Int J Biochem Mol Biol 2012;3(4):365-373
Original Article
microRNA (miRNA) speciation in Alzheimer’s disease
(AD) cerebrospinal fluid (CSF) and extracellular fluid
(ECF)
Peter N Alexandrov, Prerna Dua, James M Hill, Surjyadipta Bhattacharjee, Yuhai Zhao, Walter J Lukiw
Russian Academy of Medical Sciences, Moscow 113152, Russia; Deptartment of Health Information Management,
Louisiana State University, Ruston LA 71270 USA; LSU Neuroscience Center, Louisiana State University
Health Sciences Center, New Orleans LA 70112 USA; Department of Ophthalmology, Louisiana State University
Health Sciences Center, New Orleans LA 70112 USA
Received November 5, 2012; Accepted December 5, 2012; Epub December 24, 2012; Published December 30,
2012
Abstract: Human cerebrospinal fluid (CSF), produced by the choroid plexus and secreted into the brain ventricles
and subarachnoid space, plays critical roles in intra-cerebral transport and the biophysical and immune protection
of the brain. CSF composition provides valuable insight into soluble pathogenic bio-markers that may be diagnostic
for brain disease. In these experiments we analyzed amyloid beta (Aβ) peptide and micro RNA (miRNA) abundance
in CSF and in short post-mortem interval (PMI <2.1 hr) brain tissue-derived extracellular fluid (ECF) from Alzheimer’s
disease (AD) and age-matched control neocortex. There was a trend for decreased abundance of Aβ42 in the
CSF and ECF in AD but it did not reach statistical significance (mean age ~72 yr; N=12; p~0.06, ANOVA). The
most abundant nucleic acids in AD CSF and ECF were miRNAs, and their speciation and inducibility were studied
further. Fluorescent miRNA-array-based analysis indicated significant increases in miRNA-9, miRNA-125b, miRNA-
146a, miRNA-155 in AD CSF and ECF (N=12; p<0.01, ANOVA). Primary human neuronal-glial (HNG) cell co-cultures
stressed with AD-derived ECF also displayed an up-regulation of these miRNAs, an effect that was quenched using
the anti-NF-кB agents caffeic acid phenethyl ester (CAPE) or 1-fluoro-2-[2-(4-methoxy-phenyl)-ethenyl]-benzene
(CAY10512). Increases in miRNAs were confirmed independently using a highly sensitive LED-Northern dot-blot
assay. Several of these NF-кB-sensitive miRNAs are known to be up-regulated in AD brain, and associate with the
progressive spreading of inflammatory neurodegeneration. The results indicate that miRNA-9, miRNA-125b, miRNA-
146a and miRNA-155 are CSF- and ECF-abundant, NF-кB-sensitive pro-inflammatory miRNAs, and their enrichment
in circulating CSF and ECF suggest that they may be involved in the modulation or proliferation of miRNA-triggered
pathogenic signaling throughout the brain and central nervous system (CNS). (IJBMB1211001).
Keywords: Alzheimer’s disease (AD), CAPE (caffeic acid phenethyl ester), CAY10512 (1-fluoro-2-[2-(4-methoxyphenyl)-
ethenyl]-benzene), inflammatory signaling, micro RNA, NF-кB
Address all correspondence to:
Dr. WJ Lukiw,
Neuroscience, Ophthalmology and Human Genetics
LSU Neuroscience Center of Excellence
Louisiana State University Health Sciences Center
2020 Gravier Street, Suite 904
New Orleans LA 70112-2272 USA.
Phone: 504-5990842; Fax: 504-5685801
E-mail: wlukiw@lsuhsc.edu
Original Article
microRNA (miRNA) speciation in Alzheimer’s disease
(AD) cerebrospinal fluid (CSF) and extracellular fluid
(ECF)
Peter N Alexandrov, Prerna Dua, James M Hill, Surjyadipta Bhattacharjee, Yuhai Zhao, Walter J Lukiw
Russian Academy of Medical Sciences, Moscow 113152, Russia; Deptartment of Health Information Management,
Louisiana State University, Ruston LA 71270 USA; LSU Neuroscience Center, Louisiana State University
Health Sciences Center, New Orleans LA 70112 USA; Department of Ophthalmology, Louisiana State University
Health Sciences Center, New Orleans LA 70112 USA
Received November 5, 2012; Accepted December 5, 2012; Epub December 24, 2012; Published December 30,
2012
Abstract: Human cerebrospinal fluid (CSF), produced by the choroid plexus and secreted into the brain ventricles
and subarachnoid space, plays critical roles in intra-cerebral transport and the biophysical and immune protection
of the brain. CSF composition provides valuable insight into soluble pathogenic bio-markers that may be diagnostic
for brain disease. In these experiments we analyzed amyloid beta (Aβ) peptide and micro RNA (miRNA) abundance
in CSF and in short post-mortem interval (PMI <2.1 hr) brain tissue-derived extracellular fluid (ECF) from Alzheimer’s
disease (AD) and age-matched control neocortex. There was a trend for decreased abundance of Aβ42 in the
CSF and ECF in AD but it did not reach statistical significance (mean age ~72 yr; N=12; p~0.06, ANOVA). The
most abundant nucleic acids in AD CSF and ECF were miRNAs, and their speciation and inducibility were studied
further. Fluorescent miRNA-array-based analysis indicated significant increases in miRNA-9, miRNA-125b, miRNA-
146a, miRNA-155 in AD CSF and ECF (N=12; p<0.01, ANOVA). Primary human neuronal-glial (HNG) cell co-cultures
stressed with AD-derived ECF also displayed an up-regulation of these miRNAs, an effect that was quenched using
the anti-NF-кB agents caffeic acid phenethyl ester (CAPE) or 1-fluoro-2-[2-(4-methoxy-phenyl)-ethenyl]-benzene
(CAY10512). Increases in miRNAs were confirmed independently using a highly sensitive LED-Northern dot-blot
assay. Several of these NF-кB-sensitive miRNAs are known to be up-regulated in AD brain, and associate with the
progressive spreading of inflammatory neurodegeneration. The results indicate that miRNA-9, miRNA-125b, miRNA-
146a and miRNA-155 are CSF- and ECF-abundant, NF-кB-sensitive pro-inflammatory miRNAs, and their enrichment
in circulating CSF and ECF suggest that they may be involved in the modulation or proliferation of miRNA-triggered
pathogenic signaling throughout the brain and central nervous system (CNS). (IJBMB1211001).
Keywords: Alzheimer’s disease (AD), CAPE (caffeic acid phenethyl ester), CAY10512 (1-fluoro-2-[2-(4-methoxyphenyl)-
ethenyl]-benzene), inflammatory signaling, micro RNA, NF-кB
Address all correspondence to:
Dr. WJ Lukiw,
Neuroscience, Ophthalmology and Human Genetics
LSU Neuroscience Center of Excellence
Louisiana State University Health Sciences Center
2020 Gravier Street, Suite 904
New Orleans LA 70112-2272 USA.
Phone: 504-5990842; Fax: 504-5685801
E-mail: wlukiw@lsuhsc.edu
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