Extracellular vesicles from immortalized mesenchymal stromal cells protect against neonatal hypoxic-ischemic brain injury.

IF 5 3区 医学 Q2 IMMUNOLOGY
Nicole Labusek, Yanis Mouloud, Christian Köster, Eva Diesterbeck, Tobias Tertel, Constanze Wiek, Helmut Hanenberg, Peter A Horn, Ursula Felderhoff-Müser, Ivo Bendix, Bernd Giebel, Josephine Herz
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引用次数: 6

Abstract

Background: Human mesenchymal stromal cell (MSC)-derived extracellular vesicles (EV) revealed neuroprotective potentials in various brain injury models, including neonatal encephalopathy caused by hypoxia-ischemia (HI). However, for clinical translation of an MSC-EV therapy, scaled manufacturing strategies are required, which is challenging with primary MSCs due to inter- and intra-donor heterogeneities. Therefore, we established a clonally expanded and immortalized human MSC line (ciMSC) and compared the neuroprotective potential of their EVs with EVs from primary MSCs in a murine model of HI-induced brain injury. In vivo activities of ciMSC-EVs were comprehensively characterized according to their proposed multimodal mechanisms of action.

Methods: Nine-day-old C57BL/6 mice were exposed to HI followed by repetitive intranasal delivery of primary MSC-EVs or ciMSC-EVs 1, 3, and 5 days after HI. Sham-operated animals served as healthy controls. To compare neuroprotective effects of both EV preparations, total and regional brain atrophy was assessed by cresyl-violet-staining 7 days after HI. Immunohistochemistry, western blot, and real-time PCR were performed to investigate neuroinflammatory and regenerative processes. The amount of peripheral inflammatory mediators was evaluated by multiplex analyses in serum samples.

Results: Intranasal delivery of ciMSC-EVs and primary MSC-EVs comparably protected neonatal mice from HI-induced brain tissue atrophy. Mechanistically, ciMSC-EV application reduced microglia activation and astrogliosis, endothelial activation, and leukocyte infiltration. These effects were associated with a downregulation of the pro-inflammatory cytokine IL-1 beta and an elevated expression of the anti-inflammatory cytokines IL-4 and TGF-beta in the brain, while concentrations of cytokines in the peripheral blood were not affected. ciMSC-EV-mediated anti-inflammatory effects in the brain were accompanied by an increased neural progenitor and endothelial cell proliferation, oligodendrocyte maturation, and neurotrophic growth factor expression.

Conclusion: Our data demonstrate that ciMSC-EVs conserve neuroprotective effects of primary MSC-EVs via inhibition of neuroinflammation and promotion of neuroregeneration. Since ciMSCs can overcome challenges associated with MSC heterogeneity, they appear as an ideal cell source for the scaled manufacturing of EV-based therapeutics to treat neonatal and possibly also adult brain injury.

Abstract Image

Abstract Image

Abstract Image

永生化间充质间质细胞胞外囊泡对新生儿缺氧缺血性脑损伤的保护作用。
背景:人间充质间质细胞(MSC)来源的细胞外囊泡(EV)在各种脑损伤模型中显示出神经保护作用,包括新生儿缺氧缺血(HI)所致的脑病。然而,对于MSC-EV治疗的临床转化,规模化生产策略是必需的,由于供体间和供体内的异质性,这对于原发msc来说是具有挑战性的。因此,我们建立了一个克隆扩增和永生化的人MSC系(ciMSC),并在小鼠hi诱导脑损伤模型中比较了其ev与原代MSC的ev的神经保护潜力。根据其提出的多模态作用机制,对cimsc - ev的体内活性进行了全面表征。方法:将9日龄的C57BL/6小鼠暴露于HI,然后在HI后1、3和5天重复鼻内递送原发msc - ev或cimsc - ev。假手术的动物作为健康对照。为了比较两种EV制剂的神经保护作用,在HI后7天用甲酚紫染色评估总脑萎缩和局部脑萎缩。免疫组织化学、western blot和实时荧光定量PCR检测神经炎症和再生过程。外周血炎症介质的数量通过血清样本的多重分析来评估。结果:经鼻给药cimsc - ev和原代msc - ev可显著保护新生小鼠免受hi诱导的脑组织萎缩。在机制上,ciMSC-EV的应用减少了小胶质细胞的激活和星形胶质细胞形成,内皮细胞的激活和白细胞的浸润。这些作用与大脑中促炎细胞因子IL-1 β的下调和抗炎细胞因子IL-4和tgf - β的表达升高有关,而外周血中细胞因子的浓度不受影响。在大脑中,cimsc - ev介导的抗炎作用伴随着神经祖细胞和内皮细胞增殖、少突胶质细胞成熟和神经营养生长因子表达的增加。结论:我们的数据表明,通过抑制神经炎症和促进神经再生,cimsc - ev保留了原代msc - ev的神经保护作用。由于ciMSCs可以克服与间充质干细胞异质性相关的挑战,因此它们似乎是大规模生产以ev为基础的治疗方法的理想细胞来源,可用于治疗新生儿和成人脑损伤。
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来源期刊
CiteScore
11.10
自引率
1.20%
发文量
45
审稿时长
11 weeks
期刊介绍: Inflammation and Regeneration is the official journal of the Japanese Society of Inflammation and Regeneration (JSIR). This journal provides an open access forum which covers a wide range of scientific topics in the basic and clinical researches on inflammation and regenerative medicine. It also covers investigations of infectious diseases, including COVID-19 and other emerging infectious diseases, which involve the inflammatory responses. Inflammation and Regeneration publishes papers in the following categories: research article, note, rapid communication, case report, review and clinical drug evaluation.
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