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Objective— To gain mechanistic insights into the role ofLIPA (lipase A), the gene encoding LAL (lysosomal acid lipase) protein, in human macrophages.Approach and Results— We used CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR–associated protein 9) technology to knock outLIPA in human induced pluripotent stem cells and then differentiate to macrophage (human-induced pluripotent stem cells–derived macrophage [IPSDM]) to explore the human macrophageLIPA loss-of-function phenotypes.LIPA was abundantly expressed in monocyte-derived macrophages and was markedly induced on IPSDM differentiation to comparable levels as in human monocyte–derived macrophage. IPSDM with knockout ofLIPA (LIPA −/− ) had barely detectable LAL enzymatic activity. Control andLIPA −/− IPSDM were loaded with [3 H]-cholesteryl oleate–labeled AcLDL (acetylated low-density lipoprotein) followed by efflux to apolipoprotein A-I. Efflux of liberated [3 H]-cholesterol to apolipoprotein A-I was abolished inLIPA −/− IPSDM, indicating deficiency in LAL-mediated lysosomal cholesteryl ester hydrolysis. In cells loaded with [3 H]-cholesterol–labeled AcLDL, [3 H]-cholesterol efflux was, however, not different between control andLIPA −/− IPSDM.ABCA1 (ATP-binding cassette, subfamily A, member 1) expression was upregulated by AcLDL loading but to a similar extent between control andLIPA −/− IPSDM. In nonlipid loaded state,LIPA −/− IPSDM had high levels of cholesteryl ester mass compared with minute amounts in control IPSDM. Yet, with AcLDL loading, overall cholesteryl ester mass was increased to similar levels in both control andLIPA −/− IPSDM.LIPA −/− did not impact lysosomal apolipoprotein-B degradation or expression ofIL1B ,IL6 , andCCL5. Conclusions— LIPA −/− IPSDM reveals macrophage-specific hallmarks ofLIPA deficiency. CRISPR/Cas9 and IPSDM provide important tools to study human macrophage biology and more broadly for future studies of disease-associatedLIPA genetic variation in human macrophages.

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