title: Protein misfolding cyclic amplification (PMCA): Current status and future directions.
abstract: Transmissible spongiform encephalopathies (TSEs) most commonly known as prion diseases are invariably fatal neurological disorders that affect humans and animals. These disorders differ from other neurodegenerative conformational diseases caused by the accumulation in the brain of misfolded proteins, sometimes with amyloid properties, in their ability to infect susceptible species by various routes. While the infectious properties of amyloidogenic proteins, other than misfolded prion protein (PrP(TSE)), are currently under scrutiny, their potential to transmit from cell to cell, one of the intrinsic properties of the prion, has been recently shown in vitro and in vivo. Over the decades, various cell culture and laboratory animal models have been developed to study TSEs. These assays have been widely used in a variety of applications but showed to be time consuming and entailed elevated costs. Novel economic and fast alternatives became available with the development of in vitro assays that are based on the property of conformationally abnormal PrP(TSE) to recruit normal cellular PrP(C) to misfold. These include the cell-free conversion assay, protein misfolding cyclic amplification (PMCA) and quaking induced conversion assay (QuIC), of which the PMCA has been the only technology shown to generate infectious prions. Moreover, it allows indefinite amplification of PrP(TSE) with strain-specific biochemical and biological properties of the original molecules and under certain conditions may give rise to new spontaneously generated prions. The method also allows addressing the species barrier phenomena and assessing possible risks of animal-to-animal and animal-to-human transmission. Additionally, its unprecedented sensitivity has made possible the detection of as little as one infectious dose of PrP(TSE) and the biochemical identification of this protein in different tissues and biological fluids, including blood, cerebral spinal fluid (CSF), semen, milk, urine and saliva during the pre-clinical and clinical phases of the disease. The mechanistic similarities between TSEs and other conformational disorders have resulted in the adaptation of the PMCA to the amplification and detection of various amyloidogenic proteins. Here we provide a compelling discussion of the different applications of this technology to the study of TSEs and other neurodegenerative diseases.
authors.affiliation: Transmissible Diseases Department, American National Red Cross, Biomedical Services, Holland Laboratory, 15601 Crabbs Branch Way, Rockville, MD 20855, United States. Electronic address: Paula.Saa@redcross.org.
authors.forename: Paula
authors.initials: P
authors.lastname: Saá
authors.affiliation: Transmissible Diseases Department, American National Red Cross, Biomedical Services, Holland Laboratory, 15601 Crabbs Branch Way, Rockville, MD 20855, United States.
authors.forename: Larisa
authors.initials: L
authors.lastname: Cervenakova
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods / Q000639:trends*; D006801:Humans; D017096:Prion Diseases / Q000235:genetics / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D011328:Prions; D011506:Proteins
keywords: In vitro; Neurodegenerative diseases; Prions; Protein conformational disorders; Protein misfolding cyclic amplification; Transmissible spongiform encephalopathies