Fatty acid ethyl esters (FAEEs) are cytotoxic nonoxidative ethanol metabolites produced by esterification of fatty acids and ethanol. FAEEs are detectable in blood up to 24 h after ethanol consumption. The objective of this study was to assess the impact of gender, serum or plasma triglyceride concentration, time and temperature of specimen storage, type of alcoholic beverage ingested, and the rate of ethanol consumption on FAEE concentrations in plasma or serum.For some studies, subject were recruited volunteers; in others, residual blood samples after ethanol quantification were used. FAEEs were isolated by solid-phase extraction and quantified by gas chromatography-mass spectrometry.For weight-adjusted amounts of ethanol intake, FAEE concentrations were twofold greater for men than women (P /=24 h. The type of alcoholic beverage and rate of consumption did not affect FAEE concentrations.These studies advance plasma and serum FAEE measurements closer to implementation as a clinical test for ethanol intake.
<p>Supplementary Figure 4 - PDF file 240K, Time-dependent effects of PI3K/mTOR pathway inhibitors on the activation of AKT and MAPK signaling pathways</p>
Abstract In colorectal cancer, malignant cells are surrounded by a complex microenvironment encompassing a range of non-transformed cells, but also a diverse collection of microorganisms. A growing body of evidence demonstrates the role of particular microorganisms in modulating inflammatory environments and promoting tumor growth and metastasis. Studies by our group, and others, reveal a consistent enrichment of Fusobacterium nucleatum in human colorectal cancer, and F. nucleatum has been shown to accelerate tumorigenesis using both in vitro and in vivo preclinical models. We recently demonstrated via microbiome analysis and microbial culture that fusobacteria and its co-occurring microbiota, including Bacteroides, Prevotella and Selenomonas species, persist in liver metastasis of Fusobacterium-positive colorectal cancers. Many of the liver metastasis share the same dominant microbiome (>1% relative abundance) as the paired primary colorectal tumor. Additionally, we have cultured fusobacteria from paired primary and metastatic tumors, and following whole genome sequencing analysis reveal the same strains of Fusobacterium are present in the primary tumors and distant site metastasis, despite the tissue being resected months or even years apart. In situ hybridization analysis demonstrate that Fusobacterium is invasive in the primary tumors and distal metastasis, and is associated with cells whose morphology is consistent with malignant cells. Additionally, we demonstrate via microbiome analysis and microbial culture, that Fusobacterium and its co-occurring microbiome also persist and remain viable in patient derived xenografts of colorectal cancers. Treatment of a patent derived colon cancer xenograft harboring F. nucleatum, with an antibiotic that kills F. nucleatum reduced tumor growth, cancer cell proliferation and tumor fusobacterial load. We have isolated and sequenced the genomes of over 60 F. nucleatum strains from human colorectal cancer tumors with detailed microbiome and patient metadata. In addition to phenotypic analysis and small molecule inhibitory screens of the F. nucleatum colorectal cancer isolates, we are conducting comparative genomic analysis with F. nucleatum isolates from the oral cavity of non-cancer patients to determine colorectal cancer-specific markers and identify targetable genomic attributes. In summary, these findings suggest that the tumor microbiota are intrinsic and essential components of the cancer microenvironment and warrant further investigation into the modulation of the tumor microbiota for the treatment of Fusobacterium-associated colorectal cancer in both early and late stage disease. Citation Format: Susan Bullman, Chandra S. Pedamallu, Ewa Sicinska, Thomas Clancy, Shuji Ogino, Josep Tabernero, Charles Fuchs, William C. Hahn, Paolo Nuciforo, Matthew Meyerson. Fusobacterium and co-occurring microbes in primary and metastatic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5129.
Well-differentiated/dedifferentiated liposarcomas (WD/DDLPS) are among the most common subtypes of soft tissue sarcomas. Conventional systemic chemotherapy has limited efficacy and novel therapeutic strategies are needed to achieve better outcomes for patients. The cyclin-dependent kinase 4 (CDK4) gene is highly amplified in more than 95% of WD/DDLPS. In this study, we explored the role of CDK4 and the effects of NVP-LEE011 (LEE011), a novel selective inhibitor of CDK4/CDK6, on a panel of human liposarcoma cell lines and primary tumor xenografts. We found that both CDK4 knockdown by siRNA and inhibition by LEE011 diminished retinoblastoma (RB) phosphorylation and dramatically decreased liposarcoma cell growth. Cell-cycle analysis demonstrated arrest at G0-G1. siRNA-mediated knockdown of RB rescued the inhibitory effects of LEE011, demonstrating that LEE011 decreased proliferation through RB. Oral administration of LEE011 to mice bearing human liposarcoma xenografts resulted in approximately 50% reduction in tumor (18)F-fluorodeoxyglucose uptake with decreased tumor biomarkers, including RB phosphorylation and bromodeoxyuridine incorporation in vivo. Continued treatment inhibited tumor growth or induced regression without detrimental effects on mouse weight. After prolonged continuous dosing, reestablishment of RB phosphorylation and cell-cycle progression was noted. These findings validate the critical role of CDK4 in maintaining liposarcoma proliferation through its ability to inactivate RB function, and suggest its potential function in the regulation of survival and metabolism of liposarcoma, supporting the rationale for clinical development of LEE011 for the treatment of WD/DDLPS.
