CRISPR-Cas9 base editing of pathogenic CaMKIIδ improves cardiac function in a humanized mouse model.
CRISPR-Cas9 base editing of pathogenic CaMKIIδ improves cardiac function in a humanized mouse model.
复制标题
致病性CaMKIIδ的CRISPR-Cas9碱基编辑改善人源化小鼠模型中的心脏功能。
DOI：
10.1172/jci175164
复制
发表时间：
2024-01-02
影响因子：
15.9
通讯作者：
Olson, Eric N.
中科院分区：
文献类型：
--
作者：
Lebek, Simon;Caravia, Xurde M.;Straub, Leon G.;Alzhanov, Damir;Tan, Wei;Li, Hui;McAnally, John R.;Chen, Kenian;Xu, Lin;Scherer, Philipp E.;Liu, Ning;Bassel-Duby, Rhonda;Olson, Eric N.
心血管疾病是全球发病率和死亡率的最常见原因，突出了先进治疗策略的必要性。Ca 2 +/钙调素依赖性蛋白激酶IIδ（CaMKIIδ）是一种重要的心脏疾病诱导剂，由调节结构域中的2个氧化敏感性蛋氨酸残基介导。我们之前已经表明，通过CRISPR-Cas9碱基编辑消除CaMKIIδ氧化使心脏能够从缺血/再灌注（IR）损伤后的严重损伤中恢复功能。在这里，我们将这种治疗概念扩展到潜在的临床转化。我们产生了人源化CAMK 2D敲入小鼠模型，其中编码整个调控结构域的基因组序列被替换为人序列。这使得能够比较和优化小鼠中人类基因组的两种不同编辑策略。为了在体内编辑CAMK 2D，我们将优化的编辑组分包装到工程化的亲肌性腺相关病毒（MyoAAV 2A）中，其能够在IR损伤时以非常低的AAV剂量有效递送到人源化小鼠中。CAMK 2D编辑的小鼠恢复了心脏功能，表现出改善的运动表现，并受到心肌纤维化的保护，这在IR后的损伤对照小鼠中观察到。
Cardiovascular diseases are the most common cause of worldwide morbidity and mortality, highlighting the necessity for advanced therapeutic strategies. Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ) is a prominent inducer of various cardiac disorders, which is mediated by 2 oxidation-sensitive methionine residues within the regulatory domain. We have previously shown that ablation of CaMKIIδ oxidation by CRISPR-Cas9 base editing enables the heart to recover function from otherwise severe damage following ischemia/reperfusion (IR) injury. Here, we extended this therapeutic concept toward potential clinical translation. We generated a humanized CAMK2D knockin mouse model in which the genomic sequence encoding the entire regulatory domain was replaced with the human sequence. This enabled comparison and optimization of two different editing strategies for the human genome in mice. To edit CAMK2D in vivo, we packaged the optimized editing components into an engineered myotropic adeno-associated virus (MyoAAV 2A), which enabled efficient delivery at a very low AAV dose into the humanized mice at the time of IR injury. CAMK2D-edited mice recovered cardiac function, showed improved exercise performance, and were protected from myocardial fibrosis, which was otherwise observed in injured control mice after IR. Our findings identify a potentially effective strategy for cardioprotection in response to oxidative damage.
影响因子：
3.8
作者：
Lebek S;Wester M;Pec J;Poschenrieder F;Tafelmeier M;Fisser C;Provaznik Z;Schopka S;Debl K;Schmid C;Buchner S;Maier LS;Arzt M;Wagner S
通讯作者：
Wagner S