Highlights
- •A hPSC-derived multicellular liver culture mimicking liver composition was developed.
- •Multicellular liver culture recapitulates many key features of NAFLD development.
- •Multicellular liver cultures harbouring PNPLA3I148M enhance susceptibility to NAFLD.
- •Elevating IL-6/STAT3 signalling promotes PNPLA3I148M-induced NAFLD progression.
- •Blocking trans-signalling by sgp130Fc protects against PNPLA3I148M-induced NAFLD progression.
Background & Aims
A number of genetic polymorphisms have been associated with susceptibility to or protection
against non-alcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain
unknown. Here, we focused on the rs738409 C>G single nucleotide polymorphism (SNP),
which produces the I148M variant of patatin-like phospholipase domain-containing protein
3 (PNPLA3) and is strongly associated with NAFLD.
Methods
To enable mechanistic dissection, we developed a human pluripotent stem cell (hPSC)-derived
multicellular liver culture by incorporating hPSC-derived hepatocytes, hepatic stellate
cells, and macrophages. We first applied this liver culture to model NAFLD by utilising
a lipotoxic milieu reflecting the circulating levels of disease risk factors in affected
individuals. We then created an isogenic pair of liver cultures differing only at
rs738049 and compared NAFLD phenotype development.
Results
Our hPSC-derived liver culture recapitulated many key characteristics of NAFLD development
and progression including lipid accumulation and oxidative stress, inflammatory response,
and stellate cell activation. Under the lipotoxic conditions, the I148M variant caused
the enhanced development of NAFLD phenotypes. These differences were associated with
elevated IL-6/signal transducer and activator of transcription 3 (STAT3) activity
in liver cultures, consistent with transcriptomic data of liver biopsies from individuals
carrying the rs738409 SNP. Dampening IL-6/STAT3 activity alleviated the I148M-mediated
susceptibility to NAFLD, whereas boosting it in wild-type liver cultures enhanced
NAFLD development. Finally, we attributed this elevated IL-6/STAT3 activity in liver
cultures carrying the rs738409 SNP to increased NF-κB activity.
Conclusions
Our study thus reveals a potential causal link between elevated IL-6/STAT3 activity
and 148M-mediated susceptibility to NAFLD.
Impact and implications
An increasing number of genetic variants manifest in non-alcoholic fatty liver disease
(NAFLD) development and progression; however, the underlying mechanisms remain elusive.
To study these variants in human-relevant systems, we developed an induced pluripotent
stem cell-derived multicellular liver culture and focused on a common genetic variant
(i.e. rs738409 in PNPLA3). Our findings not only provide mechanistic insight, but also a potential therapeutic
strategy for NAFLD driven by this genetic variant in PNPLA3. Our liver culture is therefore a useful platform for exploring genetic variants
in NAFLD development.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: August 29, 2022
Accepted:
August 17,
2022
Received in revised form:
July 27,
2022
Received:
January 27,
2022
Footnotes
Author names in bold designate shared co-first authorship.
Identification
Copyright
© 2022 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
