MicroRNAs in Human Cerebrospinal Fluid as Biomarkers for Alzheimer's Disease.

BACKGROUND Currently available biomarkers of Alzheimer's disease (AD) include cerebrospinal fluid (CSF) protein analysis and amyloid PET imaging, each of which has limitations. The discovery of extracellular microRNAs (miRNAs) in CSF raises the possibility that miRNA may serve as novel biomarkers of AD. OBJECTIVE Investigate miRNAs in CSF obtained from living donors as biomarkers for AD. METHODS We profiled miRNAs in CSF from 50 AD patients and 49 controls using TaqMan® arrays. Replicate studies performed on a subset of 32 of the original CSF samples verified 20 high confidence miRNAs. Stringent data analysis using a four-step statistical selection process including log-rank and receiver operating characteristic (ROC) tests, followed by random forest tests, identified 16 additional miRNAs that discriminate AD from controls. Multimarker modeling evaluated linear combinations of these miRNAs via best-subsets logistic regression, and computed area under the ROC (AUC) curve ascertained classification performance. The influence of ApoE genotype on miRNA biomarker performance was also evaluated. RESULTS We discovered 36 miRNAs that discriminate AD from control CSF. 20 of these retested in replicate studies verified differential expression between AD and controls. Stringent statistical analysis also identified these 20 miRNAs, and 16 additional miRNA candidates. Top-performing linear combinations of 3 and 4 miRNAs have AUC of 0.80-0.82. Addition of ApoE genotype to the model improved performance, i.e., AUC of 3 miRNA plus ApoE4 improves to 0.84. CONCLUSIONS CSF miRNAs can discriminate AD from controls. Combining miRNAs improves sensitivity and specificity of biomarker performance, and adding ApoE genotype improves classification.

[1]  A. Roses,et al.  Identification of miRNA Changes in Alzheimer's Disease Brain and CSF Yields Putative Biomarkers and Insights into Disease Pathways , 2008 .

[2]  M. Folstein,et al.  Clinical diagnosis of Alzheimer's disease , 1984, Neurology.

[3]  Christine S. Siegismund,et al.  MicroRNA Profiling of CSF Reveals Potential Biomarkers to Detect Alzheimer`s Disease , 2015, PloS one.

[4]  J. Morris,et al.  The diagnosis of dementia due to Alzheimer’s disease: Recommendations from the National Institute on Aging-Alzheimer’s Association workgroups on diagnostic guidelines for Alzheimer's disease , 2011, Alzheimer's & Dementia.

[5]  H. Geekiyanage,et al.  MicroRNA-137/181c Regulates Serine Palmitoyltransferase and In Turn Amyloid β, Novel Targets in Sporadic Alzheimer's Disease , 2011, The Journal of Neuroscience.

[6]  D. Galimberti,et al.  Circulating miRNAs as potential biomarkers in Alzheimer's disease. , 2014, Journal of Alzheimer's disease : JAD.

[7]  Leo Breiman,et al.  Random Forests , 2001, Machine Learning.

[8]  Roderick J. A. Little,et al.  Statistical Analysis with Missing Data: Little/Statistical Analysis with Missing Data , 2002 .

[9]  A. Delacourte,et al.  Loss of microRNA cluster miR-29a/b-1 in sporadic Alzheimer's disease correlates with increased BACE1/β-secretase expression , 2008, Proceedings of the National Academy of Sciences.

[10]  W. Filipowicz,et al.  Mechanisms of miRNA-mediated post-transcriptional regulation in animal cells. , 2009, Current opinion in cell biology.

[11]  Yu Wang,et al.  Cerebrospinal fluid biomarkers for Parkinson disease diagnosis and progression , 2011, Annals of neurology.

[12]  G. Jicha,et al.  Blood serum miRNA: Non-invasive biomarkers for Alzheimer's disease , 2012, Experimental Neurology.

[13]  D. Salmon,et al.  Pulse pressure is associated with Alzheimer biomarkers in cognitively normal older adults , 2013, Neurology.

[14]  Giuliano Binetti,et al.  Cerebrospinal Fluid Biomarkers for Alzheimer’s Disease: The Present and the Future , 2011, Neurodegenerative Diseases.

[15]  Zoltan Dezso,et al.  Circulating miRNA Biomarkers for Alzheimer's Disease , 2013, Alzheimer's & Dementia.

[16]  Dong Hee Kim,et al.  Genetic markers for diagnosis and pathogenesis of Alzheimer's disease. , 2014, Gene.

[17]  M. Verbeek,et al.  MicroRNAs in Alzheimer's disease: differential expression in hippocampus and cell-free cerebrospinal fluid , 2014, Neurobiology of Aging.

[18]  D. Drachman,et al.  Apolipoprotein E ε4 allele and the lifetime risk of Alzheimer's disease : what physicians know, and what they should know , 1995 .

[19]  J. Quinn Biomarkers for Alzheimer's disease: showing the way or leading us astray? , 2012, Journal of Alzheimer's disease : JAD.

[20]  H. Shill,et al.  Profiles of Extracellular miRNA in Cerebrospinal Fluid and Serum from Patients with Alzheimer's and Parkinson's Diseases Correlate with Disease Status and Features of Pathology , 2014, PloS one.

[21]  D. Wong,et al.  Extracellular RNAs: development as biomarkers of human disease , 2015, Journal of extracellular vesicles.

[22]  Paul Bertone,et al.  Systematic comparison of microarray profiling, real-time PCR, and next-generation sequencing technologies for measuring differential microRNA expression. , 2010, RNA.

[23]  H. Struyfs,et al.  Validation of microRNAs in Cerebrospinal Fluid as Biomarkers for Different Forms of Dementia in a Multicenter Study. , 2016, Journal of Alzheimer's disease : JAD.

[24]  K. Hornik,et al.  Unbiased Recursive Partitioning: A Conditional Inference Framework , 2006 .

[25]  G. V. Kass An Exploratory Technique for Investigating Large Quantities of Categorical Data , 1980 .

[26]  A. Fischer,et al.  MicroRNAs as biomarkers for CNS disease , 2013, Front. Mol. Neurosci..

[27]  M. Folstein,et al.  The Mini-Mental State Examination. , 1983, Archives of general psychiatry.

[28]  Kei-Hoi Cheung,et al.  Integration of extracellular RNA profiling data using metadata, biomedical ontologies and Linked Data technologies , 2015, Journal of extracellular vesicles.

[29]  D. Bartel,et al.  The impact of microRNAs on protein output , 2008, Nature.

[30]  S. Hébert,et al.  Circulating microRNAs in Alzheimer’s disease: the search for novel biomarkers , 2013, Front. Mol. Neurosci..

[31]  Ge Li,et al.  Age and apolipoprotein E*4 allele effects on cerebrospinal fluid beta-amyloid 42 in adults with normal cognition. , 2006, Archives of neurology.

[32]  D. Bartel MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[33]  Hiroyuki Arai,et al.  MicroRNAs in plasma and cerebrospinal fluid as potential markers for Alzheimer's disease. , 2014, Journal of Alzheimer's disease : JAD.