Gumusgoz Emrah, Kasiri Sahba, Youssef Ibrahim, Verma Mayank, Chopra Rajiv, Villarreal Acha Daniel, Wu Jun, Marriam Ummay, Alao Esther, Chen Xin, Guisso Dikran R, Gray Steven J, Shah Bhavya R, Minassian Berge A
Division of Neurology, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.
Department of Chemistry, Faculty of Science, Mansoura University, Mansoura, Egypt.
Gene Ther. 2025 May;32(3):237-245. doi: 10.1038/s41434-025-00517-w. Epub 2025 Feb 1.
Because children have little temporal exposure to environment and aging, most pediatric neurological diseases are inherent, i.e. genetic. Since postnatal neurons and astrocytes are mostly non-replicating, gene therapy and genome editing present enormous promise in child neurology. Unlike in other organs, which are highly permissive to adeno-associated viruses (AAV), the mature blood-brain barrier (BBB) greatly limits circulating AAV distribution to the brain. Intrathecal administration improves distribution but to no more than 20% of brain cells. Focused ultrasound (FUS) opens the BBB transiently and safely. In the present work we opened the hippocampal BBB and delivered a Cas9 gene via AAV9 intrathecally. This allowed brain first-pass, and subsequent vascular circulation and re-entry through the opened BBB. The mouse model used was of Lafora disease, a neuroinflammatory disease due to accumulations of misshapen overlong-branched glycogen. Cas9 was targeted to the gene of the glycogen branch-elongating enzyme glycogen synthase. We show that FUS dramatically (2000-fold) improved hippocampal Cas9 distribution and greatly reduced the pathogenic glycogen accumulations and hippocampal inflammation. FUS is in regular clinical use for other indications. Our work shows that it has the potential to vastly broaden gene delivery or editing along with clearance of corresponding pathologic basis of brain disease.
由于儿童接触环境和衰老的时间较短，大多数儿科神经系统疾病是先天性的，即遗传性的。由于出生后神经元和星形胶质细胞大多不可复制，基因治疗和基因组编辑在儿童神经病学领域展现出巨大前景。与其他对腺相关病毒（AAV）高度允许的器官不同，成熟的血脑屏障（BBB）极大地限制了循环中的AAV在脑内的分布。鞘内给药可改善分布，但不超过20%的脑细胞。聚焦超声（FUS）可短暂且安全地打开血脑屏障。在本研究中，我们打开了海马体的血脑屏障，并通过鞘内注射AAV9递送了Cas9基因。这使得药物能够首次通过大脑，随后通过开放的血脑屏障进行血管循环和再次进入。所使用的小鼠模型是拉福拉病，这是一种由于畸形的超长分支糖原积累导致的神经炎症性疾病。Cas9靶向糖原分支延长酶糖原合酶的基因。我们表明，FUS显著（2000倍）改善了海马体中Cas9的分布，并大大减少了致病性糖原积累和海马体炎症。FUS已在常规临床中用于其他适应症。我们的研究表明，它有可能极大地拓宽基因递送或编辑范围，并清除脑部疾病相应的病理基础。
