Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling.
Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling.
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DOI：
10.1186/s13287-023-03533-y
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发表时间：
2023-10-26
影响因子：
7.5
通讯作者：
中科院分区：
文献类型：
--
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多能间充质干细胞(MSCs)的治疗应用面临着巨大的挑战，主要源于它们生长不足和自我更新能力有限。此外，随着MSCs的繁殖，其自我更新的能力下降，而导致这种情况的确切细胞和分子变化却知之甚少。本研究旨在揭示控制原始(P)MSCs自我更新的复杂分子机制。在低传代(Lp，P3)和高传代(Hp，P20)条件下，分别用胎牛血清(FM)和无异种血清(XM)培养PMSCs。为了评价LP和HP PMSCs，我们检测了它们的物理特性、细胞表面标志、生长速度、集落形成能力、BrdU增殖试验、端粒酶活性以及分化为三种谱系的可能性。此外，我们还进行了RNA-seq分析转录组和MNase-seq分析以研究核小体的占有率。当在FM中生长时，PMSCs的细胞形态发生了变化，变得更大、更长。伴随着CD90和CD49f表达的降低，以及CFE、增殖率和端粒酶活性的降低。此外，这些细胞显示出分化为成脂细胞系的趋势增加。然而，当在XM中生长时，PMSCs保持了其自我更新能力和分化为多个谱系的能力，同时保持了其成纤维细胞的形态。转录分析显示，XM培养的PMSCs中与自我更新、细胞周期调节和DNA复制相关的基因上调，而FM培养的PMSCs中衰老相关基因上调。进一步的分析表明，在XM和FM中生长的PMSCs在自我更新和衰老相关基因中的核小体占有率不同。这些发现被qRT-PCR分析所证实，该分析揭示了与自我更新、细胞周期调节、DNA复制、分化和衰老相关的基因的表达变化。为了了解其潜在的机制，我们通过用激动剂和拮抗剂来调节它们，研究了WNT和转化生长因子β信号通路的参与。这一实验操作导致了PMSCs自我更新基因的上调和下调，为进一步了解调控PMSCs自我更新和衰老的信号通路提供了进一步的见解。我们的研究表明，PMSCs的自我更新能力与Wnt途径有关，而衰老与转化生长因子β有关。网上版载有补充材料，可在10.1186/s13287-023-03533-y查阅。
The therapeutic application of multipotent mesenchymal stem cells (MSCs) encounters significant challenges, primarily stemming from their inadequate growth and limited self-renewal capabilities. Additionally, as MSCs are propagated, their ability to self-renew declines, and the exact cellular and molecular changes responsible for this are poorly understood. This study aims to uncover the complex molecular mechanisms that govern the self-renewal of primitive (p) MSCs. We grew pMSCs using two types of medium, fetal bovine serum (FM) and xeno-free (XM), at both low passage (LP, P3) and high passage (HP, P20). To evaluate LP and HP pMSCs, we examined their physical characteristics, cell surface markers, growth rate, colony-forming ability, BrdU assays for proliferation, telomerase activity, and potential to differentiate into three lineages. Moreover, we conducted RNA-seq to analyze their transcriptome and MNase-seq analysis to investigate nucleosome occupancies. When grown in FM, pMSCs underwent changes in their cellular morphology, becoming larger and elongated. This was accompanied by a decrease in the expression of CD90 and CD49f, as well as a reduction in CFE, proliferation rate, and telomerase activity. In addition, these cells showed an increased tendency to differentiate into the adipogenic lineage. However, when grown in XM, pMSCs maintained their self-renewal capacity and ability to differentiate into multiple lineages while preserving their fibroblastoid morphology. Transcriptomic analysis showed an upregulation of genes associated with self-renewal, cell cycle regulation, and DNA replication in XM-cultured pMSCs, while senescence-related genes were upregulated in FM-cultured cells. Further analysis demonstrated differential nucleosomal occupancies in self-renewal and senescence-related genes for pMSCs grown in XM and FM, respectively. These findings were confirmed by qRT-PCR analysis, which revealed alterations in the expression of genes related to self-renewal, cell cycle regulation, DNA replication, differentiation, and senescence. To understand the underlying mechanisms, we investigated the involvement of Wnt and TGFβ signaling pathways by modulating them with agonists and antagonists. This experimental manipulation led to the upregulation and downregulation of self-renewal genes in pMSCs, providing further insights into the signaling pathways governing the self-renewal and senescence of pMSCs. Our study shows that the self-renewal potential of pMSCs is associated with the Wnt pathway, while senescence is linked to TGFβ. The online version contains supplementary material available at 10.1186/s13287-023-03533-y.
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影响因子：
16.6
作者：
Dong, Peng;Maddali, Manoj V.;Srimani, Jaydeep K.;Thelot, Francois;Nevins, Joseph R.;Mathey-Prevot, Bernard;You, Lingchong
通讯作者：
You, Lingchong
影响因子：
7.5
作者：
Brown C;Agosta P;McKee C;Walker K;Mazzella M;Alamri A;Svinarich D;Chaudhry GR
通讯作者：
Chaudhry GR
影响因子：
4.3
作者：
Savickienė J;Baronaitė S;Zentelytė A;Treigytė G;Navakauskienė R
通讯作者：
Navakauskienė R
DOI：
10.1073/pnas.0505259102
发表时间：
2005-11-29
影响因子：
11.1
作者：
Clément-Lacroix, P;Ai, MR;Rawadi, G
通讯作者：
Rawadi, G
影响因子：
4.5
作者：
Al-Saqi, Shahla Hamza;Saliem, Mohammed;Gotherstrom, Cecilia
通讯作者：
Gotherstrom, Cecilia