Chronic wasting disease (CWD) is a relentless epidemic disorder caused by infectious prions that threatens the survival of cervid populations and raises increasing public health concerns in North America. In Europe, CWD was detected for the first time in wild Norwegian reindeer (Rangifer tarandus) and moose (Alces alces) in 2016. In this study, we aimed at comparing the strain properties of CWD prions derived from different cervid species in Norway and North America. Using a classical strain typing approach involving transmission and adaptation to bank voles (Myodes glareolus), we found that prions causing CWD in Norway induced incubation times, neuropathology, regional deposition of misfolded prion protein aggregates in the brain, and size of their protease-resistant core, different from those that characterize North American CWD. These findings show that CWD prion strains affecting Norwegian cervids are distinct from those found in North America, implying that the highly contagious North American CWD prions are not the proximate cause of the newly discovered Norwegian CWD cases. In addition, Norwegian CWD isolates showed an unexpected strain variability, with reindeer and moose being caused by different CWD strains. Our findings shed light on the origin of emergent European CWD, have significant implications for understanding the nature and the ecology of CWD in Europe, and highlight the need to assess the zoonotic potential of the new CWD strains detected in Europe. Significance Chronic wasting disease (CWD) is a highly contagious disease caused by prions that affects several cervid species and is relentlessly spreading across North America. Very recently, CWD was detected for the first time in Europe. In this study, we found that Norwegian CWD strains are distinct from those causing the epidemic in North America. Moreover, we show that Norwegian reindeer and moose are affected by different CWD strains, revealing an unprecedented prion strain variation in Norwegian wild cervid populations. These findings indicate that North American CWD prions are not the proximate cause of the newly discovered Norwegian CWD cases and have implications for CWD control strategies in Europe, as well as for the safety of humans.

The symptoms of prion infection can take years or decades to manifest following the initial exposure. Molecular markers of prion disease include accumulation of the misfolded prion protein (PrP^Sc), which is derived from its cellular precursor (PrP^C), as well as downregulation of the PrP-like Shadoo (Sho) glycoprotein. Given the overlapping cellular environments for PrP^C and Sho, we inferred that PrP^C levels might also be altered as part of a host response during prion infection. Using rodent models, we found that, in addition to changes in PrP^C glycosylation and proteolytic processing, net reductions in PrP^C occur in a wide range of prion diseases, including sheep scrapie, human Creutzfeldt-Jakob disease, and cervid chronic wasting disease. The reduction in PrP^C results in decreased prion replication, as measured by the protein misfolding cyclic amplification technique for generating PrP^Sc in vitro. While PrP^C downregulation is not discernible in animals with unusually short incubation periods and high PrP^C expression, slowly evolving prion infections exhibit downregulation of the PrP^C substrate required for new PrP^Sc synthesis and as a receptor for pathogenic signaling. Our data reveal PrP^C downregulation as a previously unappreciated element of disease pathogenesis that defines the extensive, presymptomatic period for many prion strains.

Adaptation of prions to new species is thought to reflect the capacity of the host-encoded cellular form of the prion protein (PrP^C) to selectively propagate optimized prion conformations from larger ensembles generated in the species of origin. Here we describe an alternate replicative process, termed nonadaptive prion amplification (NAPA), in which dominant conformers bypass this requirement during particular interspecies transmissions. To model susceptibility of horses to prions, we produced transgenic (Tg) mice expressing cognate PrP^C. Although disease transmission to only a subset of infected TgEq indicated a significant barrier to EqPrP^C conversion, the resulting horse prions unexpectedly failed to cause disease upon further passage to TgEq. TgD expressing deer PrP^C was similarly refractory to deer prions from diseased TgD infected with mink prions. In both cases, the resulting prions transmitted to mice expressing PrP^C from the species of prion origin, demonstrating that transmission barrier eradication of the originating prions was ephemeral and adaptation superficial in TgEq and TgD. Horse prions produced in vitro by protein misfolding cyclic amplification of mouse prions using horse PrP^C also failed to infect TgEq but retained tropism for wild-type mice. Concordant patterns of neuropathology and prion deposition in susceptible mice infected with NAPA prions and the corresponding prion of origin confirmed preservation of strain properties. The comparable responses of both prion types to guanidine hydrochloride denaturation indicated this occurs because NAPA precludes selection of novel prion conformations. Our findings provide insights into mechanisms regulating interspecies prion transmission and a framework to reconcile puzzling epidemiological features of certain prion disorders. Significance To study mechanisms of interspecies prion transmission, we used transgenic mice that recapitulate natural disease susceptibilities. Predictably, prions from a particular species inefficiently caused disease in mice expressing prion proteins from different species. However, in contrast to the expected adaptation that allows efficient prion replication in the new host, we discovered a process in which pathogenic prions, despite causing disease on primary passage, failed to adapt for sustained propagation in the new host, instead preserving the strain and host range characteristics of the original prions. Our findings provide insights into how prions replicate during species transitions, help clarify unresolved epidemiological features of prion disorders, and have considerable bearing on future disease risk assessments.

Significance: Searching for drugs to prevent conversion of host-encoded prion protein (PrP ^C ) to its infectious conformation (PrP ^Sc ) is a key strategy in the pursuit of therapies for prion disorders: fatal, transmissible epidemic diseases of unpredictable occurrence and uncertain zoonotic potential. Despite quinacrine’s ability to reduce mouse PrP ^Sc in cell models, its use to treat patients has been unsuccessful. Here, we show that quinacrine augments PrP ^Sc and intensifies replication of prions, causing chronic wasting disease of deer, elk, and moose, and that the resulting prions have altered conformational and transmission properties. Our finding, that a drug capable of restraining PrP ^Sc in one species acts to improve replicative ability and induce mutation in another, forces reexamination of current strategies to combat these diseases.

Significance: Our gene-targeting strategy addresses several critical issues relating to chronic wasting disease (CWD), a contagious, lethal neurodegenerative prion disease affecting wild as well as captive cervids, which poses an uncertain risk to humans. First, we show that residue 226, the sole primary structural difference between deer and elk prion protein, dictates the selection and propagation of different CWD strains. Because the strain properties of prions affect their host-range potential, these findings suggest that CWD from elk and deer pose distinct risks to humans and other species. Second, we show that these gene-targeted mice offer an unprecedented means to address distinctive aspects of CWD peripheral pathogenesis and horizontal transmission that are not afforded by previously generated transgenic mouse models.