scholarly journals Prion protein quantification in human cerebrospinal fluid as a tool for prion disease drug development

2019 ◽  
Vol 116 (16) ◽  
pp. 7793-7798 ◽  
Author(s):  
Sonia M. Vallabh ◽  
Chloe K. Nobuhara ◽  
Franc Llorens ◽  
Inga Zerr ◽  
Piero Parchi ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Prion Protein ◽  
Prion Disease ◽  
Protein Quantification ◽  
Blood Contamination ◽  
Human Cerebrospinal Fluid ◽  
Highly Sensitive ◽  
The Central Nervous System ◽  
Test Retest Reliability ◽  
And Storage

Reduction of native prion protein (PrP) levels in the brain is an attractive strategy for the treatment or prevention of human prion disease. Clinical development of any PrP-reducing therapeutic will require an appropriate pharmacodynamic biomarker: a practical and robust method for quantifying PrP, and reliably demonstrating its reduction in the central nervous system (CNS) of a living patient. Here we evaluate the potential of ELISA-based quantification of human PrP in human cerebrospinal fluid (CSF) to serve as a biomarker for PrP-reducing therapeutics. We show that CSF PrP is highly sensitive to plastic adsorption during handling and storage, but its loss can be minimized by the addition of detergent. We find that blood contamination does not affect CSF PrP levels, and that CSF PrP and hemoglobin are uncorrelated, together suggesting that CSF PrP is CNS derived, supporting its relevance for monitoring the tissue of interest and in keeping with high PrP abundance in brain relative to blood. In a cohort with controlled sample handling, CSF PrP exhibits good within-subject test–retest reliability (mean coefficient of variation, 13% in samples collected 8–11 wk apart), a sufficiently stable baseline to allow therapeutically meaningful reductions in brain PrP to be readily detected in CSF. Together, these findings supply a method for monitoring the effect of a PrP-reducing drug in the CNS, and will facilitate development of prion disease therapeutics with this mechanism of action.

scholarly journals Prion protein quantification in cerebrospinal fluid as a tool for prion disease drug development

2018 ◽  
Author(s):  
Sonia M Vallabh ◽  
Chloe K Nobuhara ◽  
Franc Llorens ◽  
Inga Zerr ◽  
Piero Parchi ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Prion Protein ◽  
Prion Disease ◽  
Prion Diseases ◽  
Protein Quantification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Blood Contamination ◽  
Highly Sensitive ◽  
The Central Nervous System ◽  
Test Retest Reliability

AbstractReduction of native prion protein (PrP) levels in the brain is an attractive and genetically validated strategy for the treatment or prevention of human prion diseases. However, clinical development of any PrP-reducing therapeutic will require an appropriate pharmacodynamic biomarker: a practical and robust method for quantifying PrP, and reliably demonstrating its reduction, in the central nervous system (CNS) of a living patient. Here we evaluate the potential of enzyme-linked immunosorbent assay (ELISA)-based quantification of human PrP in human cerebrospinal fluid (CSF) to serve as a biomarker for PrP-reducing therapeutics. We show that CSF PrP is highly sensitive to plastic adsorption during handling and storage, but its loss can be minimized by addition of detergent. We find that blood contamination does not affect CSF PrP levels, and that CSF PrP and hemoglobin are uncorrelated, together suggesting that CSF PrP is CNS-derived, supporting its relevance for monitoring the tissue of interest and in keeping with high PrP abundance in brain relative to blood. In a cohort with controlled sample handling, CSF PrP exhibits good within-subject test-retest reliability (mean coefficient of variation 13% in samples collected 8-11 weeks apart), a sufficiently stable baseline to allow therapeutically meaningful reductions in brain PrP to be readily detected in CSF. Together, these findings supply a method for monitoring the effect of a PrP-reducing drug in the CNS, enabling the development of prion disease therapeutics with this mechanism of action.

The neuroepidemiology of human prion disease

2020 ◽  
pp. 367-378
Author(s):  
Patrick JM Urwin ◽  
Anna M Molesworth
Keyword(s):  
Prion Protein ◽  
Prion Disease ◽  
Protein Misfolding ◽  
Prion Diseases ◽  
Prion Protein Gene ◽  
Human Prion Disease ◽  
Disease States ◽  
Jakob Disease ◽  
The Central Nervous System ◽  
Sporadic Disease

Human prion diseases comprise a number of rare and fatal neurodegenerative conditions that result from the accumulation in the central nervous system of an abnormal form of a naturally occurring protein, called the prion protein. The diseases occur in genetic, sporadic, and acquired forms: genetic disease is associated with mutations in the prion protein gene (PRNP); sporadic disease is thought to result from a spontaneous protein misfolding event; acquired disease results from transmission of infection from an animal or another human. The potential transmissibility of the prion in any of these forms, either in disease states or during the incubation period, has implications for public health. Here we focus on Creutzfeldt-Jakob Disease (CJD), including variant Creutzfeldt-Jakob Disease (vCJD), although we will also discuss other forms of human prion disease.

scholarly journals Effects of Blood Contamination and the Rostro-Caudal Gradient on the Human Cerebrospinal Fluid Proteome

PLoS ONE ◽  
2014 ◽  
Vol 9 (3) ◽  
pp. e90429 ◽  
Author(s):  
Elise Aasebø ◽  
Jill Anette Opsahl ◽  
Yngvild Bjørlykke ◽  
Kjell-Morten Myhr ◽  
Ann Cathrine Kroksveen ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Blood Contamination ◽  
Human Cerebrospinal Fluid

Creatine kinase isoenzymes in human cerebrospinal fluid and brain.

1984 ◽  
Vol 30 (11) ◽  
pp. 1804-1806 ◽  
Author(s):  
W L Chandler ◽  
K J Clayson ◽  
W T Longstreth ◽  
J S Fine
Keyword(s):  
Cerebrospinal Fluid ◽  
Creatine Kinase ◽  
Cardiac Arrest ◽  
Brain Ischemia ◽  
Blood Contamination ◽  
Global Brain Ischemia ◽  
Qualitative And Quantitative ◽  
Human Cerebrospinal Fluid ◽  
Brain Extracts ◽  
Global Brain

Abstract Extracts of normal brains obtained at autopsy and cerebrospinal fluid (CSF) from patients with global brain ischemia were analyzed for creatine kinase (CK; EC 2.7.3.2) isoenzymes. We used both qualitative and quantitative assays (electrophoresis and immunoinhibition). Brain extracts contained CK-BB isoenzyme and mitochondrial CK. In 54 CSF samples free of blood contamination and with total activities ranging from 7 to 2010 U/L (mean 202 U/L), virtually all of the CK activity was due to CK-BB, and none to CK-MM or CK-MB. We conclude that brain contains CK-BB and mitochondrial CK, but lacks CK-MM and CK-MB. After cardiac arrest, CK-BB is released into the CSF. Any CK-MM in the CSF is probably from blood contamination, in which case immunoinhibition with anti-CK-M antibodies accurately quantifies CK-BB.

scholarly journals Prion protein in the cerebrospinal fluid of healthy and naturally scrapie-affected sheep

2006 ◽  
Vol 87 (12) ◽  
pp. 3723-3727 ◽  
Author(s):  
Nicole Picard-Hagen ◽  
Véronique Gayrard ◽  
Catherine Viguié ◽  
Mohammed Moudjou ◽  
Chantal Imbs ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Physicochemical Properties ◽  
Prion Protein ◽  
Prion Disease ◽  
Genetic Variants ◽  
Proteinase K ◽  
Soluble Form ◽  
C Terminus ◽  
The Mean ◽  
Mean Concentration

The aim of this study was to characterize the cerebrospinal fluid (CSF) prion protein (PrP) of healthy and naturally scrapie-affected sheep. The soluble form of CSF PrPC immunoblotted with an anti-octarepeat and an anti-C terminus mAb showed two isoforms of approximately 33 and 26 kDa, corresponding to the biglycosylated and unglycosylated isoforms of brain PrPC. Neither the mean concentration nor the electrophoretic profile of CSF PrP differed between healthy and scrapie-affected sheep, whereas a slightly increased resistance of CSF PrP to mild proteolysis by proteinase K was evident in the CSF of scrapie-affected sheep. No difference in susceptibility to proteolysis was observed between the two ARR and VRQ genetic variants of the purified prokaryote recombinant PrP. It was concluded that the physicochemical properties of PrPC in the CSF could be altered during scrapie and that these changes might reflect the physiopathological process of prion disease.

A soluble form of prion protein in human cerebrospinal fluid: Implications for prion-related encephalopathies

1992 ◽  
Vol 184 (3) ◽  
pp. 1398-1404 ◽  
Author(s):  
Fabrizio Tagliavini ◽  
Frances Prelli ◽  
Monica Porro ◽  
Mario Salmona ◽  
Orso Bugiani ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Prion Protein ◽  
Soluble Form ◽  
Human Cerebrospinal Fluid

scholarly journals Cerebrospinal fluid and plasma biomarkers in individuals at risk for genetic prion disease

2019 ◽  
Author(s):  
Sonia M Vallabh ◽  
Eric Vallabh Minikel ◽  
Victoria J Williams ◽  
Becky C Carlyle ◽  
Alison J McManus ◽  
...  
Keyword(s):  
At Risk ◽  
Cerebrospinal Fluid ◽  
Prion Protein ◽  
Prion Disease ◽  
Neuronal Damage ◽  
Massachusetts General Hospital ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Mutation Carriers ◽  
One Year

ABSTRACTBACKGROUNDFluid biomarkers are important in the development of therapeutics to delay or prevent prion disease, but have not been systematically evaluated in pre-symptomatic individuals at risk for genetic prion disease.METHODSWe recruited pre-symptomatic individuals with pathogenic mutations in the prion protein gene (PRNP; N=27) and matched controls (N=16), to donate cerebrospinal fluid (CSF) and blood at multiple timepoints to a cohort study at Massachusetts General Hospital. We quantified total prion protein (PrP) and real-time quaking-induced conversion (RT-QuIC) prion seeding activity in CSF, and the neuronal damage markers total tau (T-tau) and neurofilament light chain (NfL) in both CSF and plasma. We compared these markers cross-sectionally between mutation carriers and controls, evaluated short-term test-retest reliability over 2-4 months, and conducted a pilot longitudinal study over 10-20 months for a subset of participants.FINDINGSCSF PrP levels were stable on test-retest with a mean coefficient of variation of 7% both over 2-4 months in N=29 participants and over 10-20 months in N=10 participants. RT-QuIC was negative in 22/23 mutation carriers. The sole individual with positive RT-QuIC seeding activity at two study visits had steady CSF PrP levels and slightly increased tau and NfL concentrations compared with others, though still within the normal range, and remained asymptomatic one year later. Overall, tau and NfL showed no significant differences between mutation carriers and controls in either CSF or plasma.INTERPRETATIONCSF PrP will be interpretable as a pharmacodynamic readout of the effects of a PrP-lowering therapeutic in pre-symptomatic individuals, and may serve as a surrogate biomarker in a “primary prevention” trial paradigm. In contrast, current markers of prion seeding activity and of neuronal damage do not reliably cross-sectionally distinguish mutation carriers from controls, arguing against the feasibility of a “secondary prevention” paradigm in which trials specifically recruit pre-symptomatic participants with prodromal evidence of pathology.

scholarly journals Domain-specific quantification of prion protein in cerebrospinal fluid by targeted mass spectrometry

2019 ◽  
Author(s):  
Eric Vallabh Minikel ◽  
Eric Kuhn ◽  
Alexandra R Cocco ◽  
Sonia M Vallabh ◽  
Christina R Hartigan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cerebrospinal Fluid ◽  
Prion Protein ◽  
Prion Disease ◽  
Multiple Reaction Monitoring ◽  
Disease Process ◽  
Brain Homogenate ◽  
Preclinical Development ◽  
Progressive Dementia ◽  
Intracellular Compartments

AbstractTherapies currently in preclinical development for prion disease seek to lower prion protein (PrP) expression in the brain. Trials of such therapies are likely to rely on quantification of PrP in cerebrospinal fluid (CSF) as a pharmacodynamic biomarker and possibly as a trial endpoint. Studies using PrP ELISA kits have reproducibly shown that CSF PrP is lowered in the symptomatic phase of disease, a potential confounder for reading out the effect of PrP-lowering drugs in symptomatic patients. To date it has been unclear whether the reduced abundance of PrP in CSF results from its incorporation into plaques, retention in intracellular compartments, downregulation as a function of the disease process, or other factors. Because misfolding or proteolytic cleavage could potentially render PrP invisible to ELISA even if its concentration were constant or increasing in disease, we sought to establish an orthogonal method for CSF PrP quantification. We developed a targeted mass spectrometry method based on multiple reaction monitoring (MRM) of nine PrP tryptic peptides quantified relative to known concentrations of isotopically labeled standards. Analytical validation experiments showed process replicate coefficients of variation below 15%, good dilution linearity and recovery, and suitable performance for both CSF and brain homogenate and across humans as well as preclinical species of interest. InN=55 CSF samples from individuals referred to prion surveillance centers with rapidly progressive dementia, all six human PrP peptides, spanning the N- and C-terminal domains of PrP, were uniformly reduced in prion disease cases compared to individuals with non-prion diagnoses. This confirms the findings from ELISA studies, demonstrating that lowered CSF PrP concentration in prion disease is a genuine result of the disease process and not merely an artifact of ELISA-based measurement. We provide a targeted mass spectrometry-based method suitable for preclinical and clinical quantification of CSF PrP as a tool for drug development.

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