School of Integrated Design Engineering, Graduate School of Science and Technology, Keio University, Yokohama, Japan.
Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, Yokohama, Japan.
Biotechnol Bioeng. 2023 Aug;120(8):2371-2377. doi: 10.1002/bit.28482. Epub 2023 Jun 27.
Adeno-associated virus (AAV)-based gene therapy holds promise as a fundamental treatment for genetic disorders. For clinical applications, it is necessary to control AAV release timing to avoid an immune response to AAV. Here we propose an ultrasound (US)-triggered on-demand AAV release system using alginate hydrogel microbeads (AHMs) with a release enhancer. By using a centrifuge-based microdroplet shooting device, the AHMs encapsulating AAV with tungsten microparticles (W-MPs) are fabricated. Since W-MPs work as release enhancers, the AHMs have high sensitivity to the US with localized variation in acoustic impedance for improving the release of AAV. Furthermore, AHMs were coated with poly-l-lysine (PLL) to adjust the release of AAV. By applying US to the AAV encapsulating AHMs with W-MPs, the AAV was released on demand, and gene transfection to cells by AAV was confirmed without loss of AAV activity. This proposed US-triggered AAV release system expands methodological possibilities in gene therapy.
腺相关病毒 (AAV) 为基础的基因治疗有望成为治疗遗传疾病的基本方法。为了临床应用，需要控制 AAV 的释放时间，以避免对 AAV 的免疫反应。在这里，我们提出了一种使用具有释放增强剂的藻酸盐水凝胶微球 (AHM) 的超声 (US) 触发按需 AAV 释放系统。通过使用基于离心的微滴喷射装置，制造了封装 AAV 和钨微颗粒 (W-MP) 的 AHM。由于 W-MP 作为释放增强剂，AHM 对局部声阻抗变化的超声具有高灵敏度，从而改善了 AAV 的释放。此外，AHM 被聚-l-赖氨酸 (PLL) 涂层以调节 AAV 的释放。通过将 US 施加到封装有 W-MP 的 AAV 的 AHM 上，按需释放 AAV，并且通过 AAV 确认了对细胞的基因转染，而不会损失 AAV 活性。这种提出的 US 触发的 AAV 释放系统扩展了基因治疗中的方法学可能性。
