Establishment of Intestinal Organoid from Rousettus leschenaultii and the Susceptibility to Bat-Associated Viruses, SARS-CoV-2 and Pteropine Orthoreovirus

Abstract

Various pathogens, such as Ebola virus, Marburg virus, Nipah virus, Hendra virus, Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and SARS-CoV-2, are threatening human health worldwide. The natural hosts of these pathogens are thought to be bats. The rousette bat, a megabat, is thought to be a natural reservoir of filoviruses, including Ebola and Marburg viruses. Additionally, the rousette bat showed a transient infection in the experimental inoculation of SARS-CoV-2. In the current study, we established and characterized intestinal organoids from Leschenault’s rousette, Rousettus leschenaultii. The established organoids successfully recapitulated the characteristics of intestinal epithelial structure and morphology, and the appropriate supplements necessary for long-term stable culture were identified. The organoid showed susceptibility to Pteropine orthoreovirus (PRV) but not to SARS-CoV-2 in experimental inoculation. This is the first report of the establishment of an expandable organoid culture system of the rousette bat intestinal organoid and its sensitivity to bat-associated viruses, PRV and SARS-CoV-2. This organoid is a useful tool for the elucidation of tolerance mechanisms of the emerging rousette bat-associated viruses such as Ebola and Marburg virus.

Antiviral response mechanisms in a Jamaican Fruit Bat intestinal organoid model of SARS-CoV-2 infection

“…Organoids from Chinese horseshoe bats, where SARS-CoV-2-like virus has been detected 7 , produced infectious SARS-CoV-2 virions at similar levels as human intestinal organoids 25 . In contrast, intestinal organoids from Leschenault's rousette bats (Rousettus leschenaultii) failed to support SARS-CoV-2 replication 23 . Interestingly, PCR analysis revealed a signi cant increase in viral and sgRNA in the JFB distal organoids at 48 and 72 hpi, which demonstrates initiation of viral replication in the organoids.…”

“…Importantly, growth conditions for GI organoids appear similar across multiple species 22 . Two previous studies have described the generation of intestinal organoid cultures from bat species 23,24 . Intestinal organoids developed from Chinese horseshoe bats, Rhinolophus sinicus, showed susceptibility to SARS-CoV-2, but lacked long-term active proliferation past 4-5 weeks 25 .…”

Generation of self-replicating airway organoids from the cave nectar bat Eonycteris spelaea as a model system for studying host–pathogen interactions in the bat airway epithelium

“…Recent studies reported the generation of intestinal organoids from Chinese horseshoe bat, Rhinolophus sinicus [ 14 ] and Leschenault's rousette, Rousettus leschenaultia [ 15 ]. To our knowledge, this is the first study to establish a sustainable approach for the propagation of bat airway organoids, and their conversion into ALI-AECs to aid infection studies.…”

“…Our findings are consistent with these environmental sampling data. Similarly, it was observed that intestinal organoids from another Pteropodid bat, R. leschenaultia , were not susceptible to SARS-CoV-2 [ 15 ], while intestinal organoids from a Rhinolophus species readily supported SARS-CoV-2 replication [ 28 ]. The observed lack of susceptibility to SARS-CoV-2 is despite the E. spelaea ACE2 receptor homologue mediating SARS-CoV-2 spike pseudovirus entry.…”

Smart/stimuli-responsive hydrogels: Cutting-edge platforms for tissue engineering and other biomedical applications

“…Likewise, they provide structural stability vital for cell self-assembly [ 459 ]. Such systems fit a broad spectrum of applications in the biomedical scope, mainly in elevated-throughput drug inspection [ 460 , 461 ], cell biology [ 15 ], cancer biology [ [462] , [463] , [464] , [465] , [466] , [467] , [468] ], disease modeling [ 469 , 470 ], and tissue engineering [ 459 ]. Collectively, hydrogel matrices not only present encouraging results for 3D cell culture but also exhibit high promising potentials for future broad range applications in the biomedical fields, chiefly tissue engineering [ 471 ].…”