CRISPR/Cas9-Mediated Gene Editing in Human iPSC-Derived Macrophage Reveals Lysosomal Acid Lipase Function in Human Macrophages—Brief Report

• DOI: 10.1161/atvbaha.117.310023
• Publication Date: 2017-09-08T01:00:27Z
• Authors: Hanrui Zhang; Jianting Shi; Melanie A. Hachet; Chenyi Xue; Robert C. Bauer; Hongfeng Jiang; Wenjun Li; Junichiro Tohyama; John Millar; Jeffrey Billheimer; Michael C. Phillips; Babak Razani; Daniel J. Rader; Muredach P. Reilly
• Keywords: Cholesteryl ester; Apolipoprotein E

To gain mechanistic insights into the role of LIPA (lipase A), gene encoding LAL (lysosomal acid lipase) protein, in human macrophages.We used CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) technology to knock out induced pluripotent stem cells and then differentiate macrophage (human-induced cells-derived [IPSDM]) explore loss-of-function phenotypes. was abundantly expressed monocyte-derived macrophages markedly on IPSDM differentiation comparable levels as macrophage. with knockout (LIPA-/-) had barely detectable enzymatic activity. Control LIPA-/- were loaded [3H]-cholesteryl oleate-labeled AcLDL (acetylated low-density lipoprotein) followed by efflux apolipoprotein A-I. Efflux liberated [3H]-cholesterol A-I abolished IPSDM, indicating deficiency LAL-mediated lysosomal cholesteryl ester hydrolysis. In [3H]-cholesterol-labeled AcLDL, was, however, not different between control IPSDM. ABCA1 (ATP-binding cassette, subfamily A, member 1) expression upregulated loading but a similar extent nonlipid state, high mass compared minute amounts Yet, loading, overall increased both did impact apolipoprotein-B degradation or IL1B, IL6, CCL5. CONCLUSIONS: reveals macrophage-specific hallmarks deficiency. CRISPR/Cas9 provide important tools study biology more broadly for future studies disease-associated genetic variation macrophages.