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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Extracell+Vesicles
2017 ; 6
(1
): 1369805
Nephropedia Template TP
gab.com Text
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English Wikipedia
Cerebrospinal fluid extracellular vesicle enrichment for protein biomarker
discovery in neurological disease; multiple sclerosis
#MMPMID28959386
Welton JL
; Loveless S
; Stone T
; von Ruhland C
; Robertson NP
; Clayton A
J Extracell Vesicles
2017[]; 6
(1
): 1369805
PMID28959386
show ga
The discovery of disease biomarkers, along with the use of "liquid biopsies" as a
minimally invasive source of biomarkers, continues to be of great interest. In
inflammatory diseases of the central nervous system (CNS), cerebrospinal fluid
(CSF) is the most obvious biofluid source. Extracellular vesicles (EVs) are also
present in CSF and are thought to be potential "biomarker treasure chests".
However, isolating these CSF-derived EVs remains challenging. This small-scale
pilot study developed and tested a protocol to enrich for CSF-EVs, both in
relapsing remitting multiple sclerosis (RRMS) CSF and controls. These were
subsequently compared, using an aptamer based proteomics array, SOMAscan?. EVs
were enriched from RRMS patient (n = 4) and non-demyelinating control (idiopathic
intracranial hypertension (IIH) (n = 3)) CSF using precipitation and mini
size-exclusion chromatography (SEC). EV-enriched fractions were selected using
pre-defined EV characteristics, including increased levels of tetraspanins. EVs
and paired CSF were analysed by SOMAscan?, providing relative abundance data for
1128 proteins. CSF-EVs were characterised, revealing exosome-like features: rich
in tetraspanins CD9 and CD81, size ~100 nm, and exosome-like morphology by TEM.
Sufficient quantities of, SOMAscan? compatible, EV material was obtained from
5 ml CSF for proteomics analysis. Overall, 348 and 580 proteins were identified
in CSF-EVs and CSF, respectively, of which 50 were found to be significantly
(t-test) and exclusively enriched in RRMS CSF-EVs. Selected proteins, Plasma
kallikrein and Apolipoprotein-E4, were further validated by western blot and
appeared increased in CSF-EVs compared to CSF. Functional enrichment analysis of
the 50 enriched proteins revealed strong associations with biological processes
relating to MS pathology and also extracellular regions, consistent with EV
enrichment. This pilot study demonstrates practicality for EV enrichment in CSF
derived from patients with MS and controls, allowing detailed analysis of protein
profiles that may offer opportunities to identify novel biomarkers and
therapeutic approaches in CNS inflammatory diseases.