Image analysis of platelet derived growth factor receptor-beta (PDGFRβ) expression to determine the grade and dynamics of myelofibrosis in bone marrow biopsy samples.

Background: Myelofibrosis (MF) is characterized by accumulation of stromal cells and extracellular matrix. Progression of fibrosis is an important clinical issue and monitoring is required for new therapeutic approaches. Currently the quantification is based on semi-quantitative evaluation of reticulin silver stained slides. We recently reported that platelet derived growth factor receptor beta (PDGFRβ) expression in fibroblasts is a useful marker of stromal activation. PDGFRβ expression based scores represent significant differences in different myelofibrosis grade which provides optimal source of quantification. In this study slide based measurements were performed in order to support correlations of PDGFRβ expression with MF grade. Methods: Scanned image tiles from 79 bone marrow samples (BM) with different myelofibrosis grades were evaluated for PDGFRβ-related IHC parameters. Following the determination of immunopositive (brown component) and total area (region of interest) of the BM, PDGFRβ related image parameters were defined and evaluated in comparison to the classical reticulin based grading. Results: Eight PDGFRβ expression related image parameters showed excellent correlation with the MF grade (correlation coefficient ranging between 0.79 and 0.83) and with PDGFRβ score (0.76-0.87). Despite the significant sample heterogeneity the parameters showed significant differences between fibrotic and non-fibrotic cases and between mild and advanced fibrosis. Distribution of values within a particular specimen emphasizes the heterogeneity of bone marrow involvement which may cause difficulties in semi-quantitative methods. Conclusions: Our results clearly demonstrated the correlation between myelofibrosis and PDGFRβ expression considering all relevant areas in BM samples. This method provides good basis for follow-up comparison of the fibrotic samples. © 2014 Clinical Cytometry Society.