Table 1.
Summary of the main proteomic and lipidomic studies on the molecular changes associated with pleiotropic effects of PCSK9.
| Sample type | Matrix | Experimental design | Analytical techniques | Pathways involved | Main proteomic and/or lipidomic targets | Reference |
|---|---|---|---|---|---|---|
| Human hepatocyte cell line HuH7 | Cells | PCSK9 gain-of-function | SILAC labelling, SDS-PAGE, LC-MS/MS | Cytoskeletal organization, vesicle transport, membrane receptor recycling, lipid and cholesterol homeostasis, cell signalling and protein folding | EH-domain binding protein 1, Rab family of GTPases, and A-kinase anchor protein-12 | Denis et al. (63) |
| Human pancreatic beta cell line EndoC-βH1, HepG2 and HEK293T, primary hepatocytes, human islets | Cells | PCSK9 knockdown, and loss- and gain-of-function experiments | Tandem Mass Tag labelling, LC-MS/MS, WB | Basal and glucose-stimulated insulin secretion, β-cell fatty acid homeostasis, cell-lymphocytes interaction | PCSK1, MHC-I complex and Golgi/ER proteins, LDLR, VLDLR, CD36, PDL1, and HLA-ABC | Saitoski et al. (75) |
| Mouse hearts and primary cardiomyocytes | Cells/ tissues | PCSK9 loss-of-function, and PCSK9 overexpression | WB | – | Hypoxia inducible factor-1 α, microtubule-associated protein 1 light chain 3, beclin-1, AMP-activated protein kinase, and ataxia-telangiectasia mutated serine/threonine kinase | Ding et al. (65) |
| Non-diabetic patients with established coronary heart disease | Plasma | PCSK9 inhibitor (monoclonal antibody RG7652) | LC-MS /MS | – | Sphingolipids, cholesteryl esters, free cholesterol, phospholipids, ceramides and lipid composition of the lipoproteins | Hilvo et al. (67) |
| Patients with a very high risk of ASCVD | Serum | PCSK9 inhibitor (Evolocumab) | UPLC-MS | – | Phospholipids, cholesteryl esters, free cholesterol, triacylglyceride, diacylglycerol, sphingolipids, LDL-C, and Lp(a) | Huang et al. (69) |
| Patients with FH | Plasma | PCSK9 inhibitor (Evolocumab) | UHPLC-MS | – | Sphingomyelin, ceramides, cholesteryl ester, phosphatidylcholine, triacylglycerol, and phosphatidylinositol | Anesi et al. (71) |
| Patients with hypercholesterolemia | Serum | PCSK9 inhibitors (Evolocumab and Alirocumab) | Isotope dilution GC-MS selected ion-monitoring | – | Total cholesterol, LDL-C, 24S-hydroxycholesterol and 27-hydroxycholesterol and their ratios to cholesterol | Lutjohann et al. (91) |
| Human T-Rex-293 stable cell line, and mouse embryonic fibroblasts | Cells | PCSK9 gain-of-function | Immunoprecipitation, WB, SDS-PAGE, LC-MS/MS, LC/MALDI/MS/MS | Ubiquitination pathway | Cellular inhibitor of apoptosis protein 1, endoplasmic reticulum-localised proteins, mitochondrial carriers, and molecular chaperones | Xu et al. (96) |
| HepG2 and Huh7 cells | Cells | PCSK9 overexpression | Immunoprecipitation, WB, and UHPLC-MS/MS | Cholesterol metabolism | Glypican-3 | Ly et al. (97) |
| Human hepatic C3A cells | Cells | PCSK9 overexpression | Affinity chromatography, WB, LC-MS/MS | – | Alpha-1-antitrypsin, alpha-1-microglobulin/bikunin precursor, and apolipoprotein H | Melendez etal. (98) |
| Healthy volunteers, patients with incident primary CVD events (defined as myocardial infarction, ischaemic stroke, or vascular death), patients with coronary artery disease, and HepG2 cells | Plasma/cells | PCSK9 overexpression | Nuclear magnetic resonance spectroscopy, size-exclusion chromatography, immuno-isolation, crosslinking MS, label-free and Tandem Mass Tag labelling, LC-MS/MS | – | Apolipoprotein C3, and apolipoprotein A1 | Burnap et al. (107) |
| Human VSMCs, endothelial cell line EAhy926, THP-1 monocytes and THP-1-derived macrophages, J774 macrophages, and zebrafish embryos | Cells/EV | PCSK9 overexpression | WB, LC-MSE | Extracellular matrix composition, lipoprotein particle receptor binding, very-low-density lipoprotein particle receptor binding, signalling receptor binding, adhesion, inflammation, immune response, immune effector processes | -– | Greco et al. (114) |
ASCVD, atherosclerotic cardiovascular disease; CVD, cardiovascular disease; EVs, extracellular vesicles; FH, Familial Hypercholesterolemia; GC-MS, gas chromatography-mass spectrometry; GOF, gain-of-function; HLA-ABC, human leukocyte antigen-ABC; LC-MS/MS, liquid chromatography-tandem mass spectrometry; LC-MSE, liquid chromatography-mass spectrometry in data-independent analysis mode; LDL-C, LDL cholesterol; LDLR, low-density lipoprotein receptor; Lp(a), lipoprotein(a); MALDI, Matrix-Assisted Laser Desorption/Ionization; PCSK1, proprotein convertase subtilisin/kexin type 1; PDL1, programmed death-ligand 1; SDS-PAGE, Sodium Dodecyl Sulphate-PolyAcrylamide Gel Electrophoresis; SILAC, stable isotope labelling by amino acids in cell culture; UHPLC, ultra-high-performance LC; UPLC, ultra-performance LC; VLDLR, very low-density lipoprotein receptor; VSMCs, vascular smooth muscle cells.