High-Throughput Microscopy Characterization of Rare LDLR Variants

Rafael Graça; Magdalena Zimon; Ana C Alves; Rainer Pepperkok; Mafalda Bourbon

doi:10.1016/j.jacbts.2023.03.013

High-Throughput Microscopy Characterization of Rare LDLR Variants

JACC Basic Transl Sci. 2023 Jun 28;8(8):1010-1021. doi: 10.1016/j.jacbts.2023.03.013. eCollection 2023 Aug.

Authors

Rafael Graça^{1

2}, Magdalena Zimon^{3

4}, Ana C Alves^{1

2}, Rainer Pepperkok^{4

5}, Mafalda Bourbon^{1

2}

Affiliations

¹ Unidade de Investigação e Desenvolvimento, Grupo de Investigação Cardiovascular, Departamento de Promoção da Saúde e Prevenção de Doenças Não Transmissíveis, Instituto Nacional de Saúde Doutor Ricardo Jorge, Lisbon, Portugal.
² BioISI-Biosystems and Integrative Sciences Institute, Faculdade de Ciências, Universidade de Lisboa, Lisbon, Portugal.
³ Roche Diabetes Care GmbH, Mannheim, Germany.
⁴ Cell Biology and Cell Biophysics Unit, European Molecular Biology Laboratory (EMBL) Heidelberg, Heidelberg, Germany.
⁵ Advanced Light Microscopy Facility, EMBL Heidelberg, Heidelberg, Germany.

Abstract

Familial hypercholesterolemia (FH) is the most common inherited life-threatening disorder of lipid metabolism. Early diagnosis and treatment are the key to reduce the cumulative life-long cardiovascular burden of patients with FH. The high number of LDLR variants described as variants of unknown significance is the largest obstacle to achieve a definitive FH diagnosis. This study established a time- and cost-effective high-throughput cell-based assay to functionally profile LDLR variants, which allowed us to discriminate disruptive rare variants from silent ones. This work generated a valuable resource for systematic functional characterization of LDLR variants solving 1 of the major issues to achieve a definitive FH diagnosis.

Keywords: ACMG classification; LDLR; familial hypercholesterolemia; high-throughput characterization.