LDLR missense variants disturb structural conformation and LDLR activity in T-lymphocytes of Familial hypercholesterolemia patients

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Almendros Barbosa, Thais Kristini ; Crespo Hirata, Rosario Dominguez ; Ferreira, Glaucio Monteiro ; Borges, Jessica Bassani ; de Oliveira, Victor Fernandes ; Gorjao, Renata ; da Silva Marcal, Elisangela Rodrigues ; Goncalves, Rodrigo Marques ; Faludi, Andre Arpad ; Costa de Freitas, Renata Caroline ; Dagli-Hernandez, Carolina ; Bortolin, Raul Hernandes ; Bastos, Gisele Medeiros ; Pithon-Curi, Tania Cristina ; Nader, Helena Bonciani ; Hirata, Mario Hiroyuki

• Document type: Journal article
• Source: Gene; v. 853, p. 14-pg., 2022-12-12.

Abstract

Familial hypercholesterolemia (FH) is caused by deleterious mutations in the LDLR that increase markedly lowdensity lipoprotein (LDL) cholesterol and cause premature atherosclerotic cardiovascular disease. Functional effects of pathogenic LDLR variants identified in Brazilian FH patients were assessed using in vitro and in silico studies. Variants in LDLR and other FH-related genes were detected by exon-target gene sequencing. T-lymphocytes were isolated from 26 FH patients, and 3 healthy controls and LDLR expression and activity were assessed by flow cytometry and confocal microscopy. The impact of LDLR missense variants on protein structure was assessed by molecular modeling analysis. Ten pathogenic or likely pathogenic LDLR variants (six missense, two stop-gain, one frameshift, and one in splicing region) and six non-pathogenic variants were identified. Carriers of pathogenic and non-pathogenic variants had lower LDL binding and uptake in activated T-lymphocytes compared to controls (p < 0.05), but these variants did not influence LDLR expression on cell surface. Reduced LDL binding and uptake was also observed in carriers of LDLR null and defective variants. Modeling analysis showed that p.(Ala431Thr), p.(Gly549Asp) and p.(Gly592Glu) disturb intramolecular interactions of LDLR, and p.(Gly373Asp) and p.(Ile488Thr) reduce the stability of the LDLR protein. Docking and molecular interactions analyses showed that p.(Cys184Tyr) and p.(Gly373Asp) alter interaction of LDLR with Apolipoprotein B (ApoB). In conclusion, LDLR null and defective variants reduce LDL binding capacity and uptake in activated T-lymphocytes of FH patients and LDLR missense variants affect LDLR conformational stability and dissociation of the LDLR-ApoB complex, having a potential role in FH pathogenesis. (AU)

• FAPESP's process: 16/12899-6 - Genomics, epigenomics and pharmacogenomics characterization of familial hypercholesterolemia in the Brazilian population
• Grantee: Mario Hiroyuki Hirata
• Support Opportunities: Research Projects - Thematic Grants