Two ongoing clinical trials were completed, patient samples have been collected from the vitamin K intervention trial (vitaVasK) to be tested on smooth muscle cells (SMCs). Endothelial cells and SMCs in vitro models have successfully been established for testing these patients samples on their atherogeneity (ref: Jaminon et al., Cells 2021). The Biohybrid calcification screening assay was used to isolate and identify novel regulators of calcification using SMCs as cellular sensor of vascular disease. Patient samples from the VitaVasK and VitaK-CAC study will be tested soon. Moreover, proteins involved in calcification inhibition with a high intrinsic affinity for calcified lesions, i.e. Fetuin-A, Matrix Gla-protein and protein S were used to detect and prevent calcification progression. Proof of concept has been carried out in vitro, and preclinical models are now used to test this in vivo. Complementary to this, first endogenous peptides that arrest vascular calcification have been identified and are tested in vitro (ref Orth-Alampour S et al., BRC 2021). In vitro models using high glucose as model of diabetic SMC calcification are established and the scientific paper for this is being written. The novel role of PRG4 in SMC differentiation towards osteo/chondrogenic cells, thereby facilitating vascular calcification has been established and the paper is published (Seime et al., Cells 2021). Finally, the relation between regulatory mechanisms of atherosclerotic microcalcification favoring inflammation are being carried out. An extensive review on the role of macrophages in vascular calcification has been published (Waring et al., Cardiovascular Res 2021). Additionally, the role of platelet, endothelial cells and SMCs in the generation of calcification inducing extracellular vesicles has been set up and a position paper between groups has been published (Schurgers et al., Frontiers in Cardiovascular Medicine, 2018). Novel cell lines containing Cre-LoxP are in progress to further investigate the role of EVs, work that is still ongoing. The generation of iSMCs derived from iPSCs to generate SMCs of different embryonic origins and filed the first three iPSC lines. Additionally, as a result of INTRICARE, an iPSC core-facility has been founded at CARIM, Maastricht University.
Phosphorylcholine (PC) and malonedialdehyde (MDA) were identified as antigens and measures levels of natural antibodies IgM antiPC/antiMDA and IgG antiPC/antiMDA in different cohorts of atherosclerosis. This work has been extended to patients suffering from severe and mild COVID symptoms. A novel pathway on the regulation of SMC calcification was discovered, and the Wnt-PPARg signaling was confirmed as crucial in this pathway. The impact of microcalcification on SMC switching towards an osteochondrogenic VSMC phenotype has been investigated using novels models for biochemical initiation of atherogenesis and seven iPSC lines have been registered. Next to iPSC derived iSMCs, primary SMC isolates from different patients have been screened as SMC donors for the use of metabolomics screening. Within the INTRICARE consortium, novel regulatory mechanisms of atherosclerotic microcalcification favouring inflammation have been unravelled. This is also including marker identification and imaging. Fractions from bovine adrenal glands were fractionated and revealed novel mediators of calcification. Co-culturing naïve and activated macrophages and contractile and synthetic SMCs was successfully performed. Next, in vitro data are compared with in vivo imaging of novel cellular regulators of plaque vulnerability established. A first manuscript (accepted) on the correlation of CT with carotid plaque transcriptomes and association of calcification to lesion-stabilisation. PET motion correction using MR data, BOOST sequence for carotid arteries and performance and evaluation of a new dedicated carotid PET/MR coil compared to the Siemens and body coils is done.
