Correction of Hair Shaft Defects through Allele-Specific Silencing of Mutant Krt75.
Correction of Hair Shaft Defects through Allele-Specific Silencing of Mutant Krt75.
复制标题
通过突变 Krt75 的等位基因特异性沉默来纠正毛干缺陷。
DOI：
10.1038/jid.2015.375
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发表时间：
2016-01
期刊：
影响因子：
--
通讯作者：
Chen J
中科院分区：
文献类型：
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作者：
Liu Y;Snedecor ER;Zhang X;Xu Y;Huang L;Jones EC;Zhang L;Clark RA;Roop DR;Qin C;Chen J
角蛋白基因的显性突变可引起许多遗传性皮肤病，其特征在于表皮内起泡、表皮角化过度或皮肤附属物异常，如甲板营养不良和毛发结构缺陷。通过RNA干扰使突变角蛋白等位基因特异性沉默是一种有前途的治疗方法，用于抑制表皮中突变角蛋白和相关表型的表达。然而，其对皮肤附属物的有效性仍有待体内证实。在这项研究中，我们开发了等位基因特异性siRNA，能够选择性地抑制突变Krt75的表达，这会导致小鼠毛干结构缺陷，其特征在于沿毛干沿着发育水泡。从突变Krt75小鼠模型分离的表皮角质形成细胞祖细胞再生的毛发在体内移植时再现了起泡表型。相反，用表达突变Krt75特异性shRNA的慢病毒载体操纵的突变细胞持续抑制这种表型。表型校正与皮肤移植物中突变Krt75 mRNA的显著减少相关。因此，从这项研究中获得的数据证明了利用RNA干扰通过突变角蛋白基因的等位基因特异性沉默来实现毛发结构表型的持久校正的可行性。
Dominant mutations in keratin genes can cause a number of inheritable skin disorders characterized by intraepidermal blistering, epidermal hyperkeratosis, or abnormalities in skin appendages, such as nail plate dystrophy and structural defects in hair. Allele-specific silencing of mutant keratins through RNA interference is a promising therapeutic approach for suppressing the expression of mutant keratins and related phenotypes in the epidermis. However, its effectiveness on skin appendages remains to be confirmed in vivo. In this study, we developed allele specific siRNAs capable of selectively suppressing the expression of a mutant Krt75, which causes hair shaft structural defects characterized by the development of blebs along the hair shaft in mice. Hair regenerated from epidermal keratinocyte progenitor cells isolated from mutant Krt75 mouse models reproduced the blebbing phenotype when grafted in vivo. In contrast, mutant cells manipulated with a lentiviral vector expressing mutant Krt75-specific shRNA persistently suppressed this phenotype. The phenotypic correction was associated with significant reduction of mutant Krt75 mRNA in the skin grafts. Thus, data obtained from this study demonstrated the feasibility of utilizing RNA interference to achieve durable correction of hair structural phenotypes through allele-specific silencing of the mutant keratin genes.