Two-dimensional vascularized liver organoid on extracellular matrix with defined stiffness for modeling fibrotic and normal tissues.

IF 6.7 1区 工程技术 Q1 CELL & TISSUE ENGINEERING
Journal of Tissue Engineering Pub Date : 2024-08-10 eCollection Date: 2024-01-01 DOI:10.1177/20417314241268344
Lei Ma, Lin Yin, Hai Zhu, Jing Li, Dong Wang
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引用次数: 0

Abstract

Antifibrotic drug screening requires evaluating the inhibitory effects of drug candidates on fibrotic cells while minimizing any adverse effects on normal cells. It is challenging to create organ-specific vascularized organoids that accurately model fibrotic and normal tissues for drug screening. Our previous studies have established methods for culturing primary microvessels and epithelial cells from adult tissues. In this proof-of-concept study, we used rats as a model organism to create a two-dimensional vascularized liver organoid model that comprised primary microvessels, epithelia, and stellate cells from adult livers. To provide appropriate substrates for cell culture, we engineered ECMs with defined stiffness to mimic the different stages of fibrotic tissues and normal tissues. We examined the effects of two TGFβ signaling inhibitors, A83-01 and pirfenidone, on the vascularized liver organoids on the stiff and soft ECMs. We found that A83-01 inhibited fibrotic markers while promoting epithelial genes of hepatocytes and cholangiocytes. However, it inhibited microvascular genes on soft ECM, indicating a detrimental effect on normal tissues. Furthermore, A83-01 significantly promoted the expression of markers of stem cells and cancers, increasing the potential risk of it being a carcinogen. In contrast, pirfenidone, an FDA-approved compound for antifibrotic treatments, did not significantly affect all the genes examined on soft ECM. Although pirfenidone had minor effects on most genes, it did reduce the expression of collagens, the major components of fibrotic tissues. These results explain why pirfenidone can slow fibrosis progression with minor side effects in clinical trials. In conclusion, our study presents a method for creating vascularized liver organoids that can accurately mimic fibrotic and normal tissues for drug screening. Our findings provide valuable insights into the potential risks and benefits of using A83-01 and pirfenidone as antifibrotic drugs. This method can be applied to other organs to create organ-specific vascularized organoids for drug development.

细胞外基质上的二维血管化肝器官模型,具有确定的硬度,可用于纤维化和正常组织建模。
抗纤维化药物筛选需要评估候选药物对纤维化细胞的抑制作用,同时尽量减少对正常细胞的不利影响。创建器官特异性血管化器官组织,准确模拟纤维化和正常组织进行药物筛选是一项挑战。我们之前的研究已经建立了从成人组织中培养原始微血管和上皮细胞的方法。在这项概念验证研究中,我们以大鼠为模型生物,创建了一个二维血管化肝脏类器官模型,该模型由来自成人肝脏的原代微血管、上皮细胞和星状细胞组成。为了给细胞培养提供合适的基质,我们设计了具有特定硬度的 ECM,以模拟纤维化组织和正常组织的不同阶段。我们研究了两种 TGFβ 信号抑制剂 A83-01 和吡非尼酮对硬质和软质 ECM 上的血管化肝组织细胞的影响。我们发现,A83-01 可抑制纤维化标志物,同时促进肝细胞和胆管细胞的上皮基因。然而,它抑制了软 ECM 上的微血管基因,这表明它对正常组织有不利影响。此外,A83-01 还能明显促进干细胞和癌症标志物的表达,增加了其成为致癌物的潜在风险。相比之下,美国食品和药物管理局(FDA)批准用于抗纤维化治疗的化合物吡非尼酮并没有对软 ECM 上的所有基因产生明显影响。虽然吡非尼酮对大多数基因的影响较小,但它确实降低了纤维化组织的主要成分胶原蛋白的表达。这些结果解释了为什么在临床试验中,吡非尼酮可以减缓纤维化的进展,而且副作用很小。总之,我们的研究提出了一种创建血管化肝器官组织的方法,这种方法可以准确模拟纤维化组织和正常组织进行药物筛选。我们的研究结果为了解使用 A83-01 和吡非尼酮作为抗纤维化药物的潜在风险和益处提供了宝贵的见解。这种方法可应用于其他器官,以创建器官特异性血管化器官组织用于药物开发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Tissue Engineering
Journal of Tissue Engineering Engineering-Biomedical Engineering
CiteScore
11.60
自引率
4.90%
发文量
52
审稿时长
12 weeks
期刊介绍: The Journal of Tissue Engineering (JTE) is a peer-reviewed, open-access journal dedicated to scientific research in the field of tissue engineering and its clinical applications. Our journal encompasses a wide range of interests, from the fundamental aspects of stem cells and progenitor cells, including their expansion to viable numbers, to an in-depth understanding of their differentiation processes. Join us in exploring the latest advancements in tissue engineering and its clinical translation.
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