New methodologies to accurately assess circulating active transforming growth factor-β1 levels: implications for evaluating heart failure and the impact of left ventricular assist devices

Transforming growth factor-beta 1 (TGF-beta 1) has been used as a biomarker in disorders associated with pathologic fibrosis. However, plasma TGF-beta 1 assessment is confounded by the significant variation in reported normal values, likely reflecting variable release of the large pool of platelet TGF-beta 1 after blood drawing. Moreover, current assays measure only total TGF-beta 1, which is dominated by the latent form of TGF-beta 1 rather than the biologically active form. To address these challenges, we developed methodologies to prevent ex vivo release of TGF-beta 1 and to quantify active TGF-beta 1. We then used these techniques to measure TGF-beta 1 in healthy controls and patients with heart failure (HF) before and after insertion of left ventricular assist devices (LVAD). Total plasma TGF-beta 1 was 1.0 +/- 0.60 ng/mL in controls and 3.76 +/- 1.55ng/mL in subjects with HF (P < 0.001), rising to 5.2 +/- 2.3 ng/mL following LVAD placement (P = 0.006). These results were paralleled by the active TGF-beta 1 values; controls had 3-16 pg/mL active TGF-beta 1, whereas levels were 2.7-fold higher in patients with HF before, and 4.2-fold higher after, LVAD implantation. Total TGF-beta 1 correlated with levels of the platelet-derived protein thrombospondin-1 (r = 0.87; P < 0.001), suggesting that plasma TGF-beta 1 may serve as a surrogate indicator of in vivo platelet activation. von Willebrand factor high molecular weight multimers correlated inversely with TGF-beta 1 levels (r = -0.63; P = 0.023), suggesting a role for shear forces in loss of these multimers and platelet activation. In conclusion, accurate assessment of circulating TGF-beta 1 may provide a valuable biomarker for in vivo platelet activation and thrombotic disorders.