Division of Oncology, Department of Medical and Surgical Sciences for Children & Adults, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
Rigenerand srl, Modena, Italy.
Cancer Gene Ther. 2019 Feb;26(1-2):11-16. doi: 10.1038/s41417-018-0034-1. Epub 2018 Jun 29.
Cellular therapies based on mesenchymal stromal/stem cells (MSC) are promising strategies in regenerative medicine and oncology. Despite encouraging results, there is still some level of concerns on inoculating MSC in cancer patients. To face this issue, one possibility resides in engineering MSC by incorporating a suicide gene in order to control their fate once infused. Strategies based on Herpes Simplex Virus Thymidine Kinase (HSV-TK) and the Cytosine Deaminase genes have been developed and more recently a novel suicide gene, namely, iCasp9, has been proposed. This approach is based on a variant of human Caspase9 that binds with high affinity to a synthetic, bioinert small molecule (AP20187) leading to cell death. Based on this technology so far marginally applied to MSC, we tested the suitability of iCasp9 suicide strategy in MSC to further increase their safety. MSC have been transfected by a lentiviral vector carrying iCasp9 gene and then tested for viability after AP20187 treatment in comparison with mock-transfected cells. Moreover, accounting our anti-tumor approaches based on MSC expressing potent anti-cancer ligand TNF-Related Apoptosis-Inducing Ligand (TRAIL), we generated adipose MSC co-expressing iCasp9 and TRAIL successfully targeting an aggressive sarcoma type. These data show that anti-cancer and suicide mechanisms can coexist without affecting cells performance and hampering the tumoricidal activity mediated by TRAIL. In conclusion, this study originally indicates the suitability of combining a MSC-based anti-cancer gene approach with iCasp9 demonstrating efficiency and specificity.
基于间充质基质/干细胞（MSC）的细胞疗法是再生医学和肿瘤学中很有前途的策略。尽管取得了令人鼓舞的结果，但在癌症患者中接种 MSC 仍存在一定程度的担忧。为了应对这一问题，一种可能性是通过将自杀基因整合到 MSC 中，从而在注入后控制其命运。已经开发了基于单纯疱疹病毒胸苷激酶（HSV-TK）和胞嘧啶脱氨酶基因的策略，最近还提出了一种新的自杀基因，即 iCasp9。这种方法基于人类 Caspase9 的一种变体，该变体与一种合成的、生物惰性的小分子（AP20187）具有高亲和力，导致细胞死亡。基于迄今为止这项技术仅在 MSC 中得到了有限的应用，我们测试了 iCasp9 自杀策略在 MSC 中的适用性，以进一步提高其安全性。MSC 被携带 iCasp9 基因的慢病毒载体转染，然后与 mock 转染细胞进行比较，在 AP20187 处理后测试其存活率。此外，考虑到我们基于表达强效抗癌配体 TNF 相关凋亡诱导配体（TRAIL）的 MSC 的抗肿瘤方法，我们成功地生成了共表达 iCasp9 和 TRAIL 的脂肪 MSC，靶向一种侵袭性肉瘤类型。这些数据表明，抗癌和自杀机制可以共存，而不会影响细胞性能并阻碍 TRAIL 介导的杀肿瘤活性。总之，这项研究最初表明了将基于 MSC 的抗癌基因方法与 iCasp9 相结合的适用性，证明了其效率和特异性。
