Efficient cochlear gene transfection in guinea-pigs with adeno-associated viral vectors by partial digestion of round window membrane.
Efficient cochlear gene transfection in guinea-pigs with adeno-associated viral vectors by partial digestion of round window membrane.
复制标题
通过部分消化圆窗膜，用腺相关病毒载体高效转染豚鼠耳蜗基因。
作者：
内耳的听觉部分，耳蜗，由于其相对隔离，是用于局部基因转染的理想器官。已经测试了用于耳蜗基因转染的各种载体。迄今为止，病毒载体似乎具有比非病毒机制高得多的转染功效。在这些载体中，重组腺相关病毒（rAV）载体具有几个优点，例如非致病性以及在各种细胞类型中产生延长基因表达的能力。然而，rAAV载体不能穿过完整的圆窗膜（RWM），否则是进入人内耳的非常有吸引力的方法。在这项研究中，在豚鼠中进行，我们描述了一种方法，以增加RWM的rAAV载体的部分消化与胶原酶溶液的渗透性。rAAV通过部分消化的RWM的升高递送将转染功效增加到令人满意的水平，即使它仍然低于通过直接耳蜗造口注射实现的转染功效。功能测试（听觉脑干反应）表明，如果应用得当，这种酶操纵不会导致永久性听力损失。形态学观察表明，部分消化引起的RWM损伤在4周内愈合。总之，这些发现表明，部分消化RWM是一种安全有效的方法，用于增加耳蜗感觉细胞的转染rAAV。
The auditory portion of the inner ear, the cochlea, is an ideal organ for local gene transfection owing to its relative isolation. Various carriers have been tested for cochlear gene transfection. To date, viral vectors appear to have much higher transfection efficacy than non-viral mechanisms. Among these vectors, recombinant adeno-associated virus (rAAV) vectors have several advantages such as being non-pathogenic and the ability to produce prolonged gene expression in various cell types. However, rAAV vectors cannot pass through the intact round window membrane (RWM), otherwise a very attractive approach to access the human inner ear. In this study, performed in guinea-pigs, we describe a method to increase the permeability of RWM to rAAV vectors by partial digestion with collagenase solution. Elevated delivery of rAAV across the partially digested RWM increased transfection efficacy to a satisfactory level, even though it was still lower than that achieved by direct cochleostomy injection. Functional tests (auditory brainstem responses) showed that this enzymatic manipulation did not cause permanent hearing loss if applied appropriately. Morphological observations suggested that the damage to RWM caused by partial digestion healed within four weeks. Taken together, these findings suggest that partial digestion of the RWM is a safe and effective method for increasing the transfection of cochlear sensory cells with rAAV.
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影响因子：
1.7
作者：
Cotanche, Douglas A.
通讯作者：
Cotanche, Douglas A.
DOI：
10.1073/pnas.87.6.2211
发表时间：
1990-03-01
影响因子：
11.1
作者：
KOTIN, RM;SINISCALCO, M;BERNS, KI
通讯作者：
BERNS, KI
影响因子：
12.4
作者：
Petrs-Silva, Hilda;Dinculescu, Astra;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
影响因子：
3.5
作者：
Boëda, B;Weil, D;Petit, C
通讯作者：
Petit, C
DOI：
10.1159/000218208
发表时间：
2009-01-01
期刊：
GENE THERAPY OF COCHLEAR DEAFNESS: PRESENT CONCEPTS AND FUTURE ASPECTS
影响因子：
--
作者：
Kesser, Bradley W.;Lalwani, Anil K.
通讯作者：
Lalwani, Anil K.