Miyagawa Yoshitaka, Maruyama Motoyo, Sakai Atsushi, Sato Yuriko, Shimizu Masumi, Kuroda Seiji, Hayashita-Kinoh Hiromi, Yamamoto Motoko, Hashizume Ryotaro, Suzuki Hidenori, Cohen Justus B, Glorioso Joseph C, Okada Takashi
Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo, Japan.
Department of Pharmacology, Nippon Medical School, Tokyo, Japan.
Mol Ther Methods Clin Dev. 2025 Aug 20;33(3):101573. doi: 10.1016/j.omtm.2025.101573. eCollection 2025 Sep 11.
This report describes the distribution and transgene expression of two non-cytotoxic, replication-defective (rd) herpes simplex virus (HSV) vectors, JΔNI7 and JΔNI8, following intraperitoneal delivery to newborn mice. The two vectors are functionally defective for all immediate-early genes, and JΔNI8 is further deleted for the UL41 endonuclease (). Both vectors were engineered to express a red luciferase gene from the LAT locus to track vector distribution and gene expression . A comparison of reporter gene activities under the control of four different promoters in JΔNI7 showed that the strongest expression was achieved with the CAG promoter. Distribution analysis at 1 week post-injection showed transgene expression in multiple tissues, but at 4 weeks, high-level expression was limited to the spinal cord, skin, and muscles. JΔNI8 showed rapid clearance of vector DNA in most tissues, suggesting a role for the gene in vector stability. Compared with wild-type KOS strain injections, JΔNI7-based non-cytotoxic rdHSV did not induce substantial CD45 immune-cell infiltration or tissue destruction, suggesting that our rdHSV vectors are safe. Taken together, these results demonstrate tissue-specific, durable transgene expression following systemic delivery of rdHSV vectors, suggesting their potential for systemic gene therapy for newborns with skin or neuromuscular diseases.
本报告描述了两种无细胞毒性、复制缺陷型（rd）单纯疱疹病毒（HSV）载体JΔNI7和JΔNI8经腹腔注射给新生小鼠后的分布及转基因表达情况。这两种载体的所有立即早期基因均功能缺陷，且JΔNI8进一步缺失了UL41核酸内切酶基因。两种载体均经改造，可从LAT基因座表达红色荧光素酶基因，以追踪载体分布和基因表达。对JΔNI7中受四种不同启动子控制的报告基因活性进行比较，结果表明CAG启动子可实现最强表达。注射后1周的分布分析显示转基因在多个组织中表达，但在4周时，高水平表达仅限于脊髓、皮肤和肌肉。JΔNI8在大多数组织中显示载体DNA快速清除，提示该基因在载体稳定性方面发挥作用。与野生型KOS株注射相比，基于JΔNI7的无细胞毒性rdHSV未诱导大量CD45免疫细胞浸润或组织破坏，表明我们的rdHSV载体是安全的。综上所述，这些结果证明了rdHSV载体经全身给药后可实现组织特异性、持久的转基因表达，提示其在治疗患有皮肤或神经肌肉疾病的新生儿全身性基因治疗方面具有潜力。
