Engineered red blood cells carrying PCSK9 inhibitors persistently lower LDL and prevent obesity

• Published in: PloS one Vol. 16; no. 11; p. e0259353
• Main Authors: Deshycka, Rhogerry, Sudaryo, Valentino, Huang, Nai-Jia, Xie, Yushu, Smeding, Liyan Y, Choi, Moon Kyung, Ploegh, Hidde L, Lodish, Harvey F, Pishesha, Novalia
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 03.11.2021; Public Library of Science (PLoS)
• Subjects: Animals; Antibodies; Apolipoprotein E; Apolipoproteins; Bioengineering; Biology and Life Sciences; Biomedical research; Blood; Blood cells; Body Weight; Bone marrow; Bone marrow transplantation; Cardiovascular disease; Cardiovascular diseases; Cells, Cultured; Cholesterol; Cholesterol, LDL - blood; Density; Diagnosis; Diet, High-Fat - adverse effects; Domains; Down-Regulation; Engineering; Epidermal growth factor; Erythrocytes; Erythrocytes - metabolism; Female; Flow cytometry; Gene therapy; Genetic Engineering; Glycophorins - chemistry; Glycophorins - genetics; Growth factors; Health aspects; Health risks; HEK293 Cells; Hematopoietic stem cells; High fat diet; Humans; Hyperlipidemia; Hyperlipidemias - chemically induced; Hyperlipidemias - metabolism; Hyperlipidemias - prevention & control; Kexin; Lipid metabolism; Lipids; Lipoprotein receptors; Lipoproteins; Liver; Low density lipoprotein; Low density lipoprotein receptors; Low density lipoproteins; Medicine; Medicine and Health Sciences; Metabolism; Mice; Mutation; Obesity; Peptides; Plasma; Plasma levels; Plasmids; Pregnancy; Progenitor cells; Proprotein Convertase 9 - blood; Proprotein convertases; Proteins; Receptors; Receptors, LDL - chemistry; Receptors, LDL - genetics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Risk factors; Risk management; RNA-mediated interference; Stem Cell Transplantation; Stem cells; Subtilisin; Transduction, Genetic; Transplantation; United States; United States > US; Massachusetts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0259353

Low plasma levels of Proprotein Convertase Subtilisin/Kexin 9 (PCSK9) are associated with decreased low-density lipoprotein (LDL) cholesterol and a reduced risk of cardiovascular disease. PCSK9 binds to the epidermal growth factor-like repeat A (EGFA) domain of LDL receptors (LDLR), very low-density lipoprotein receptors (VLDLR), apolipoprotein E receptor 2 (ApoER2), and lipoprotein receptor-related protein 1 (LRP1) and accelerates their degradation, thus acting as a key regulator of lipid metabolism. Antibody and RNAi-based PCSK9 inhibitor treatments lower cholesterol and prevent cardiovascular incidents in patients, but their high-cost hampers market penetration. We sought to develop a safe, long-term and one-time solution to treat hyperlipidemia. We created a cDNA encoding a chimeric protein in which the extracellular N- terminus of red blood cells (RBCs) specific glycophorin A was fused to the LDLR EGFA domain and introduced this gene into mouse bone marrow hematopoietic stem and progenitor cells (HSPCs). Following transplantation into irradiated mice, the animals produced RBCs with the EGFA domain (EGFA-GPA RBCs) displayed on their surface. These animals showed significantly reduced plasma PCSK9 (66.5% decrease) and reduced LDL levels (40% decrease) for as long as 12 months post-transplantation. Furthermore, the EGFA- GPA mice remained lean for life and maintained normal body weight under a high-fat diet. Hematopoietic stem cell gene therapy can generate red blood cells expressing an EGFA-glycophorin A chimeric protein as a practical and long-term strategy for treating chronic hyperlipidemia and obesity.