Department of Dermatology and Allergy, HTCC - Skin Cancer Center Charité, Charité-Universitätsmedizin Berlin.
Exp Dermatol. 2010 Aug;19(8):e56-66. doi: 10.1111/j.1600-0625.2009.00977.x.
The high mortality of melanoma demands the development of new strategies, and gene therapy may be considered provided improvements in efficacy and selectivity. Overexpression of the death ligand CD95L/FasL has been shown in previous studies as highly effective for apoptosis induction in melanoma cells. For efficient and selective targeting of melanoma, a conditional replication-competent adenoviral vector was constructed (Ad5-FFE-02), which drives CD95L expression by a tetracycline-inducible promoter. For restricting its replication to melanoma cells, the adenoviral E1A gene is controlled by a tyrosinase-derived promoter. Furthermore, adenoviral E1B was deleted and a mutated E1A was used to preferentially support replication in tumor cells. Proving its high selectivity and efficiency, strong expression of E1A and doxycycline-dependent induction of CD95L were characteristic for tyrosinase-positive melanoma cells after Ad5-FFE-02 transduction, whereas absent in non-melanoma cell lines. Importantly, Ad5-FFE-02-mediated cell lysis was restricted to melanoma cells, and induction of apoptosis was found only in tyrosinase and CD95 expressing cells. Finally, the combination of adenoviral replication and CD95L-mediated apoptosis resulted in an enhanced repression of melanoma cell growth. This new adenoviral vector may provide a basis for an efficient targeting of melanoma.
黑色素瘤的高死亡率要求开发新策略，并且可以考虑基因治疗，只要能提高疗效和选择性。在先前的研究中已经表明，死亡配体 CD95L/FasL 的过表达对黑色素瘤细胞的凋亡诱导非常有效。为了有效地、有选择性地针对黑色素瘤，构建了一种条件复制型腺病毒载体（Ad5-FFE-02），该载体通过四环素诱导的启动子驱动 CD95L 的表达。为了将其复制限制在黑色素瘤细胞中，腺病毒 E1A 基因受酪氨酸酶衍生启动子的控制。此外，腺病毒 E1B 被删除，并使用突变的 E1A 优先支持肿瘤细胞中的复制。Ad5-FFE-02 转导后，强表达 E1A 和四环素依赖性 CD95L 的表达是酪氨酸酶阳性黑色素瘤细胞的特征，而在非黑色素瘤细胞系中则不存在，证明了其高度的选择性和效率。重要的是，Ad5-FFE-02 介导的细胞裂解仅限于黑色素瘤细胞，并且仅在表达酪氨酸酶和 CD95 的细胞中发现诱导凋亡。最后，腺病毒复制和 CD95L 介导的凋亡的组合导致黑色素瘤细胞生长受到更强的抑制。这种新的腺病毒载体可能为黑色素瘤的有效靶向提供基础。
