Genetic and environmental factors that increase the risk of late-onset Alzheimer disease are now well recognized but the cause of variable progression rates and phenotypes of sporadic Alzheimer’s disease is largely unknown. We aimed to investigate the relationship between diverse structural assemblies of amyloid-β and rates of clinical decline in Alzheimer’s disease. Using novel biophysical methods, we analysed levels, particle size, and conformational characteristics of amyloid-β in the posterior cingulate cortex, hippocampus and cerebellum of 48 cases of Alzheimer’s disease with distinctly different disease durations, and correlated the data with APOE gene polymorphism. In both hippocampus and posterior cingulate cortex we identified an extensive array of distinct amyloid-β₄₂ particles that differ in size, display of N-terminal and C-terminal domains, and conformational stability. In contrast, amyloid-β₄₀ present at low levels did not form a major particle with discernible size, and both N-terminal and C- terminal domains were largely exposed. Rapidly progressive Alzheimer’s disease that is associated with a low frequency of APOE e4 allele demonstrates considerably expanded conformational heterogeneity of amyloid-β₄₂, with higher levels of distinctly structured amyloid-β₄₂ particles composed of 30–100 monomers, and fewer particles composed of < 30 monomers. The link between rapid clinical decline and levels of amyloid-β₄₂ with distinct structural characteristics suggests that different conformers may play an important role in the pathogenesis of distinct Alzheimer’s disease phenotypes. These findings indicate that Alzheimer’s disease exhibits a wide spectrum of amyloid-β₄₂ structural states and imply the existence of prion-like conformational strains.

Sporadic Creutzfeldt-Jakob disease (sCJD), the most common human prion disease, is transmissible through iatrogenic routes due to abundant infectious prions [misfolded forms of the prion protein (PrP^Sc)] in the central nervous system (CNS). Some epidemiological studies have associated sCJD risk with non-CNS surgeries. We explored the potential prion seeding activity and infectivity of skin from sCJD patients. Autopsy or biopsy skin samples from 38 patients [21 sCJD, 2 variant CJD (vCJD), and 15 non-CJD] were analyzed by Western blotting and real-time quaking-induced conversion (RT-QuIC) for PrP^Sc. Skin samples from two patients were further examined for prion infectivity by bioassay using two lines of humanized transgenic mice. Western blotting revealed dermal PrP^Sc in one of five deceased sCJD patients and one of two vCJD patients. However, the more sensitive RT-QuIC assay detected prion seeding activity in skin from all 23 CJD decedents but not in skin from any non-CJD control individuals (with other neurological conditions or other diseases) during blinded testing. Although sCJD patient skin contained ~10³- to 10⁵-fold lower prion seeding activity than did sCJD patient brain tissue, all 12 mice from two transgenic mouse lines inoculated with sCJD skin homogenates from two sCJD patients succumbed to prion disease within 564 days after inoculation. Our study demonstrates that the skin of sCJD patients contains both prion seeding activity and infectivity, which raises concerns about the potential for iatrogenic sCJD transmission via skin.

The mammalian prions replicate by converting cellular prion protein (PrP^C) into pathogenic conformational isoform (PrP^Sc). Variations in prions, which cause different disease phenotypes, are referred to as strains. The mechanism of high-fidelity replication of prion strains in the absence of nucleic acid remains unsolved. We investigated the impact of different conformational characteristics of PrP^Sc on conversion of PrP^C in vitro using PrP^Sc seeds from the most frequent human prion disease worldwide, the Creutzfeldt-Jakob disease (sCJD). The conversion potency of a broad spectrum of distinct sCJD prions was governed by the level, conformation, and stability of small oligomers of the protease-sensitive (s) PrP^Sc. The smallest most potent prions present in sCJD brains were composed only of∼20 monomers of PrP^Sc. The tight correlation between conversion potency of small oligomers of human sPrP^Sc observed in vitro and duration of the disease suggests that sPrP^Sc conformers are an important determinant of prion strain characteristics that control the progression rate of the disease. Mammalian prion diseases were originally characterized by accumulation of protease-resistant prion protein (PrP^Sc), often forming large amyloid deposits and fibrils. However, the apparent absence of protease-resistant PrP^Sc or amyloid fibrils in growing number of prion diseases raised several fundamental questions; specifically, whether presumably protease-sensitive forms of PrP^Sc exist as distinct conformers; and whether they comprise the initial steps in prion replication or are related to the alternative misfolding pathway generating noninfectious aggregates. We investigated the conformational characteristics of protease sensitive conformers of PrP^Sc and their role in the pathogenesis of sporadic Creutzfeldt-Jakob disease (sCJD). Using two different in vitro prion protein (PrP^C) conversion techniques in tandem with biophysical methods, we identified small oligomers of protease sensitive PrP^Sc present in sCJD brains as the most potent initiators of PrP^C conversion. Their concentration and conformational stability determine the distinctly different replication potency of PrP^Sc in individual isolates of sCJD and each of these characteristics correlates tightly with duration of the disease. These features argue for a broad range of distinct prion strains causing the sCJD and imply that small oligomers of protease sensitive conformers of pathogenic prion protein are encoding incubation time and progression rate of the disease.

The origin, range, and structure of prions causing the most common human prion disease, sporadic Creutzfeldt-Jakob disease (sCJD), are largely unknown. To investigate the molecular mechanism responsible for the broad phenotypic variability of sCJD, we analyzed the conformational characteristics of protease-sensitive and protease-resistant fractions of the pathogenic prion protein (PrP^Sc) using novel conformational methods derived from a conformation-dependent immunoassay (CDI). In 46 brains of patients homozygous for polymorphisms in the PRNP gene and exhibiting either Type 1 or Type 2 western blot pattern of the PrP^Sc, we identified an extensive array of PrP^Sc structures that differ in protease sensitivity, display of critical domains, and conformational stability. Surprisingly, in sCJD cases homozygous for methionine or valine at codon 129 of the PRNP gene, the concentration and stability of protease-sensitive conformers of PrP^Sc correlated with progression rate of the disease. These data indicate that sCJD brains exhibit a wide spectrum of PrP^Sc structural states, and accordingly argue for a broad spectrum of prion strains coding for different phenotypes. The link between disease duration, levels, and stability of protease-sensitive conformers of PrP^Sc suggests that these conformers play an important role in the pathogenesis of sCJD. Sporadic Creutzfeldt-Jakob disease (sCJD) is the most common human prion disease worldwide. This neurodegenerative disease, which is transmissible and invariably fatal, is characterized by the accumulation of an abnormally folded isoform (PrP^Sc) of a host-encoded protein (PrP^C), predominantly in the brain. Most researchers believe that PrP^Sc is the infectious agent and five or six subtypes of sCJD have been identified. Whether or not these subtypes represent distinct strains of sCJD prions is debated in the context of the extraordinary variability of sCJD phenotypes, frequent co-occurrence of different PrP^Sc fragments in the same brain, and the fact that up to 90% of protease-sensitive PrP^Sc eludes the conventional analysis because it is destroyed by protease treatment. Using novel conformational methods, we identified within each clinical and pathological category an array of PrP^Sc structures that differ in protease-sensitivity, display of critical domains, and conformational stability. Each of these features offers evidence of a distinct conformation. The link between the rate at which the disease progresses, on the one hand, and the concentration and stability of protease-sensitive conformers of PrP^Sc on the other, suggests that these conformers play an important role in how the disease originates and progresses.

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.