Engineering of Trophoblast Extracellular Vesicle-Delivering Hydrogels for Localized Tolerance Induction in Cell Transplantation.
Engineering of Trophoblast Extracellular Vesicle-Delivering Hydrogels for Localized Tolerance Induction in Cell Transplantation.
复制标题
用于细胞移植中局部耐受诱导的滋养层细胞外囊泡递送水凝胶的工程。
DOI：
10.1007/s12195-023-00778-8
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发表时间：
2023
影响因子：
2.8
通讯作者：
Weaver,JessicaD
中科院分区：
文献类型：
--
作者：
Hiremath,ShivaniC;Weaver,JessicaD
慢性全身性免疫抑制对移植患者有许多急性风险，这仍然是许多细胞疗法（如用于治疗1型糖尿病的胰岛素分泌细胞）翻译的主要限制。滋养细胞是胎盘的专业耐受性细胞，它们分泌一系列可溶性因子来诱导怀孕期间对异体胎儿组织的抗原特异性耐受性，包括细胞外囊泡。在这里，我们开发了一种滋养层细胞外囊泡递送水凝胶，用于在移植部位持续、局部递送耐受性因子，以诱导对移植物的局部耐受性。方法我们设计了一种基于聚乙二醇的合成水凝胶系统来系住细胞外囊泡以持续递送，并将该系统与海藻酸盐水凝胶系统中的被动囊泡包裹进行比较。我们对滋养层细胞外囊泡的大小和形态进行了表征，并通过蛋白质组学分析评估了囊泡的耐受性蛋白含量。我们通过扫描电子和受激发射耗尽显微镜验证了水凝胶系统中细胞外囊泡的保留和束缚，并测量了囊泡随时间的释放率。最后，我们评估了滋养细胞胞外囊泡对体外自然杀伤细胞活化的影响。结果滋养细胞胞外囊泡的分离和蛋白质组学证实了耐受性因子的存在。我们证实了水凝胶递送载体中细胞外囊泡的存在，并且相对于被动水凝胶系统，合成水凝胶延长了细胞外囊泡的释放。最后，细胞外囊泡在体外降低了自然杀伤细胞的激活，证实了水凝胶递送的细胞外囊泡的耐受性。结论：这种设计用于移植部位内递送的耐受性细胞外囊泡递送水凝胶平台可以作为细胞移植中全身免疫抑制的替代方案，潜在地降低与细胞治疗相关的风险，并扩大符合条件的患者群体。
PurposeThe need for chronic systemic immunosuppression, which presents a host of acute risks to transplantation patients, remains the primary limitation for the translation of many cell therapies, such as insulin secreting cells for the treatment of type 1 diabetes. Trophoblasts are the professional tolerogenic cells of the placenta, and they secrete a range of soluble factors to induce antigen specific tolerance toward allogeneic fetal tissue during pregnancy, including extracellular vesicles. Here we develop a trophoblast extracellular vesicle-delivering hydrogel designed for sustained, localized tolerogenic factor delivery within a transplant site to induce localized tolerance toward cell grafts.MethodsWe engineer a synthetic poly(ethylene glycol)-based hydrogel system to tether extracellular vesicles for sustained delivery, and compare this system to passive vesicle entrapment within an alginate hydrogel system. We characterize trophoblast extracellular vesicles for size and morphology, and evaluate vesicle tolerogenic protein content via proteomic analysis. We validate the retention and tethering of extracellular vesicles within the hydrogel systems via scanning electron and stimulated emission depletion microscopy, and measure vesicle release rate over time. Finally, we evaluate trophoblast extracellular vesicle influence on natural killer cell activation in vitro.ResultsWe isolated trophoblast extracellular vesicles and proteomics confirmed the presence of tolerogenic factors. We confirmed the presence of extracellular vesicles within hydrogel delivery vehicles, and synthetic hydrogels extended extracellular vesicle release relative to a passive hydrogel system. Finally, extracellular vesicles reduced natural killer cell activation in vitro, confirming the tolerogenic potential of hydrogel-delivered extracellular vesicles.ConclusionsThis tolerogenic extracellular vesicle-delivering hydrogel platform designed for delivery within a transplant site could serve as an alternative to systemic immunosuppression in cell transplantation, potentially reducing the risks associated with cell therapies and widening the eligible patient population.