bluuebunny/crossref_metadata_2025 huggingface dataset:
Ventilatory, metabolic, and thermal responses to hypercapnia in female rats: effects of estrous cycle, ovariectomy, and hormonal replacement
Authors: Marques Danuzia A. , de Carvalho Débora , da Silva Glauber S. F. , Szawka Raphael E. , Anselmo-Franci Janete A. , Bícego Kênia C. , Gargaglioni Luciane H.
Published: May 2015

The aim of this study was to examine how estrous cycle, ovariectomy, and hormonal replacement affect the respiratory [ventilation (V̇e), tidal volume, and respiratory frequency], metabolic (V̇o2), and thermoregulatory (body temperature) responses to hypercapnia (7% CO2) in female Wistar rats. The parameters were measured in rats during different phases of the estrous cycle, and also in ovariectomized (OVX) rats supplemented with 17β-estradiol (OVX+E2), with a combination of E2and progesterone (OVX+E2P), or with corn oil (OVX+O, vehicle). All experiments were conducted on day 8 after ovariectomy. The intact animals did not present alterations during normocapnia or under hypercapnia in V̇e, tidal volume, respiratory frequency, V̇o2, and V̇e/V̇o2in the different phases of the estrous cycle. However, body temperature was higher in female rats on estrus. Hormonal replacement did not change the ventilatory, thermoregulatory, or metabolic parameters during hypercapnia, compared with the OVX animals. Nevertheless, OVX+E2, OVX+E2P, and OVX+O presented lower hypercapnic ventilatory responses compared with intact females on the day of estrus. Also, rats in estrus showed higher V̇e and V̇e/V̇o2during hypercapnia than OVX animals. The data suggest that other gonadal factors, besides E2and P, are possibly involved in these responses.
Metabolic reprogramming and inflammation act in concert to control vascular remodeling in hypoxic pulmonary hypertension
Authors: Stenmark Kurt R. , Tuder Rubin M. , El Kasmi Karim C.
Published: May 2015

Pulmonary hypertension (PH) is a complex, multifactorial syndrome that remains poorly understood despite decades of research. PH is characterized by profound pulmonary artery (PA) remodeling that includes significant fibro-proliferative and inflammatory changes of the PA adventitia. In line with the emerging concept that PH shares key features with cancer, recent work centers on the idea that PH results from a multistep process driven by reprogramming of gene-expression patterns that govern changes in cell metabolism, inflammation, and proliferation. Data demonstrate that in addition to PA endothelial cells and smooth muscle cells, adventitial fibroblasts from animals with experimental hypoxic PH and from humans with PH (hereafter, termed PH-Fibs) exhibit proinflammatory activation, increased proliferation, and apoptosis resistance, all in the context of metabolic reprogramming to aerobic glycolysis. PH-Fibs can also recruit, retain, and activate naïve macrophages (Mϕ) toward a proinflammatory/proremodeling phenotype through secretion of chemokines, cytokines, and glycolytic metabolites, among which IL-6 and lactate play key roles. Furthermore, these fibroblast-activated Mϕ (hereafter, termed FAMϕ) exhibit aerobic glycolysis together with high expression of arginase 1, Vegfa, and I1lb, all of which require hypoxia-inducible factor 1α and STAT3 signaling. Strikingly, in situ, the adventitial Mϕ phenotype in the remodeled PA closely resembles the Mϕ phenotype induced by fibroblasts in vitro (FAMϕ), suggesting that FAMϕ crosstalk involving metabolic and inflammatory signals is a critical, pathogenetic component of vascular remodeling. This review discusses metabolic and inflammatory changes in fibroblasts and Mϕ in PH with the goal of raising ideas about new interventions to abrogate remodeling in hypoxic forms of PH.
Skipping vs. running as the bipedal gait of choice in hypogravity
Authors: Pavei Gaspare , Biancardi Carlo M. , Minetti Alberto E.
Published: May 2015

Hypogravity challenges bipedal locomotion in its common forms. However, as previously theoretically and empirically suggested, humans can rely on “skipping,” a less common gait available as a functional analog (perhaps a vestigium) of quadrupedal gallop, to confidently move when gravity is much lower than on Earth. We set up a 17-m-tall cavaedium (skylight shaft) with a bungee rubber body-suspension system and a treadmill to investigate the metabolic cost and the biomechanics of low-gravity (Mars, Moon) locomotion. Although skipping is never more metabolically economical than running, the difference becomes marginal at lunar gravities, with both bouncing gaits approaching values of walking on Earth (cost ≈2 J·kg−1·m−1). Nonmetabolic factors may thus be allowed to dominate the choice of skipping on the Moon. On the basis of center of pressure measurements and body segments kinetics, we can speculate that these factors may include a further reduction of mechanical work to move the limbs when wearing space suits and a more effective motor control during the ground (regoliths)-boot interaction.
NOD-Like Receptors: Guardians of Intestinal Mucosal Barriers
Authors: Claes Anne-Kathrin , Zhou Jun Yu , Philpott Dana J.
Published: May 2015

The NOD-like receptors (NLRs) are cytosolic pattern-recognition receptors, which are critically involved in mucosal immune defense. The association of the NLR, NOD2, with inflammatory bowel disease first pointed to the NLRs potential function as guardians of the intestinal barrier. Since then, several studies have emphasized the importance of NLRs in maintaining gut homeostasis and intestinal infections, and in shaping the microbiota. In this review, we will highlight the function of NLRs in intestinal inflammation.
Ecophysiology of Omega Fatty Acids: A Lid for Every Jar
Authors: Arnold Walter , Giroud Sylvain , Valencak Teresa G. , Ruf Thomas
Published: May 2015

Omega fatty acids affect various physiological functions, such as locomotion, cardiac function, and thermogenesis. We highlight evidence from animal models that points to pathways by which specific omega fatty acids exert differential effects. We suggest that optimizing the omega fatty acid composition of tissues involves trade-offs between costs and benefits of specific fatty acids.
DNA Damage in Cancer Therapeutics: A Boon or a Curse?
Authors: Khanna Anchit
Published: May 2015

Millions of DNA-damaging lesions occur every day in each cell of our bodies due to various stresses. The failure to detect and accurately repair these lesions can give rise to cells with high levels of endogenous DNA damage, deleterious mutations, or genomic aberrations. Such genomic instability can lead to the activation of specific signaling pathways, including the DNA damage response (DDR) pathway. Constitutive activation of DDR proteins has been observed in human tumor specimens from different cancer stages, including precancerous and metastatic cancers, although not in normal tissues. The tumor-suppressive role of DDR activity during the premalignant stage has been studied, and strong evidence is emerging for an oncogenic role for DDR proteins such as DNA-PK and CHK1 during the later stages of tumor development. However, the majority of current cancer therapies induce DNA damage, potentially exacerbating protumorigenic genomic instability and enabling the development of resistance. Therefore, elucidating the molecular basis of DNA damage–mediated genomic instability and its role in tumorigenesis is critical. Finally, I discuss the potential existence of distinct DNA damage thresholds at various stages of tumorigenesis and what the ramifications of such thresholds would be, including the ambiguous role of the DDR pathway in human cancers, therapy-induced malignancies, and enhanced therapies. Cancer Res; 75(11); 2133–8. ©2015 AACR.
Lifetime Physical Activity and the Risk of Non-Hodgkin Lymphoma
Authors: Boyle Terry , Gallagher Richard P. , Gascoyne Randy D. , Connors Joseph M. , Le Nhu D. , Spinelli John J.
Published: May 2015

Research regarding the association between physical activity and the risk of non-Hodgkin lymphoma (NHL) is limited and inconsistent, and few studies have investigated whether the intensity and timing of physical activity influence the association. A case–control study of NHL was conducted in British Columbia, Canada, in 2000 to 2004. Data were collected on various NHL risk factors, including moderate-intensity and vigorous-intensity physical activity performed over the lifetime. Logistic regression was used to estimate the association between physical activity and the risk of NHL. This analysis included 818 controls and 749 cases. Lifetime vigorous-intensity physical activity was inversely associated with NHL risk. Participants in the second, third, and fourth quartiles of lifetime vigorous-intensity physical activity had an approximately 25% to 30% lower risk of NHL than those in the lowest quartile [adjusted odds ratios, 0.69 (95% confidence interval [CI], 0.52–0.93); 0.68 (95% CI, 0.50–0.92); and 0.75 (95% CI, 0.55–1.01), respectively]. No consistent associations were observed for total or moderate-intensity physical activity. There were no apparent age periods in which physical activity appeared to confer a greater risk reduction. In this study, we found that lifetime vigorous-intensity physical activity was associated with a significantly reduced risk of NHL. Given this finding, more research on physical activity intensity and timing in relation to NHL risk is warranted. Cancer Epidemiol Biomarkers Prev; 24(5); 873–7. ©2015 AACR.
Statin Use and Survival from Lung Cancer: A Population-Based Cohort Study
Authors: Cardwell Chris R. , Mc Menamin Úna , Hughes Carmel M. , Murray Liam J.
Published: May 2015

Background: Preclinical evidence from lung cancer cell lines and animal models suggest that statins could have anticancer properties. We investigated whether statin users had reduced risk of cancer-specific mortality in a population-based cohort of lung cancer patients.

Methods: Newly diagnosed lung cancer patients, from 1998 to 2009, were identified from English cancer registry data and linked to the UK Clinical Practice Research Datalink, providing prescription records, and to Office of National Statistics mortality data up to 2012. Cox regression models were used to calculate HRs for cancer-specific mortality and 95% confidence intervals (CI) by statin use before and after diagnosis, and to adjust these HRs for potential confounders.

Results: In 3,638 lung cancer patients, there was some evidence that statin use after diagnosis was associated with reduced lung cancer–specific mortality (adjusted HR, 0.89; 95% CI, 0.78–1.02; P = 0.09). Associations were more marked after 12 prescriptions (adjusted HR, 0.81; 95% CI, 0.67–0.98; P = 0.03) and when lipophilic statins were investigated (adjusted HR, 0.81; 95% CI, 0.70–0.94; P = 0.01), but were attenuated in some sensitivity analyses. Furthermore, in 11,051 lung cancer patients, statin use before diagnosis was associated with reduced lung cancer–specific mortality (adjusted HR, 0.88; 95% CI, 0.83–0.93; P < 0.001).

Conclusions: There was some evidence that lung cancer patients who used statins, and particularly simvastatin, had reduced rates of cancer-specific mortality.

Impact: These findings should first be confirmed in observational studies, but provide some support for conducting randomized controlled trials of simvastatin as adjuvant cancer therapy in lung cancer patients. Cancer Epidemiol Biomarkers Prev; 24(5); 833–41. ©2015 AACR.
The Role of Gastroesophageal Reflux and Other Factors during Progression to Esophageal Adenocarcinoma
Authors: Hazelton William D. , Curtius Kit , Inadomi John M. , Vaughan Thomas L. , Meza Rafael , Rubenstein Joel H. , Hur Chin , Luebeck E. Georg
Published: May 2015

Background: U.S. esophageal adenocarcinoma (EAC) incidence increased over 5-fold between 1975 and 2009. Symptomatic gastroesophageal reflux disease (sGERD) elevates the risk for EAC. However, a simple calculation suggests that changes in sGERD prevalence can explain at most approximately 16% of this trend. Importantly, a mechanistic understanding of the influence of sGERD and other factors (OF) on EAC is lacking.

Methods: A multiscale model was developed to estimate temporal trends for sGERD and OF, and their mechanistic role during carcinogenesis. Model calibration was to Surveillance, Epidemiology, and End Results (SEER) incidence and age-dependent sGERD data using maximum likelihood and Markov chain Monte Carlo (MCMC) methods.

Results: Among men, 77.8% [95% credibility interval (CI), 64.9%–85.6%] of the incidence trend is attributable to OF, 13.4% (95% CI, 11.4%–17.3%) to sGERD, and 8.8% (95% CI, 4.2%–13.7%) to sGERD–OF interactions. Among women, 32.6% (95% CI, 27.0%–39.9%) of the trend is attributable to OF, 13.6% (95% CI, 12.5%–15.9%) to sGERD, and 47.4% (95% CI, 30.7%–64.6%) to interactions. The predicted trends were compared with historical trends for obesity, smoking, and proton pump inhibitor use. Interestingly, predicted OF cohort trends correlated most highly with median body mass index (BMI) at age 50 (r = 0.988 for men; r = 0.998 for women).

Conclusions: sGERD and OF mechanistically increase premalignant cell promotion, which increases EAC risk exponentially with exposure duration.

Impact: Surveillance should target individuals with long-duration sGERD and OF exposures. Cancer Epidemiol Biomarkers Prev; 24(7); 1012–23. ©2015 AACR.
CCR 20th Anniversary Commentary: Setting the Stage for Nanoparticle Albumin-Bound Paclitaxel—How Far Science Has Come
Authors: Bashour Sami I. , Ibrahim Nuhad K.
Published: May 2015

In this phase I pharmacokinetic study of ABI-007, which was published in the May 1, 2002, issue of Clinical Cancer Research, Ibrahim and colleagues provided the framework needed for subsequent studies to confirm the benefits of ABI-007 over solvent-based formulations. Since the study's publication, experiments have highlighted the importance of drug-delivery systems, the immune system in cancer biology, and immunoregulatory properties of taxane compounds. Clin Cancer Res; 21(9); 1975–7. ©2015 AACR.

See related article by Ibrahim et al., Clin Cancer Res 2002;8(5) May 2002;1038–44
Targeting Notch Signaling with a Notch2/Notch3 Antagonist (Tarextumab) Inhibits Tumor Growth and Decreases Tumor-Initiating Cell Frequency
Authors: Yen Wan-Ching , Fischer Marcus M. , Axelrod Fumiko , Bond Christopher , Cain Jennifer , Cancilla Belinda , Henner William R. , Meisner Rene , Sato Aaron , Shah Jalpa , Tang Tracy , Wallace Breanna , Wang Min , Zhang Chun , Kapoun Ann M. , Lewicki John , Gurney Austin , Hoey Timothy
Published: May 2015

Purpose: The Notch pathway plays an important role in both stem cell biology and cancer. Dysregulation of Notch signaling has been reported in several human tumor types. In this report, we describe the development of an antibody, OMP-59R5 (tarextumab), which blocks both Notch2 and Notch3 signaling.

Experimental Design: We utilized patient-derived xenograft tumors to evaluate antitumor effect of OMP-59R5. Immunohistochemistry, RNA microarray, real-time PCR, and in vivo serial transplantation assays were employed to investigate the mechanisms of action and pharmacodynamic readouts.

Results: We found that anti-Notch2/3, either as a single agent or in combination with chemotherapeutic agents was efficacious in a broad spectrum of epithelial tumors, including breast, lung, ovarian, and pancreatic cancers. Notably, the sensitivity of anti-Notch2/3 in combination with gemcitabine in pancreatic tumors was associated with higher levels of Notch3 gene expression. The antitumor effect of anti-Notch2/3 in combination with gemcitabine plus nab-paclitaxel was greater than the combination effect with gemcitabine alone. OMP-59R5 inhibits both human and mouse Notch2 and Notch3 function and its antitumor activity was characterized by a dual mechanism of action in both tumor and stromal/vascular cells in xenograft experiments. In tumor cells, anti-Notch2/3 inhibited expression of Notch target genes and reduced tumor-initiating cell frequency. In the tumor stroma, OMP-59R5 consistently inhibited the expression of Notch3, HeyL, and Rgs5, characteristic of affecting pericyte function in tumor vasculature.

Conclusions: These findings indicate that blockade of Notch2/3 signaling with this cross-reactive antagonist antibody may be an effective strategy for treatment of a variety of tumor types. Clin Cancer Res; 21(9); 2084–95. ©2015 AACR.
Pembrolizumab for Treatment of Patients with Advanced or Unresectable Melanoma
Authors: Sullivan Ryan J. , Flaherty Keith T.
Published: May 2015

From Coley's toxin to combination immune checkpoint inhibition, strategies to activate the immune system and generate anticancer immunity have been ongoing for well over a century. Over the past decade, the so-called immune checkpoint inhibitors, generally monoclonal antibodies that target key regulators of T-cell activation, emerged as the most effective immune-targeted agents. Pembrolizumab is the first anti-programmed death 1 (PD-1) antibody approved by the FDA for the treatment of metastatic melanoma. With responses seen in 25% to 40% of patients, depending on dose and setting (i.e., before or after ipilimumab), pembrolizumab specifically and anti–PD-1 antibodies generally are revolutionizing the treatment of melanoma. However, in the setting of other recent advances in the field, a number of practical issues are emerging that need to be addressed to optimize the care of patients with melanoma. First, the optimal sequencing of therapy (first-line immunotherapy over molecular targeted therapy, ipilimumab versus pembrolizumab as initial immune checkpoint inhibitor) is unknown and must be evaluated through randomized trials. Second, there is a strong rationale to combine immune checkpoint inhibitors (i.e., anti–PD-1 with ipilimumab) and to combine immune therapies with targeted therapy agents, so determining whether combination therapy is better than direct sequencing is another critical issue that needs to be addressed in carefully carried out studies. Clin Cancer Res; 21(13); 2892–7. ©2015 AACR.
Abstract 1016: Brain-derived neurotrophic factor increases vascular endothelial growth factor expression and subsequently enhances angiogenesis in human chondrosarcoma cells
Authors: Lin Chih-Yang , Tang Chih-Hsin
Published: May 2015

Chondrosarcomas are a type of primary malignant bone cancer, with a potent capacity for local invasion and distant metastasis. Brain-derived neurotrophic factor (BDNF) is commonly upregulated during neurogenesis. The aim of the present study was to examine the mechanism involved in BDNF-mediated vascular endothelial growth factor (VEGF) expression and angiogenesis in human chondrosarcoma cells. Here, we knocked down BDNF expression in chondrosarcoma cells and assessed their capacity to control VEGF expression and angiogenesis in vitro and in vivo. We found knockdown of BDNF decreased VEGF expression and abolished chondrosarcoma conditional medium-mediated angiogenesis in vitro as well as angiogenesis effects in vivo in the chick chorioallantoic membrane and Matrigel plug nude mouse models. In addition, in the xenograft tumor angiogenesis model, the knockdown of BDNF significantly reduced tumor growth and tumor-associated angiogenesis. BDNF increased VEGF expression and angiogenesis through the TrkB receptor, PLCγ, PKCα, and the HIF-1α signaling pathway. Finally, we analyzed samples from chondrosarcoma patients by immunohistochemical staining. The expression of BDNF and VEGF protein in 56 chondrosarcoma patients was significantly higher than in normal cartilage. In addition, the high level of BDNF expression correlated strongly with VEGF expression and tumor stage. Taken together, our results indicate that BDNF increases VEGF expression and enhances angiogenesis through a signal transduction pathway that involves the TrkB receptor, PLCγ, PKCα, and the HIF-1α. Therefore, BDNF may represent a novel target for anti-angiogenic therapy for human chondrosarcoma.

Citation Format: Chih-Yang Lin, Chih-Hsin Tang. Brain-derived neurotrophic factor increases vascular endothelial growth factor expression and subsequently enhances angiogenesis in human chondrosarcoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1016. doi:10.1158/1538-7445.AM2014-1016
Abstract 1018: The diarylpentanoid curcumin analog GO-Y078 can effectively inhibit tumor angiogenesis through actin disorganization
Authors: sugiyama shunsuke , Ishioka Chikashi , Shibata Hiroyuki
Published: May 2015

Background

Curcumin is a dietary pigment with anti-tumor potential. However, its bioavailability is low, and its numerous analogs are being developed. Diarylpentanoid analogs have approximately 50 times higher tumor suppression potential than curcumin and more effectively regulate various targets including β-catenin, nuclear factor-κB, and caspase-3.

Methods

Analogs were screened using endothelial cell sprouting and Xenopus vasculogenesis assays. Growth inhibition, apoptosis induction, wound healing, and invasion analyses as well as western blotting and cDNA microarrays were also performed. Stress fiber structures and focal adhesion (FA) were examined by immunohistochemistry. Subcellular analog localization was examined using a biotinylated analog.

Results

Sprouting and Xenopus assays indicated that GO-Y078 was comparable to the clinically approved angiogenesis inhibitors sorafenib and sunitinib. Inhibition against endothelial cell growth and migration and apoptosis induction of GO-Y078 were similar to or at most 20 times higher than those of sorafenib and sunitinib. However, inhibition of vascular endothelial growth factor (VEGF) signaling was much less than that observed with sorafenib and sunitinib. GO-Y078 inhibited actin stress fiber formation more effectively than sorafenib and sunitinib. It also decreased FA and induced detachment, resulting in anoikis. These potentials were suggested to be the result of mislocalization of heat shock protein 27 and actin as well as downregulation of fibronectin 1.

Conclusion

GO-Y078 is a new class of angiogenesis inhibitor that suppresses actin organization and vascular endothelial cell FA. Inhibition by these analogs could also be effective for redundancy due to complex angiogenesis signaling.

Citation Format: shunsuke sugiyama, Chikashi Ishioka, Hiroyuki Shibata. The diarylpentanoid curcumin analog GO-Y078 can effectively inhibit tumor angiogenesis through actin disorganization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1018. doi:10.1158/1538-7445.AM2014-1018
Abstract 1021: JFD, a novel small molecule for inhibiting vascular endothelial growth factor receptor-mediated angiogenesis
Authors: Dhandayuthapani Sivanesan , Rathinavelu Appu
Published: May 2015

Angiogenesis is the process of generating new capillary of blood vessels from an existing vasculature that plays a vital role in tumor growth and metastasis. Binding of vascular endothelial growth factor (VEGF) to tyrosine kinase receptor (VEGFR) is an important step for initiating vasculogenesis via. transphosphorylation of VEGF receptor associated tyrosine kinase. Therefore, blocking VEGFR or associated kinases has become a common strategy for arresting tumor angiogenesis. The newly discovered JFD is a novel anti-angiogenic agent that can limit the angiogenic capacity of endothelial cells and block tumor angiogenesis. The In vitro anti-angiogenesis potential of JFD was examined by ECMatrigelTM assay with Human umbilical vein endothelial cells (HUVEC) using different concentrations of JFD in the range of 0 - 10 µM. Among the different concentrations used, 1µM JFD significantly inhibited the angiogenesis when compared to control and a complete inhibition was observed at 10 µM concentration. Competitive binding of JFD against Fluorokine labeled VEGF for the same VEGFR was studied at 0.001 - 1.0 µM concentrations of JFD. A dose dependent inhibition of Fluorokine conjugated VEGF binding to its specific receptor in HUVEC cells was confirmed in our study. In the competition binding assay 1.0 µM concentration of JFD caused 93.5% inhibition of conjugated VEGF binding. The same 1.0 µM concentration of JFD was able to inhibit VEGF (10 ng/ml) induced phosphorylation of VEGFR2 in HUVEC cells. Furthermore, the potency of JFD as inhibitor of angiogenesis was experimentally proved by treating athymic nude mice that was implanted with GI-101A (human breast adenocarcinoma) xenograft tumor. Intraperitoneal injection of JFD at 12 mg/kg of body weight was able to significantly inhibit the tumor growth compared to the control group at different time periods. At the end of the treatment period 70% inhibition of tumor growth was observed in JFD treated animals along with prolongation of the survival. The plasma concentrations of JFD was also measured in Balb-C mice at different time periods (0 min - 180 min), using HPLC method. The data from both in vitro and in vivo study provide a strong basis for JFD to be used as a potent anti-angiogenic agent. (The financial support from the Royal Dames of Cancer Research Inc., Ft. Lauderdale, Florida is gratefully acknowledged).

Citation Format: Sivanesan Dhandayuthapani, Appu Rathinavelu. JFD, a novel small molecule for inhibiting vascular endothelial growth factor receptor-mediated angiogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1021. doi:10.1158/1538-7445.AM2014-1021
Abstract 105: Establishment of imaging-based orthotopic tumor models for pharmacological evaluation of anticancer agents
Authors: Zhang Ning , Wang Wei , Meng Dan , Pan Jinjin , Jin Jing , Fan Yanxia , Jin Wenhao , Zhang Shuo , Chen Ze , Yang Xueqin , Wang Hongjun
Published: May 2015

Since its emergence in the mid 90's, the in vivo bioluminescent imaging (BLI) technology has been widely accepted by academia and pharmaceutical industry for research and pre-clinical drug development. The non-invasiveness and high sensitivity nature of BLI enables tumor lesions to be detectable far before they are palpable, allowing longitudinal tracking of tumors from their early development stages. BLI is particularly advantageous for monitoring systemic and orthotopic tumors that are not accessible by caliper measurement. Current preclinical evaluation of anti-cancer agents relies heavily on subcutaneous xenograft models. However, ectopically developed tumors in the subcutaneous tissue differ drastically in tumor stromata in aspects of cell contents and drug permeability, thus will often poorly predict pharmacological efficacy of the test agents. The notion that orthotopic models can better recapitulate the clinical tumor development and metastasis, thus better predict the clinical outcome of the anticancer drugs has gradually gained acceptance. Towards the development of better mouse models for pre-clinical drug development, we have established over twenty imaging based orthotopic tumor cell lines, including U87MG and AM38 brain tumors, HCT116, LoVo and SW620 colorectal cancers, A549 lung cancer, MSTO-211H mesothelioma, Raji and Namalwa lymphoma, MiaPaca-3 and Panc-1 pancreatic cancer, HepG2 liver cancer, and DU145 prostatic cancer. These tumor cells are extremely bright in luminescence emission, ranging from 1085 to 6397 photons/second/cell, thus allowing very sensitive detection in orthotopic tissues even with a few hundred cells. The tumorigenicity of the luciferase labeled cells has been validated in the pertinent orthotopic models. Evaluation of the responsiveness of some of the models to standard therapeutic agents has been conducted. In the U87MG orthotopic glioma model, we observed tumor regression in mice treated with temozolomide in a 5 mg/kg, po, q2d x 5 regimen. Current study of the orthotopic MSTO-211H mesothelioma model in response to standard clinical treatment drugs, such as pemetrexed, carboplatin and paclitaxel is in progress. It is expected that these imaging based orthotopic models will improve the pre-clinical pharmacological evaluation of anti-cancer agents and therefore better bridge the preclinical to clinical translation.

Citation Format: Ning Zhang, Wei Wang, Dan Meng, Jinjin Pan, Jing Jin, Yanxia Fan, Wenhao Jin, Shuo Zhang, Ze Chen, Xueqin Yang, Hongjun Wang. Establishment of imaging-based orthotopic tumor models for pharmacological evaluation of anticancer agents. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 105. doi:10.1158/1538-7445.AM2014-105
Abstract 1050: Alvespimycin (17-DMAG) blocks TGFβ-induced EMT and migration in A549 lung cancer cells
Authors: Gordian Edna , Welsh Eric , Muñoz-Antonia Teresita
Published: May 2015

Background: Transforming Growth Factor β (TGFβ) can induce Epithelial-to-Messenchymal Transition (EMT) in Non-Small Cell Lung Cancer (NSCLC) leading to increased potential to migrate and metastasize. A unique Tumor Promoting signature (TGFβ-TP97) with genes highly associated with TGFβ-induced EMT was derived from human lung cancer cell line microarray experiments. The TGFβ-TP97 EMT signature was then searched against the Connectivity Map (CMAP) dataset for drugs that induce gene expression changes opposite to those of the TGFβ-TP97 signature, and identified 17-DMAG as a compound that could potentially inhibit EMT in NSCLC cells. 17-DMAG is an Hsp90 inhibitor recently described to have a stronger antiproliferative effect in NSCLC harboring EGFR mutations over EGFR wild type lung cancers cells. Furthermore, 17-DMAG has been recently shown to prevent TGFβ induced fibrosis. Methods: A549 and A549T (A549 cells grown in the presence of TGFβ for over two weeks, that have acquired a mesenchymal/fibroblast-like phenotype) were treated with increasing concentrations of 17-DMAG, both in the absence and presence of 5ng/mL TGFβ, and proliferation was measured through GLO assay after 72 hours. The effects of 17-DMAG on TGFβ signaling was tested by Western Blotting for phosphorylated Smads and markers of TGFβ-induced EMT. 17-DMAG effects on morphology and migration were also determined through wound healing and invasion assays. Results: 1) A549T cells grown in TGFβ are more sensitive to the effects of 17-DMAG than cells grown without TGFβ; 2) Pre-treatment of A549 cells with 17-DMAG blocks the induction of EMT markers such as Snail and the downregulation of Ecadherin; 3) 17-DMAG blocks TGFβ-induced phosphorylation of SMAD2, SMAD3; and 4) 17-DMAG inhibits migration and decreases motility of A549 cells in the presence of TGFβ. Conclusion: These results show that 17-DMAG is a potent inhibitor of the effects of TGFβ-induced EMT in A549 cells. It interferes with the regulated process of TGFβ signaling by its ability to block Smad activation and induction of EMT proteins. These findings suggest that 17-DMAG can potentially be used in the treatment of NSCLC cells to prevent disease dissemination and invasion.

Citation Format: Edna Gordian, Eric Welsh, Teresita Muñoz-Antonia. Alvespimycin (17-DMAG) blocks TGFβ-induced EMT and migration in A549 lung cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1050. doi:10.1158/1538-7445.AM2014-1050
Abstract 1051: Metastasis suppressor CD82 functions as a negative regulator in the adhesion-dependent epithelial-to-mesenchymal transition in prostate cancer cells
Authors: Byun Hee-Jung , Lee Yong-Sun , Jin Young-June , Lee Jae-Sub , Kim Young-Myeong , Lee Hansoo
Published: May 2015

Tetraspanin CD82 suppresses cancer metastasis by inhibiting cell motility and invasiveness. In an effect to reveal the mechanism underlying the metastasis-suppressing role of CD82, we here examined the effect of CD82 on epithelial-mesenchymal transition (EMT), a critical event of cancer cells that promotes migration and invasion. In human prostate cancer cell lines, an increase in CD82 expression levels by exogenous gene delivery elevated E-cadherin levels, but decreased the levels of mesenchymal cell proteins including Snail, Twist, vimentin, N-cadhherin, and α-smooth muscle actin, suggesting EMT-suppressing role of CD82. Also, increased CD82 expression resulted in decreased motility and invasiveness of prostate cancer cells. In contrast, CD82 knockdown decreased E-cadherin levels, but increased the levels of mesenchymal cell proteins, together with cell morphology change to fibroblast-like shape. CD82 also interfered with TGF-β1- and Wnt-induced EMT. Additionally, CD82 decreased expression of β-catenin target genes such as c-myc and cyclin D1 by inhibiting nuclear translocation of β-catenin. Notably, the negative effect of CD82 on EMT was apparent on cells adhered to fibronectin. Fibronectin-binding α3β1/α5β1 integrins were involved in CD82-mediated inhibition of EMT. Importantly, human prostate cancer tissue specimens displayed a positive correlation of CD82 with E-cadherin expression levels, along with a negative correlation of CD82 with vimentin levels. Moreover, both CD82 and E-cadherin levels were inversely correlated with the malignant progression stages of human prostate cancers. Taken together, these results strongly suggest that CD82 suppresses motility and invasiveness of prostate cancer cells by inhibiting EMT in an α3β1/α5β1 integrin-dependent manner.

Citation Format: Hee-Jung Byun, Yong-Sun Lee, Young-June Jin, Jae-Sub Lee, Young-Myeong Kim, Hansoo Lee. Metastasis suppressor CD82 functions as a negative regulator in the adhesion-dependent epithelial-to-mesenchymal transition in prostate cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1051. doi:10.1158/1538-7445.AM2014-1051
Abstract 1091: CCR4+ regulatory T cells progressively accumulate in the presence of leukocyte-derived CCL22/CCL17 in an experimental model of glioblastoma multiforme
Authors: Chang Alan L. , Wainwright Derek A. , Dey Mahua , Han Yu , Lesniak Maciej S.
Published: May 2015

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor in adults. One hallmark of GBM is the accumulation of infiltrating regulatory T cells (Tregs), a highly immunosuppressive T cell subset that suppresses T cell-mediated GBM rejection. Previous work has demonstrated that the Treg-recruiting chemokine, CCL22, is expressed by patient-resected GBM. Importantly, the number of Tregs expressing CCR4, the cognate chemokine receptor for CCL22 and CCL17, is increased in the peripheral blood of GBM patients. To investigate the therapeutic potential of targeting the CCR4 chemokine-receptor axis in brain tumors, we performed FACS analysis to determine the level of CCR4-expressing Tregs and other T cell populations. In addition, we analyzed the mRNA expression levels of the Treg-recruiting chemokines CCL22 and CCL17 in different cellular populations by Percoll gradient isolation followed by qRT-PCR. Using the orthotopic GL261 model of GBM, we found a progressive accumulation of CCR4+ Tregs in the brain tumor. Coincidently, the expression of CCL17 and CCL22 was predominantly localized to the tumor-infiltrating leukocytes, rather than the tumor tissue itself. Finally, we tested the potential impact of a CCR4 antagonist on Treg recruitment and CD8+/Foxp3+ T cell ratio. We found that CCR4 antagonist treatment was able to increase the CD8+/Foxp3+ T cell ratio during the early time points post-GL261 injection. These results implicate the chemokine receptor CCR4 as a therapeutic target for further investigation as a potential strategy to target Treg trafficking to GBM.

Citation Format: Alan L. Chang, Derek A. Wainwright, Mahua Dey, Yu Han, Maciej S. Lesniak. CCR4+ regulatory T cells progressively accumulate in the presence of leukocyte-derived CCL22/CCL17 in an experimental model of glioblastoma multiforme. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1091. doi:10.1158/1538-7445.AM2014-1091
Abstract 1099: Sorafenib increases numbers and functions of tumor-infiltrated T cells and enhances therapeutic outcomes of adoptive T cell therapy by modifying tumor microenvironment
Authors: Hwang Jeng-Jong , Chuang Hui-Yen , Chang Ya-Fang
Published: May 2015

Purpose: Many chemotherapeutic agents are known to have immunomodulatory activity to improve the effectiveness of adoptive T-cell therapy. We hypothesized that sorafenib could reverse the immunosuppression of tumor microenvironment to enhance T-cell mediated tumor regression.

Experimental design: OVA-specific CD8+ T cells were co-cultured with sorafenib-pretreated OVA-overexpressing E.G7 mouse lymphoma cells to evaluate their function using flow cytometry, 51Cr-release assay, and transwell assay. The underlying mechanism of sorafenib-mediated immunomodulation was also explored by Western blotting. In animal studies, the activation of OVA-specific CD8+ T cells transduced with a granzyme B promoter-driven imaging reporter was monitored by bioluminescence imaging and the levels of regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) were assessed by flow cytometry.

Results: We found that sorafenib suppressed the expression of critical immunosuppressive components, including TGF-β , IL-10, VEGF, indoleamine 2,3-dioxygenase (IDO) and CCL2/MCP-1, in E.G7 cells through the inhibition of STAT3 signaling pathway and decreased the percentage of Tregs and MDSCs in tumors. Surprisingly, the anti-tumor effect of small numbers of CD8+ T cells was substantially enhanced when combined with sorafenib in vivo.

Citation Format: Jeng-Jong Hwang, Hui-Yen Chuang, Ya-Fang Chang. Sorafenib increases numbers and functions of tumor-infiltrated T cells and enhances therapeutic outcomes of adoptive T cell therapy by modifying tumor microenvironment. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1099. doi:10.1158/1538-7445.AM2014-1099
Abstract 1101: Acidosis-induced NF-κB promotes cell invasion in breast cancer
Authors: Gupta Subash C. , Mo Yin-Yuan
Published: May 2015

Although acidic microenvironment has been previously shown to promote breast cancer growth and invasion, the underlying mechanism remains largely unknown. Because inflammation is a major culprit in cancer progression, we hypothesize that pro-inflammatory transcription factor nuclear factor (NF)-κB in the acidic microenvironment plays a major role in the invasion and metastasis of breast cancer. In this study we showed that acidosis (pH 6.6) activates NF-κB but it has no effect on STAT3 activity in breast cancer cells. Mechanistically, the activation of NF-κB is mediated through phosphorylation and degradation of IκBα; nuclear translocation, and phosphorylation of p65, leading to enhanced cell invasion. Upstream to NF-κB signaling, AKT, PDK1, and ERK1/2 are also activated under acidic condition. Moreover, acidosis induces reactive oxygen species (ROS) which can be suppressed by ROS scavengers such as glutathione and N-acetyl-L-cysteine, reversing the acidosis-induced activation of AKT, ERK1/2 and NF-κB, and invasiveness. In addition, the NADPH oxidase (NOX) inhibitor diphenyleneiodonium can suppressed acidosis-induced ROS. Importantly, we observed all of these effects only in cancer cells, but not in non-malignant cells. Together, these results suggest that production of ROS, activation of AKT, ERK1/2 and NF-κB may constitute key acidosis-induced signaling cascades in breast cancer cells.

Citation Format: Subash C. Gupta, Yin-Yuan Mo. Acidosis-induced NF-κB promotes cell invasion in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1101. doi:10.1158/1538-7445.AM2014-1101
Abstract 1136: p53-induced miR-15a/16-1 and AP4 form a double-negative feedback loop to regulate epithelial-mesenchymal transition and metastasis in colorectal cancer
Authors: Shi Lei , Jackstadt Rene , Siemens Helge , Li Huihui , Kirchner Thomas , Hermeking Heiko
Published: May 2015

The transcription factor AP4 mediates epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) but its control in this setting is not fully understood. Here we report the definition of a double-negative feedback loop involving AP4 and miR-15a/16-1 that regulates EMT and metastatic progression. In CRC cells, AP4 was downregulated by DNA damage in a p53-dependent manner. AP4 downregulation by p53 was mediated indirectly by the tumor suppressive microRNAs miR-15a and miR-16-1, which targeted the 3′-UTR of AP4 mRNA, induced mesenchymal-epithelial transition (MET) and inhibited CRC cell migration and invasion. The downregulation of AP4 was necessary for MET induction by miR-15a/16-1. In tumor xenoplants, ectopic miR-15a/16-1 suppressed formation of lung metastases. Furthermore, AP4 directly suppressed expression of miR-15a/16-1. In clinical specimens of colorectal cancer, miR-15a levels inversely correlated with AP4 protein levels shown previously to correlate with distant metastasis and poor survival. In summary, our results define a double-negative feedback loop involving miR-15a/16-1 and AP4 that stabilizes epithelial and mesenchymal states, respectively, that may determine metastatic prowess.

Citation Format: Lei Shi, Rene Jackstadt, Helge Siemens, Huihui Li, Thomas Kirchner, Heiko Hermeking. p53-induced miR-15a/16-1 and AP4 form a double-negative feedback loop to regulate epithelial-mesenchymal transition and metastasis in colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1136. doi:10.1158/1538-7445.AM2014-1136
Abstract 1141: p53-dependent regulation of epithelial-to-mescenchymal transition by nf-κb in head and neck squamous cell carcinoma
Authors: Lin Yuan , Luo Jie , Abemayor Elliot , Sharma Sherven , Dubinett Steven , John Maie S.
Published: May 2015

PURPOSE:

In head and neck squamous cell carcinoma (HNSCC), mutations of p53 usually coexist with aberrant activation of NF-kappaB (NF-κB) and other transcription factors which promote tumor pathogenesis. However, how these factors mediate oncogenesis and metastasis is not fully understood. Herein we investigate the mechanisms by which p53 and NF-κB contribute to Epithelial-Mesenchymal Transition (EMT) in HNSCC.

EXPERIMENTAL DESIGN:

We evaluated the effects of p65 mutation on the molecular events of EMT in surgical specimens and HNSCC cell lines. We examined the correlation with tumor histologic features.

RESULTS:

In HNSCC cell lines with mutant p53, p65 knockdown promotes EMT. In contrast, in normal oral epithelial cells or HNSCC cells with WT p53, p65 over-expression, instead of knockdown, promotes EMT. Ablation of p53 in normal oral epithelial cells blocks p65 induced EMT. These findings are functionally corroborated herein in three-dimensional cell culture, anchorage-independent growth assays, and tumor cell migration assays.

CONCLUSIONS:

This is the first report indicating the role of p53 status in the NF-κB -induced promotion of EMT in HNSCC. Bortezomib-based regimens have been shown to have minimal activity in recurrent or metastatic head and neck cancer. Our new findings are important in understanding a possible basis for these clinical findings as well as defining important implications for targeted chemoprevention and therapy.

Citation Format: Yuan Lin, Jie Luo, Elliot Abemayor, Sherven Sharma, Steven Dubinett, Maie St John. p53-dependent regulation of epithelial-to-mescenchymal transition by nf-κb in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1141. doi:10.1158/1538-7445.AM2014-1141
Abstract 1143: Induction of epithelial-mesenchymal transition (EMT) in human gastric cancer cell lines induced by Tipα of H. pylori mediated through surface nucleolin
Authors: Fujiki Hirota , Watanabe Tatsuro , Takahashi Atsushi , Suzuki Kaori , Kurusu-Kanno Miki , Yamaguchi Kensei , Suganuma Masami
Published: May 2015

(a) Introduction: Tumor necrosis factor-α (TNF-α)-inducing protein (Tipα) of H. pylori acts as a carcinogenic factor in the gastric epithelium: it binds to surface nucleolin on human and mouse gastric cancer cell lines, resulting in cancer progression. We studied various phenotypes of epithelial-mesenchymal transition (EMT) to examine the function of surface nucleolin in gastric cancer cell line.

(b) Experimental procedures :

1) Induction of cell migration and elongation: After human gastric cancer cell line MKN-1 was transfected with nucleolin-targeted siRNAs (siRNA-n), cell migration and elongation induced by Tipα were determined using Transwell assay and cell morphology.

2) Formation of filopodia in MKN-1 cells was determined by staining filamentous actin with AF488-phalloidin.

3) Phosphorylation of 89 cancer-related proteins was determined using Cancer Signaling Phosphoantibody Array.

4) Expression of EMT markers was examined with Western blotting.

5) Cell stiffness was determined from Young's modulus using atomic force microscope (AFM).

(c) A summary of the new data:

1) The transfected cells with siRNA-n showed a reduced amount of surface nucleolin in membrane fraction, but not in cytosol or nuclear fractions. Knockdown of nucleolin inhibited cell migration and elongation induced by Tipα.

2) Tipα induced the formation of filopodia in MKN-1 cells, which suggests the invasive morphological changes.

3) Tipα enhanced the phosphorylation of 11 cancer-related proteins in serine, threonine and tyrosine residues, indicating activation of MEK-ERK signal cascade.

4) Tipα induced the expression of vimentin, a significant marker of EMT, even though down-regulation of E-cadherin was not found in MKN-1 cells.

5) Tipα reduced the Young's modulus of MKN-1 cells as determined by AFM: The results showed that lower cell stiffness and increased cell motility were induced by Tipα.

(d) Conclusions:

This is the first report that Tipα is a new inducer of EMT in human gastric cancer cells, and that EMT was induced through the binding of Tipα to surface nucleolin.

Citation Format: Hirota Fujiki, Tatsuro Watanabe, Atsushi Takahashi, Kaori Suzuki, Miki Kurusu-Kanno, Kensei Yamaguchi, Masami Suganuma. Induction of epithelial-mesenchymal transition (EMT) in human gastric cancer cell lines induced by Tipα of H. pylori mediated through surface nucleolin. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1143. doi:10.1158/1538-7445.AM2014-1143