Mechanisms involved in the protective effect of short-chain fatty acids against Clostridium difficile-associated colitis

• Grant number: 17/06577-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: August 01, 2017
• End date: May 31, 2021
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Marco Aurélio Ramirez Vinolo
• Grantee: José Luís Fachi
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 12/10653-9 - Role of short chain fatty acids and their receptor (GPR43) in the immune response to anaerobic bacteria in vivo and in vitro, AP.JP
• Associated scholarship(s): 18/10165-0 - Role of microbiota-derived metabolites, the short-chain fatty acids, on innate lymphoid cells, BE.EP.DR

Abstract

Under physiological conditions, commensal intestinal bacteria restrict colonization by Clostridium difficile, a bacillus resistant to various antimicrobial agents. After the use of antibiotics, there is an imbalance of the microbiota and proliferation of the C.difficile, causing diverses pathological disease. It is known that the products of bacterial metabolism, short chain fatty acids (SCFAs), are produced during the fermentation process of dietary fibers and present a protective effect against intestinal infections, such as that caused by C. difficile. However, the molecular mechanisms behind these effects are not known. This work aims to investigate the participation of AGCCs activated pathways in the protection observed in murine model of acute C. difficile colitis. Specifically, we will examine the relevance of the G protein coupled receptor, FFAR2/GPR43, and the hypoxia-induced transcription factor 1 (HIF-1), as well as the possible involvement of miRNAs over the gene expression modifications relevant to the cure. For this, we will use in vivo models (FFAR2 whole-body knockout animals, tissue specific deletion of HIF-1± and/or Dicer, an enzyme involved in the maturation of miRNA) and in vitro (HCT-116 cell line or intestinal organoids). We will perform molecular analyzes including Western blotting, immunofluorescence, RT-qPCR for specific genes and miRNAs and RNA sequencing and histopathological analyzes including analysis of stained colonies of H&E, identification of leukocyte populations in the lamina propria and mesenteric lymph nodes by flow cytometry, as well as analysis of inflammatory mediators by ELISA and epithelial permeability by immunohistochemical labeling of proteins relevant to the process and of bacterial and FITC-dextran translocation after infection. With this study, we intend to advance in the understanding of the interactions between the microbiota and the development of intestinal infections and to identify new molecular targets for therapeutic interventions in the disease caused by C.difficile. (AU)

Scientific publications

(9)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

FACHI, JOSE LUIS; SECCA, CRISTIANE; RODRIGUES, PATRICIA BRITO; PINHEIRO DE MATO, FELIPE CEZAR; DI LUCCIA, BLANDA; FELIPE, JAQUELINE DE SOUZA; PRAL, LAIS PASSARIELLO; RUNGUE, MARCELLA; ROCHA, VICTOR DE MELO; SATO, FABIO TAKEO; et al. Acetate coordinates neutrophil and ILC3 responses against C. difficile through FFAR2. JOURNAL OF EXPERIMENTAL MEDICINE, v. 217, n. 3, MAR 2020. (17/06577-9, 19/14342-7, 12/10653-9, 18/02208-1, 17/16280-3)

FACHI, JOSE L.; DE OLIVEIRA, SARAH; GILFILLAN, SUSAN; ANTONOVA, ALINA ULEZKO; HOU, JINCHAO; VINOLO, MARCO A. R.; COLONNA, MARCO. NKp46+ILC3s promote early neutrophil defense against Clostridioides difficile infection through GM-CSF secretion. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 121, n. 45, p. 10-pg., 2024-11-05. (21/09155-3, 17/06577-9, 18/15313-8, 23/00393-4)

PRAL, LAIS P.; FACHI, JOSE L.; CORREA, RENAN O.; COLONNA, MARCO; VINOLO, MARCO A. R.. Hypoxia and HIF-1 as key regulators of gut microbiota and host interactions. TRENDS IN IMMUNOLOGY, v. 42, n. 7, p. 604-621, JUL 2021. (16/23142-3, 17/06577-9, 18/02208-1, 18/15313-8)

FACHI, JOSE LUIS; FELIPE, JAQUELINE DE SOUZA; PRAL, LAIS PASSARIELLO; DA SILVA, BRUNA KARADI; CORREA, RENAN OLIVEIRA; PEREIRA DE ANDRADE, MIRELLA CRISTINY; DA FONSECA, DENISE MORAIS; BASSO, PAULO JOSE; SARAIVA CAMARA, NIELS OLSEN; DE SALES E SOUZA, ERICKA LORENNA; et al. Butyrate Protects Mice from Clostridium difficile-Induced Colitis through an HIF-1-Dependent Mechanism. CELL REPORTS, v. 27, n. 3, p. 750+, APR 16 2019. (17/01451-7, 12/10653-9, 16/23142-3, 13/06810-4, 14/03002-7, 15/01507-7, 18/01753-6, 15/06134-4, 17/25679-7, 17/06577-9, 18/02208-1, 17/26366-2, 17/16280-3)

FACHI, J. L.; PRAL, L. P.; DOS SANTOS, J. A. C.; CODO, A. C.; DE OLIVEIRA, S.; FELIPE, J. S.; ZAMBOM, F. F. F.; CAMARA, N. O. S.; VIEIRA, P. M. M. M.; COLONNA, M.; et al. Hypoxia enhances ILC3 responses through HIF-1 alpha-dependent mechanism. MUCOSAL IMMUNOLOGY, v. 14, n. 4, JAN 2021. (15/15626-8, 19/11662-0, 18/22505-0, 18/15313-8, 18/02208-1, 17/06577-9, 19/06372-3, 17/16280-3)

ANTUNES, KRIST HELEN; FACHI, JOSE LUIS; DE PAULA, ROSEMEIRE; DA SILVA, EMANUELLE FRAGA; PRAL, LAIS PASSARIELLO; DOS SANTOS, ADARA AUREA; MALAQUIAS DIAS, GREICY BRISA; VARGAS, JOSE EDUARDO; PUGA, RENATO; MAYER, FABIANA QUOOS; et al. Microbiota-derived acetate protects against respiratory syncytial virus infection through a GPR43-type 1 interferon response. NATURE COMMUNICATIONS, v. 10, JUL 22 2019. (13/06810-4, 17/06577-9, 12/10653-9)

CORREA, RENAN OLIVEIRA; CASTRO, POLLYANA RIBEIRO; FACHI, JOSE LUIS; NIRELLO, VINICIUS DIAS; EL-SAHHAR, SALMA; IMADA, SHINYA; PEREIRA, GABRIEL VASCONCELOS; PRAL, LAIS PASSARIELLO; PEREIRA ARAUJO, NATHALIA VITORIA; FERNANDES, MARIANE FONT; et al. Inulin diet uncovers complex diet-microbiota-immune cell interactions remodeling the gut epithelium. MICROBIOME, v. 11, n. 1, p. 25-pg., 2023-04-26. (16/23142-3, 21/06572-2, 19/06372-3, 19/16113-5, 19/02640-3, 18/15313-8, 20/02919-5, 17/16280-3, 20/14071-0, 21/00393-9, 17/06577-9, 20/13689-0, 21/05269-4, 20/02312-3, 18/10165-0, 20/00311-0)

FACHI, JOSE L.; DE OLIVEIRA, SARAH; TRSAN, TIHANA; PENATI, SILVIA; GILFILLAN, SUSAN; CAO, SIYAN; CASTRO, POLLYANA RIBEIRO; FERNANDES, MARIANE FONT; HYRC, KRZYSZTOF L.; LIU, XIULI; et al. Article Fiber- and acetate-mediated modulation of MHC-II expression on intestinal epithelium protects from Clostridioides difficile infection. CELL HOST & MICROBE, v. 33, n. 2, p. 25-pg., 2025-02-12. (21/09155-3, 18/15313-8, 23/00393-4, 17/06577-9)

FACHI, JOSE L.; PRAL, LAIS. P.; ASSIS, HELDER C.; OLIVEIRA, SARAH; RODOVALHO, VINICIUS R.; DOS SANTOS, JEFFERSON A. C.; FERNANDES, MARIANE F.; MATHEUS, VALQUIRIA A.; SESTI-COSTA, RENATA; BASSO, PAULO J.; et al. Hyperbaric oxygen augments susceptibility to C. difficile infection by impairing gut microbiota ability to stimulate the HIF-1α-IL-22 axis in ILC3. GUT MICROBES, v. 16, n. 1, p. 26-pg., 2024-12-31. (17/06577-9, 18/15313-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

FACHI, José Luís. Mechanisms involved to the protective effect of short-chain fatty acids during Clostridioides difficile-associated colitis. 2021. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.