Sporadic Creutzfeldt–Jakob disease infected human cerebral organoids retain the original human brain subtype features following transmission to humanized transgenic mice

Abstract

Human cerebral organoids (COs) are three-dimensional self-organizing cultures of cerebral brain tissue differentiated from induced pluripotent stem cells. We have recently shown that COs are susceptible to infection with different subtypes of Creutzfeldt–Jakob disease (CJD) prions, which in humans cause different manifestations of the disease. The ability to study live human brain tissue infected with different CJD subtypes opens a wide array of possibilities from differentiating mechanisms of cell death and identifying neuronal selective vulnerabilities to testing therapeutics. However, the question remained as to whether the prions generated in the CO model truly represent those in the infecting inoculum. Mouse models expressing human prion protein are commonly used to characterize human prion disease as they reproduce many of the molecular and clinical phenotypes associated with CJD subtypes. We therefore inoculated these mice with COs that had been infected with two CJD subtypes (MV1 and MV2) to see if the original subtype characteristics (referred to as strains once transmitted into a model organism) of the infecting prions were maintained in the COs when compared with the original human brain inocula. We found that disease characteristics caused by the molecular subtype of the disease associated prion protein were similar in mice inoculated with either CO derived material or human brain material, demonstrating that the disease associated prions generated in COs shared strain characteristics with those in humans. As the first and only in vitro model of human neurodegenerative disease that can faithfully reproduce different subtypes of prion disease, these findings support the use of the CO model for investigating human prion diseases and their subtypes.

Advanced Cellular Models for Rare Disease Study: Exploring Neural, Muscle and Skeletal Organoids

“…These 3D models can be infected with different subtypes of Creutzfeldt-Jakob disease (CJD) prions, which in humans cause different manifestations of the disease, maintaining the original subtype characteristics of the infecting prions. Mouse models expressing human prion proteins inoculated either with cerebral organoids that had been infected with two CJD subtypes (MV1 and MV2) or with the original human brain material present similar disease characteristics, demonstrating that this in vitro model can faithfully reproduce different subtypes of prion disease [68,113].…”

Efficacy of Wex-cide 128 disinfectant against multiple prion strains

“…Tg33 mouse brains were screened by RT-QuIC assay (methods below) for the presence of prion seeding activity. Most tg66 mouse brains were screened for neuropathology (H&E sections) and IHC demonstration of prion deposition consistent with prion disease using antiprion antibody 3F4 as described [32]. A small subset of tg66 mice that did not have formalin fixed tissue available were screened by RT-QuIC assay (methods below) for prion seeding activity rather than neuropathology.…”

Neural cell engraftment therapy for sporadic Creutzfeldt-Jakob disease restores neuroelectrophysiological parameters in a cerebral organoid model

“…We have previously shown that human cerebral organoids (COs) can be infected with sCJD prions and faithfully propagate the original inoculum [ 21 , 22 ]. The infection could be reduced by treatment with PPS either administered before and during infection or after establishment of infection, supporting the use of the organoid model for screening putative therapeutic strategies [ 23 ].…”