Expression data from LAM-associated fibroblasts and normal human lung fibroblasts co-cultured with TSC2-/- 621-101 cells

Patients with Lymphangioleiomyomatosis (LAM) develop nodules of cells within the lung parenchyma. These lesions contain both TSC2-/- LAM cells, and wild type fibroblasts. We investigated the effect of TSC2-/- cells on the transcriptional profile of both normal fibroblasts and fibroblasts isolated from LAM patient tissue (LAFs) using Affymetrix Gene Chip arrays, to identify upregulated pathways which could contribute to LAM pathology. We used Affymetrix microarrays to detail the changes in the global profile of gene expression in fibroblasts following co-culture with 621-101 cells, and identified upregulation of several chemokines, indicating that the fibroblasts had adopted an inflammatory phenotype. LAFs derived from three donors (LAMFB1, 2, 3) and normal humal lung fibroblasts (NHLFs) derived from three donors (NORMFB1, 2, 3) at matched passages, were used for this analysis. NHLFs and LAFs were cultured for 96 hours +/- 621-101 cells in a non-contact Boyden chamber co-culture system, in serum free DMEM-F12 medium with 10nM β-oestradiol at 37˚C and 5% CO2. Fibroblasts were harvested, RNA extracted and microarray hybridisation performed.