Tanya M Monaghan1, Anna M Seekatz2, Nicholas O Markham3, Tung On Yau4, Maria Hatziapostolou4, Tahseen Jilani5, Niki Christodoulou4, Brandi Roach6, Eleni Birli4, Odette Pomenya4, Thomas Louie7, D Borden Lacy8, Peter Kim9, Christine Lee10, Dina Kao11, Christos Polytarchou12. 1. National Institute for Health Research Nottingham Biomedical Research Centre, University of Nottingham, Nottingham, United Kingdom; Nottingham Digestive Diseases Centre, School of Medicine, University of Nottingham, Nottingham, United Kingdom. Electronic address: tanya.monaghan@nottingham.ac.uk. 2. Department of Biological Sciences, Clemson University, Clemson, South Carolina, USA. 3. Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA; Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, Tennessee, USA. 4. Department of Biosciences, John van Geest Cancer Research Centre, Centre for Health Aging and Understanding Disease, School of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom. 5. Advanced Data Analysis Centre, School of Computer Science, University of Nottingham, Nottingham, United Kingdom. 6. Department of Medicine, University of Alberta, Edmonton, Alberta, Canada. 7. Department of Microbiology and infectious Diseases, University of Calgary, Calgary, Alberta, Canada. 8. Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA; Veterans Affairs Tennessee Valley Healthcare System, Nashville, Tennessee, USA. 9. Department of Mathematics and Statistics, University of Guelph, Ontario, Canada. 10. Vancouver Island Health Authority, Victoria, British Columbia, Canada; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada. 11. Department of Medicine, University of Alberta, Edmonton, Alberta, Canada. Electronic address: dkao@ualberta.ca. 12. Department of Biosciences, John van Geest Cancer Research Centre, Centre for Health Aging and Understanding Disease, School of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom. Electronic address: christos.polytarchou@ntu.ac.uk.
Abstract
BACKGROUND AND AIMS: The molecular mechanisms underlying successful fecal microbiota transplantation (FMT) for recurrent Clostridioides difficile infection (rCDI) remain poorly understood. The primary objective of this study was to characterize alterations in microRNAs (miRs) following FMT for rCDI. METHODS: Sera from 2 prospective multicenter randomized controlled trials were analyzed for miRNA levels with the use of the Nanostring nCounter platform and quantitative reverse-transcription (RT) polymerase chain reaction (PCR). In addition, rCDI-FMT and toxin-treated animals and ex vivo human colonoids were used to compare intestinal tissue and circulating miRs. miR inflammatory gene targets in colonic epithelial and peripheral blood mononuclear cells were evaluated by quantitative PCR (qPCR) and 3'UTR reporter assays. Colonic epithelial cells were used for mechanistic, cytoskeleton, cell growth, and apoptosis studies. RESULTS: miRNA profiling revealed up-regulation of 64 circulating miRs 4 and 12 weeks after FMT compared with screening, of which the top 6 were validated in the discovery cohort by means of RT-qPCR. In a murine model of relapsing-CDI, RT-qPCR analyses of sera and cecal RNA extracts demonstrated suppression of these miRs, an effect reversed by FMT. In mouse colon and human colonoids, C difficile toxin B (TcdB) mediated the suppressive effects of CDI on miRs. CDI dysregulated DROSHA, an effect reversed by FMT. Correlation analyses, qPCR ,and 3'UTR reporter assays revealed that miR-23a, miR-150, miR-26b, and miR-28 target directly the 3'UTRs of IL12B, IL18, FGF21, and TNFRSF9, respectively. miR-23a and miR-150 demonstrated cytoprotective effects against TcdB. CONCLUSIONS: These results provide novel and provocative evidence that modulation of the gut microbiome via FMT induces alterations in circulating and intestinal tissue miRs. These findings contribute to a greater understanding of the molecular mechanisms underlying FMT and identify new potential targets for therapeutic intervention in rCDI.
BACKGROUND AND AIMS: The molecular mechanisms underlying successful fecal microbiota transplantation (FMT) for recurrent Clostridioides difficile infection (rCDI) remain poorly understood. The primary objective of this study was to characterize alterations in microRNAs (miRs) following FMT for rCDI. METHODS: Sera from 2 prospective multicenter randomized controlled trials were analyzed for miRNA levels with the use of the Nanostring nCounter platform and quantitative reverse-transcription (RT) polymerase chain reaction (PCR). In addition, rCDI-FMT and toxin-treated animals and ex vivo human colonoids were used to compare intestinal tissue and circulating miRs. miR inflammatory gene targets in colonic epithelial and peripheral blood mononuclear cells were evaluated by quantitative PCR (qPCR) and 3'UTR reporter assays. Colonic epithelial cells were used for mechanistic, cytoskeleton, cell growth, and apoptosis studies. RESULTS: miRNA profiling revealed up-regulation of 64 circulating miRs 4 and 12 weeks after FMT compared with screening, of which the top 6 were validated in the discovery cohort by means of RT-qPCR. In a murine model of relapsing-CDI, RT-qPCR analyses of sera and cecal RNA extracts demonstrated suppression of these miRs, an effect reversed by FMT. In mouse colon and human colonoids, C difficile toxin B (TcdB) mediated the suppressive effects of CDI on miRs. CDI dysregulated DROSHA, an effect reversed by FMT. Correlation analyses, qPCR ,and 3'UTR reporter assays revealed that miR-23a, miR-150, miR-26b, and miR-28 target directly the 3'UTRs of IL12B, IL18, FGF21, and TNFRSF9, respectively. miR-23a and miR-150 demonstrated cytoprotective effects against TcdB. CONCLUSIONS: These results provide novel and provocative evidence that modulation of the gut microbiome via FMT induces alterations in circulating and intestinal tissue miRs. These findings contribute to a greater understanding of the molecular mechanisms underlying FMT and identify new potential targets for therapeutic intervention in rCDI.
Authors: Roni Nowarski; Ruaidhrí Jackson; Nicola Gagliani; Marcel R de Zoete; Noah W Palm; Will Bailis; Jun Siong Low; Christian C D Harman; Morven Graham; Eran Elinav; Richard A Flavell Journal: Cell Date: 2015-12-03 Impact factor: 41.582
Authors: Gang Chen; Yuan Feng; Xuezheng Li; Zhe Jiang; Bei Bei; Lin Zhang; Yueqing Han; Yanwu Li; Ning Li Journal: Front Genet Date: 2019-06-19 Impact factor: 4.599
Authors: Tanya Monaghan; Benjamin H Mullish; Jordan Patterson; Gane Ks Wong; Julian R Marchesi; Huiping Xu; Tahseen Jilani; Dina Kao Journal: Gut Microbes Date: 2018-09-05
Authors: Tanya M Monaghan; Niharika A Duggal; Elisa Rosati; Ruth Griffin; Jamie Hughes; Brandi Roach; David Y Yang; Christopher Wang; Karen Wong; Lynora Saxinger; Maja Pučić-Baković; Frano Vučković; Filip Klicek; Gordan Lauc; Paddy Tighe; Benjamin H Mullish; Jesus Miguens Blanco; Julie A K McDonald; Julian R Marchesi; Ning Xue; Tania Dottorini; Animesh Acharjee; Andre Franke; Yingrui Li; Gane Ka-Shu Wong; Christos Polytarchou; Tung On Yau; Niki Christodoulou; Maria Hatziapostolou; Minkun Wang; Lindsey A Russell; Dina H Kao Journal: Cells Date: 2021-11-19 Impact factor: 6.600