Adenosine is a modulator of neuronal activity with anticonvulsant and neuroprotective properties. Conversely, focal deficiency in adenosine contributes to ictogenesis. Thus, focal reconstitution of adenosine within an epileptogenic brain region constitutes a rational therapeutic approach, whereas systemic augmentation of adenosine is precluded by side effects. To meet the therapeutic goal of focal adenosine augmentation, genetic disruption of the adenosine metabolizing enzyme, adenosine kinase (ADK) in rodent cells was used as a molecular strategy to induce adenosine release from cellular brain implants, which demonstrated antiepileptic and neuroprotective properties. Currently, the second generation of adenosine-releasing cells is under development based on the rationale to use human stem cell-derived brain implants to avoid xenotransplantation. To effectively engineer human stem cells to release adenosine, a lentiviral vector was constructed to express inhibitory micro-RNA directed against ADK. Lentiviral knockdown of ADK induced therapeutic adenosine release in human mesenchymal stem cells, which reduced acute injury and seizures, as well as chronic seizures, when grafted into the mouse hippocampus. The therapeutic potential of this approach suggests the feasibility to engineer autologous adenosine-releasing stem cells derived from a patient. Human embryonic stem cells (hESCs) have a high proliferative capacity and can be subjected to specific cellular differentiation pathways. hESCs, differentiated in vitro into neuroepithelial cells and grafted into the mouse brain, displayed intrahippocampal location and neuronal morphology. Using the same lentiviral micro-RNA vector, we demonstrated knockdown of ADK in hESCs. New developments and therapeutic challenges in using human mesenchymal stem cells and hESCs for epilepsy therapy will be critically evaluated.

中文翻译：

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用于癫痫治疗的工程化腺苷释放细胞：人类间充质干细胞和人类胚胎干细胞。

腺苷是神经元活动的调节剂，具有抗惊厥和神经保护特性。相反，腺苷的局灶性缺乏有助于 ictogenesis。因此，致癫痫脑区域内腺苷的局灶性重建构成了一种合理的治疗方法，而腺苷的全身性增加被副作用排除。为了实现局灶性腺苷增强的治疗目标，啮齿动物细胞中腺苷代谢酶、腺苷激酶 (ADK) 的遗传破坏被用作诱导细胞脑植入物释放腺苷的分子策略，这证明了抗癫痫和神经保护特性。目前，基于使用人类干细胞衍生的大脑植入物来避免异种移植的基本原理，正在开发第二代腺苷释放细胞。为了有效地改造人类干细胞以释放腺苷，构建了一种慢病毒载体来表达针对 ADK 的抑制性微 RNA。ADK 的慢病毒敲低诱导人间充质干细胞中治疗性腺苷释放，当移植到小鼠海马体时，可减少急性损伤和癫痫发作以及慢性癫痫发作。这种方法的治疗潜力表明设计源自患者的自体腺苷释放干细胞的可行性。人类胚胎干细胞 (hESCs) 具有很高的增殖能力，可以进行特定的细胞分化途径。hESC，分化的 ADK 的慢病毒敲低诱导人间充质干细胞中治疗性腺苷释放，当移植到小鼠海马体时，可减少急性损伤和癫痫发作以及慢性癫痫发作。这种方法的治疗潜力表明设计源自患者的自体腺苷释放干细胞的可行性。人类胚胎干细胞 (hESCs) 具有很高的增殖能力，可以进行特定的细胞分化途径。hESC，分化的 ADK 的慢病毒敲低诱导人间充质干细胞中治疗性腺苷释放，当移植到小鼠海马体时，可减少急性损伤和癫痫发作以及慢性癫痫发作。这种方法的治疗潜力表明设计源自患者的自体腺苷释放干细胞的可行性。人类胚胎干细胞 (hESCs) 具有很高的增殖能力，可以进行特定的细胞分化途径。hESC，分化的 人类胚胎干细胞 (hESCs) 具有很高的增殖能力，可以进行特定的细胞分化途径。hESC，分化的 人类胚胎干细胞 (hESCs) 具有很高的增殖能力，可以进行特定的细胞分化途径。hESC，分化的在体外进入神经上皮细胞并移植到小鼠大脑中，显示海马内的位置和神经元形态。使用相同的慢病毒 micro-RNA 载体，我们证明了 hESC 中 ADK 的敲低。将严格评估使用人类间充质干细胞和 hESC 进行癫痫治疗的新进展和治疗挑战。