Cong Lin, Qi Benxiang, Chen Shijiu, Liu Ruiling, Li Suxia, Zhou Qingjun, Cao Yihai, Zhang Bi Ning, Xie Lixin
Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China.
School of Ophthalmology, Shandong First Medical University, Qingdao, China.
Diabetes. 2025 Jan 1;74(1):22-35. doi: 10.2337/db24-0393.
Diabetic keratopathy (DK) is a common chronic metabolic disorder that causes ocular surface complications. Among various therapeutic approaches, local delivery of nerve growth factor (NGF) remains the most effective treatment of DK. However, achieving a sustained therapeutic effect with NGF and the frequent drug delivery burden remain challenging during clinical practice. Here, we developed a novel adeno-associated virus (AAV)-based NGF delivery system that achieved 1-year-long-lasting effects by a single injection. We refined the corneal stromal injection technique, resulting in reduced corneal edema and improved AAV distribution homogeneity. AAV serotype AAV.rh10 exhibited high tropism and specificity to corneal nerves. A dose of 2 × 109 vector genomes was determined to achieve efficient Ngf gene expression without inducing corneal immune responses. Moreover, NGF protein was highly expressed in trigeminal ganglion through a retrograde transport mechanism, indicating the capacity for repairing corneal nerve damage at both the root and corneal nerve endings. In a mouse DK model, a single injection of AAV-Ngf into the corneal stroma led to marked corneal nerve regeneration for over 5 months. Together, we provide a novel therapeutic paradigm for long-term effective treatment of DK, and this therapeutic approach is superior to current DK therapies.
糖尿病性角膜病变（DK）是一种常见的慢性代谢紊乱疾病，可导致眼表并发症。在各种治疗方法中，局部递送神经生长因子（NGF）仍然是DK最有效的治疗方法。然而，在临床实践中，实现NGF的持续治疗效果以及频繁的给药负担仍然具有挑战性。在此，我们开发了一种新型的基于腺相关病毒（AAV）的NGF递送系统，通过单次注射可实现长达1年的持久效果。我们改进了角膜基质注射技术，减少了角膜水肿并提高了AAV分布的均匀性。AAV血清型AAV.rh10对角膜神经表现出高嗜性和特异性。确定2×10⁹载体基因组的剂量可实现有效的Ngf基因表达，而不会诱导角膜免疫反应。此外，NGF蛋白通过逆行转运机制在三叉神经节中高表达，表明其具有修复角膜神经根部和角膜神经末梢损伤的能力。在小鼠DK模型中，向角膜基质单次注射AAV-Ngf可导致角膜神经显著再生超过5个月。总之，我们为DK的长期有效治疗提供了一种新的治疗范例，这种治疗方法优于目前的DK治疗方法。
