Key Laboratory of Biomaterials and Biofabrication for Tissue Engineering in Jiangxi Province, Gannan Medical University, Ganzhou, People's Republic of China.
Center for Molecular Diagnosis and Precision Medicine, and Department of Clinical Laboratory, The First Affiliated Hospital of Nanchang University, Nanchang, People's Republic of China.
Stem Cells Transl Med. 2024 Feb 14;13(2):151-165. doi: 10.1093/stcltm/szad076.
Hepatocyte-like cells (HLCs) that are differentiated from mesenchymal stem cells (MSCs) provide a valuable resource for drug screening and cell-based regeneration therapy. Differentiating HLCs into 3D spheroids enhances their phenotypes and functions. However, the molecular mechanisms underlying MSCs hepatogenic differentiation are not fully understood. In this study, we generated HLCs from human adipose-derived mesenchymal stem cells (hADMSCs) in both 2D and 3D cultures. We performed an acetyl-proteomics assay on the HLCs derived from both 2D and 3D differentiation and identified a differential change in H3K56 acetylation between the 2 differentiated cells. Our findings revealed that 3D differentiation activated ALB gene transcription by increasing the acetylation level of H3K56, thereby enhancing the phenotypes and functions of HLCs and further promoting their maturation. Notably, inhibiting p300 reduced the acetylation level of H3K56 during hepatogenic differentiation, leading to decreased phenotypes and functions of HLCs, whereas activation of p300 promoted hepatogenic differentiation, suggesting that p300 plays a critical role in this process. In summary, our study demonstrates a potential mechanism through which 3D spheroids differentiation facilitates hADMSCs differentiation into HLCs by promoting p300-mediated H3K56 acetylation, which could have significant clinical applications in liver regeneration and disease modeling.
肝细胞样细胞（HLCs）由间充质干细胞（MSCs）分化而来，为药物筛选和基于细胞的再生治疗提供了有价值的资源。将 HLCs 分化为 3D 球体可增强其表型和功能。然而，MSCs 向肝细胞分化的分子机制尚不完全清楚。在这项研究中，我们在 2D 和 3D 培养中从人脂肪间充质干细胞（hADMSCs）中生成 HLCs。我们对 2D 和 3D 分化而来的 HLCs 进行了乙酰化蛋白质组学分析，发现 2 种分化细胞之间 H3K56 乙酰化存在差异变化。我们的研究结果表明，3D 分化通过增加 H3K56 的乙酰化水平激活 ALB 基因转录，从而增强 HLCs 的表型和功能，并进一步促进其成熟。值得注意的是，抑制 p300 会降低肝向分化过程中 H3K56 的乙酰化水平，导致 HLCs 的表型和功能下降，而激活 p300 则促进肝向分化，表明 p300 在这一过程中起着关键作用。总之，我们的研究表明，3D 球体分化通过促进 p300 介导的 H3K56 乙酰化促进 hADMSCs 分化为 HLCs，这可能在肝脏再生和疾病建模方面具有重要的临床应用价值。
