PDF(Pubmed)
Abstract：
X-linked adrenoleukodystrophy (ALD), an inherited neurometabolic disorder caused by mutations in ABCD1, which encodes the peroxisomal ABC transporter, mainly affects the brain, spinal cord, adrenal glands, and testes. In ALD patients, very-long-chain fatty acids (VLCFAs) fail to enter the peroxisome and undergo subsequent β-oxidation, resulting in their accumulation in the body. It has not been tested whether in vivo base editing or prime editing can be harnessed to ameliorate ALD. We developed a humanized mouse model of ALD by inserting a human cDNA containing the pathogenic variant into the mouse Abcd1 locus. The humanized ALD model showed increased levels of VLCFAs. To correct the mutation, we tested both base editing and prime editing and found that base editing using ABE8e(V106W) could correct the mutation in patient-derived fibroblasts at an efficiency of 7.4%. Adeno-associated virus (AAV)-mediated systemic delivery of NG-ABE8e(V106W) enabled robust correction of the pathogenic variant in the mouse brain (correction efficiency: ∼5.5%), spinal cord (∼5.1%), and adrenal gland (∼2%), leading to a significant reduction in the plasma levels of C26:0/C22:0. This established humanized mouse model and the successful correction of the pathogenic variant using a base editor serve as a significant step toward treating human ALD disease.
摘要：
X-连锁肾上腺脑白质营养不良(ALD),由ABCD1突变引起的遗传性神经代谢紊乱，编码过氧化物酶体ABC转运体，主要影响大脑，脊髓,肾上腺,和睾丸。在ALD患者中，超长链脂肪酸（VLCFAs）无法进入过氧化物酶体并随后进行β-氧化，导致它们在体内的积累。尚未测试是否可以利用体内碱基编辑或主编辑来改善ALD。我们通过将含有致病性变体的人cDNA插入小鼠Abcd1基因座中，开发了ALD的人源化小鼠模型。人源化ALD模型显示VLCFA水平增加。为了纠正突变，我们测试了碱基编辑和初免编辑,发现使用ABE8e(V106W)进行碱基编辑可以纠正患者来源的成纤维细胞中的突变,效率为7.4%.腺相关病毒（AAV）介导的NG-ABE8e（V106W）的全身递送能够对小鼠大脑中的致病变异进行稳健的校正（校正效率：~5.5％），脊髓(~5.1%)，和肾上腺(~2%)，导致血浆C26：0/C22：0水平显着降低。这种建立的人源化小鼠模型和使用碱基编辑器成功校正致病变体作为治疗人类ALD疾病的重要步骤。
