Biosafety Research Institute and Laboratory of Veterinary Pathology, College of Veterinary Medicine, Jeonbuk National University, Iksan 54596, Korea.
Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Korea; College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Korea.
Mol Ther. 2022 May 4;30(5):1994-2004. doi: 10.1016/j.ymthe.2022.01.010. Epub 2022 Jan 7.
Adeno-associated virus (AAV)-mediated gene delivery holds great promise for gene therapy. However, the non-invasive delivery of AAV for lung tissues has not been adequately established. Here, we revealed that the intratracheal administration of an appropriate amount of AAV2/8 predominantly targets lung tissue. AAV-mediated gene delivery that we used in this study induced the expression of the desired protein in lung parenchymal cells, including alveolar type II cells. We harnessed the technique to develop severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-susceptible mice. Three kinds of immune function-relevant gene knockout (KO) mice were transduced with AAV encoding human angiotensin-converting enzyme 2 (hACE2) and then injected with SARS-CoV-2. Among these mice, type I interferon receptor (IFNAR) KO mice showed increased viral titer in the lungs compared to that in the other KO mice. Moreover, nucleocapsid protein of SARS-CoV-2 and multiple lesions in the trachea and lung were observed in AAV-hACE2-transduced, SARS-CoV-2-infected IFNAR KO mice, indicating the involvement of type I interferon signaling in the protection of SARS-CoV-2. In this study, we demonstrate the ease and rapidness of the intratracheal administration of AAV for targeting lung tissue in mice, and this can be used to study diverse pulmonary diseases.
腺相关病毒(AAV)介导的基因传递在基因治疗中具有巨大的应用前景。然而,AAV 的非侵入性肺部给药尚未得到充分确立。在这里,我们揭示了适量的 AAV2/8 通过气管内给药主要靶向肺部组织。我们在这项研究中使用的 AAV 介导的基因传递诱导了肺实质细胞(包括肺泡 II 型细胞)中所需蛋白的表达。我们利用该技术开发了对严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)敏感的小鼠。三种免疫功能相关基因敲除(KO)小鼠被转导了编码人血管紧张素转换酶 2(hACE2)的 AAV,然后注射了 SARS-CoV-2。在这些小鼠中,与其他 KO 小鼠相比,I 型干扰素受体(IFNAR)KO 小鼠肺部的病毒滴度更高。此外,在接受 AAV-hACE2 转导和 SARS-CoV-2 感染的 IFNAR KO 小鼠中观察到 SARS-CoV-2 的核衣壳蛋白和气管及肺部的多处病变,表明 I 型干扰素信号通路参与了对 SARS-CoV-2 的保护。在这项研究中,我们证明了在小鼠中通过气管内给药靶向肺部组织的简便性和快速性,这可用于研究多种肺部疾病。