Engineering adeno-associated viral vectors to evade innate immune and inflammatory responses.
Engineering adeno-associated viral vectors to evade innate immune and inflammatory responses.
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DOI：
10.1126/scitranslmed.abd3438
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发表时间：
2021-02-10
影响因子：
17.1
通讯作者：
Church GM
中科院分区：
文献类型：
--
作者：
Chan YK;Wang SK;Chu CJ;Copland DA;Letizia AJ;Costa Verdera H;Chiang JJ;Sethi M;Wang MK;Neidermyer WJ Jr;Chan Y;Lim ET;Graveline AR;Sanchez M;Boyd RF;Vihtelic TS;Inciong RGCO;Slain JM;Alphonse PJ;Xue Y;Robinson-McCarthy LR;Tam JM;Jabbar MH;Sahu B;Adeniran JF;Muhuri M;Tai PWL;Xie J;Krause TB;Vernet A;Pezone M;Xiao R;Liu T;Wang W;Kaplan HJ;Gao G;Dick AD;Mingozzi F;McCall MA;Cepko CL;Church GM
核酸被用于包括基因治疗在内的许多治疗方式，但它们在体内触发宿主免疫反应的能力可能会导致安全性和有效性降低。就腺相关病毒(AAV)载体而言，研究表明，载体的基因组激活Toll样受体9(TLR9)，TLR9是一种感知外源DNA的模式识别受体。在这里，我们通过将拮抗TLR9激活的短DNA寡核苷酸直接整合到载体基因组中，使AAV载体具有内在较低的免疫原性。在临床相关的小鼠和猪模型中，这些工程载体显著降低了先天免疫和T细胞反应，并增强了不同组织(包括肝脏、肌肉和视网膜)的基因表达。视网膜下注射大剂量AAV可导致光感受器病理改变，并伴有小胶质细胞和T细胞的浸润。在注射了基因工程载体的同一动物的对侧眼睛中，这些不利的发现被避免了。然而，将更高剂量的AAV注射到猕猴的玻璃体内，这是一种更具免疫原性的给药途径，结果表明，工程载体延迟了临床葡萄膜炎的发生，但并未预防临床葡萄膜炎，这表明除了TLR9之外，其他免疫因素可能对该模型的眼内炎症起到了作用。我们的结果表明，将特定的免疫调节非编码序列与更长的治疗性核酸联系起来，可以在多个(但不是所有)模型中“掩盖”载体诱导不需要的免疫反应。这种“偶联免疫调节”策略可能会拓宽AAV治疗以及其他基于DNA的基因转移方法的治疗窗口。在多种动物模型中，将TLR9抑制序列整合到AAV载体基因组中可抑制免疫原性并增强转基因表达。
Nucleic acids are utilized in many therapeutic modalities, including gene therapy, but their ability to trigger host immune responses in vivo can lead to decreased safety and efficacy. In the case of adeno-associated viral (AAV) vectors, studies have shown that the genome of the vector activates Toll-like receptor 9 (TLR9), a pattern recognition receptor that senses foreign DNA. Here, we engineered AAV vectors to be intrinsically less immunogenic by incorporating short DNA oligonucleotides that antagonize TLR9 activation directly into the vector genome. The engineered vectors elicited markedly reduced innate immune and T cell responses and enhanced gene expression in clinically relevant mouse and pig models across different tissues, including liver, muscle and retina. Subretinal administration of higher-dose AAV in pigs resulted in photoreceptor pathology with microglia and T cell infiltration. These adverse findings were avoided in the contralateral eyes of the same animals that were injected with the engineered vectors. However, intravitreal injection of higher-dose AAV in macaques, a more immunogenic route of administration, showed that the engineered vector delayed but did not prevent clinical uveitis, suggesting that other immune factors in addition to TLR9 may contribute to intraocular inflammation in this model. Our results demonstrate that linking specific immunomodulatory non-coding sequences to much longer therapeutic nucleic acids can “cloak” the vector from inducing unwanted immune responses in multiple, but not all, models. This “coupled immunomodulation” strategy may widen the therapeutic window for AAV therapies as well as other DNA-based gene transfer methods. Incorporating TLR9-inhibitory sequences in the AAV vector genome inhibits immunogenicity and enhances transgene expression in multiple animal models.
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影响因子：
3.7
作者：
Chu CJ;Herrmann P;Carvalho LS;Liyanage SE;Bainbridge JW;Ali RR;Dick AD;Luhmann UF
通讯作者：
Luhmann UF
影响因子：
4.4
作者：
Gursel, I;Gursel, M;Klinman, DM
通讯作者：
Klinman, DM
DOI：
10.1016/j.omtm.2018.12.011
发表时间：
2019-03-15
影响因子：
4.7
作者：
Collaud, Fanny;Bortolussi, Giulia;Mingozzi, Federico
通讯作者：
Mingozzi, Federico
影响因子：
13.5
作者：
Hoesel, Marianna;Broxtermann, Mathias;Buening, Hildegard
通讯作者：
Buening, Hildegard
影响因子：
4.4
作者：
de Castro, Juan P. Fernandez;Scott, Patrick A.;McCall, Maureen A.
通讯作者：
McCall, Maureen A.