Unknown,Transcriptomics,Genomics,Proteomics

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Effect of live Mycobacterium leprae infection on global gene expression of primary human Schwann cells at 24 hours

ABSTRACT: In an attempt to understand M. leprae interaction with the human host, Stanford Genomics HEEBO Arrays were use to identify modulated genes in primary human Schwann cells infected with live M. leprae at a MOI of 100:1 for 24 hours. Dual channel competitive hybridizations between M. leprae infected and non infected Schwann cells including 3 independent biological replicates and a technical replicate in the form of dye swap.

ORGANISM(S): Homo sapiens

SUBMITTER: Anna Beatriz Ferreira

PROVIDER: E-GEOD-40950 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Hereditary Motor and Sensory Neuropathies (HMSN) are the most common inherited peripheral neuropathies and comprise a group of genetically heterogeneous diseases. We study the genetic defects causing three demyelinating disorders: HMSN1A, HMSN-LOM and congenital cateracts facial dysmophism neuropathy (CCFDN) syndrome, which are caused by mutations in PMP22, NDRG1 and CTDP1, respectively. The function and patho-mechanisms of these genes is still unknown. To identify effects downstream of the mutations, thereby possibly involved in the disease process, we used Serial expression of gene expression (SAGE) and microarray analysis. We analyzed human Schwann cell lines from three normal subjects, one HMSN-LOM patient, two HMSN-1A patients and two CCFDN patients and compared our data to a recent dataset of human Schwann cells (4 normal subjects passage 1 vs. 3, GDS1869/GSE4030 M.B. Bunge, 2006). We excluded genes that were not consistently regulated between biological and technical replicates and genes that were found to be influenced by passage number (p<0.05). Comparing expression profiles of demyelinating neuropathies with different mutations we identified: 1. expression signatures specific for each genotype. 2. Genes commonly (dys)regulated in all screened neuropathies. Interestingly, these include genes that play a role in reorganization of sodium channels in central and peripheral nerves (S100A10, CD90), or extracellular matrix remodeling (e.g. TGFBI, SERPINE1, THBS1). We also found high up-regulation of genes involved in Schwann cell lineage and in particular, genes that are associated with an immature or pre-myelinating Schwann cell phenotype (i.e. Sox2, Sox10, COL18A1, TFAP2A, Pax3, CDH19). Our study shows that many genes, possibly contributing to nerve pathology can be identified by this approach. Experiment Overall Design: Experimental design: in total 12 (two-color) arrays were run. All neuropathy samples were in duplicate (technical replicates). Two CCFDN and two HMSN1A patients were also analyzed as biological replicates. Three healthy subjects were included from which one was in duplicate. All samples (Cy3) were compared to a common reference pool (Cy5) which consisted of a mix of all samples.

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Effect of live Mycobacterium leprae infection on global gene expression of primary human Schwann cells at 24 hours

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