Engineered human hepatocyte organoids enable CRISPR-based target discovery and drug screening for steatosis

TY - JOUR

T1 - Engineered human hepatocyte organoids enable CRISPR-based target discovery and drug screening for steatosis

AU - Hendriks, Delilah

AU - Brouwers, Jos F

AU - Hamer, Karien

AU - Geurts, Maarten H

AU - Luciana, Léa

AU - Massalini, Simone

AU - López-Iglesias, Carmen

AU - Peters, Peter J

AU - Rodríguez-Colman, Maria J

AU - Chuva de Sousa Lopes, Susana

AU - Artegiani, Benedetta

AU - Clevers, Hans

N1 - © 2023. The Author(s).

PY - 2023/11

Y1 - 2023/11

N2 - The lack of registered drugs for nonalcoholic fatty liver disease (NAFLD) is partly due to the paucity of human-relevant models for target discovery and compound screening. Here we use human fetal hepatocyte organoids to model the first stage of NAFLD, steatosis, representing three different triggers: free fatty acid loading, interindividual genetic variability (PNPLA3 I148M) and monogenic lipid disorders (APOB and MTTP mutations). Screening of drug candidates revealed compounds effective at resolving steatosis. Mechanistic evaluation of effective drugs uncovered repression of de novo lipogenesis as the convergent molecular pathway. We present FatTracer, a CRISPR screening platform to identify steatosis modulators and putative targets using APOB-/- and MTTP-/- organoids. From a screen targeting 35 genes implicated in lipid metabolism and/or NAFLD risk, FADS2 (fatty acid desaturase 2) emerged as an important determinant of hepatic steatosis. Enhancement of FADS2 expression increases polyunsaturated fatty acid abundancy which, in turn, reduces de novo lipogenesis. These organoid models facilitate study of steatosis etiology and drug targets.

AB - The lack of registered drugs for nonalcoholic fatty liver disease (NAFLD) is partly due to the paucity of human-relevant models for target discovery and compound screening. Here we use human fetal hepatocyte organoids to model the first stage of NAFLD, steatosis, representing three different triggers: free fatty acid loading, interindividual genetic variability (PNPLA3 I148M) and monogenic lipid disorders (APOB and MTTP mutations). Screening of drug candidates revealed compounds effective at resolving steatosis. Mechanistic evaluation of effective drugs uncovered repression of de novo lipogenesis as the convergent molecular pathway. We present FatTracer, a CRISPR screening platform to identify steatosis modulators and putative targets using APOB-/- and MTTP-/- organoids. From a screen targeting 35 genes implicated in lipid metabolism and/or NAFLD risk, FADS2 (fatty acid desaturase 2) emerged as an important determinant of hepatic steatosis. Enhancement of FADS2 expression increases polyunsaturated fatty acid abundancy which, in turn, reduces de novo lipogenesis. These organoid models facilitate study of steatosis etiology and drug targets.

KW - Humans

KW - Non-alcoholic Fatty Liver Disease/drug therapy

KW - Drug Evaluation, Preclinical

KW - Hepatocytes/metabolism

KW - Lipid Metabolism

KW - Apolipoproteins B/metabolism

KW - Liver/metabolism

U2 - 10.1038/s41587-023-01680-4

DO - 10.1038/s41587-023-01680-4

M3 - Article

C2 - 36823355

SN - 1087-0156

VL - 41

SP - 1567

EP - 1581

JO - Nature Biotechnology

JF - Nature Biotechnology

IS - 11

ER -