Chronic Thrombo-embolic Pulmonary Hypertension (CTEPH) is a progressive lung disease, characterized by progressive thrombotic occlusion of the pulmonary vasculature. Recent studies have stressed the importance of inflammation and endothelial injury in the development and progression of CTEPH. In addition, inflammation driven thrombosis in the pulmonary vasculature is also seen in other diseases including Acute Respiratory Distress Syndrome (ARDS) and COVID. The endothelium plays a major role in the control of thrombosis, however, it remains largely unknown how activation or injury of the pulmonary endothelium contributes to CTEPH and other forms of inflammatory thrombosis.
The Amsterdam UMC is a central referral center for CTEPH patients, and patients undergo surgery for CTEPH at a regular basis, and we have an established pipeline for isolating cells from patient material. Following the transcriptomic and proteomic analysis of CTEPH endothelial cells in in last years, we have identified a number of candidate genes/proteins which contribute to in situ thrombosis in CTEPH.About your role
The work will mainly involve laboratory work using endothelial cells. The Department of Pulmonary Disease hosts a large preclinical laboratory, which directly links fundamental cell biology to daily patient care. As a result, a wide variety of patient tissues, plasma and cells are at your disposal. For the current project, you will be working with state-of-the art cell culture techniques, including 3D culture systems, a thrombosis-on-a-chip mode, flow/shear stress set-ups as well as lentivirus-based gene editing systems.Your core business
- Isolation and culture of endothelial cells from patient tissue;
- Endothelial cell culture, using 3D flow models;
- Genetic editing with CRISPR/Cas9, short hairpin RNA and or lentiviral overexpression models;
- Perform Chromatic Immunoprecipitation (ChIP);
- qPCR, Western Blot and immunofluorescence;
- Data analysis;
- Scientific writing.