ACE inhibition and cardiometabolic risk factors, lung ACE2 and TMPRSS2 gene expression, and plasma ACE2 levels: a Mendelian randomization study

Dipender Gill; Marios Arvanitis; Paul Carter; Ana I Hernandez Cordero; Brian Jo; Ville Karhunen; Susanna C Larsson; Xuan Li; Sam M Lockhart; Amy M Mason; Evanthia Pashos; Ashis Saha; Vanessa Tan; Verena Zuber; Yohan Bosse; Sarah Fahle; Ke Hao; Tao Jiang; Philippe Joubert; Alan C Lunt; Willem hendrik Ouwehand; David J Roberts; Wim Timens; Maarten van den Berge; Nicholas A Watkins; Alexis Battle; Adam S Butterworth; John Danesh; Barbara E Engelhard; James E Peters; Don Sin; Stephen Burgess

Genetic association estimates with circulating plasma ACE2 levels were obtained in a subcohort of 4,998 blood donors enrolled in the INTERVAL BioResource (1). Plasma ACE2 levels were measured using a multiplex proximity extension immunoassay (Cardiovascular 2 panel, Olink Bioscience, Uppsala, Sweden). A total of 4,947 samples passed quality control. The data were pre-processed using standard Olink workflows including applying median centring normalization across plates, where the median is centred to the overall median for all plates, followed by log2 transformation to provide normalized protein levels (NPX). NPX values were regressed on age, sex, plate, time from blood draw to processing (in days), and season. The residuals were then rank-inverse normalized. Genotype data was processed as described previously (43) . Genome-wide pQTL analysis was performed by linear regression of the rank-inverse normalized residuals on genotype in SNPTEST (44) , with the first three components of multi-dimensional scaling as covariates to adjust for ancestry.