School of Life Sciences, Tianjin University, Tianjin, 300072, China; Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, Tianjin, 300072, China.
Center for Biosafety Research and Strategy, Tianjin University, Tianjin, 300072, China; Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin, 300072, China.
Biomaterials. 2023 Mar;294:122005. doi: 10.1016/j.biomaterials.2023.122005. Epub 2023 Jan 20.
Microbes regulate brain function through the gut-brain axis, deriving the technology to modulate the gut-brain axis in situ by engineered probiotics. Optogenetics offers precise and flexible strategies for controlling the functions of probiotics in situ. However, the poor penetration of most frequently used short wavelength light has limited the application of optogenetic probiotics in the gut. Herein, a red-light optogenetic gut probiotic was applied for drug production and delivery and regulation of the host behaviors. Firstly, a Red-light Optogenetic E. coli Nissle 1917 strain (ROEN) that could respond to red light and release drug product by light-controlled lysis was constructed. The remaining optical power of red light after 3 cm tissue was still able to initiate gene expression of ROEN and produce about approximately 3-fold induction efficiency. To give full play to the in vivo potential of ROEN, its responsive ability of the penetrated red light was tested, and its encapsulation was realized by PH-sensitive alginate microcapsules for further oral administration. The function of ROEN for gut-brain regulation was realized by releasing Exendin-4 fused with anti-neonatal Fc receptor affibody. Neuroprotection and behavioral regulation effects were evaluated in the Parkinson's disease mouse model, after orally administration of ROEN delivering Exendin-4 under optogenetic control in the murine gut. The red-light optogenetic probiotic might be a perspective platform for in situ drug delivery and gut-brain axis regulation.
微生物通过肠脑轴调节大脑功能，通过工程益生菌在原位调节肠脑轴的技术。光遗传学为控制益生菌在原位的功能提供了精确和灵活的策略。然而，大多数常用的短波长光的穿透性差，限制了光遗传益生菌在肠道中的应用。本文应用红光光遗传肠道益生菌进行药物生产和递送以及宿主行为调控。首先，构建了能够响应红光并通过光控裂解释放药物产物的红光光遗传大肠杆菌 Nissle 1917 菌株（ROEN）。在经过 3 cm 组织后，红光的剩余光功率仍然能够启动 ROEN 的基因表达，并产生约 3 倍的诱导效率。为了充分发挥 ROEN 的体内潜力，测试了其穿透红光的响应能力，并通过 pH 敏感的海藻酸钠微胶囊对其进行封装，以进一步进行口服给药。通过释放与抗新生 Fc 受体亲和体融合的 Exendin-4，实现了 ROEN 对肠道-大脑调节的功能。在帕金森病小鼠模型中，通过口服给予 ROEN 并在小鼠肠道中进行光遗传控制下的 Exendin-4 递送，评估了其神经保护和行为调节作用。红光光遗传益生菌可能是一种有前途的原位药物递送和肠道-大脑轴调节平台。
