Nilles Jacob P, Roberts Darby, Salmon Jacklyn H, Song Liujiang, O'Dea Carly, Marjoram Lindsay T, Bower Jacquelyn J, Hirsch Matthew L, Gilger Brian C
Clinical Sciences, North Carolina State University, Raleigh, NC, USA.
Ophthalmology, University of North Carolina, Chapel Hill, NC, USA.
Mol Ther Methods Clin Dev. 2023 Mar 24;29:227-235. doi: 10.1016/j.omtm.2023.03.012. eCollection 2023 Jun 8.
Ocular graft versus host disease (OGvHD) develops after allogeneic hematopoietic stem cell transplantation (HSCT) and manifests as ocular surface inflammatory disease. This study evaluated the efficacy of adeno-associated virus (AAV) gene therapy encoding human leukocyte antigen G (HLA-G) to inhibit OGvHD. A major histocompatibility mismatch chronic OGvHD murine model was evaluated. 7 days after HSCT, mice were dosed subconjunctivally with scAAV8-HLA-G1/5 (1 x 10 vg/eye), topical cyclosporine (twice daily), or left untreated. Body weights and tear production (red thread test) were recorded, and eyelid, corneal opacity, and corneal fluorescein retention were scored through day 44 after HSCT. Tissues were collected for vector biodistribution, ocular histology, and immunofluorescence. Compared with untreated HSCT eyes, those dosed with scAAV8-HLA-G1/5 had significantly reduced clinical inflammatory signs of OGvHD. On histology, eyes that received scAAV8-HLA-G1/5 or cyclosporine had a significantly lower mean limbal mononuclear cell count when compared with non-treated HSCT eyes. HLA-G immunofluorescence was detected in the subconjunctiva and peripheral cornea in HSCT animals treated with scAAV8-HLA-G1/5. Vector genomes were detected in the lacrimal gland, but not in the other tested organs. These results provide evidence that subconjunctival AAV targets ocular surface and corneal disease and support that HLA-G-based gene therapy may be an effective treatment for OGvHD.
眼移植物抗宿主病（OGvHD）在异基因造血干细胞移植（HSCT）后发生，表现为眼表炎性疾病。本研究评估了编码人白细胞抗原G（HLA-G）的腺相关病毒（AAV）基因疗法抑制OGvHD的疗效。评估了一种主要组织相容性不匹配的慢性OGvHD小鼠模型。HSCT后7天，小鼠结膜下注射scAAV8-HLA-G1/5（1×10病毒基因组/眼）、局部应用环孢素（每日两次）或不进行治疗。记录体重和泪液分泌（红线试验），并在HSCT后44天内对眼睑、角膜混浊和角膜荧光素潴留进行评分。收集组织用于载体生物分布、眼部组织学和免疫荧光检测。与未治疗的HSCT眼相比，注射scAAV8-HLA-G1/5的眼OGvHD的临床炎症体征明显减轻。组织学检查显示，与未治疗的HSCT眼相比，接受scAAV8-HLA-G1/5或环孢素治疗的眼平均角膜缘单核细胞计数显著降低。在接受scAAV8-HLA-G1/5治疗的HSCT动物的结膜下和周边角膜中检测到HLA-G免疫荧光。在泪腺中检测到载体基因组，但在其他测试器官中未检测到。这些结果证明结膜下AAV可靶向眼表和角膜疾病，并支持基于HLA-G的基因疗法可能是治疗OGvHD的有效方法。
