Modelling Cryptosporidium infection in human small intestinal and lung organoids
Inha Heo, Devanjali Dutta, Deborah A Schaefer, Nino Iakobachvili, Benedetta Artegiani, Norman Sachs, Kim E Boonekamp, Gregory Bowden, Antoni P A Hendrickx, Robert J L Willems, Peter J Peters, Michael W Riggs, Roberta O'Connor, Hans Clevers, Inha Heo, Devanjali Dutta, Deborah A Schaefer, Nino Iakobachvili, Benedetta Artegiani, Norman Sachs, Kim E Boonekamp, Gregory Bowden, Antoni P A Hendrickx, Robert J L Willems, Peter J Peters, Michael W Riggs, Roberta O'Connor, Hans Clevers
Abstract
Stem-cell-derived organoids recapitulate in vivo physiology of their original tissues, representing valuable systems to model medical disorders such as infectious diseases. Cryptosporidium, a protozoan parasite, is a leading cause of diarrhoea and a major cause of child mortality worldwide. Drug development requires detailed knowledge of the pathophysiology of Cryptosporidium, but experimental approaches have been hindered by the lack of an optimal in vitro culture system. Here, we show that Cryptosporidium can infect epithelial organoids derived from human small intestine and lung. The parasite propagates within the organoids and completes its complex life cycle. Temporal analysis of the Cryptosporidium transcriptome during organoid infection reveals dynamic regulation of transcripts related to its life cycle. Our study presents organoids as a physiologically relevant in vitro model system to study Cryptosporidium infection.
Conflict of interest statement
Conflict of interest
N.S. and H.C. are inventors on patents/patent applications related to organoid technology.
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References
- Clevers H. Modeling Development and Disease with Organoids. Cell. 2016;165:1586–1597.
- Sato T, et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology. 2011;141:1762–1772.
- Bouzid M, Hunter PR, Chalmers RM, Tyler KM. Cryptosporidium pathogenicity and virulence. Clinical microbiology reviews. 2013;26:115–134.
- Thompson RC, et al. Cryptosporidium and cryptosporidiosis. Advances in parasitology. 2005;59:77–158.
- Current WL, Garcia LS. Cryptosporidiosis. Clinics in laboratory medicine. 1991;11:873–897.
- Hunter PR, Nichols G. Epidemiology and clinical features of Cryptosporidium infection in immunocompromised patients. Clinical microbiology reviews. 2002;15:145–154.
- Checkley W, et al. A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for cryptosporidium. The Lancet Infectious diseases. 2015;15:85–94.
- Liu L, et al. Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet. 2012;379:2151–2161.
- Sponseller JK, Griffiths JK, Tzipori S. The evolution of respiratory Cryptosporidiosis: evidence for transmission by inhalation. Clinical microbiology reviews. 2014;27:575–586.
- Mor SM, et al. Expectoration of Cryptosporidium Parasites in Sputum of Human Immunodeficiency Virus-Positive and -Negative Adults. The American journal of tropical medicine and hygiene. 2018
- Kotloff KL, et al. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet. 2013;382:209–222.
- Amadi B, et al. Effect of nitazoxanide on morbidity and mortality in Zambian children with cryptosporidiosis: a randomised controlled trial. Lancet. 2002;360:1375–1380.
- Amadi B, et al. Reduced production of sulfated glycosaminoglycans occurs in Zambian children with kwashiorkor but not marasmus. The American journal of clinical nutrition. 2009;89:592–600.
- Striepen B. Parasitic infections: Time to tackle cryptosporidiosis. Nature. 2013;503:189–191.
- Lendner M, Daugschies A. Cryptosporidium infections: molecular advances. Parasitology. 2014;141:1511–1532.
- Karanis P, Aldeyarbi HM. Evolution of Cryptosporidium in vitro culture. International journal for parasitology. 2011;41:1231–1242.
- Karanis P. The truth about in vitro culture of Cryptosporidium species. Parasitology. 2017:1–10.
- Varughese EA, Bennett-Stamper CL, Wymer LJ, Yadav JS. A new in vitro model using small intestinal epithelial cells to enhance infection of Cryptosporidium parvum. Journal of microbiological methods. 2014;106:47–54.
- Castellanos-Gonzalez A, Cabada MM, Nichols J, Gomez G, White AC., Jr Human primary intestinal epithelial cells as an improved in vitro model for Cryptosporidium parvum infection. Infection and immunity. 2013;81:1996–2001.
- Morada M, et al. Continuous culture of Cryptosporidium parvum using hollow fiber technology. International journal for parasitology. 2016;46:21–29.
- DeCicco RePass MA, et al. Novel Bioengineered Three-Dimensional Human Intestinal Model for Long-Term Infection of Cryptosporidium parvum. Infection and immunity. 2017;85
- Dutta D, Heo I, Clevers H. Disease Modeling in Stem Cell-Derived 3D Organoid Systems. Trends in molecular medicine. 2017;23:393–410.
- McCracken KW, et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature. 2014;516:400–404.
- Ettayebi K, et al. Replication of human noroviruses in stem cell-derived human enteroids. Science. 2016;353:1387–1393.
- Umemiya R, Fukuda M, Fujisaki K, Matsui T. Electron microscopic observation of the invasion process of Cryptosporidium parvum in severe combined immunodeficiency mice. The Journal of parasitology. 2005;91:1034–1039.
- Aldeyarbi HM, Karanis P. The fine structure of sexual stage development and sporogony of Cryptosporidium parvum in cell-free culture. Parasitology. 2016;143:749–761.
- Aldeyarbi HM, Karanis P. Electron microscopic observation of the early stages of Cryptosporidium parvum asexual multiplication and development in in vitro axenic culture. European journal of protistology. 2016;52:36–44.
- Aldeyarbi HM, Karanis P. The Ultra-Structural Similarities between Cryptosporidium parvum and the Gregarines. The Journal of eukaryotic microbiology. 2016;63:79–85.
- Fayer R, Xiao L. Cryptosporidium and cryptosporidiosis. 2nd Edition 2008.
- Ernest JA, Blagburn BL, Lindsay DS, Current WL. Infection dynamics of Cryptosporidium parvum (Apicomplexa: Cryptosporiidae) in neonatal mice (Mus musculus) The Journal of parasitology. 1986;72:796–798.
- Shahiduzzaman M, Dyachenko V, Obwaller A, Unglaube S, Daugschies A. Combination of cell culture and quantitative PCR for screening of drugs against Cryptosporidium parvum. Veterinary parasitology. 2009;162:271–277.
- Riggs MW, Perryman LE. Infectivity and neutralization of Cryptosporidium parvum sporozoites. Infection and immunity. 1987;55:2081–2087.
- Sachs N, et al. Long-term expanding human airway organoids for disease modelling. BioRxiv. 2018 doi: 10.1101/318444.
- Beiting DP. Protozoan parasites and type I interferons: a cold case reopened. Trends in parasitology. 2014;30:491–498.
- Barakat FM, McDonald V, Foster GR, Tovey MG, Korbel DS. Cryptosporidium parvum infection rapidly induces a protective innate immune response involving type I interferon. The Journal of infectious diseases. 2009;200:1548–1555.
- Frenal K, Soldati-Favre D. Role of the parasite and host cytoskeleton in apicomplexa parasitism. Cell host & microbe. 2009;5:602–611.
- Elliott DA, et al. Cryptosporidium parvum infection requires host cell actin polymerization. Infection and immunity. 2001;69:5940–5942.
- Rayamajhi M, Humann J, Penheiter K, Andreasen K, Lenz LL. Induction of IFN-alphabeta enables Listeria monocytogenes to suppress macrophage activation by IFN-gamma. The Journal of experimental medicine. 2010;207:327–337.
- O'Connell RM, et al. Type I interferon production enhances susceptibility to Listeria monocytogenes infection. The Journal of experimental medicine. 2004;200:437–445.
- In J, et al. Enterohemorrhagic Escherichia coli reduce mucus and intermicrovillar bridges in human stem cell-derived colonoids. Cellular and molecular gastroenterology and hepatology. 2016;2:48–62 e43.
- Vinayak S, et al. Genetic modification of the diarrhoeal pathogen Cryptosporidium parvum. Nature. 2015;523:477–480.
- Blokzijl F, et al. Tissue-specific mutation accumulation in human adult stem cells during life. Nature. 2016;538:260–264.
- Zhou R, Gong AY, Eischeid AN, Chen XM. miR-27b targets KSRP to coordinate TLR4-mediated epithelial defense against Cryptosporidium parvum infection. PLoS pathogens. 2012;8:e1002702.
- Chen XM, Splinter PL, O'Hara SP, LaRusso NF. A cellular micro-RNA, let-7i, regulates Toll-like receptor 4 expression and contributes to cholangiocyte immune responses against Cryptosporidium parvum infection. The Journal of biological chemistry. 2007;282:28929–28938.
- Faas FG, et al. Virtual nanoscopy: generation of ultra-large high resolution electron microscopy maps. The Journal of cell biology. 2012;198:457–469.
- Riggs MW, et al. Protective monoclonal antibody defines a circumsporozoite-like glycoprotein exoantigen of Cryptosporidium parvum sporozoites and merozoites. Journal of immunology. 1997;158:1787–1795.
- Grun D, et al. Single-cell messenger RNA sequencing reveals rare intestinal cell types. Nature. 2015;525:251–255.
- Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25:1754–1760.
- Heiges M, et al. CryptoDB: a Cryptosporidium bioinformatics resource update. Nucleic acids research. 2006;34:D419–422.
Source: PubMed