PO.CL05.02 · 临床研究
经促炎细胞因子预激的沃顿胶间充质干细胞：一种对抗胰腺癌的强效免疫调节策略
Pro-inflammatory cytokine primed Wharton's jelly mesenchymal stem cells: A potent immunomodulatory strategy to combat pancreatic cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景与目的：来源于脐带（UC）的间充质干细胞（MSC）由于其高增殖率、多谱系分化潜能和免疫耐受特性，在损伤性、免疫相关及炎症性疾病领域具有潜在的治疗应用价值。虽然临床试验已明确证明其具有良好的安全记录，但 MSC 的疗效仍然有限且不一致。基于给药前对 MSC 进行预激可提高其治疗疗效的假设，本研究考察了通过细胞因子预激对 MSC 行为的调控，并评估其在促进肿瘤凋亡方面的有效性。蛋白质组学分析显示细胞因子预激后成分发生变化。
方法：将沃顿胶来源的 MSC（hWJMSC）用肿瘤坏死因子-α（TNF-alpha；25 ng/ml）和干扰素-γ（IFN-gamma；25 ng/ml）进行预激，在 37℃/5%CO2 条件下孵育 24 小时，检测其形态学改变、多谱系分化潜能（成骨细胞/软骨细胞/脂肪细胞）、免疫表型（CD90+73+105+44+34-45-19- HLADR-）、黏附分子（ICAM、VCAM、NCAM）和迁移能力。对鉴定出的 2400 多种蛋白进行了无标记定量 LCMS（nLC/MS/MS 联用 Orbitrap fusion Tribrid 质谱仪）分析。此外，正在使用健康人类供体的 PBMNC 检测其免疫原性潜能。
结果：经 TNF-alpha 预激的 hWJMSC（hWJMSC-T）/IFN-gamma 预激的 hWJMSC（HWJMSC-I）显示 CD90+73+105+44+ 的表达率 >90%。显微镜下用鬼笔环肽/DAPI 染色观察到的形态学差异显示，hWJMSC-T 呈现为拉长和扁平的细胞，且补体与凝血级联和促血管生成蛋白质组特征丰富，提示其在固有免疫稳态中的作用。另一方面，IFN-gamma 诱导显示出感染/炎症的免疫调节蛋白特征，尤其是鸟苷酸结合蛋白。差异表达的蛋白特征指向 NOD 样受体信号通路和细胞黏附分子通路的调控。
结论：本文数据提供了证据，表明干细胞的再生能力特性得以保留，在炎症条件下不受损害。此外，预激的 hWJMSC 能够调节或预防感染，很可能是通过防御系统级联之间的相互作用实现的。因此，提示其作为调节炎症和/或重塑炎症微环境的替代策略的潜在作用。
关键词：间充质干细胞、促炎细胞因子、炎症、免疫调节
查看英文原文 English abstract
Background & Aim: Mesenchymal stem cells (MSCs) from umbilical cord (UC) have the potential therapeutic applications in context of injured, immune related & inflammatory diseases due to high proliferation rate, multilineage differentiation potential and immune tolerance properties. While clinical trials have clearly demonstrated strong safety records, efficacy of MSCs is still modest and inconsistent. Hypothesizing that priming MSCs prior to administration can improve their therapeutic efficacy, present study investigates the modulation of MSC behaviour through cytokine priming and assesses their effectiveness in ameliorating tumor apoptosis. Proteomic analysis shows compositional changes post cytokine priming.
Method: Wharton's jelly derived MSCs (hWJMSCs) were primed with tumor necrosis factor-alpha (TNF-alpha; 25ng/ml) and interferon-gamma (IFN-gamma; 25ng/ml), incubated for 24 hrs at 37°C/5%CO2, their morphological alterations, multi-lineage differentiation potentials (osteocytes/chondrocytes/adipocytes), immunophenotypying (CD90+73+105+44+34-45-19- HLADR-), Adhesion molecules (ICAM, VCAM, NCAM) migration capabilities were examined.Label-free quantitative LCMS (nLC/MS/MS coupled to Orbitrap fusion Tribrid mass-spectrometer) analysis was performed on more than 2400 proteins identified. Further, immunogenic potentials are being tested using PBMNCs from healthy human donors.
Result: hWJMSCs primed with TNF-alpha (hWJMSC-T)/IFN-gamma (HWJMSC-I) showed >90% expression of CD90 + 73 + 105 + 44 + . Microscopically morphological distinction stained with phyalloidin/DAPI observed showed hWJMSC-T as elongated and flattened cells and Complement & coagulation cascade and Pro-angiogenic proteome signature in abundance, suggesting their role in innate immunity homeostasis. On the other hand, IFN-gamma induction showed immune-modulating protein signature of infection/inflammation especially, Guanylate-binding proteins. Differentially expressed protein signature points towards modulation of NOD-like receptor signaling and cell adhesion molecule pathways.
Conclusion: Data herein provides the evidences that stem cell properties of regenerative capability remains conserved and do not compromise under inflammatory conditions. Additionally, primed hWJMSCs can modulate or prevent infection most likely via cross-talk between cascades of defense systems. Thus, suggesting a potential role as alternate strategy to regulate inflammation and/or reshape inflammatory microenvironment.
Keywords: Mesenchymal stem cells, pro-inflammatory cytokines, inflammation, immunomodulation
利益披露 Disclosure
S. Choudhury, None..
P. Yadav, None..
M. Saad, None..
V. Singh, None..
R. Tamang, None.