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Development of a new therapy of severe ichthyosis caused by ABCA12 mutations

ABCA12突变引起的严重鱼鳞病新疗法的开发

基本信息

批准号：
18390310
负责人：
MASASHI Akiyama
金额：
$ 11.07万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390310/
关键词：
ichthyosis ABCA12 gene therapy mutation harlequin ichthyosis skin barrier 皮膚バリア

项目摘要

Serious defects in the epidermal keratinocyte lipid transporter ABCA12 are known to result in a deficient skin lipid barrier, leading to severe ichthyosis. In this study, we intended to develop a gene therapy method of severe icththyosis by an ABCA12 corrective gene transfer system for hair follicle epithelial stem cells. For persistent and stable transgene expression in hair follicle stem cells, we transferred retroviral vectors encoding reporter genes into cultured hair follicle stem cells. We performed gene transfection experiments into the bulge stem cells in vitro. We dissected bulge areas from mice vibrissa hair follicles and established primary cultures. The ABCA12 transfected cells were mixed with cultured dermal papilla cells and transplanted on to immunodeficient mice. We succeeded in reconstituting hair follciles and their appendages from the cells harboring a transgene reporter. The transgene expression was observed in all skin epithelial compartments including the hair follicle epithelium, sebaceous gland and epidermis. In addition, we performed gene transfection experiments into the bulge stem cells in vivo using the methods that had been established from in vitro studies.Furthermore, we performed transfection experiments of gene constructs for gene therapy. In detail, we cloned normal ABCA12 cDNA construct with the transfection vector and the reporter genes selected from the results of previous experiments. We obtained keratinocyte cultures form severe ichthyosis patients harboring ABCA12 mutations after fully informed consents. Epidermis showing characteristic ichthyosis phenotype was reconstituted from these cultured keratinocytes form the patients on the back of nude mice. Targetting the reconstructed ichthyosis skin lesions, we tried gene therapy experiments using normal ABCA12 gene constructs. This hair follicle stem cell targeted ABCA12 corrective gene transfer system provided reliable gene therapy.

已知表皮角质形成细胞脂质转运蛋白 ABCA12 的严重缺陷会导致皮肤脂质屏障缺陷，从而导致严重的鱼鳞病。在这项研究中，我们打算通过针对毛囊上皮干细胞的ABCA12校正基因转移系统开发一种严重鱼鳞病的基因治疗方法。为了在毛囊干细胞中持久稳定地表达转基因，我们将编码报告基因的逆转录病毒载体转移到培养的毛囊干细胞中。我们在体外对隆起干细胞进行了基因转染实验。我们解剖了小鼠触毛毛囊的隆起区域并建立了原代培养物。将ABCA12转染的细胞与培养的真皮乳头细胞混合并移植到免疫缺陷小鼠体内。我们成功地从含有转基因报告基因的细胞中重建了毛囊及其附属物。在所有皮肤上皮区室中观察到转基因表达，包括毛囊上皮、皮脂腺和表皮。此外，我们使用体外研究建立的方法对隆起干细胞进行体内基因转染实验。此外，我们进行了用于基因治疗的基因构建体的转染实验。具体来说，我们使用转染载体和从先前实验结果中选择的报告基因克隆了正常的ABCA12 cDNA构建体。在充分知情同意后，我们从携带 ABCA12 突变的严重鱼鳞病患者中获得了角质形成细胞培养物。显示特征性鱼鳞病表型的表皮是由来自裸鼠背部的患者的这些培养的角质形成细胞重建的。针对重建的鱼鳞病皮肤病变，我们尝试使用正常的 ABCA12 基因构建体进行基因治疗实验。这种毛囊干细胞靶向ABCA12校正基因转移系统提供了可靠的基因治疗。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Unique mutations in the filaggrin gene in Japanese patients with ichthyosis vulgaris and atopic dermatitis

DOI：
10.1016/j.jaci.2006.12.646
发表时间：
2007-02-01
期刊：
JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY
影响因子：
14.2
作者：
Nomura, Toshifumi;Sandilands, Aileen;Shimizu, Hiroshi
通讯作者：
Shimizu, Hiroshi

J Allergy Clin Immunol

过敏与临床免疫学杂志

DOI：
发表时间：
2007
期刊：
119
影响因子：
0
作者：
Nomura T;Sandilands A;Akiyama M;Sakai K;Ota M;Sugiura H;Yamamoto K;Sato H;Smith FJD;McLean WHI;Shimizu H.
通讯作者：
Shimizu H.

J Invest Dermatol

J Invest Dermatol 杂志

DOI：
发表时间：
2007
期刊：
127
影响因子：
0
作者：
Akiyama M;Titeux M;Sakai K;McMillan JR;Tonasso L;Calvas P;Jossic F;Hovnanian A;Shimizu H.
通讯作者：
Shimizu H.

Novel ALDH3A2 heterozygous mutations in a Japanese family of Sjogren-Larsson syndrome

日本干燥综合征家族中的新 ALDH3A2 杂合突变

DOI：
发表时间：
2006
期刊：
J Invest Dermatol 126・11
影响因子：
0
作者：
Sakai K;Akiyama M;Watanabe T;Sanayama K;Sugita K;Takahashi M;Suehiro K;Yorifuji K;Shibaki A;Shimizu H
通讯作者：
Shimizu H

DNA-based prenatal diagnosis of harlequin ichthyosis and characterization of ABCA12 mutation consequences