Quantifying miRNA Deregulation in Alzheimer's Disease

Methods Mol Biol. 2018:1750:307-319. doi: 10.1007/978-1-4939-7704-8_21.

Abstract

Analysis of miRNA expression in circulating immune cells, such as monocytes, using qRT-PCR arrays, allows the quantification of a wide range of miRNAs in easily accessible biosamples from Alzheimer's disease patients. This technique enables the identification of differentially expressed miRNAs and provides important clues for the discovery of new miRNA-based biomarkers. Here we describe how to isolate a specific lymphocyte population from human blood samples, CD14+ monocytes, and how to extract total RNA, containing short RNAs, from these cells, transcribe the RNA into cDNA and quantify a pre-set of specific miRNAs using customizable PCR plates of 96 or 384 wells.

Keywords: Alzheimer’s disease; Biomarkers; Blood samples; Immune cells; Inflammation; Monocytes; miRNAs; qRT-PCR arrays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / blood
  • Alzheimer Disease / diagnosis*
  • Alzheimer Disease / genetics*
  • Biomarkers / analysis
  • Gene Expression Regulation*
  • Humans
  • Lipopolysaccharide Receptors / metabolism
  • Lymphocyte Subsets
  • MicroRNAs / genetics*
  • Monocytes
  • Real-Time Polymerase Chain Reaction / methods*

Substances

  • Biomarkers
  • Lipopolysaccharide Receptors
  • MicroRNAs