# Human Liver Organoids as an Experimental Tool to Investigate Lipocalin-2 in Hepatic Inflammation

## TL;DR

Human liver organoids were developed to study lipocalin-2 (LCN2) in liver inflammation, showing they respond to inflammatory signals like IL-1β and TNF-α.

## Contribution

Patient-derived liver organoids are introduced as a novel in vitro model for studying LCN2 in hepatic inflammation.

## Key findings

- Liver organoids mimic hepatocyte architecture and show self-renewal potential.
- LCN2 expression is significantly increased in organoids under IL-1β and TNF-α stimulation.
- LCN2 upregulation occurs via NF-κB-dependent signaling but not with IL-6 or LPS.

## Abstract

What are the main findings?
Patient-derived liver organoids successfully mimic key architectural features of hepatocytes and exhibit self-renewing potential, making them a valuable in vitro model for studying lipocalin-2 (LCN2) in hepatic inflammation.LCN2 expression is significantly upregulated in these organoids under inflammatory conditions, particularly through stimulation with interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α), indicating their responsiveness to pro-inflammatory signals.

Patient-derived liver organoids successfully mimic key architectural features of hepatocytes and exhibit self-renewing potential, making them a valuable in vitro model for studying lipocalin-2 (LCN2) in hepatic inflammation.

LCN2 expression is significantly upregulated in these organoids under inflammatory conditions, particularly through stimulation with interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α), indicating their responsiveness to pro-inflammatory signals.

What are the implications of the main findings?
The establishment of human liver organoids as a model provides new opportunities for investigating the role of LCN2 in liver pathologies, potentially aiding in the development of patient-specific treatment strategies for diseases such as metabolic dysfunction-associated steatotic liver disease (MASLD) and hepatocellular carcinoma (HCC).Understanding the signaling pathways involved in LCN2 regulation may contribute to therapeutic advancements targeting inflammatory processes in liver diseases, enhancing personalized medicine approaches.

The establishment of human liver organoids as a model provides new opportunities for investigating the role of LCN2 in liver pathologies, potentially aiding in the development of patient-specific treatment strategies for diseases such as metabolic dysfunction-associated steatotic liver disease (MASLD) and hepatocellular carcinoma (HCC).

Understanding the signaling pathways involved in LCN2 regulation may contribute to therapeutic advancements targeting inflammatory processes in liver diseases, enhancing personalized medicine approaches.

The 25 kDa glycoprotein lipocalin-2 (LCN2) is widely expressed and has diverse functions, ranging from physiological to pathophysiological processes. In the liver, LCN2 is primarily associated with inflammatory processes and is considered a potential biomarker in metabolic disorders. However, a significant challenge is the absence of a suitable human in vitro model for studying LCN2 and its associated signaling pathways. Therefore, we have successfully generated patient-derived liver organoids of both male and female origin, providing a novel in vitro model for LCN2 research. Our data show that the self-renewing organoids mimic essential architectural features of hepatocytes, as demonstrated by electron microscopy and F-actin staining. Consistent with the expression profile observed in liver tissue, the isolated 3D organoids exhibit minimal endogenous LCN2 levels. Next, the LCN2 expression was studied at the protein and mRNA levels under inflammatory conditions by treating the organoids with various cytokines and lipopolysaccharides (LPS). Our results show that LCN2 expression is significantly upregulated by IL-1β and TNF-α in an NF-κB-dependent manner, but remains unchanged with IL-6 or LPS. In conclusion, we have established human patient-derived liver organoids as a valuable model for investigating LCN2 signaling mechanisms. This study lays the foundation for future research on the role of LCN2 in liver pathologies, aiding in disease progression understanding and facilitating patient-specific treatment predictions.