A model of corrective gene transfer in X-linked ichthyosis.
A model of corrective gene transfer in X-linked ichthyosis.
复制标题
X连锁鱼鳞病纠正性基因转移模型。
DOI：
10.1093/hmg/6.6.927
复制
发表时间：
1997
影响因子：
3.5
通讯作者：
Khavari,PA
中科院分区：
文献类型：
--
作者：
Freiberg,RA;Choate,KA;Deng,H;Alperin,ES;Shapiro,LJ;Khavari,PA
单基因隐性遗传性皮肤病为治疗性皮肤基因治疗的发展提供了诱人的原型。我们利用X连锁鱼鳞病(XLI)，其特征是类固醇硫酸酯酶芳基硫酸酯酶C(STS)功能丧失，建立了一种在活体内向人类皮肤传递校正基因的模型。构建了一种新的逆转录病毒表达载体，将STS基因转移到XLI患者的原代角质形成细胞中。转导与全长STS蛋白表达的恢复以及类固醇硫酸酯酶活性与XLI细胞中前病毒整合的数量成比例相关。然后，将转导和未矫正的XLI角质形成细胞与正常对照一起移植到免疫缺陷小鼠身上，以再生全层人表皮。未经修饰的XLI角质形成细胞再生出缺乏STS表达且皮肤屏障功能缺陷的角化过度的表皮，有效地再现了体内人类的疾病。然而，来自同一患者的转导XLI角质形成细胞，再生的表皮在组织学上与来自正常皮肤的角质形成细胞形成的表皮没有区别。移植后5周，转导XLI的表皮通过免疫组织化学染色显示STS在活体中表达，组织学外观正常化。此外，转导的XLI表皮显示屏障功能参数恢复正常。这些发现证明了XLI患者皮肤组织在分子和功能水平上的正确基因传递，并为人类皮肤基因治疗提供了一个模型。
Single gene recessive genetic skin disorders offer attractive prototypes for the development of therapeutic cutaneous gene delivery. We have utilized X-linked ichthyosis (XLI), characterized by loss of function of the steroid sulfatase arylsulfatase C (STS), to develop a model of corrective gene delivery to human skinin vivo. A new retroviral expression vector was produced and utilized to effect STS gene transfer to primary keratinocytes from XLI patients. Transduction was associated with restoration of full-length STS protein expression as well as steroid sulfatase enzymatic activity in proportion to the number of proviral integrations in XLI cells. Transduced and uncorrected XLI keratinocytes, along with normal controls, were then grafted onto immunodeficient mice to regenerate full thickness human epidermis. Unmodified XLI keratinocytes regenerated a hyperkeratotic epidermis lacking STS expression with defective skin barrier function, effectively recapitulating the human diseasein vivo. Transduced XLI keratinocytes from the same patients, however, regenerated epidermis histologically indistinguishable from that formed by keratinocytes from patients with normal skin. Transduced XLI epidermis demonstrated STS expressionin vivoby immunostaining as well as a normalization of histologic appearance at 5 weeks post-grafting. In addition, transduced XLI epidermis demonstrated a return of barrier function parameters to normal. These findings demonstrate corrective gene delivery in human XLI patient skin tissue at both molecular and functional levels and provide a model of human cutaneous gene therapy.
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影响因子：
82.9
作者：
Choate, KA;Medalie, DA;Khavari, PA
通讯作者：
Khavari, PA
影响因子：
6.5
作者：
MURPHY, GF;FLYNN, TC;PINKUS, GS
通讯作者：
PINKUS, GS
转基因皮肤治疗疾病：潜力和局限性。
DOI：
10.1038/jid.1994.14
发表时间：
1994
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
G. Krueger;J. Morgan;C. M. Jorgensen;L. Schmidt;H. L. Li;M. Kwan;S. Boyce;H. Wiley;J. Kaplan;M. Petersen
通讯作者：
M. Petersen
特定人类和细菌 DNA 序列的多聚体在人类细胞中的自主复制
影响因子：
5.3
作者：
P. Krysan;J. G. Smith;M. Calos
通讯作者：
M. Calos
影响因子：
5.4
作者：
HOEBEN, RC;MIGCHIELSEN, AAJ;VANDEREB, AJ
通讯作者：
VANDEREB, AJ