Derivation and characterisation of endothelial cells from patients with chronic thromboembolic pulmonary hypertension

Material obtained at pulmonary endarterectomy (PEA) offers the unique opportunity to unravel pathophysiological mechanisms underlying CTEPH. Vascular changes occurring in CTEPH might be linked to metabolic dysregulation of CTEPH endothelial cells (EC). The aim of this study was the development of an in vitro model of EC pathology using primary cultures of patient derived EC to assay cell metabolism in CTEPH.

Like previously showed, isolated cells from specimens collected at PEA (N=12) were confirmed as being EC. Cell migration was evaluated using scratch assay. Metabolic changes in CTEPH-EC are being studied using RT-PCR, Western-Blot and colorimetric assays.

CTEPH-EC showed a reduced capacity to recover from a scratch when compared with human pulmonary arterial endothelial cells (HPAE) (1-way anova, p<0.0001). Lactate dehydrogenase (LDH) activity, measured in the supernatant of cultured cells, was higher for CETPH-EC when compared with controls (77.57±12.11 vs 53.82±3.70; p<0.001). The amount of lactate in the supernatant of CETPH-EC was higher when compared to HPAE cells, whereas glucose was lower in comparison with controls (Lactate: 6.80±2.19 vs 4.49±2.29; p<0.05, Glucose: 48.31±21.89 vs 60±20.80; p>0.05).

Conclusion: CTEPH-EC show functional impairment. Preliminary data suggest an increase in LDH activity. Variations in lactate and glucose support the presence of a metabolic pathophenotype in CTEPH-EC.