Laboratório de Vetores Virais, Centro de Investigação Translacional em Oncologia, Instituto Do Câncer Do Estado de São Paulo, Faculdade de Medicina, Universidade de São Paulo, Av. Dr. Arnaldo, 251, 8th floor, São Paulo, SP, Brazil.
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
Sci Rep. 2020 Oct 21;10(1):17893. doi: 10.1038/s41598-020-74826-y.
Since melanomas often retain wild type p53, we developed an adenoviral vector, AdRGD-PG, which provides robust transduction and transgene expression in response to p53. Previously, this vector was used for interferon-β gene transfer in mouse models of melanoma, resulting in control of tumor progression, but limited cell killing. Here, the AdRGD-PG-hIFNβ vector encoding the human interferon-β cDNA (hIFNβ) was used to transduce human melanoma cell lines SK-MEL-05 and SK-MEL-147 (both wild type p53). In vitro, cell death was induced in more than 80% of the cells and correlated with elevated annexinV staining and caspase 3/7 activity. Treatment with hIFNβ promoted cell killing in neighboring, non-transduced cells, thus revealing a bystander effect. In situ gene therapy resulted in complete inhibition of tumor progression for SK-MEL-147 when using nude mice with no evidence of hepatotoxicity. However, the response in Nod-Scid mice was less robust. For SK-MEL-05, tumor inhibition was similar in nude and Nod-Scid mice and was less efficient than seen for SK-MEL-147, indicating both cell type and host specific responses. The AdRGD-PG-hIFNβ vector provides extensive killing of human melanoma cells in vitro and a potent anti-tumor effect in vivo. This study provides a critical advance in the development of our melanoma gene therapy approach.
由于黑色素瘤通常保留野生型 p53,我们开发了一种腺病毒载体 AdRGD-PG,它可以响应 p53 进行强大的转导和转基因表达。以前,该载体曾用于黑色素瘤小鼠模型中的干扰素-β基因转移,导致肿瘤进展得到控制,但细胞杀伤有限。在这里,编码人干扰素-β cDNA(hIFNβ)的 AdRGD-PG-hIFNβ 载体被用于转导人黑色素瘤细胞系 SK-MEL-05 和 SK-MEL-147(均为野生型 p53)。在体外,超过 80%的细胞诱导细胞死亡,与 AnnexinV 染色和 caspase 3/7 活性升高相关。用 hIFNβ 处理可促进邻近未转导细胞的细胞杀伤,从而显示出旁观者效应。原位基因治疗导致裸鼠中 SK-MEL-147 肿瘤进展完全抑制,无肝毒性证据。然而,Nod-Scid 小鼠的反应不那么强烈。对于 SK-MEL-05,裸鼠和 Nod-Scid 小鼠中的肿瘤抑制作用相似,且不如 SK-MEL-147 有效,这表明存在细胞类型和宿主特异性反应。AdRGD-PG-hIFNβ 载体在体外对人黑色素瘤细胞具有广泛的杀伤作用,并在体内具有强大的抗肿瘤作用。这项研究为我们的黑色素瘤基因治疗方法的发展提供了重要进展。