Periodic Reporting for period 2 - INTRICARE (International Network for Training on Risks of vascular Intimal Calcification And roads to Regression of cardiovascular diseasE)
Période du rapport: 2019-03-01 au 2021-08-31
Cardiovascular disease remains one of the major challenges of contemporary society, being responsible for a staggering 47% of deaths in Europe. Most cardiovascular diseases arise from thrombotic rupture of an atherosclerotic plaque, the pathologic thickening of (coronary and carotid) artery segment and subsequent distal ischemia in heart or brain (infarct, cardiac arrest and stroke). In fact, 20% of European deaths are directly attributable to thrombotic rupture of a vulnerable plaque; a clear indication that current treatments and preventive measures are insufficient. Recent reports demonstrate that intimal vascular calcification, especially punctuated and spotty calcifications (microcalcifications) in the plaque are life-threatening. Microcalcifications are particularly prominent in the vulnerable atherosclerotic plaque that is at high risk of rupture. Interestingly, a high level of vascular calcification frequently occurs in two major cardiovascular comorbidities, type 2 diabetes and chronic kidney diseases, identifying vascular calcification as a shared culprit in profoundly elevated risk of cardiovascular diseases and as an excellent new target for intervention.
The objective of INTRICARE is to train a new generation of scientist that will be excellently skilled to expedite our understanding of vulnerable plaque formation, with a particular focus on microcalcification, and to develop innovative solutions for the early prevention, treatment and diagnosis of atherosclerosis.
The objective of INTRICARE is to train a new generation of scientist that will be excellently skilled to expedite our understanding of vulnerable plaque formation, with a particular focus on microcalcification, and to develop innovative solutions for the early prevention, treatment and diagnosis of atherosclerosis.
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.
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.
To attain maximum scientific and socio-economic impact, INTRICARE created awareness about the results amongst relevant stakeholders across the globe; the scientific community, industry, policy makers and reimbursement agencies with a particular focus on areas with high incidences of acute myocardial infarction AMI. For this, we contributed to conferences via special sessions on vascular calcification (i.e. ECTH 2019 Glasgow). We published in high-impact journals (i.e. Circulation Research) to reach out to both scientists and medical specialists. We established external collaborations with other H2020 ITNs (such as EVOluTION, CaReSyAn and TICARDIO) as well as with industry (such as NattoPharma). Moreover, the close collaboration with the H2020 ITN CaReSyAn resulted in the establishment of the AMICARE institute (Aachen – Maastricht Institute for Cardio Renal research). Finally, INTRICARE has made use of press-releases to create optimal socio-economic impact. To implement the communication activities, the consortium exploited and build on communications expertise available at all partners participating in the consortium.
INTRICARE research activities gave rise to results that can generate IP, mainly patents, and which have a significant potential for commercial exploitation. We envisage that our research will lead to IP on novel drugs targeting vulnerable plaque formation. Moreover, novel imaging tools/compounds will be developed. As the consortium actively trained its ESRs on valorisation and encouraged the creation of a spin-off company to exploit INTRICARE’s results, ESR13 has founded the company InflaNova in Stockholm that exploits vaccination against phosphatidylcholine to prevent atherosclerosis (https://www.linkedin.com/in/shailesh-kumar-samal).
In sum, INTRICARE research and efforts have led to 59 papers of which were published in open access journals or made open access by the beneficiary including one paper published in the top tier journal “Nature”.
INTRICARE research activities gave rise to results that can generate IP, mainly patents, and which have a significant potential for commercial exploitation. We envisage that our research will lead to IP on novel drugs targeting vulnerable plaque formation. Moreover, novel imaging tools/compounds will be developed. As the consortium actively trained its ESRs on valorisation and encouraged the creation of a spin-off company to exploit INTRICARE’s results, ESR13 has founded the company InflaNova in Stockholm that exploits vaccination against phosphatidylcholine to prevent atherosclerosis (https://www.linkedin.com/in/shailesh-kumar-samal).
In sum, INTRICARE research and efforts have led to 59 papers of which were published in open access journals or made open access by the beneficiary including one paper published in the top tier journal “Nature”.