PDF(Pubmed)
Abstract：
Gene-editing technologies promise to create a new class of therapeutics that can achieve permanent correction with a single intervention. Besides eliminating mutant alleles in familial disease, gene-editing can also be used to favorably manipulate upstream pathophysiologic events and alter disease-course in wider patient populations, but few such feasible therapeutic avenues have been reported. Here we use CRISPR-Cas9 to edit the last exon of amyloid precursor protein (App), relevant for Alzheimer\'s disease (AD). Our strategy effectively eliminates an endocytic (YENPTY) motif at APP C-terminus, while preserving the N-terminus and compensatory APP-homologues. This manipulation favorably alters events along the amyloid-pathway - inhibiting toxic APP-β-cleavage fragments (including Aβ) and upregulating neuroprotective APP-α-cleavage products. AAV-driven editing ameliorates neuropathologic, electrophysiologic, and behavioral deficits in an AD knockin mouse model. Effects persist for many months, and no abnormalities are seen in WT mice even after germline App-editing; underlining overall efficacy and safety. Pathologic alterations in the glial-transcriptome of App-KI mice, as seen by single nuclei RNA-sequencing (sNuc-Seq), are also normalized by App C-terminus editing. Our strategy takes advantage of innate transcriptional rules that render terminal exons insensitive to nonsense-decay, and the upstream manipulation is expected to be effective for all forms of AD. These studies offer a path for a one-time disease-modifying treatment for AD.
摘要：
基因编辑技术有望创造一种新的治疗方法，通过单一干预即可实现永久性矫正。除了消除家族性疾病中的突变等位基因，基因编辑还可用于有利地操纵上游病理生理事件，并在更广泛的患者群体中改变病程，但很少有这样可行的治疗途径被报道。在这里，我们使用CRISPR-Cas9来编辑淀粉样前体蛋白（App）的最后一个外显子，与阿尔茨海默病（AD）有关。我们的策略有效地消除了APPC末端的内吞（YENPTY）基序，同时保留N端和补偿性APP同源物。这种操作有利地改变了沿着淀粉样蛋白途径的事件-抑制毒性APP-β-裂解片段(包括Aβ)和上调神经保护性APP-α-裂解产物。AAV驱动的编辑改善神经病理学，电生理,和AD敲入小鼠模型中的行为缺陷。效果持续数月，并且即使在种系App编辑后,在WT小鼠中也没有观察到异常;强调总体功效和安全性。App-KI小鼠神经胶质转录组的病理学改变，如单核RNA测序（sNuc-Seq）所示，也通过AppC末端编辑进行归一化。我们的策略利用了先天的转录规则，使末端外显子对无义衰变不敏感，上游操作预计对所有形式的AD都有效。这些研究为AD的一次性疾病修饰治疗提供了途径。
