RESUMEN
Can the course of fatal prion diseases be changed by removing the protein before it goes bad?
Asunto(s)
Enfermedades por Prión , Priones , Humanos , Enfermedades por Prión/genética , Enfermedades por Prión/prevención & control , Priones/antagonistas & inhibidores , Priones/genética , Mutación , Dedos de ZincRESUMEN
A distinctive signature of the prion diseases is the accumulation of the pathogenic isoform of the prion protein, PrPSc, in the central nervous system of prion-affected humans and animals. PrPSc is also found in peripheral tissues, raising concerns about the potential transmission of pathogenic prions through human food supplies and posing a significant risk to public health. Although muscle tissues are considered to contain levels of low prion infectivity, it has been shown that myotubes in culture efficiently propagate PrPSc. Given the high consumption of muscle tissue, it is important to understand what factors could influence the establishment of a prion infection in muscle tissue. Here we used in vitro myotube cultures, differentiated from the C2C12 myoblast cell line (dC2C12), to identify factors affecting prion replication. A range of experimental conditions revealed that PrPSc is tightly associated with proteins found in the systemic extracellular matrix, mostly fibronectin (FN). The interaction of PrPSc with FN decreased prion infectivity, as determined by standard scrapie cell assay. Interestingly, the prion-resistant reserve cells in dC2C12 cultures displayed a FN-rich extracellular matrix while the prion-susceptible myotubes expressed FN at a low level. In agreement with the in vitro results, immunohistopathological analyses of tissues from sheep infected with natural scrapie demonstrated a prion susceptibility phenotype linked to an extracellular matrix with undetectable levels of FN. Conversely, PrPSc deposits were not observed in tissues expressing FN. These data indicate that extracellular FN may act as a natural barrier against prion replication and that the extracellular matrix composition may be a crucial feature determining prion tropism in different tissues.
Asunto(s)
Fibronectinas , Enfermedades por Prión , Priones , Scrapie , Animales , Humanos , Línea Celular , Fibronectinas/uso terapéutico , Enfermedades por Prión/tratamiento farmacológico , Enfermedades por Prión/prevención & control , Priones/metabolismo , Scrapie/metabolismo , OvinosRESUMEN
Numerous studies have investigated the cellular prion protein (PrPC) since its discovery. These investigations have explained that its structure is predominantly composed of alpha helices and short beta sheet segments, and when its abnormal scrapie isoform (PrPSc) is infected, PrPSc transforms the PrPC, leading to prion diseases, including Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy in cattle. Given its ubiquitous distribution across a variety of cellular types, the PrPC manifests a diverse range of biological functions, including cell-cell adhesion, neuroprotection, signalings, and oxidative stress response. PrPC is also expressed in immune tissues, and its functions in these tissues include the activation of immune cells and the formation of secondary lymphoid tissues, such as the spleen and lymph nodes. Moreover, high expression of PrPC in immune cells plays a crucial role in the pathogenesis of prion diseases. In addition, it affects inflammation and the development and progression of cancer via various mechanisms. In this review, we discuss the studies on the role of PrPC from various immunological perspectives. [BMB Reports 2023; 56(12): 645-650].
Asunto(s)
Encefalopatía Espongiforme Bovina , Enfermedades por Prión , Priones , Humanos , Animales , Bovinos , Proteínas Priónicas/química , Proteínas Priónicas/metabolismo , Enfermedades por Prión/patología , Enfermedades por Prión/prevención & control , Encefalopatía Espongiforme Bovina/metabolismo , Encefalopatía Espongiforme Bovina/prevención & control , Sistema Inmunológico/metabolismoRESUMEN
Prion diseases are fatal infectious neurodegenerative disorders and prototypic conformational diseases, caused by the conformational conversion of the normal cellular prion protein (PrPC) into the pathological PrPSc isoform. Examples are scrapie in sheep and goat, bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in cervids, and Creutzfeldt-Jacob disease (CJD) in humans. There are no therapies available, and animal prion diseases like BSE and CWD can negatively affect the economy, ecology, animal health, and possibly human health. BSE is a confirmed threat to human health, and mounting evidence supports the zoonotic potential of CWD. CWD is continuously expanding in North America in numbers and distribution and was recently identified in Scandinavian countries. CWD is the only prion disease occurring both in wild and farmed animals, which, together with extensive shedding of infectivity into the environment, impedes containment strategies. There is currently a strong push to develop vaccines against CWD, including ones that can be used in wildlife. The immune system does not develop a bona fide immune response against prion infection, as PrPC and PrPSc share an identical protein primary structure, and prions seem not to represent a trigger for immune responses. This asks for alternative vaccine strategies, which focus on PrPC-directed self-antibodies or exposure of disease-specific structures and epitopes. Several groups have established a proof-of-concept that such vaccine candidates can induce some levels of protective immunity in cervid and rodent models without inducing unwanted side effects. This review will highlight the most recent developments and discuss progress and challenges remaining.
Asunto(s)
Ciervos , Encefalopatía Espongiforme Bovina , Enfermedades por Prión , Priones , Vacunas , Enfermedad Debilitante Crónica , Animales , Bovinos , Humanos , Ovinos , Objetivos , Enfermedades por Prión/prevención & control , Enfermedades por Prión/metabolismo , Priones/metabolismo , Encefalopatía Espongiforme Bovina/metabolismo , Enfermedad Debilitante Crónica/prevención & control , Enfermedad Debilitante Crónica/metabolismo , Ciervos/metabolismo , CabrasRESUMEN
Transmissible spongiform encephalopathies are incurable neurodegenerative diseases, associated with the conversion of the physiological prion protein to its disease-associated counterpart. Even though immunization against transmissible spongiform encephalopathies has shown great potential, immune tolerance effects impede the use of active immunization protocols for successful prophylaxis. In this study, we evaluate the use of trypanosomes as biological platforms for the presentation of a prion antigenic peptide to the host immune system. Using the engineered trypanosomes in an immunization protocol without the use of adjuvants led to the development of a humoral immune response against the prion protein in wild type mice, without the appearance of adverse reactions. The immune reaction elicited with this protocol displayed in vitro therapeutic potential and was further evaluated in a bioassay where immunized mice were partially protected in a representative murine model of prion diseases. Further studies are underway to better characterize the immune reaction and optimize the immunization protocol.
Asunto(s)
Enfermedades por Prión , Priones , Trypanosoma , Animales , Inmunización , Ratones , Enfermedades por Prión/prevención & control , Proteínas Priónicas , Priones/genética , VacunaciónRESUMEN
In Japan, 156 cases of dura mater-transplanted Creutzfeldt-Jakob disease(dCJD)with a history of Lyodura transplantation have been confirmed until February 2022, with only a few new cases still being identified. The history of Lyodura transplantation is one involving a neurosurgical procedure. The cumulative global number of cases of bovine spongiform encephalopathy-related variant CJD(BSE-related vCJD), which has shaken societies around the world, is 232 as of 2019. Thus, the impact of dCJD on the society in Japan needs no explanation. Thanks to the world's concerted efforts in research and countermeasures, medically induced prion diseases are finally becoming a thing of the past. However, due to the extremely long incubation period of CJD and the difficulty of tracing the source of infection, immediate action in the event of an outbreak is not possible, and efforts must focus on preventing disease outbreaks. Independent of this, approximately 200 cases of solitary and hereditary prion diseases occur annually in Japan. If neurosurgery must be performed on such patients, secondary transmission of prion disease by neurosurgical instruments must be prevented. Therefore, sterilization methods for neurosurgical instruments are critical, and various measures including sterilization methods have been determined and published by a research group designated by the Japanese Ministry of Health, Labour and Welfare. The sterilization of neurosurgical instruments should comply with the latest guidelines that are published by this study group.
Asunto(s)
Síndrome de Creutzfeldt-Jakob , Neurocirugia , Enfermedades por Prión , Priones , Animales , Bovinos , Colágeno , Síndrome de Creutzfeldt-Jakob/prevención & control , Síndrome de Creutzfeldt-Jakob/cirugía , Humanos , Procedimientos Neuroquirúrgicos , Enfermedades por Prión/epidemiología , Enfermedades por Prión/prevención & control , Enfermedades por Prión/cirugíaRESUMEN
Nestled within the Rocky Mountain National Forest, 114 scientists and students gathered at Colorado State University's Mountain Campus for this year's 21st annual Rocky Mountain National Virology Association meeting. This 3-day retreat consisted of 31 talks and 30 poster presentations discussing advances in research pertaining to viral and prion diseases. The keynote address provided a timely discussion on zoonotic coronaviruses, lessons learned, and the path forward towards predicting, preparing, and preventing future viral disease outbreaks. Other invited speakers discussed advances in SARS-CoV-2 surveillance, molecular interactions involved in flavivirus genome assembly, evaluation of ethnomedicines for their efficacy against infectious diseases, multi-omic analyses to define risk factors associated with long COVID, the role that interferon lambda plays in control of viral pathogenesis, cell-fusion-dependent pathogenesis of varicella zoster virus, and advances in the development of a vaccine platform against prion diseases. On behalf of the Rocky Mountain Virology Association, this report summarizes select presentations.
Asunto(s)
Virología , Animales , Interacciones Huésped-Patógeno , Humanos , Pandemias/prevención & control , Enfermedades por Prión/diagnóstico , Enfermedades por Prión/prevención & control , Priones/inmunología , Priones/aislamiento & purificación , Priones/patogenicidad , Vacunas , Virología/organización & administración , Virosis/diagnóstico , Virosis/epidemiología , Virosis/prevención & control , Virosis/virología , Virus/clasificación , Virus/inmunología , Virus/aislamiento & purificación , Virus/patogenicidadRESUMEN
The study of prions and the discovery of candidate therapeutics for prion disease have been facilitated by the ability of prions to replicate in cultured cells. Paradigms in which prion proteins from different species are expressed in cells with low or no expression of endogenous prion protein (PrP) have expanded the range of prion strains that can be propagated. In these systems, cells stably expressing a PrP of interest are typically generated via coexpression of a selectable marker and treatment with an antibiotic. Here, we report the unexpected discovery that the aminoglycoside G418 (Geneticin) interferes with the ability of stably transfected cultured cells to become infected with prions. In G418-resistant lines of N2a or CAD5 cells, the presence of G418 reduced levels of protease-resistant PrP following challenge with the RML or 22L strains of mouse prions. G418 also interfered with the infection of cells expressing hamster PrP with the 263K strain of hamster prions. Interestingly, G418 had minimal to no effect on protease-resistant PrP levels in cells with established prion infection, arguing that G418 selectively interferes with de novo prion infection. As G418 treatment had no discernible effect on cellular PrP levels or its localization, this suggests that G418 may specifically target prion assemblies or processes involved in the earliest stages of prion infection.
Asunto(s)
Gentamicinas/farmacología , Proteínas Priónicas/efectos de los fármacos , Priones/antagonistas & inhibidores , Aminoglicósidos/metabolismo , Aminoglicósidos/farmacología , Animales , Línea Celular , Línea Celular Tumoral , Gentamicinas/metabolismo , Ratones , Proteínas PrPC/efectos de los fármacos , Proteínas PrPC/metabolismo , Proteínas PrPSc/efectos de los fármacos , Proteínas PrPSc/metabolismo , Enfermedades por Prión/prevención & control , Proteínas Priónicas/metabolismo , Priones/metabolismo , Inhibidores de la Síntesis de la ProteínaRESUMEN
Prions are infectious agents causing prion diseases, which include Creutzfeldt-Jakob disease (CJD) in humans. Several cases have been reported to be transmitted through medical instruments that were used for preclinical CJD patients, raising public health concerns on iatrogenic transmissions of the disease. Since preclinical CJD patients are currently difficult to identify, medical instruments need to be adequately sterilized so as not to transmit the disease. In this study, we investigated the sterilizing activity of two oxidizing agents, ozone gas and vaporized hydrogen peroxide, against prions fixed on stainless steel wires using a mouse bioassay. Mice intracerebrally implanted with prion-contaminated stainless steel wires treated with ozone gas or vaporized hydrogen peroxide developed prion disease later than those implanted with control prion-contaminated stainless steel wires, indicating that ozone gas and vaporized hydrogen peroxide could reduce prion infectivity on wires. Incubation times were further elongated in mice implanted with prion-contaminated stainless steel wires treated with ozone gas-mixed vaporized hydrogen peroxide, indicating that ozone gas mixed with vaporized hydrogen peroxide reduces prions on these wires more potently than ozone gas or vaporized hydrogen peroxide. These results suggest that ozone gas mixed with vaporized hydrogen peroxide might be more useful for prion sterilization than ozone gas or vaporized hydrogen peroxide alone.
Asunto(s)
Peróxido de Hidrógeno/química , Ozono/química , Priones , Acero Inoxidable , Animales , Encéfalo/metabolismo , Encéfalo/patología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Peróxido de Hidrógeno/farmacología , Ratones , Ozono/farmacología , Proteínas PrPC/antagonistas & inhibidores , Proteínas PrPC/química , Enfermedades por Prión/etiología , Enfermedades por Prión/prevención & control , Acero Inoxidable/químicaRESUMEN
Brain organoids, or brainoids, have shown great promise in the study of central nervous system (CNS) infection. Modeling Zika virus (ZIKV) infection in brain organoids may help elucidate the relationship between ZIKV infection and microcephaly. Brain organoids have been used to study the pathogenesis of SARS-CoV-2, human immunodeficiency virus (HIV), HSV-1, and other viral infections of the CNS. In this review, we summarize the advances in the development of viral infection models in brain organoids and their potential application for exploring mechanisms of viral infections of the CNS and in new drug development. The existing limitations are further discussed and the prospects for the development and application of brain organs are prospected.
Asunto(s)
Encéfalo/citología , Organoides/citología , Técnicas de Cultivo de Tejidos/métodos , Animales , Encéfalo/patología , Encéfalo/virología , Humanos , Organoides/virología , Enfermedades por Prión/patología , Enfermedades por Prión/prevención & control , Enfermedades por Prión/terapia , Técnicas de Cultivo de Tejidos/economía , Virosis/patología , Virosis/prevención & control , Virosis/terapiaRESUMEN
Transmissible spongiform encephalopathies or prion diseases describe a number of different human disorders that differ in their clinical phenotypes, which are nonetheless united by their transmissible nature and common pathology. Clinical variation in the absence of a conventional infectious agent is believed to be encoded by different conformations of the misfolded prion protein. This misfolded protein is the target of methods designed to prevent disease transmission in a surgical setting and reduction of the misfolded seed or preventing its continued propagation have been the focus of therapeutic strategies. It is therefore possible that strain variation may influence the efficacy of prevention and treatment approaches. Historically, an understanding of prion disease transmission and pathogenesis has been focused on research tools developed using agriculturally relevant strains of prion disease. However, an increased understanding of the molecular biology of human prion disorders has highlighted differences not only between different forms of the disease affecting humans and animals but also within diseases such as Creutzfeldt-Jakob Disease (CJD), which is represented by several sporadic CJD specific conformations and an additional conformation associated with variant CJD. In this chapter we will discuss whether prion strain variation can affect the efficacy of methods used to decontaminate prions and whether strain variation in pre-clinical models of prion disease can be used to identify therapeutic strategies that have the best possible chance of success in the clinic.
Asunto(s)
Enfermedades por Prión/prevención & control , Enfermedades por Prión/terapia , Priones/metabolismo , Humanos , Enfermedades por Prión/transmisiónRESUMEN
Prions are a self-propagating misfolded conformation of a cellular protein. Prions are found in several eukaryotic organisms with mammalian prion diseases encompassing a wide range of disorders. The first recognized prion disease, the transmissible spongiform encephalopathies (TSEs), affect several species including humans. Alzheimer's disease, synucleinopathies, and tauopathies share a similar mechanism of self-propagation of the prion form of the disease-specific protein reminiscent of the infection process of TSEs. Strain diversity in prion disease is characterized by differences in the phenotype of disease that is hypothesized to be encoded by strain-specific conformations of the prion form of the disease-specific protein. Prion therapeutics that target the prion form of the disease-specific protein can lead to the emergence of drug-resistant strains of prions, consistent with the hypothesis that prion strains exist as a dynamic mixture of a dominant strain in combination with minor substrains. To overcome this obstacle, therapies that reduce or eliminate the template of conversion are efficacious, may reverse neuropathology, and do not result in the emergence of drug resistance. Recent advancements in preclinical diagnosis of prion infection may allow for a combinational approach that treats the prion form and the precursor protein to effectively treat prion diseases.
Asunto(s)
Enfermedades por Prión/terapia , Priones/metabolismo , Animales , Humanos , Proteínas PrPSc/metabolismo , Enfermedades por Prión/prevención & control , Enfermedades por Prión/transmisiónAsunto(s)
Investigación Biomédica , Demencia/prevención & control , Pruebas Genéticas , Oligonucleótidos Antisentido/uso terapéutico , Enfermedades por Prión/prevención & control , Proteínas Priónicas/genética , Conflicto de Intereses , Demencia/genética , Femenino , Humanos , Enfermedades por Prión/genética , Proteínas Priónicas/efectos de los fármacosRESUMEN
Incidences of iatrogenic Creutzfeldt-Jakob disease (iCJD) are caused by transplantation of prion-contaminated hormones, cornea and dura mater as well as contact with prion- contaminated medical devices, such as stereotactic electrodes, used in neurosurgery. Because prions are highly resistant and difficult to inactivate, prion contamination is a severe risk when medical instruments are reused after surgical procedures involving suspicious and confirmed cases of patients with prion diseases. Therefore, when high-risk procedures such as cerebral surgery, craniotomy surgery, orthopaedic spinal surgery and ophthalmic surgery are performed for high-risk patients or individuals with prion diseases, it is neces- sary to appropriately treat the medical devices using scientifically proven prion inactivation methods. In this chapter, we introduce fundamental aspects of prion inactivation methods, looking specifically at the practical issues involved in their implementation.
Asunto(s)
Enfermedades por Prión/prevención & control , Enfermedades por Prión/transmisión , Priones/antagonistas & inhibidores , Animales , Detergentes/química , Humanos , Enfermedades por Prión/sangre , Enfermedades por Prión/etiología , Priones/sangre , Priones/efectos de los fármacos , Factores de Riesgo , Equipo QuirúrgicoRESUMEN
Prion disease dementias are currently not practically treatable. However, a proposed treatment approach using specifically targeted dual-peptide ligand masks can mask prion surface proteins and treat specific prion diseases. Different approaches might be used to treat these prion diseases. One treatment introduces genetically modified cells into the gastrointestinal tract or other locations to produce dual-peptide ligand masks; and another treatment introduces only the dual-peptide ligand masks into the center of prion infections to mask prion surface proteins. An independent group introduced genetically modified therapeutic bacteria into large numbers of mammals, including several human volunteers, with safe and effective experimental results, without long-term colonization by the bacteria, which experimentally supports the feasibility of the first treatment. These approaches offer several advantages compared with other potential treatments against prion diseases in humans.
Asunto(s)
Demencia/tratamiento farmacológico , Péptidos/uso terapéutico , Enfermedades por Prión/tratamiento farmacológico , Animales , Demencia/inmunología , Demencia/prevención & control , Humanos , Memoria Inmunológica , Ligandos , Microorganismos Modificados Genéticamente , Enfermedades por Prión/diagnóstico , Enfermedades por Prión/inmunología , Enfermedades por Prión/prevención & control , Proteínas Priónicas , VacunasRESUMEN
OBJECTIVE: To determine whether preventive trials in genetic prion disease could be designed to follow presymptomatic mutation carriers to onset of disease. METHODS: We assembled age at onset or death data from 1,094 individuals with high penetrance mutations in the prion protein gene (PRNP) in order to generate survival and hazard curves and test for genetic modifiers of age at onset. We used formulae and simulations to estimate statistical power for clinical trials. RESULTS: Genetic prion disease age at onset varies over several decades for the most common mutations and neither sex, parent's age at onset, nor PRNP codon 129 genotype provided additional explanatory power to stratify trials. Randomized preventive trials would require hundreds or thousands of at-risk individuals in order to be statistically powered for an endpoint of clinical onset, posing prohibitive cost and delay and likely exceeding the number of individuals available for such trials. CONCLUSION: The characterization of biomarkers suitable to serve as surrogate endpoints will be essential for the prevention of genetic prion disease. Parameters such as longer trial duration, increased enrollment, and the use of historical controls in a postmarketing study could provide opportunities for subsequent determination of clinical benefit.
Asunto(s)
Edad de Inicio , Enfermedades por Prión/prevención & control , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Ensayos Clínicos como Asunto , Femenino , Genotipo , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Mutación , Penetrancia , Enfermedades por Prión/genética , Proteínas Priónicas/genética , Modelos de Riesgos Proporcionales , Proyectos de Investigación , Adulto JovenRESUMEN
Recently, the problem of neurodegenerative diseases in the medical community has become increasingly relevant. This is due to many factors: from insufficiently studied mechanisms of development of some nosological units to low awareness of medical workers. Among neurodegenerative diseases in humans, prions constitute a very specific group, which are infectious protein particles with a unique morphological structure and capable of causing a number of incurable diseases. Despite years of research, no optimal remedy has yet been found to treat them. This review examines the already studied aspects of prion diseases as a class, including small historical background, features of ethiology, pathogenesis, course and outcome of the most common of them, as well as existing research on experimental methods of diagnostics, treatment and prevention of prion infections.
Asunto(s)
Humanos , Enfermedad de Gerstmann-Straussler-Scheinker/terapia , Síndrome de Creutzfeldt-Jakob/terapia , Enfermedades por Prión/prevención & control , Enfermedades por Prión/terapia , Insomnio Familiar Fatal/terapia , Kuru/terapiaRESUMEN
Prion-related encephalopathies or transmissible spongiform encephalopathies (TSEs) are a group of rare progressive neurodegenerative disorders that are invariably fatal with often only six months elapsing from diagnosis to patient death. This makes the development of effective therapeutic strategies challenging. Nonetheless, compounds have been identified in animal models of TSE that prolong survival and, in some instances, eradicate the disease. These have been tested in the clinic, although with modest or negative outcomes. While little progress has been made over the last decade, new findings that include the ability to identify prion aggregates at low levels in biological fluids and cells may lead to the development of early-stage biomarkers for TSE. An increased focus on immunotherapeutic approaches to TSE may result in the development of novel preventive approaches for TSE.
Asunto(s)
Enfermedades por Prión/terapia , Animales , Doxiciclina/uso terapéutico , Humanos , Inmunoterapia , Enfermedades por Prión/diagnóstico , Enfermedades por Prión/prevención & controlRESUMEN
Based on previous studies reporting the anti-prion activity of poly-L-lysine and poly-L-arginine, we investigated cationic poly-L-ornithine (PLO), poly-L-histidine (PLH), anionic poly-L-glutamic acid (PLE) and uncharged poly-L-threonine (PLT) in cultured cells chronically infected by prions to determine their anti-prion efficacy. While PLE and PLT did not alter the level of PrPSc, PLO and PLH exhibited potent PrPSc inhibition in ScN2a cells. These results suggest that the anti-prion activity of poly-basic amino acids is correlated with the cationicity of their functional groups. Comparison of anti-prion activity of PLO and PLH proposes that the anti-prion activity of poly-basic amino acids is associated with their acidic cellular compartments.
Asunto(s)
Endopeptidasas/efectos de los fármacos , Histidina/antagonistas & inhibidores , Péptidos/antagonistas & inhibidores , Proteínas PrPSc/efectos de los fármacos , Proteínas PrPSc/metabolismo , Aminoácidos Básicos , Péptidos Catiónicos Antimicrobianos/antagonistas & inhibidores , Línea Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Humanos , Fragmentos de Péptidos/antagonistas & inhibidores , Enfermedades por Prión/prevención & control , Priones/efectos de los fármacos , Priones/patogenicidadRESUMEN
Prion diseases are fatal neurodegenerative diseases caused by misfolding of the prion protein (PrP). These conditions affect humans and animals, including endemic forms in sheep and deer. Bovine, rodents, and many zoo mammals also developed prion diseases during the "mad-cow" epidemic in the 1980's. Interestingly, rabbits, horses, and dogs show unusual resistance to prion diseases, suggesting that specific sequence changes in the corresponding endogenous PrP prevents the accumulation of pathogenic conformations. In vitro misfolding assays and structural studies have identified S174, S167, and D159 as the key residues mediating the stability of rabbit, horse, and dog PrP, respectively. Here, we expressed the WT forms of rabbit, horse, and dog PrP in transgenic Drosophila and found that none of them is toxic. Replacing these key residues with the corresponding amino acids in hamster PrP showed that mutant horse (S167D) and dog (D159N) PrP are highly toxic, whereas mutant rabbit (S174â¯N) PrP is not. These results confirm the impact of S167 and D159 in local and long-range structural features in the globular domain of PrP that increase its stability, while suggesting the role of additional residues in the stability of rabbit PrP. Identifying these protective amino acids and the structural features that stabilize PrP can contribute to advance the field towards the development of therapies that halt or reverse the devastating effects of prion diseases.