Formation and abundance of 5-hydroxymethylcytosine in RNA.

RNA methylation is emerging as a regulatory RNA modification that could have important roles in the control and coordination of gene transcription and protein translation. Herein, we describe an in vivo isotope-tracing methodology to demonstrate that the ribonucleoside 5-methylcytidine (m(5)C) is subject to oxidative processing in mammals, forming 5-hydroxymethylcytidine (hm(5)C) and 5-formylcytidine (f(5)C). Furthermore, we have identified hm(5)C in total RNA from all three domains of life and in polyA-enriched RNA fractions from mammalian cells. This suggests m(5)C oxidation is a conserved process that could have critical regulatory functions inside cells.This work was supported by the Cambridge PhD Training Programme in Chemical Biology and Molecular Medicine and the Wellcome Trust (grant number 099232/Z/12/Z). Dr. Donna Bond is thanked for the provision of A. thaliana total RNA and Drs. Santiago Uribe-Lewis and Adele Murrell are acknowledged for the labelled mouse tissue supply.This is the final published version. It first appeared at http://onlinelibrary.wiley.com/doi/10.1002/cbic.201500013/abstract;jsessionid=61B9B3D8937FE50CFA4954A4C4B445B6.f02t04

2-Hydroxyglutarate Metabolism Is Altered in an in vivo Model of LPS Induced Endotoxemia.

The metabolic response to endotoxemia closely mimics those seen in sepsis. Here, we show that the urinary excretion of the metabolite 2-hydroxyglutarate (2HG) is dramatically suppressed following lipopolysaccharide (LPS) administration in vivo, and in human septic patients. We further show that enhanced activation of the enzymes responsible for 2-HG degradation, D- and L-2-HGDH, underlie this effect. To determine the role of supplementation with 2HG, we carried out co-administration of LPS and 2HG. This co-administration in mice modulates a number of aspects of physiological responses to LPS, and in particular, protects against LPS-induced hypothermia. Our results identify a novel role for 2HG in endotoxemia pathophysiology, and suggest that this metabolite may be a critical diagnostic and therapeutic target for sepsis

Fumarate metabolic signature for the detection of Reed Syndrome in humans

Purpose: Inherited pathogenic variants in genes encoding the metabolic enzymes succinate dehydrogenase (SDH) and fumarate hydratase (FH) predispose to tumour development through accumulation of oncometabolites (succinate and fumarate respectively) (1). Non-invasive in vivo detection of tumour succinate by proton magnetic resonance spectroscopy (1H-MRS) has been reported in SDH-deficient tumours but the potential utility of this approach in the management of patients with hereditary leiomyomatosis and renal cell cancer syndrome or Reed syndrome is unknown. Experimental design: Magnetic resonance spectroscopy (1H-MRS) was performed on three cases and correlated with germline genetic results and tumour immunohistochemistry when available. Results: Here, we have demonstrated a proof-of-principle that 1H-MRS can provide a non-invasive diagnosis of hereditary leiomyomatosis and renal cell cancer syndrome or Reed syndrome through detection of fumarate accumulation in vivo. Conclusion: This study demonstrates that in vivo detection of fumarate could be employed as a functional biomarkerGIST Support UK (RC), NIHR (RC), Cambridge Experimental Cancer Medicine Centre, Addenbrooke’s Charitable Trust, National Institute for Health Research (NIHR) Cambridge Biomedical Research Centre, Cancer Research UK CRUK (FAG), CRUK Cambridge Centre (MM, FAG), the University of Cambridge, and Hutchison Whampoa Ltd (MM), NIHR Senior Investigator Award (ERM), European Research Council Advanced Researcher Award (ERM), CRUK and Engineering and Physical Sciences Research Council (EPSRC) Imaging Centre in Cambridge and Manchester (FAG). The University of Cambridge has received salary support in respect of EM from the NHS in the East of England through the Clinical Academic Reserve

Divinylpyrimidine reagents generate antibody-drug conjugates with excellent in vivo efficacy and tolerability.

The development of divinylpyrimidine (DVP) reagents for the synthesis of antibody-drug conjugates (ADCs) with in vivo efficacy and tolerability is reported. Detailed structural characterisation of the synthesised ADCs was first conducted followed by in vitro and in vivo evaluation of the ADCs' ability to safely and selectively eradicate target-positive tumours