aim was to identify appropriate planes of acquisition and lines of measurement and record corresponding normal values.

Methods and Results—

We studied 120 healthy volunteers, 10 of each gender in each decile between 20 and 80 years, by using a 1.5-T cardiovascular magnetic resonance system. Steady-state free precession cine acquisitions aligned with the left ventricular outflow tract in oblique sagittal and coronal orientations were used to locate 2 sinus planes that transected the root at its widest point in its maximally expanded systolic and at its end diastolic positions. We measured the cusp-cusp and the cusp-commissure dimensions in these cine planes, each as the average of 3. Diastolic cusp-commissure dimensions were smaller than diastolic cusp-cusp dimensions (32.0�3.5 mm versus 34.6�4.0 mm in men, 28.4�2.8 mm versus 30.7�3.3 mm in women,
P
<0.001 for both). The diastolic cusp-commissure dimensions increased by 0.9 mm per decade in men and 0.7 mm per decade in women (
P
<0.001 for both) and gave higher
R
2
values with respect to age and body surface area (0.40 for men, 0.27 for women) than diastolic cusp-cusp, systolic cusp-commissure, or sinus measurements made in the left ventricular outflow tract planes.

Conclusions—

The results indicate the importance of consistent methods for measurement of the aortic root by cardiovascular magnetic resonance. We recommend diastolic cusp-commissure measurements, which yielded favorable
R
2
values with respect to age and body surface area and were found to correspond closely with reference echocardiographic root measurements recorded in the Framingham cohort. We recorded reference values for these and other possible aortic root measurements by cardiovascular magnetic resonance.
Additional Value of Myocardial Perfusion Imaging During Dobutamine Stress Magnetic Resonance for the Assessment of Coronary Artery Disease
Authors: Gebker Rolf , Jahnke Cosima , Manka Robert , Hamdan Ashraf , Schnackenburg Bernhard , Fleck Eckart , Paetsch Ingo
Published: August 2008

Background—

Dobutamine stress magnetic resonance (DSMR) imaging has emerged as a valuable tool for the detection of inducible wall motion abnormalities. The role of perfusion imaging during DSMR is not well defined. We examined whether the addition of myocardial perfusion imaging during DSMR provides incremental benefit for the evaluation of coronary artery disease.

Methods and Results—

DSMR was combined with perfusion imaging in 455 consecutive patients who were scheduled for clinically indicated invasive coronary angiography. Perfusion images were acquired in 3 standard short-views at rest and during maximum dobutamine-atropine stress. Wall motion and perfusion images were interpreted sequentially, blinded to other data. Significant (≥70%) stenoses were present in 285 patients on invasive coronary angiography. The use of DSMR combined with perfusion imaging versus DSMR increased sensitivity (91% versus 85%,
P
=0.001), but not specificity (70% versus 82%,
P
=0.001), resulting in identical overall diagnostic accuracy (84% versus 84%,
P
=NS; Youden index 0.61 versus 0.67). DSMR combined with perfusion imaging enabled the correct diagnosis of coronary artery disease in an additional 13% of DSMR-negative patients at the cost of 11% more false-positive cases.

Conclusion—

The addition of perfusion imaging during DSMR improves sensitivity for the diagnosis of coronary artery disease but does not enhance overall diagnostic accuracy because of a concomitant decrease in specificity.