Stem Cell-Derived Human Intestinal Organoids as an Infection Model for Rotaviruses

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>Directed differentiation of stem cell lines into intestine-like tissue called induced human intestinal organoids (iHIOs) is now possible (J. R. Spence, C. N. Mayhew, S. A. Rankin, M. F. Kuhar, J. E. Vallance, K. Tolle, E. E. Hoskins, V. V. Kalinichenko, S. I. Wells, A. M. Zorn, N. F. Shroyer, and J. M. Wells, Nature 470:105-109, 2011). We tested iHIOs as a new model to cultivate and study fecal viruses. Protocols for infection of iHIOs with a laboratory strain of rotavirus, simian SA11, were developed. Proof-of-principle analyses showed that iHIOs support replication of a gastrointestinal virus, rotavirus, on the basis of detection of nonstructural viral proteins (nonstructural protein 4 [NSP4] and NSP2) by immunofluorescence, increased levels of viral RNA by quantitative reverse transcription-PCR (qRT-PCR), and production of infectious progeny virus. iHIOs were also shown to support replication of 12/13 clinical rotavirus isolates directly from stool samples. An unexpected finding was the detection of rotavirus infection not only in the epithelial cells but also in the mesenchymal cell population of the iHIOs. This work demonstrates that iHIOs offer a promising new model to study rotaviruses and other gastrointestinal viruses.</jats:p><jats:p><jats:bold>IMPORTANCE</jats:bold>Gastrointestinal viral infections are a major cause of illness and death in children and adults. The ability to fully understand how viruses interact with human intestinal cells in order to cause disease has been hampered by insufficient methods for growing many gastrointestinal viruses in the laboratory. Induced human intestinal organoids (iHIOs) are a promising new model for generating intestine-like tissue. This is the first report of a study using iHIOs to cultivate any microorganism, in this case, an enteric virus. The evidence that both laboratory and clinical rotavirus isolates can replicate in iHIOs suggests that this model would be useful not only for studies of rotaviruses but also potentially of other infectious agents. Furthermore, detection of rotavirus proteins in unexpected cell types highlights the promise of this system to reveal new questions about pathogenesis that have not been previously recognized or investigated in other intestinal cell culture models.</jats:p>

Human Enteroids/Colonoids and Intestinal Organoids Functionally Recapitulate Normal Intestinal Physiology and Pathophysiology

“…To date, human enteroids have been shown to model human RV infection, cholera toxin effect on transport, and several aspects of enterohemorrhagic Escherichia coli-related human diarrhea (9). RV replicates and produces infectious virus in human enteroids and organoids, with viral replication increasing over 96 h (13,36). Rotavirus infection causes very rapid inhibition of NHE3, which might be a significant contributor to RV-induced diarrhea, the pathophysiology of which remains poorly understood (37).…”

Enteroviruses infect human enteroids and induce antiviral signaling in a cell lineage-specific manner

“…Lgr5 + intestinal crypt stem cells differentiate into the various epithelial cell subtypes found in the human small intestine in vivo, resulting in the production of enteroid structures over 4 to 5 days (11). Others have shown that human enteroids can serve as models for the study of enteric infections by human rotavirus (12,13) and norovirus (14).…”

Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids

“…This is even more apparent in PhoP mutants . Gastrointestinal viruses cause a large part of the yearly estimated 1.7 billion cases of acute diarrhea (Finkbeiner et al, 2012). Next to norovirus, rotavirus, a double-stranded RNA virus of the family Reoviridae, is the most common cause of diarrhea in young children.…”