Isogenic human liver organoids with LIPA loss reproduced lipid accumulation and multicellular inflammatory and biliary responses with increased fibrogenic gene expression. Restoring LIPA in hepatocyte-like cells attenuated these molecular signals.
Wolman disease is a severe form of lysosomal acid lipase deficiency caused by loss of LIPA function. The enzyme normally breaks down cholesteryl esters and triglycerides in lysosomes. To study the resulting liver pathology in human tissue, researchers generated LIPA-deficient induced pluripotent stem cells (iPSCs) and an isogenic control, then differentiated both into liver organoids containing hepatocyte-like, biliary-like and stromal populations. Both genotypes formed similarly sized organoids, providing a matched setting for comparing disease responses across cell types.
The edited cells retained pluripotency markers, while enzyme measurements confirmed the intended deficiency. LIPA-deficient organoids accumulated cholesteryl esters and triglycerides and showed expanded lysosomal compartments. Lipidomics also found higher levels of steryl esters, ceramides and broader sphingolipid classes. IL6 and IL8 gene expression rose alongside fibrogenic markers including CTGF, CYR61 and TGFB1.
Mutant organoids showed changes in lipid classes as well as increased expression of genes involved in lipid synthesis and handling. Those RNA changes were interpreted as part of the disease response; they did not directly measure the rate of lipid production or breakdown.
Single-cell analysis identified hepatocyte-like metabolic changes, stromal upregulation of ACTA2, TGFB1 and matrix genes, and a biliary-like state consistent with ductular reaction, a reactive biliary-cell gene program within the diseased tissue. Liver samples from people with Wolman disease showed KRT7-positive ductular remodeling, and Lipa-deficient mice showed a related response. The organoid single-cell experiment pooled multiple organoids into one sample per genotype; separate differentiation experiments supported selected molecular findings.
Isolated biliary organoids provided a contrast. LIPA loss caused lysosomal changes in these cultures but did not reproduce the ductular-reaction gene program or a clear growth change. Analysis of mouse livers from a previous hepatocyte-directed LIPA treatment experiment showed less ductular expansion. Together, these comparisons point to a tissue response linked to injured hepatocytes rather than a biliary-cell effect alone.
The team then restored LIPA expression preferentially in hepatocyte-like cells of the mutant liver organoids. Expression of genes associated with metabolic stress, inflammation, fibrogenesis and the reactive biliary state declined across independent differentiation experiments.
This cross-compartment response supports using the model to follow an intervention beyond the directly targeted hepatocyte-like cells. The organoids lacked mature vascular and immune compartments, limiting questions about systemic delivery or immune-mediated injury. The authors also report no formal randomization or blinding of the experiments.
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