肝样细胞的体外诱导及小分子化合物在肝样细胞诱导中的应用

唐薇; 王冬梅

doi:10.3969/j.issn.1001-5256.2023.04.037

肝样细胞的体外诱导及小分子化合物在肝样细胞诱导中的应用

DOI: 10.3969/j.issn.1001-5256.2023.04.037

唐薇^, ,,
王冬梅

福建师范大学生命科学学院，福建省发育与神经生物学重点实验室，福州 350117

基金项目:

福建省自然科学基金项目 (2020J05038);

福建省教育厅项目 (JAT190090)

利益冲突声明：所有作者均声明不存在利益冲突。

作者贡献声明：唐薇参与文献调研和文章撰写；王冬梅参与文章的文献查阅与内容修改。

详细信息

通信作者:
唐薇，40720007@qq.com (ORCID: 0000-0001-8841-4125)

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出版历程
- 收稿日期: 2022-09-05
- 录用日期: 2022-10-12
- 出版日期: 2023-04-20

Advances in in vitro induction of hepatocyte-like cells and the application of small-molecule compounds in inducing hepatocyte-like cells

Wei TANG^{,
,},
Dongmei WANG

Fujian Key Laboratory of Development and Neurobiology, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China

Research funding:

Natural Science Foundation of Fujian Province (2020J05038);

Fujian Education Department (JAT190090)

More Information

Corresponding author: TANG Wei, 40720007@qq.com (ORCID: 0000-0001-8841-4125)

摘要

摘要: 体外诱导肝样细胞(HLC)是获取大量具有应用价值的肝细胞的有效途径之一，这些HLC可以用于构建疾病模型、药物设计和药物毒理学评价等。目前，HLC的体外诱导主要通过引入外源转录因子、细胞因子或小分子化合物组合的处理。小分子化合物因其结构多样性、时间和剂量的可控性及操作上的便利和安全等优势，让科学家们致力于筛选小分子化合物来取代外源转录因子和细胞因子，其在再生医学领域的应用前景广阔。本文主要对体外诱导多能干细胞及其他成体细胞分化成HLC的研究进行概述，并总结了小分子化合物在体外诱导HLC中的应用，以期为HLC的体外诱导研究提供思路和借鉴。
- 肝样细胞 /
- 小分子化合物 /
- 再生医学
Abstract: The induction of hepatocyte-like cells (HLCs) in vitro is one of the effective ways to obtain a large number of useful hepatocyte, and these HLCs can be used in disease modeling, drug design, and toxicological evaluation. At present, the induction of HLCs in vitro is mainly achieved by introducing exogenous transcription factors, cytokines or small-molecule compounds. Since small-molecule compounds have the advantages of structural diversity, controllable time and dose, and convenient and safe operation, scientists are devoted to screening out the small-molecule compounds to replace exogenous transcription factors and cytokines, and such compounds have a promising application prospect in the field of regenerative medicine. This article reviews the studies on the in vitro induction of HLCs from pluripotent stem cells and other adult stem cells and summarizes the application of small-molecule compounds in the in vitro induction of HLCs, in order to provide ideas and references for the in vitro induction of HLCs.
- Hepatocyte-Like Cells /
- Small Molecules /
- Regenerative Medicine

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表 1 hPSC向HLC分化的不同阶段所涉及的主要生长因子和信号通路总结

Table 1. Summary of common growth factors and target signaling pathways during the differentiation of hepatocyte-like cells from hPSC

分化阶段	生长因子	相关信号通路
内胚层定向分化	Activin A，Wnt3α	Activin/Nodal信号通路
		Wnt/β-catenin信号通路
肝向分化	BMP2，FGF4	MAPK信号通路
		PI3K信号通路
		TGFβ信号通路
肝的成熟	OSM，HGF	OSM/gp130信号通路
		HGF信号通路
注：Activin A，激活素A；MAPK，丝裂原活化蛋白激酶；PI3K，磷脂酰肌醇3激酶；gp130，糖蛋白130。

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表 2 hPSC来源的肝疾病模型

Table 2. hPSC-deroved liver disease models

疾病	建模策略	应用
家族性高胆甾醇血症	HLC由患者来源的hiPSC诱导	高通量药物筛选^[20]；临床前药物疗效评价^[21]
肝豆状核变性	HLC由基因编辑过的hPSC诱导(ATP7B基因突变)	构建疾病特征模型^[22]；临床前药物疗效评价^[23]
脂肪肝	肝类器官由hiPSC诱导	在芯片系统上构建脂肪肝特征模型^[24]；抗脂肪变性药物筛选^[25]
mtDNA缺失综合征	HLC由基因编辑过的hiPSC诱导(DGUOK缺失)	高通量药物筛选^[26]
尿素循环障碍	肝类器官由患者来源的hiPSC诱导	基因编辑修复治疗^[27]
先天性肝纤维化	HLC由基因编辑过的hiPSC诱导(PKHD1敲除)	疾病机制研究^[28]

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表 3 小分子化合物在促进肝向分化中的应用

Table 3. Application of small molecules in facilitating hepatic conversion

小分子化合物	起始细胞	目的细胞	参考文献
LY294002、BIO、DMSO、SB431542	hPSC	HLC	[35]
CHIR99021、DMSO、Dihexa	hPSC	HLC	[36]
CHIR99021、DMSO、A83-01、SB、FH1、FPH1	hPSC	HLC	[37]
CHIR99021、RepSox、VPA、Parnate、TTNPB、DZNep	小鼠成纤维细胞	HLC	[38]
A-83-01、CHIR99021	小鼠成纤维细胞	HLC	[40]
CHIR99021、RepSox、VPA、Parnate、TTNPB、DZNep	人尿细胞	HLC	[39]
IDE1、CHIR99021、LY294002、FH1	间充质干细胞	HLC	[42]
DMSO、TSA、5-AZA	人脐带来源间充质干细胞	HLC	[43]

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表 4 小分子化合物在HLC诱导中的潜在机制

Table 4. Small molecules and possible mechanisms in HLC formation

诱导阶段	小分子化合物	机制	参考文献
定形内胚层的形成	CHIR99021	GSK-3抑制剂，激活Wnt/β-catenin通路，促进内胚层标志性基因表达	[35-37]
	IDE1	可取代Activin A，介导Smad2磷酸化，促使细胞向内胚层分化	[42, 46]
	LY294002	PI3K信号通路抑制剂，促进干细胞向内胚层分化	[35]
肝向分化与肝样细胞的成熟	DMSO	促进内胚层肝向分化，能够促进HLC的成熟	[35-37]
	SB、VPA、TSA	组蛋白去乙酰化抑制剂，促进肝细胞代表性基因表达，减少细胞死亡	[41, 43, 47]
	Dihexa	HGF受体激动剂，促进HLC的成熟	[36]
	SB431542	TGFβ抑制剂，促进肝前体细胞分化为HLC	[35]
	A83-01、RepSox	抑制TGFβ，促进细胞MET进程，有利于肝向分化	[8, 37, 48]
	Parnate、DZNep、5-AZA	表观遗传调控分子，转录水平上促进细胞肝向分化	[8, 43, 49]
	TTNPB	激活视黄酸受体参与调控肝核受体介导的信号通路	[50]
	FH1和FPH1	FH1可以替代HGF，FPH1可以替代OSM促进HLC成熟	[37]

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