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.

感音神经性听力损失（SNHL）是一种常见的残疾，但目前的治疗选择仅限于人工耳蜗和助听器。因此，正在进行研究，以提供生物治疗的替代手段，包括基因治疗。需要开发安全有效的耳蜗基因递送方法，以促进这些听力损失治疗方法的临床应用。在这项研究中，我们研究了细胞基因治疗的潜力与非病毒载体的治疗分子进入耳蜗。采用脂质体法将脑源性神经营养因子（BDNF）基因转染NIH3T3细胞，然后将其移植到小鼠内耳。免疫组织化学和蛋白质印迹法证明移植细胞在耳蜗中的存活长达四周。移植过程未引起明显的听力损失。BDNF特异性酶联免疫吸附试验显示，移植工程细胞后，内耳中BDNF的产生显著增加。这些发现表明，细胞基因传递与非病毒载体可能适用于局部，持续交付的治疗分子进入耳蜗。我们研究了骨髓基质细胞（BMSC）作为载体将基因或其产物递送到内耳组织中的潜力。将增强型绿色荧光蛋白标记的骨髓间充质干细胞注入小鼠正常耳蜗周围淋巴间隙。移植后2周的组织学分析表明，移植细胞定居在耳蜗组织，特别是在螺旋韧带，虽然大多数移植位于外淋巴间隙。这些发现表明骨髓间充质干细胞在螺旋韧带中传递基因的潜力。

项目成果

Age-dependent degeneration of the stria vascularis in human cochleae

• DOI: 10.1097/01.mlg.0000234940.33569.39
• 发表时间: 2006-10-01
• 期刊: LARYNGOSCOPE
• 影响因子: 2.6
• 作者: Suzuki, Teruhisa;Nomoto, Yukio;Omori, Koichi
• 通讯作者: Omori, Koichi

Effects of bone-morphogenetic protein 4 on differentiation of embryonic stem cells into myosin VIIa-positive cells.

骨形态发生蛋白 4 对胚胎干细胞分化为肌球蛋白 VIIa 阳性细胞的影响。

• 发表时间: 2007
• 期刊: Acta Otolaryngol Suppl 557
• 作者: Higashi T;Nakagawa T;et al.
• 通讯作者: et al.

Effects of bone-morphogenetic protein 4 on differentiation of embryonic stem cells into myosin Vila-positive cells.

骨形态发生蛋白 4 对胚胎干细胞分化为肌球蛋白 Vila 阳性细胞的影响。

• 发表时间: 2007
• 期刊: Acta Otolaryngol 557
• 作者: Higashi T;Nakagawa T;et al.
• 通讯作者: et al.

Aging-effects on vestibule-ocular responses in C57B/6 mice : comparison with alteration in auditory function.

衰老对 C57B/6 小鼠前庭眼反应的影响：与听觉功能改变的比较。

• 发表时间: 2005
• 期刊: Audiol Neurootol 10・2
• 作者: Shiga A;Nakagawa T;et al.
• 通讯作者: et al.

Mechanisms of age-related hearing loss and strategies for protection and restoration of hearing

年龄相关性听力损失的机制及听力保护和恢复策略

• 发表时间: 2004
• 期刊: Japanese Journal of Geriatrics 41
• 作者: Lee JE;Nakagawa T;et al.;Nakagawa T.
• 通讯作者: Nakagawa T.