Gene delivery to the inner ear by transplantation of ex vivo gene-manipulated cells

通过离体基因操作细胞移植将基因递送至内耳

• 批准号: 16390488
• 负责人: NAKAGAWA Takayuki
• 金额: $ 8.26万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390488/
• 关键词: Cell transplantation; Gene therapy; inner ear; Neurotrophic factor; Bone marrow stromal cell; Spiral ligament; 再生医療; 感音難聴; 骨髄由来細胞

Sensorineural hearing loss (SNHL) is a common disability, but treatment options are currently limited to cochlear implants and hearing aids. Studies are therefore being conducted to provide alternative means of biological therapy, including gene therapy. Safe and effective methods of gene delivery to the cochlea need to be developed to facilitate the clinical application of these therapeutic treatments for hearing loss. In this study, we examined the potential of cell-gene therapy with non-viral vectors for delivery of therapeutic molecules into the cochlea. NIH3T3 cells were transfected with the brain-derived neurotrophic factor (BDNF) gene using lipofection and then transplanted into the mouse inner ear. Immunohistochemistry and western blotting demonstrated the survival of grafted cells in the cochlea for up to four weeks after transplantation. No significant hearing loss was induced by the transplantation procedure. A BDNF-specific enzyme-linked immunosorbent assay revealed a significant increase in BDNF production in the inner ear following transplantation of engineered cells. These findings indicate that cell-gene delivery with non-viral vectors may be applicable for the local, sustained delivery of therapeutic molecules into the cochlea. We examined the potential of bone-marrow stromal cell (BMSC) as a vehicle to deliver genes or their products into inner ear tissues. BMSCs labelled with enhanced green fluorescent protein were injected into the periplymphatic space of normal cochleae in mice. Histological analysis 2 weeks after transplantation demonstrated that transplanted cells settled within the cochlear tissues, especially in the spiral ligament, although most transplants were located in the perilymphatic space. These findings indicate the potential of BMSCs for delivering genes in the spiral ligament.

感官听力损失（SNHL）是常见的残疾，但治疗方案目前仅限于人工耳蜗和助听器。因此，正在进行研究以提供包括基因治疗在内的生物疗法的替代方法。需要开发出安全有效的基因输送方法，以促进这些治疗治疗的临床应用，以实现听力损失。在这项研究中，我们检查了用非病毒载体将治疗分子递送到耳蜗的潜力。使用lipofection用脑衍生的神经营养因子（BDNF）基因转染NIH3T3细胞，然后将其移植到小鼠内耳中。免疫组织化学和蛋白质印迹证明了移植后最多四个星期的移植细胞在耳蜗中的存活。移植程序未引起明显的听力损失。 BDNF特异性酶联免疫吸附测定法显示，在移植工程细胞后，内耳的BDNF产生显着增加。这些发现表明，用非病毒载体的细胞 - 基因递送可能适用于局部持续的治疗分子递送到耳蜗中。我们研究了骨髓基质细胞（BMSC）作为将基因或其产物输送到内耳组织的载体的潜力。将用增强的绿色荧光蛋白标记的BMSC注射到小鼠正常耳蜗的外膜片空间中。移植后2周，组织学分析表明，移植细胞定居在人工耳蜗内，尤其是在螺旋韧带中，尽管大多数移植都位于围绕围绕膜的空间中。这些发现表明了BMSC在螺旋韧带中输送基因的潜力。

项目成果

暮らしの科学25 難聴Q＆A

生命科学 25 听力损失问答

• 发表时间: 2005
• 作者: 伊藤壽一;中川隆之
• 通讯作者: 中川隆之

Experimental models for inner ear disorders.

内耳疾病的实验模型。

• 发表时间: 2005
• 期刊: JOHNS 21
• 作者: Iwai K;Nakaagwa T.
• 通讯作者: Nakaagwa T.

The potential use of bone marrow strormal cells for cochlear cell therapy.

骨髓基质细胞用于耳蜗细胞治疗的潜在用途。

• 发表时间: 2007
• 期刊: Neuroreport 18
• 作者: Sharif S;Nakagawa T;et al.
• 通讯作者: et al.

Cell therapy for inner ear diseases. Proceeding of the 5th International Symposium Meniere's Disease & Inner Ear Homeostasis Disorders.

内耳疾病的细胞疗法。

• 发表时间: 2006
• 作者: Ito J;Nakagawa T.
• 通讯作者: Nakagawa T.

A novel strategy for treatment of inner ears using a biodegradable gel.

使用可生物降解凝胶治疗内耳的新策略。

• 发表时间: 2005
• 期刊: Laryngoscope 115・11
• 作者: Endo T;Nakagawa t;Kita T;Iguchi F;Kim TS;Tamura T;Iwai K;Tabata Y;Ito J.
• 通讯作者: Ito J.