Evanthia E. Pashos, YoSon Park, Xiao Wang, Avanthi Raghavan, Wenli Yang et al.
Genome-wide association studies have struggled to identify functional genes and variants underlying complex phenotypes. We recruited a multi-ethnic cohort of healthy volunteers (n = 91) and used their tissue to generate induced pluripotent stem cells (iPSCs) and hepatocyte-like cells (HLCs) for genome-wide mapping of expression quantitative trait loci (eQTLs) and allele-specific expression (ASE). We identified many eQTL genes (eGenes) not observed in the comparably sized Genotype-Tissue Expression project’s human liver cohort (n = 96). Focusing on blood lipid-associated loci, we performed massively parallel reporter assays to screen candidate functional variants and used genome-edited stem cells, CRISPR interference, and mouse modeling to establish rs2277862-CPNE1, rs10889356-DOCK7, rs10889356-ANGPTL3, and rs10872142-FRK as functional SNP-gene sets. We demonstrated HLC eGenes CPNE1, VKORC1, UBE2L3, and ANGPTL3 and HLC ASE gene ACAA2 to be lipid-functional genes in mouse models. These findings endorse an iPSC-based experimental framework to discover functional variants and genes contributing to complex human traits.
Evanthia E. Pashos, YoSon Park, Xiao Wang, Avanthi Raghavan, Wenli Yang, Deepti Abbey, Derek T. Peters, Juan Arbelaez, Mayda Hernandez, Nicolas Kuperwasser, Wenjun Li, Zhaorui Lian, Ying Liu, Wenjian Lv, Stacey L. Lytle-Gabbin, Dawn H. Marchadier, Peter Rogov, Jianting Shi, Katherine J. Slovik, Ioannis M. Stylianou, Li Wang, Ruilan Yan, Xiaolan Zhang, Sekar Kathiresan, Stephen A. Duncan, Tarjei S. Mikkelsen, Edward E. Morrisey, Daniel J. Rader, Christopher D. Brown, Kiran Musunuru. Large, Diverse Population Cohorts of hiPSCs and Derived Hepatocyte-like Cells Reveal Functional Genetic Variation at Blood Lipid-Associated Loci. 2017-04-06. https://doi.org/10.1016/j.stem.2017.03.017
A pooled episomal reporter library tested major and minor alleles of candidate variants at the CPNE1, ANGPTL3, and FRK blood-lipid-associated loci. Each allele was placed in a 145-bp tile centered on the SNP or shifted by 40 nt (C/L/R) and linked to approximately 22 barcodes; the two independent transfection experiments are reported side by side.
Original supplemental and deposited inputs retained for this study. Download files individually or together as a ZIP; nested folders are preserved. Source reuse terms apply, and sequencing reads may be omitted.
Download all 2 files (ZIP)table_S10_oligonucleotides.xlsxtable_S9_mpra_results.xlsx