The gut microbiota is in continuous interaction with the innermost layer of the gut, namely the epithelium. One of the various functions of the gut epithelium, is to keep the microbes at bay to avoid overstimulation of the underlying mucosa immune cells. To do so, the gut epithelia secrete a variety of antimicrobial peptides, such as chromogranin A (CgA) peptide catestatin (CST: hCgA352-372). As a defense mechanism, gut microbes have evolved antimicrobial resistance mechanisms to counteract the killing effect of the secreted peptides. To this end, we treated wild-type mice and CST knockout (CST-KO) mice (where only the 63 nucleotides encoding CST have been deleted) with CST for 15 consecutive days. CST treatment was associated with a shift in the diversity and composition of the microbiota in the CST-KO mice. This effect was less prominent in WT mice. Levels of the microbiota-produced short-chain fatty acids, in particular, butyrate and acetate were significantly increased in CST-treated CST-KO mice but not the WT group. Both CST-treated CST-KO and WT mice showed a significant increase in microbiota-harboring phosphoethanolamine transferase-encoding genes, which facilitate their antimicrobial resistance. Finally, we show that CST was degraded by Escherichia coli via an omptin-protease and that the abundance of this gene was significantly higher in metagenomic datasets collected from patients with Crohn’s disease but not with ulcerative colitis. Overall, this study illustrates how the endogenous antimicrobial peptide, CST, shapes the microbiota composition in the gut and primes further research to uncover the role of bacterial resistance to CST in disease states such as inflammatory bowel disease.
This is a preview of subscription content
Subscribe to Journal
Get full journal access for 1 year
only $9.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
All data generated or analyzed during this study are included in this manuscript and its supplementary information files. The 16S rRNA gene amplicon sequence data were deposited under BioProject number PRJNA741992.
Kåhrström CT, Pariente N, Weiss U. Intestinal microbiota in health and disease. Nature 2016;535:47–47.
El Aidy S, van Baarlen P, Derrien M, Lindenbergh-Kortleve DJ, Hooiveld G, Levenez F, et al. Temporal and spatial interplay of microbiota and intestinal mucosa drive establishment of immune homeostasis in conventionalized mice. Mucosal Immunol. 2012;5:567–79.
Okumura R, Takeda K. Roles of intestinal epithelial cells in the maintenance of gut homeostasis. Exp Mol Med. 2017;49:e338–e338.
Mahata SK, O’Connor DT, Mahata M, Yoo SH, Taupenot L, Wu H, et al. Novel autocrine feedback control of catecholamine release. A discrete chromogranin a fragment is a noncompetitive nicotinic cholinergic antagonist. J Clin Invest. 1997;100:1623–33.
Briolat J, Wu SD, Mahata SK, Gonthier B, Bagnard D, Chasserot-Golaz S, et al. New antimicrobial activity for the catecholamine release-inhibitory peptide from chromogranin A. Cell Mol Life Sci. 2005;62:377–85.
Lugardon K, Raffner R, Goumon Y, Corti A, Delmas A, Bulet P, et al. Antibacterial and antifungal activities of vasostatin-1, the N-terminal fragment of chromogranin A. J Biol Chem. 2000;275:10745–53.
Aslam R, Atindehou M, Lavaux T, Haïkel Y, Schneider F, Metz-Boutigue M-H. Chromogranin A-derived peptides are involved in innate immunity. Curr Med Chem. 2012;19:4115–23.
El-Salhy M, Patcharatrakul T, Hatlebakk JG, Hausken T, Gilja OH, Gonlachanvit S. Chromogranin A cell density in the large intestine of Asian and European patients with irritable bowel syndrome. Scand J Gastroenterol. 2017;52:691–7.
Bartolomucci A, Possenti R, Mahata SK, Fischer-Colbrie R, Loh YP, Salton SR. The extended granin family: structure, function, and biomedical implications. Endocr Rev. 2011;32:755–97.
Mahata SK, Corti A. Chromogranin A and its fragments in cardiovascular, immunometabolic, and cancer regulation. Ann N Y Acad Sci. 2019;1455:34–58.
Corti A, Marcucci F, Bachetti T. Circulating chromogranin A and its fragments as diagnostic and prognostic disease markers. Pflugers Archiv Eur J Physiol. 2018;470:199–210.
Mahata SK, Mahata M, Fung MM, O’Connor DT. Catestatin: a multifunctional peptide from chromogranin A. Regul Pept. 2010;162:33–43.
Ying W, Mahata S, Bandyopadhyay GK, Zhou Z, Wollam J, Vu J, et al. Catestatin inhibits obesity-induced macrophage infiltration and inflammation in the liver and suppresses hepatic glucose production, leading to improved insulin sensitivity. Diabetes. 2018;67:841–8.
Mahata SK, Kiranmayi M, Mahapatra NR. Catestatin: a master regulator of cardiovascular functions. Curr Med Chem. 2018;25:1352–74.
Muntjewerff EM, Tang K, Lutter L, Christoffersson G, Nicolasen MJT, Gao H, et al. Chromogranin A regulates gut permeability via the antagonistic actions of its proteolytic peptides. Acta Physiol. 2021;232:e13655.
Rabbi MF, Munyaka PM, Eissa N, Metz-Boutigue MH, Khafipour E, Ghia JE. Human catestatin alters gut microbiota composition in mice. Front Microbiol. 2017;7:1–12.
Radek KA, Lopez-Garcia B, Hupe M, Niesman IR, Elias PM, Taupenot L, et al. The neuroendocrine peptide catestatin is a cutaneous antimicrobial and induced in the skin after injury. J Invest Dermatol. 2008;128:1525–34.
Bevins CL, Salzman NH. Paneth cells, antimicrobial peptides and maintenance of intestinal homeostasis. Nat Rev Microbiol. 2011;9:356–68.
Dupont A, Heinbockel L, Brandenburg K, Hornef MW. Antimicrobial peptides and the enteric mucus layer act in concert to protect the intestinal mucosa. Gut Microbes. 2014;5:761–5.
Tsukuda N, Yahagi K, Hara T, Watanabe Y, Matsumoto H, Mori H, et al. Key bacterial taxa and metabolic pathways affecting gut short-chain fatty acid profiles in early life. ISME J. 2021;15:2574–90.
Nuri R, Shprung T, Shai Y. Defensive remodeling: How bacterial surface properties and biofilm formation promote resistance to antimicrobial peptides. Biochim Biophys Acta Biomembr. 2015;1848:3089–100.
Jakobsson HE, Rodríguez‐Piñeiro AM, Schütte A, Ermund A, Boysen P, Bemark M, et al. The composition of the gut microbiota shapes the colon mucus barrier. EMBO Rep. 2015;16:164–77.
Samantha A, Vrielink A. Lipid A Phosphoethanolamine Transferase: regulation, structure and immune response. J Mol Biol. 2020;432:5184–96.
Gottesman S. Proteases and their targets in Escherichia coli. Annu Rev Genet. 1996;30:465–506.
Mirsepasi-Lauridsen HC, Vallance BA, Krogfelt KA, Petersen AM. Escherichia coli pathobionts associated with inflammatory bowel disease. Clin Microbiol Rev. 2019;32:1–16.
Nayfach S, Fischbach MA, Pollard KS. MetaQuery: a web server for rapid annotation and quantitative analysis of specific genes in the human gut microbiome. Bioinformatics. 2015;31:3368–70.
Ying W, Tang K, Avolio E, Schilling JM, Pasqua T, Liu MA, et al. Immunosuppression of macrophages underlies the cardioprotective effects of CST (Catestatin). Hypertension. 2021;77:1670–82.
Stojanov S, Berlec A, Štrukelj B. The influence of probiotics on the firmicutes/bacteroidetes ratio in the treatment of obesity and inflammatory bowel disease. Microorganisms. 2020;8:1–16.
Indiani CMDSP, Rizzardi KF, Castelo PM, Ferraz LFC, Darrieux M, Parisotto TM. Childhood obesity and firmicutes/bacteroidetes ratio in the gut microbiota: a systematic review. Child Obes. 2018;14:501–9.
Lam YY, Ha CWY, Campbell CR, Mitchell AJ, Dinudom A, Oscarsson J, et al. Increased gut permeability and microbiota change associate with mesenteric fat inflammation and metabolic dysfunction in diet-induced obese mice. PLoS One. 2012;7:1–10.
Herp S, Durai Raj AC, Salvado Silva M, Woelfel S, Stecher B. The human symbiont Mucispirillum schaedleri: causality in health and disease. Med Microbiol Immunol. 2021;210:173–9.
Parker BJ, Wearsch PA, Veloo ACM, Rodriguez-Palacios A. The genus alistipes: gut bacteria with emerging implications to inflammation, cancer, and mental health. Front Immunol. 2020;11:1–15.
Hiippala K, Barreto G, Burrello C, Diaz-Basabe A, Suutarinen M, Kainulainen V, et al. Novel Odoribacter splanchnicus strain and its outer membrane vesicles exert immunoregulatory effects in vitro. Front Microbiol. 2020;11:1–14.
McPhee JB, Small CL, Reid-Yu SA, Brannon JR, Moual H LE, Coombes BK. Host defense peptide resistance contributes to colonization and maximal intestinal pathology by Crohn’s disease-associated adherent-invasive Escherichia coli. Infect Immun. 2014;82:3383–93.
Xu Y, Wei W, Lei S, Lin J, Srinivas S, Feng Y. An evolutionarily conserved mechanism for intrinsic and transferable polymyxin resistance. MBio. 2018;9:1–18.
Thomassin JL, Brannon JR, Gibbs BF, Gruenheid S, Le Moual H. OmpT outer membrane proteases of enterohemorrhagic and enteropathogenic Escherichia coli contribute differently to the degradation of human LL-37. Infect Immun. 2012;80:483–92.
Desloges I, Taylor JA, Leclerc JM, Brannon JR, Portt A, Spencer JD, et al. Identification and characterization of OmpT-like proteases in uropathogenic Escherichia coli clinical isolates. Microbiologyopen. 2019;8:1–36.
McCarter JD, Stephens D, Shoemaker K, Rosenberg S, Kirsch JF, Georgiou G. Substrate specificity of the Escherichia coli outer membrane protease OmpT. J Bacteriol. 2004;186:5919–25.
Kulkarni HM, Nagaraj R, Jagannadham MV. Protective role of E. coli outer membrane vesicles against antibiotics. Microbiol Res. 2015;181:1–7.
Muntjewerff EM, Dunkel G, Nicolasen MJT, Mahata SK, van den Bogaart G. Catestatin as a Target for Treatment of Inflammatory Diseases. Front Immunol. 2018;9:2199.
Santella RM. Approaches to DNA/RNA extraction and whole genome amplification: table 1. Cancer Epidemiol Biomark Prev. 2006;15:1585–7.
Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods. 2016;13:581–3.
R Core Team. R: a language and environment for statistical computing. Vienna, Austria; 2019. https://www.r-project.org/.
Lahti L, Shetty S. microbiome R package. http://microbiome.github.io.
McMurdie PJ, Holmes S. Phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS One. 2013;8:e61217.
Paulson JN, Colin Stine O, Bravo HC, Pop M. Differential abundance analysis for microbial marker-gene surveys. Nat Methods. 2013;10:1200–2.
Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, et al. Metagenomic biomarker discovery and explanation. Genome Biol. 2011;12:R60.
Douglas GM, Maffei VJ, Zaneveld JR, Yurgel SN, Brown JR, Taylor CM, et al. PICRUSt2 for prediction of metagenome functions. Nat Biotechnol. 2020;38:685–8.
Parks DH, Tyson GW, Hugenholtz P, Beiko RG. STAMP: Statistical analysis of taxonomic and functional profiles. Bioinformatics. 2014;30:3123–4.
Beresford-Jones BS, Forster SC, Stares MD, Notley G, Viciani E, Browne HP, et al. The Mouse Gastrointestinal Bacteria Catalogue enables translation between the mouse and human gut microbiotas via functional mapping. Cell Host Microbe. 2022;30:124–138.e8.
Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Mol Biol Evol. 2013;30:772–80.
Price MN, Dehal PS, Arkin AP. FastTree 2 - approximately maximum-likelihood trees for large alignments. PLoS One. 2010;5:e9490.
Menardo F, Loiseau C, Brites D, Coscolla M, Gygli SM, Rutaihwa LK, et al. Treemmer: a tool to reduce large phylogenetic datasets with minimal loss of diversity. BMC Bioinforma. 2018;19:1–8.
Haider SR, Reid HJ, Sharp BL. Tricine-SDS-PAGE. In: Kurien B., Scofield R. editors. Protein electrophoresis. Methods in Molecular Biology (Methods and Protocols). Totowa, NJ: Humana Press; 2012. p. 81–91.
Schägger H. Tricine-SDS-PAGE. Nat Protoc. 2006;1:16–22.
Zoetendal EG, Booijink CCGM, Klaassens ES, Heilig HGHJ, Kleerebezem M, Smidt H, et al. Isolation of RNA from bacterial samples of the human gastrointestinal tract. Nat Protoc. 2006;1:954–9.
Zhou K, Zhou L, Lim Q, Zou R, Stephanopoulos G, Too HP. Novel reference genes for quantifying transcriptional responses of Escherichia coli to protein overexpression by quantitative PCR. BMC Mol Biol. 2011;12:18.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25:402–8.
PG-D thanks the National Council of Science and Technology in Mexico (CONACyt) for the Ph.D. grant assigned to CVU 690069. We thank Dr. Matthias Heinemann of Department of Molecular Systems Biology, University of Groningen, the Netherlands, for providing us ΔeptA and ΔompT E. coli BW25113 mutant strain; Dr. Greet Vandermeulen of Department of chronic diseases and metabolism, Faculty of Medicine, KU Leuven, Belgium for the help with SCFAs analysis.
SKM is supported by a Merit Review Grant (I01 BX003934) from the Department of Veterans Affairs, USA. SEA is supported by a Rosalind Franklin Fellowship, co-funded by the European Union and the University of Groningen, The Netherlands.
The authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
González-Dávila, P., Schwalbe, M., Danewalia, A. et al. Catestatin selects for colonization of antimicrobial-resistant gut bacterial communities. ISME J (2022). https://doi.org/10.1038/s41396-022-01240-9