Gene expression profiling of brains from bovine spongiform encephalopathy (BSE)-infected cynomolgus macaques

BACKGROUND: Prion diseases are fatal neurodegenerative disorders whose pathogenesis mechanisms are not fully understood. In this context, the analysis of gene expression alterations occurring in prion-infected animals represents a powerful tool that may contribute to unravel the molecular basis of prion diseases and therefore discover novel potential targets for diagnosis and therapeutics. Here we present the first large-scale transcriptional profiling of brains from BSE-infected cynomolgus macaques, which are an excellent model for human prion disorders. RESULTS: The study was conducted using the GeneChip\uae Rhesus Macaque Genome Array and revealed 300 transcripts with expression changes greater than twofold. Among these, the bioinformatics analysis identified 86 genes with known functions, most of which are involved in cellular development, cell death and survival, lipid homeostasis, and acute phase response signaling. RT-qPCR was performed on selected gene transcripts in order to validate the differential expression in infected animals versus controls. The results obtained with the microarray technology were confirmed and a gene signature was identified. In brief, HBB and HBA2 were down-regulated in infected macaques, whereas TTR, APOC1 and SERPINA3 were up-regulated. CONCLUSIONS: Some genes involved in oxygen or lipid transport and in innate immunity were found to be dysregulated in prion infected macaques. These genes are known to be involved in other neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. Our results may facilitate the identification of potential disease biomarkers for many neurodegenerative diseases

Whole transcriptome analysis in brains from BSE-infected macaques

Prion diseases are neurodegenerative disorders that affect both humans and animals. The molecular mechanisms underlying prion replication and subsequent degeneration of the central nervous system are still poorly understood. In an attempt to identify molecules that are putatively involved in the etiology of these diseases, we conducted a whole transcriptome analysis with brain tissue from BSE-infected and uninfected cynomolgus macaques (M. fascicularis). Total RNA from the gyrus frontalis region of seven BSE-infected and five noninfected control macaques was isolated. The integrity of the RNA was assessed using an Agilent 2100 Bioanalyzer. The RNA was reverse-transcribed, labeled and analyzed on an GeneChip\uae Rhesus Macaque Genome Array (Affymetrix) containing 52,024 Macaca mulatta probe sets to monitor the gene expression of approximately 47,000 transcripts. Bioinformatic analysis revealed that about 100 transcripts were significantly up-or down-regulated more than twofold. Beside others, we found up-regulation of \u3b11-antichymotrypsin, which has also been described in scrapie-infected mice and Alzheimer\u2019s disease. We are currently validating the most interesting candidates using quantitative RT-PCR. Our approach may help to identify genes that are crucial to prion disease processes and may become potential targets for diagnostic and therapeutic strategies