Secretion of functional α1-antitrypsin is cell type dependent: Implications for intramuscular delivery for gene therapy.
Secretion of functional α1-antitrypsin is cell type dependent: Implications for intramuscular delivery for gene therapy.
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功能性α1-抗胰蛋白酶的分泌是细胞类型依赖性的：对基因治疗肌内给药的影响。
DOI：
10.1073/pnas.2206103119
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发表时间：
2022-08-02
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
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作者：
非天然细胞类型中的蛋白质表达对于成功的基因治疗至关重要。在这里，我们探索α1-抗胰蛋白酶（AAT）的表达，基因治疗的目标，在各种类型的细胞，并评估化学方法，以提高其表达的能力。虽然AAT是有效地分泌肝细胞，它的天然宿主，它是在成肌细胞和肌管表达差，虽然它的分泌可以通过处理与蛋白质稳态调节剂辛二酰苯胺异羟肟酸，组蛋白去乙酰化酶抑制剂增强。因此，使用蛋白质稳态调节剂提供了一种潜在的策略，以优化蛋白质表达的各种治疗。蛋白质的异源表达广泛用于生物制剂的生物合成，其中许多是从细胞分泌的。此外，基因治疗和信使RNA（mRNA）疫苗经常指导分泌蛋白的表达到非天然宿主细胞。因此，了解蛋白质在一系列宿主细胞（包括肌细胞）中的成熟和运输至关重要，肌细胞是一种受欢迎的治疗靶点，因为肌内注射易于接近。在此，我们分析了中国仓鼠卵巢细胞（通常用于生物素生产）和成肌细胞（肌肉细胞的胚胎祖细胞）中α1-抗胰蛋白酶（AAT）的生产效率，并将其与主要天然细胞（肝细胞）中的生产进行了比较。AAT是基因治疗的靶蛋白，以解决与天然AAT活性或产生的不稳定性相关的病理学。AAT分泌和成熟在肝细胞中最有效。成肌细胞是测试的细胞类型中最差的;然而，通过用蛋白质稳态调节剂辛二酰苯胺异羟肟酸（一种组蛋白去乙酰化酶抑制剂）治疗，成肌细胞中活性AAT的分泌显著增强。这些发现在肌管（成熟肌细胞）中得到扩展和验证，其中使用基因治疗临床试验中使用的腺相关病毒衣壳转导方法转导AAT。总的来说，我们的研究揭示了一种可能的机制，以提高AAT的基因治疗方法的疗效，而且，可能有影响的蛋白质的生产从mRNA疫苗，这依赖于表达的病毒糖蛋白在非天然宿主细胞肌肉注射后。
Protein expression in nonnative cell types is critical for successful gene therapy. Here, we explore the expression of α1-antrypsin (AAT), a gene therapy target, in a variety of cell types and assess the ability of chemical methods to boost its expression. While AAT was efficiently secreted by hepatocytes, its native host, it was poorly expressed in myoblasts and myotubes, although its secretion could be augmented by treatment with the proteostasis regulator suberoylanilide hydroxamic acid, a histone deacetylase inhibitor. The use of proteostasis regulators thus provides a potential strategy to optimize protein expression for a variety of therapeutics. Heterologous expression of proteins is used widely for the biosynthesis of biologics, many of which are secreted from cells. In addition, gene therapy and messenger RNA (mRNA) vaccines frequently direct the expression of secretory proteins to nonnative host cells. Consequently, it is crucial to understand the maturation and trafficking of proteins in a range of host cells including muscle cells, a popular therapeutic target due to the ease of accessibility by intramuscular injection. Here, we analyzed the production efficiency for α1-antitrypsin (AAT) in Chinese hamster ovary cells, commonly used for biotherapeutic production, and myoblasts (embryonic progenitor cells of muscle cells) and compared it to the production in the major natural cells, liver hepatocytes. AAT is a target protein for gene therapy to address pathologies associated with insufficiencies in native AAT activity or production. AAT secretion and maturation were most efficient in hepatocytes. Myoblasts were the poorest of the cell types tested; however, secretion of active AAT was significantly augmented in myoblasts by treatment with the proteostasis regulator suberoylanilide hydroxamic acid, a histone deacetylase inhibitor. These findings were extended and validated in myotubes (mature muscle cells) where AAT was transduced using an adeno-associated viral capsid transduction method used in gene therapy clinical trials. Overall, our study sheds light on a possible mechanism to enhance the efficacy of gene therapy approaches for AAT and, moreover, may have implications for the production of proteins from mRNA vaccines, which rely on the expression of viral glycoproteins in nonnative host cells upon intramuscular injection.
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DOI：
10.1016/j.ymthe.2017.09.020
发表时间：
2017-11-01
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
Borel F;Tang Q;Gernoux G;Greer C;Wang Z;Barzel A;Kay MA;Shultz LD;Greiner DL;Flotte TR;Brehm MA;Mueller C
通讯作者：
Mueller C
DOI：
10.15252/embj.2020107240
发表时间：
2021-08-02
期刊：
The EMBO journal
影响因子：
--
作者：
Fregno I;Fasana E;Soldà T;Galli C;Molinari M
通讯作者：
Molinari M
影响因子：
3.7
作者：
Fu YL;Han DY;Wang YJ;Di XJ;Yu HB;Mu TW
通讯作者：
Mu TW
影响因子：
4.8
作者：
Bouchecareilh, Marion;Hutt, Darren M.;Balch, William E.
通讯作者：
Balch, William E.
DOI：
10.1513/pats.201001-016aw
发表时间：
2010-11-01
期刊：
Proceedings of the American Thoracic Society
影响因子：
--
作者：
Bouchecareilh, Marion;Conkright, Juliana J;Balch, William E
通讯作者：
Balch, William E