Proteomic Characterization of Human Neural Stem Cells and Their Secretome During in vitro Differentiation.
Proteomic Characterization of Human Neural Stem Cells and Their Secretome During in vitro Differentiation.
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人神经干细胞体外分化过程中的蛋白质组学特征及其分泌组学研究。
DOI：
10.3389/fncel.2020.612560
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发表时间：
2020
影响因子：
5.3
通讯作者：
Vodička P
中科院分区：
文献类型：
--
作者：
Červenka J;Tylečková J;Kupcová Skalníková H;Vodičková Kepková K;Poliakh I;Valeková I;Pfeiferová L;Kolář M;Vaškovičová M;Pánková T;Vodička P
细胞疗法是减缓目前无法治疗的神经退行性疾病（如阿尔茨海默病、帕金森病或肌萎缩侧索硬化症）进展以及支持脊髓损伤后功能性神经回路重建的一种有希望的方法。在这种治疗中，移植细胞可以在功能上融入受损组织，部分取代死亡或受损细胞，调节炎症反应，减少组织损伤，或通过分泌细胞因子、生长因子和营养因子来支持神经元存活。在移植前对细胞及其增殖潜力、分化状态和群体纯度进行全面表征对于预防安全风险至关重要，例如，未分化干细胞的增殖导致肿瘤生长。我们研究了人神经干细胞（NSCs）在自发（EGF/FGF2退出）分化和BDNF/GDNF营养支持下的分化过程中蛋白质组和分泌组的变化。我们使用LC-MS/MS在SWATH-MS模式下进行全球细胞蛋白质组分析，并定量了近3000种细胞蛋白质。我们的分析发现，在NSC分化的早期阶段，超过三分之一的受调节蛋白质（包括已知的神经元和NSC多能性标志物）存在实质性的蛋白质差异，并揭示BDNF/GDNF支持对NSC分化的后期阶段影响更大。在自发和BDNF/GDNF分化过程中被激活的途径包括HIF-1信号通路、Wnt信号通路和VEGF信号通路。我们使用Luminex多重免疫分析法进行的后续分泌组分析显示，在NSC分化过程中，VEGF和IL-6的分泌发生了显著变化。我们的研究结果进一步证明了neuropilin-1和catenin β-1的表达增加，这两种已知参与VEGF信号的调节，并表明VEGF- a亚型121 (VEGF121)，特别是诱导增殖和支持分化细胞的存活。
Cell therapies represent a promising approach to slow down the progression of currently untreatable neurodegenerative diseases (e.g., Alzheimer's and Parkinson's disease or amyotrophic lateral sclerosis), as well as to support the reconstruction of functional neural circuits after spinal cord injuries. In such therapies, the grafted cells could either functionally integrate into the damaged tissue, partially replacing dead or damaged cells, modulate inflammatory reaction, reduce tissue damage, or support neuronal survival by secretion of cytokines, growth, and trophic factors. Comprehensive characterization of cells and their proliferative potential, differentiation status, and population purity before transplantation is crucial to preventing safety risks, e.g., a tumorous growth due to the proliferation of undifferentiated stem cells. We characterized changes in the proteome and secretome of human neural stem cells (NSCs) during their spontaneous (EGF/FGF2 withdrawal) differentiation and differentiation with trophic support by BDNF/GDNF supplementation. We used LC-MS/MS in SWATH-MS mode for global cellular proteome profiling and quantified almost three thousand cellular proteins. Our analysis identified substantial protein differences in the early stages of NSC differentiation with more than a third of all the proteins regulated (including known neuronal and NSC multipotency markers) and revealed that the BDNF/GDNF support affected more the later stages of the NSC differentiation. Among the pathways identified as activated during both spontaneous and BDNF/GDNF differentiation were the HIF-1 signaling pathway, Wnt signaling pathway, and VEGF signaling pathway. Our follow-up secretome analysis using Luminex multiplex immunoassay revealed significant changes in the secretion of VEGF and IL-6 during NSC differentiation. Our results further demonstrated an increased expression of neuropilin-1 as well as catenin β-1, both known to participate in the regulation of VEGF signaling, and showed that VEGF-A isoform 121 (VEGF121), in particular, induces proliferation and supports survival of differentiating cells.
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影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
3.4
作者：
Fathi, Ali;Pakzad, Mohammad;Salekdeh, Ghasem Hosseini
通讯作者：
Salekdeh, Ghasem Hosseini
影响因子：
3.3
作者：
Cai, Y;Wu, P;Liu, M
通讯作者：
Liu, M
影响因子：
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作者：
Bohaciakova, Dasa;Hruska-Plochan, Marian;Marsala, Martin
通讯作者：
Marsala, Martin
影响因子：
3.3
作者：
Fathi, Ali;Hatami, Maryam;Salekdeh, Ghasem Hosseini
通讯作者：
Salekdeh, Ghasem Hosseini