Biology subjects

Graham, M.

Publications and source records attributed to Graham, M..

5 recordsLinked to original sources

bioRxiv2018

Microbiota-metabolites interactions in non-human primate gastrointestinal tract

BackgroundThe microbiota has been recognised as an important part for maintaining human health. Perturbation to its structure has been implicated in many diseases, such as obesity and cancers. The microbiota is highly metabolically active and fills in many niche metabolic pathways absent from the human host. Diseases such as obesity, cardiovascular disease and colorectal cancer has been linked to altered microbiota metabolism. However, there is a gap in the current knowledge on how mucosal-associated microbiota and colon mucosa interact. Here we performed an integrated analysis between the mucosal-associated microbiota and the mucosal tissue metabolites in healthy non-human primates.\n\nResultsWe found that the overall microbiota composition is influenced by both the tissue location as well as the host. We also identified bacteria signatures for different intestinal locations. The distal colon bacterial signature includes Ruminococcaceae, Bacteroidales, Christensenellaceae, Clostridiales, Sphaerochaeta, Victivallaceae, GMD14H09, CF231, ML615J-28, RF39 and R4-45B taxa. In the cecum, the signatures include Prevotella, Anaerovibrio, Roseburia, and Anaerostipes. Desulfovibrionaceae family is the only taxon that may be a signature for the duodenum. We also found an intricate global relationship between the microbiota and the host tissue metabolome that is mainly driven by the distal colon. Most importantly, we found microbial-centric tissue metabolites clusters that may have potential implications to studying host-microbiota metabolic interactions.

genomics↗

bioRxiv2018

An integrated workflow for cross-linking/mass spectrometry

We present a concise workflow to enhance the mass spectrometric detection of cross-linked peptides by introducing sequential digestion and the cross-link identification software Xi. Sequential digestion enhances peptide detection by shortening long tryptic peptides while avoiding over-digestion. We demonstrate our simple 12-fraction protocol for cross-linked multi-protein complexes and cell lysates, quantitative analysis, and high-density cross-linking, without requiring specific cross-linker features. This overall approach reveals dynamic protein-protein interaction sites, which are accessible, have fundamental functional relevance and are therefore ideally suited for the development of small molecule inhibitors.

biochemistry↗

bioRxiv2018

ALOXE3 is a hepatic fasting-responsive lipoxygenase that enhances insulin sensitivity via hepatic PPARγ

ABSTARCTThe hepatic glucose fasting response is gaining traction as a therapeutic pathway to enhance hepatic and whole-host metabolism. However, the mechanisms underlying these metabolic effects remain unclear. Here, we demonstrate the lipoxygenase, ALOXE3, is a novel effector of the thepatic fasting response. We show that ALOXE3 is activated during fasting, glucose withdrawal, and trehalose/trehalose analogue treatment. Hepatocyte-specific ALOXE3 expression reduced weight gain and hepatic steatosis in dietaryand genetically obese (db/db) models. ALOXE3 expression moreover enhanced basal thermogenesis and abrogated insulin resistance in db/db diabetic mice. Targeted metabolomics demonstrated accumulation of the PPAR{gamma} ligand, 12-KETE in hepatocytes overexpressing ALOXE3. Strikingly, PPAR{gamma} inhibition reversed hepatic ALOXE3-mediated insulin sensitization, suppression of hepatocellular ATP production and oxygen consumption, and gene induction of PPAR{gamma} coactivator-1a (PGC1) expression. Moreover, hepatocyte-specific PPAR{gamma} deletion reversed the therapeutic effect of hepatic ALOXE3 expression on diet-induced insulin intolerance. ALOXE3 is therefore a novel effector of the hepatocellular fasting response that leverages both PPAR{gamma}-mediated and pleiotropic effects to augment hepatic and whole-host metabolism, and is thus a promising target to ameliorate metabolic disease.

physiology↗

bioRxiv2017

Novel full-length major histocompatibility complex class I allele discovery and haplotype definition in pig-tailed macaques

Pig-tailed macaques (Macaca nemestrina, Mane) are important models for human immunodeficiency virus (HIV) studies. Their infectability with minimally modified HIV makes them a uniquely valuable animal model to mimic human infection with HIV and progression to acquired immunodeficiency syndrome (AIDS). However, variation in the pig-tailed macaque major histocompatibility complex (MHC) and the impact of individual transcripts on the pathogenesis of HIV and other infectious diseases is understudied compared to rhesus and cynomolgus macaques. In this study, we used Pacific Biosciences single-molecule real-time circular consensus sequencing to describe full-length MHC class I (MHC-I) transcripts for 194 pig-tailed macaques from three breeding centers. We then used the full-length sequences to infer Mane-A and Mane-B haplotypes containing groups of MHC-I transcripts that co-segregate due to physical linkage. In total, we characterized full-length open reading frames (ORFs) for 313 Mane-A, Mane-B, and Mane-I sequences that defined 86 Mane-A and 106 Mane-B MHC-I haplotypes. Pacific Biosciences technology allows us to resolve these Mane-A and Mane-B haplotypes to the level of synonymous allelic variants. The newly defined haplotypes and transcript sequences containing full-length ORFs provide an important resource for infectious disease researchers as certain MHC haplotypes have been shown to provide exceptional control of simian immunodeficiency virus (SIV) replication and prevention of AIDS-like disease in nonhuman primates. The increased allelic resolution provided by Pacific Biosciences sequencing also benefits transplant research by allowing researchers to more specifically match haplotypes between donors and recipients to the level of nonsynonymous allelic variation, thus reducing the risk of graft-versus-host disease.

genetics↗

bioRxiv2016

SNVPhyl: A Single Nucleotide Variant Phylogenomics pipeline for microbial genomic epidemiology

MotivationThe recent widespread application of whole-genome sequencing (WGS) for microbial disease investigations has spurred the development of new bioinformatics tools, including a notable proliferation of phylogenomics pipelines designed for infectious disease surveillance and outbreak investigation. Transitioning the use of WGS data out of the research lab and into the front lines of surveillance and outbreak response requires user-friendly, reproducible, and scalable pipelines that have been well validated.\n\nResultsSNVPhyl (Single Nucleotide Variant Phylogenomics) is a bioinformatics pipeline for identifying high-quality SNVs and constructing a whole genome phylogeny from a collection of WGS reads and a reference genome. Individual pipeline components are integrated into the Galaxy bioinformatics framework, enabling data analysis in a user-friendly, reproducible, and scalable environment. We show that SNVPhyl can detect SNVs with high sensitivity and specificity and identify and remove regions of high SNV density (indicative of recombination). SNVPhyl is able to correctly distinguish outbreak from non-outbreak isolates across a range of variant-calling settings, sequencing-coverage thresholds, or in the presence of contamination.\n\nAvailabilitySNVPhyl is available as a Galaxy workflow, Docker and virtual machine images, and a Unix-based command-line application. SNVPhyl is released under the Apache 2.0 license and available at http://snvphyl.readthedocs.io/ or at https://github.com/phac-nml/snvphyl-galaxy.

bioinformatics↗