Development of new adeno-associated virus capsid variants for targeted gene delivery to human cardiomyocytes.
Development of new adeno-associated virus capsid variants for targeted gene delivery to human cardiomyocytes.
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DOI：
10.1016/j.omtm.2023.08.010
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发表时间：
2023-09-14
期刊：
影响因子：
--
通讯作者：
Kizana, Eddy
中科院分区：
文献类型：
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作者：
Kok, Cindy Y.;Tsurusaki, Shinya;Cabanes-Creus, Marti;Igoor, Sindhu;Rao, Renuka;Skelton, Rhys;Liao, Sophia H. Y.;Ginn, Samantha L.;Knight, Maddison;Scott, Suzanne;Mietzsch, Mario;Fitzsimmons, Rebecca;Miller, Jessica;Mohamed, Tamer M. A.;McKenna, Robert;Chong, James J. H.;Hill, Adam P.;Hudson, James E.;Alexander, Ian E.;Lisowski, Leszek;Kizana, Eddy
重组腺相关病毒（rAAV）已成为最有前途的基因治疗载体之一，已成功地用于心脏病的临床前模型。然而，由于rAAV转导效率的物种差异，这并没有很好地转化为人类。因此，寻找人类的心脏营养衣壳是当代的一个重大挑战。我们使用captain改组的rAAV文库在人iPSC衍生的心肌细胞（hiPSC-CM）中进行定向进化。出现了五种候选物，其中四种呈现与AAV 6的高序列同一性，而第五种趋异变体与AAV 3b相关。使用hiPSC-CM、心脏类器官、人心脏切片、非人灵长类动物和猪心脏切片以及体内小鼠心脏和肝脏在体外进行了变体的功能分析。我们发现细胞进入不是转基因表达效率的最佳预测因子。新变体rAAV.KK04是基于人类的筛选平台中性能最好的载体，超过了基准rAAV 6。没有一种新的衣壳在体内表现出显著的肝脏转导。所使用的实验模型范围揭示了在人类特异性、真实、心肌和感兴趣的细胞类型条件下测试向性差异的价值。Kizana及其同事使用临床相关模型开发和筛选了心脏特异性AAV衣壳，这些模型从人iPSC衍生的心肌细胞和类器官到由原代心脏组织（人，非人灵长类动物和猪）产生的心脏切片。新型变体rAAV.KK04显示出最有希望将基因有效递送至人类心肌细胞。
Recombinant adeno-associated viruses (rAAVs) have emerged as one of the most promising gene therapy vectors that have been successfully used in pre-clinical models of heart disease. However, this has not translated well to humans due to species differences in rAAV transduction efficiency. As a result, the search for human cardiotropic capsids is a major contemporary challenge. We used a capsid-shuffled rAAV library to perform directed evolution in human iPSC-derived cardiomyocytes (hiPSC-CMs). Five candidates emerged, with four presenting high sequence identity to AAV6, while a fifth divergent variant was related to AAV3b. Functional analysis of the variants was performed in vitro using hiPSC-CMs, cardiac organoids, human cardiac slices, non-human primate and porcine cardiac slices, as well as mouse heart and liver in vivo. We showed that cell entry was not the best predictor of transgene expression efficiency. The novel variant rAAV.KK04 was the best-performing vector in human-based screening platforms, exceeding the benchmark rAAV6. None of the novel capsids demonstrate a significant transduction of liver in vivo. The range of experimental models used revealed the value of testing for tropism differences under the conditions of human specificity, bona fide, myocardium and cell type of interest. Kizana and colleagues have developed and screened cardiac-specific AAV capsids using clinically relevant models ranging from human iPSC-derived cardiomyocytes and organoids to cardiac slices generated from primary heart tissue (human, non-human primate, and pig). The novel variant rAAV.KK04 showed the most promise for efficient gene delivery to human cardiomyocytes.
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影响因子：
--
作者：
Gabisonia K;Recchia FA
通讯作者：
Recchia FA
DOI：
10.1016/j.omtm.2021.11.011
发表时间：
2022-03-10
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
Cabanes-Creus M;Navarro RG;Zhu E;Baltazar G;Liao SHY;Drouyer M;Amaya AK;Scott S;Nguyen LH;Westhaus A;Hebben M;Wilson LOW;Thrasher AJ;Alexander IE;Lisowski L
通讯作者：
Lisowski L
影响因子：
20.1
作者：
Ou, Qinghui;Jacobson, Zoe;Mohamed, Tamer M. A.
通讯作者：
Mohamed, Tamer M. A.
影响因子：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
11.1
作者：
Buening, Hildegard
通讯作者：
Buening, Hildegard