旋转培养通过 YAP 易位调节 HepaRG 来源的肝类器官的形成。

Rotating culture regulates the formation of HepaRG-derived liver organoids via YAP translocation.

机构信息

Center for Biomechanics and Bioengineering, Beijing Key Laboratory of Engineered Construction and Mechanobiology and Key Laboratory of Microgravity (National Microgravity Laboratory), Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190, China.

Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China.

出版信息

BMC Biol. 2024 Nov 15;22(1):262. doi: 10.1186/s12915-024-02062-1.

DOI:10.1186/s12915-024-02062-1

PMID:39548509

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11568593/

Abstract

BACKGROUND

Liver organoid serves as an alternative model for liver pathophysiology in carbohydrate or lipid metabolism and xenobiotic metabolism transformation. Biomechanical cues including spaceflight mission can affect liver organoid construction and their related functions, but their underlying mechanisms are not fully understood yet. Here, a rotating cell culture device, namely Rotating Flat Chamber (RFC), was specifically designed for adhering cells or cell aggregated to elucidate the effects of altered gravity vector on HepaRG-derived liver organoids construction.

RESULTS

The organoids so formed under RFC presented the fast growth rate and large projection area. Meanwhile, the expressions of two pluripotency markers of SOX9 and CD44 were enhanced. This finding was positively correlated with the increased YAP expression and nuclear translocation as well as the elevated αβ-integrin expression. Inhibition of YAP expression and nuclear translocation decreased the expression of SOX9 and CD44 under RFC, thereby attenuating the pluripotency of HepaRG-derived liver organoids.

CONCLUSIONS

In conclusion, we proposed a novel liver organoid construction method using rotating culture, by which the pluripotency of liver organoids so constructed is mediated by αβ-integrin and YAP translocation. This work furthered the understanding in how the gravity vector orientation affects the construction of liver organoids and the related mechanotransductive pathways.

摘要

背景

类器官可作为研究碳水化合物或脂质代谢及外源性化合物代谢转化中肝生理病理的替代模型。包括航天任务在内的生物力学线索会影响类器官的构建及其相关功能，但其中的机制尚未完全阐明。本研究中，我们专门设计了一种旋转细胞培养装置（Rotating Flat Chamber，RFC），用于黏附细胞或细胞聚集体，以阐明改变重力矢量对 HepaRG 来源的肝类器官构建的影响。

结果

在 RFC 下形成的类器官具有快速的生长速度和较大的投影面积。同时，两种多能性标志物 SOX9 和 CD44 的表达增强。这一发现与 YAP 表达和核易位的增加以及αβ-整联蛋白表达的升高呈正相关。在 RFC 下抑制 YAP 的表达和核易位，会降低 SOX9 和 CD44 的表达，从而减弱 HepaRG 来源的肝类器官的多能性。