Biochemistry and Pharmacology - Research Publications

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    Global expansion of Mycobacterium tuberculosis lineage 4 shaped by colonial migration and local adaptation
    Brynildsrud, OB ; Pepperell, CS ; Suffys, P ; Grandjean, L ; Monteserin, J ; Debech, N ; Bohlin, J ; Alfsnes, K ; Pettersson, JO-H ; Kirkeleite, I ; Fandinho, F ; da Silva, MA ; Perdigao, J ; Portugal, I ; Viveiros, M ; Clark, T ; Caws, M ; Dunstan, S ; Phan, VKT ; Lopez, B ; Ritacco, V ; Kitchen, A ; Brown, TS ; van Soolingen, D ; O'Neill, MB ; Holt, KE ; Feil, EJ ; Mathema, B ; Balloux, F ; Eldholm, V (AMER ASSOC ADVANCEMENT SCIENCE, 2018-10-01)
    On the basis of population genomic and phylogeographic analyses of 1669 Mycobacterium tuberculosis lineage 4 (L4) genomes, we find that dispersal of L4 has been completely dominated by historical migrations out of Europe. We demonstrate an intimate temporal relationship between European colonial expansion into Africa and the Americas and the spread of L4 tuberculosis (TB). Markedly, in the age of antibiotics, mutations conferring antimicrobial resistance overwhelmingly emerged locally (at the level of nations), with minimal cross-border transmission of resistance. The latter finding was found to reflect the relatively recent emergence of these mutations, as a similar degree of local restriction was observed for susceptible variants emerging on comparable time scales. The restricted international transmission of drug-resistant TB suggests that containment efforts at the level of individual countries could be successful.
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    Trajectories of childhood immune development and respiratory health relevant to asthma and allergy.
    Tang, HH ; Teo, SM ; Belgrave, DC ; Evans, MD ; Jackson, DJ ; Brozynska, M ; Kusel, MM ; Johnston, SL ; Gern, JE ; Lemanske, RF ; Simpson, A ; Custovic, A ; Sly, PD ; Holt, PG ; Holt, KE ; Inouye, M (eLife Sciences Publications, Ltd, 2018-10-15)
    Events in early life contribute to subsequent risk of asthma; however, the causes and trajectories of childhood wheeze are heterogeneous and do not always result in asthma. Similarly, not all atopic individuals develop wheeze, and vice versa. The reasons for these differences are unclear. Using unsupervised model-based cluster analysis, we identified latent clusters within a prospective birth cohort with deep immunological and respiratory phenotyping. We characterised each cluster in terms of immunological profile and disease risk, and replicated our results in external cohorts from the UK and USA. We discovered three distinct trajectories, one of which is a high-risk 'atopic' cluster with increased propensity for allergic diseases throughout childhood. Atopy contributes varyingly to later wheeze depending on cluster membership. Our findings demonstrate the utility of unsupervised analysis in elucidating heterogeneity in asthma pathogenesis and provide a foundation for improving management and prevention of childhood asthma.
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    FastSpar: rapid and scalable correlation estimation for compositional data
    Watts, SC ; Ritchie, SC ; Inouye, M ; Holt, KE ; Stegle, O (OXFORD UNIV PRESS, 2019-03-15)
    A common goal of microbiome studies is the elucidation of community composition and member interactions using counts of taxonomic units extracted from sequence data. Inference of interaction networks from sparse and compositional data requires specialized statistical approaches. A popular solution is SparCC, however its performance limits the calculation of interaction networks for very high-dimensional datasets. Here we introduce FastSpar, an efficient and parallelizable implementation of the SparCC algorithm which rapidly infers correlation networks and calculates P-values using an unbiased estimator. We further demonstrate that FastSpar reduces network inference wall time by 2–3 orders of magnitude compared to SparCC.
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    Dynamics of antimicrobial resistance in intestinal Escherichia coli from children in community settings in South Asia and sub-Saharan Africa
    Ingle, DJ ; Levine, MM ; Kotloff, KL ; Holt, KE ; Robins-Browne, RM (NATURE PUBLISHING GROUP, 2018-09-01)
    The dynamics of antimicrobial resistance (AMR) in developing countries are poorly understood, especially in community settings, due to a sparsity of data on AMR prevalence and genetics. We used a combination of phenotyping, genomics and antimicrobial usage data to investigate patterns of AMR amongst atypical enteropathogenic Escherichia coli (aEPEC) strains isolated from children younger than five years old in seven developing countries (four in sub-Saharan Africa and three in South Asia) over a three-year period. We detected high rates of AMR, with 65% of isolates displaying resistance to three or more drug classes. Whole-genome sequencing revealed a diversity of known genetic mechanisms for AMR that accounted for >95% of phenotypic resistance, with comparable rates amongst aEPEC strains associated with diarrhoea or asymptomatic carriage. Genetic determinants of AMR were associated with the geographic location of isolates, not E. coli lineage, and AMR genes were frequently co-located, potentially enabling the acquisition of multi-drug resistance in a single step. Comparison of AMR with antimicrobial usage data showed that the prevalence of resistance to fluoroquinolones and third-generation cephalosporins was correlated with usage, which was higher in South Asia than in Africa. This study provides much-needed insights into the frequency and mechanisms of AMR in intestinal E. coli in children living in community settings in developing countries.
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    Diversity-Generating Machines: Genetics of Bacterial Sugar-Coating
    Mostowy, RJ ; Holt, KE (ELSEVIER SCI LTD, 2018-12-01)
    Bacterial pathogens and commensals are surrounded by diverse surface polysaccharides which include capsules and lipopolysaccharides. These carbohydrates play a vital role in bacterial ecology and interactions with the environment. Here, we review recent rapid advancements in this field, which have improved our understanding of the roles, structures, and genetics of bacterial polysaccharide antigens. Genetic loci encoding the biosynthesis of these antigens may have evolved as bacterial diversity-generating machines, driven by selection from a variety of forces, including host immunity, bacteriophages, and cell-cell interactions. We argue that the high adaptive potential of polysaccharide antigens should be taken into account in the design of polysaccharide-targeting medical interventions like conjugate vaccines and phage-based therapies.
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    Population genomics of hypervirulent Klebsiella pneumoniae clonal-group 23 reveals early emergence and rapid global dissemination
    Lam, MMC ; Wyres, KL ; Duchene, S ; Wick, RR ; Judd, LM ; Gan, Y-H ; Hoh, C-H ; Archuleta, S ; Molton, JS ; Kalimuddin, S ; Koh, TH ; Passet, V ; Brisse, S ; Holt, KE (NATURE PUBLISHING GROUP, 2018-07-13)
    Severe liver abscess infections caused by hypervirulent clonal-group CG23 Klebsiella pneumoniae have been increasingly reported since the mid-1980s. Strains typically possess several virulence factors including an integrative, conjugative element ICEKp encoding the siderophore yersiniabactin and genotoxin colibactin. Here we investigate CG23's evolutionary history, showing several deep-branching sublineages associated with distinct ICEKp acquisitions. Over 80% of liver abscess isolates belong to sublineage CG23-I, which emerged in ~1928 following acquisition of ICEKp10 (encoding yersiniabactin and colibactin), and then disseminated globally within the human population. CG23-I's distinguishing feature is the colibactin synthesis locus, which reportedly promotes gut colonisation and metastatic infection in murine models. These data show circulation of CG23 K. pneumoniae decades before the liver abscess epidemic was first recognised, and provide a framework for future epidemiological and experimental studies of hypervirulent K. pneumoniae. To support such studies we present an open access, completely sequenced CG23-I human liver abscess isolate, SGH10.
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    Morphological, genomic and transcriptomic responses of Klebsiella pneumoniae to the lastline antibiotic colistin
    Cain, AK ; Boinett, CJ ; Barquist, L ; Dordel, J ; Fookes, M ; Mayho, M ; Ellington, MJ ; Goulding, D ; Pickard, D ; Wick, RR ; Holt, KE ; Parkhill, J ; Thomson, NR (NATURE PUBLISHING GROUP, 2018-06-29)
    Colistin remains one of the few antibiotics effective against multi-drug resistant (MDR) hospital pathogens, such as Klebsiella pneumoniae. Yet resistance to this last-line drug is rapidly increasing. Characterized mechanisms of colR in K. pneumoniae are largely due to chromosomal mutations in two-component regulators, although a plasmid-mediated colR mechanism has recently been uncovered. However, the effects of intrinsic colistin resistance are yet to be characterized on a whole-genome level. Here, we used a genomics-based approach to understand the mechanisms of adaptive colR acquisition in K. pneumoniae. In controlled directed-evolution experiments we observed two distinct paths to colistin resistance acquisition. Whole genome sequencing identified mutations in two colistin resistance genes: in the known colR regulator phoQ which became fixed in the population and resulted in a single amino acid change, and unstable minority variants in the recently described two-component sensor crrB. Through RNAseq and microscopy, we reveal the broad range of effects that colistin exposure has on the cell. This study is the first to use genomics to identify a population of minority variants with mutations in a colR gene in K. pneumoniae.
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    Antimicrobial-Resistant Klebsiella pneumoniae Carriage and Infection in Specialized Geriatric Care Wards Linked to Acquisition in the Referring Hospital
    Gorrie, CL ; Mirceta, M ; Wick, RR ; Judd, LM ; Wyres, KL ; Thomson, NR ; Strugnell, RA ; Pratt, NF ; Garlick, JS ; Watson, KM ; Hunter, PC ; McGloughlin, SA ; Spelman, DW ; Jenney, AWJ ; Holt, KE (OXFORD UNIV PRESS INC, 2018-07-15)
    Background: Klebsiella pneumoniae is a leading cause of extended-spectrum β-lactamase (ESBL)-producing hospital-associated infections, for which elderly patients are at increased risk. Methods: We conducted a 1-year prospective cohort study, in which a third of patients admitted to 2 geriatric wards in a specialized hospital were recruited and screened for carriage of K. pneumoniae by microbiological culture. Clinical isolates were monitored via the hospital laboratory. Colonizing and clinical isolates were subjected to whole-genome sequencing and antimicrobial susceptibility testing. Results: K. pneumoniae throat carriage prevalence was 4.1%, rectal carriage 10.8%, and ESBL carriage 1.7%, and the incidence of K. pneumoniae infection was 1.2%. The isolates were diverse, and most patients were colonized or infected with a unique phylogenetic lineage, with no evidence of transmission in the wards. ESBL strains carried blaCTX-M-15 and belonged to clones associated with hospital-acquired ESBL infections in other countries (sequence type [ST] 29, ST323, and ST340). One also carried the carbapenemase blaIMP-26. Genomic and epidemiological data provided evidence that ESBL strains were acquired in the referring hospital. Nanopore sequencing also identified strain-to-strain transmission of a blaCTX-M-15 FIBK/FIIK plasmid in the referring hospital. Conclusions: The data suggest the major source of K. pneumoniae was the patient's own gut microbiome, but ESBL strains were acquired in the referring hospital. This highlights the importance of the wider hospital network to understanding K. pneumoniae risk and infection prevention. Rectal screening for ESBL organisms on admission to geriatric wards could help inform patient management and infection control in such facilities.
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    Airway Microbiota Dynamics Uncover a Critical Window for Interplay of Pathogenic Bacteria and Allergy in Childhood Respiratory Disease
    Teo, SM ; Tang, HHF ; Mok, D ; Judd, LM ; Watts, SC ; Pham, K ; Holt, BJ ; Kusel, M ; Serralha, M ; Troy, N ; Bochkov, YA ; Grindle, K ; Lemanske, RF ; Johnston, SL ; Gern, JE ; Sly, PD ; Holt, PG ; Holt, KE ; Inouye, M (CELL PRESS, 2018-09-12)
    Repeated cycles of infection-associated lower airway inflammation drive the pathogenesis of persistent wheezing disease in children. In this study, the occurrence of acute respiratory tract illnesses (ARIs) and the nasopharyngeal microbiome (NPM) were characterized in 244 infants through their first five years of life. Through this analysis, we demonstrate that >80% of infectious events involve viral pathogens, but are accompanied by a shift in the NPM toward dominance by a small range of pathogenic bacterial genera. Unexpectedly, this change frequently precedes the detection of viral pathogens and acute symptoms. Colonization of illness-associated bacteria coupled with early allergic sensitization is associated with persistent wheeze in school-aged children, which is the hallmark of the asthma phenotype. In contrast, these bacterial genera are associated with "transient wheeze" that resolves after age 3 years in non-sensitized children. Thus, to complement early allergic sensitization, monitoring NPM composition may enable early detection and intervention in high-risk children.
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    Multidrug-Resistant Salmonella enterica, Serotype Typhi, Gulf of Guinea Region, Africa
    Baltazar, M ; Ngandjio, A ; Holt, KE ; Lepillet, E ; Pardos de la Gandara, M ; Collard, J-M ; Bercion, R ; Nzouankeu, A ; Le Hello, S ; Dougan, G ; Fonkoua, M-C ; Weill, F-X (CENTERS DISEASE CONTROL, 2015-04-01)
    We identified 3 lineages among multidrug-resistant (MDR) Salmonella enterica serotype Typhi isolates in the Gulf of Guinea region in Africa during the 2000s. However, the MDR H58 haplotype, which predominates in southern Asia and Kenya, was not identified. MDR quinolone-susceptible isolates contained a 190-kb incHI1 pST2 plasmid or a 50-kb incN pST3 plasmid.