Division of Molecular Pharmaceutics, UNC Eshelman School of Pharmacy, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Mol Ther. 2020 Jul 8;28(7):1614-1627. doi: 10.1016/j.ymthe.2020.04.022. Epub 2020 Apr 29.
The conversion of human fibroblasts into personalized induced neural stem cells (iNSCs) that actively seek out tumors and deliver cytotoxic agents is a highly promising approach for treating various types of cancer. However, the ability to generate iNSCs from the skin of cancer patients has not been explored. Here, we take an important step toward clinical application by generating iNSCs from skin biopsies of human patients undergoing treatment for the aggressive brain cancer, glioblastoma (GBM). We then utilized a panel of functional and genomic studies to investigate the efficacy and tumor-homing capacity of these patient-derived cells, as well as genomic analysis, to characterize the impact of interpatient variability on this personalized cell therapy. From the skin-tissue biopsies, we established fibroblasts and transdifferentiated the cells into iNSCs. Genomic and functional testing revealed marked variability in growth rates, therapeutic agent production, and gene expression during fibroblast-to-iNSC conversion among patient lines. In vivo testing showed patient-derived iNSCs home to tumors, yet rates and expression of homing-related pathways varied among patients. With the use of surgical-resection mouse models of invasive human cluster of differentiation 133 (CD133) GBM cells and serial kinetic imaging, we found that "high-performing" patient-derived iNSC lines reduced the volume of GBM cells 60-fold and extended survival from 28 to 45 days. Treatment with "low-performing" patient lines had minimal effect on tumor growth, but the anti-tumor effect could be rescued by increasing the intracavity dose. Together, these data show, for the first time, that tumor-homing iNSCs can be generated from the skin of cancer patients and efficaciously suppress tumor growth. We also begin to define genetic markers that could be used to identify cells that will contain the most effective attributes for tumor homing and kill in human patients, including high gene expression of the semaphorin-3B (SEMA3B), which is known to be involved in neuronal cell migration. These studies should serve as an important guide toward clinical GBM therapy, where the personalized nature of optimized iNSC therapy has the potential to avoid transplant rejection and maximize treatment durability.
将人类成纤维细胞转化为能够主动寻找肿瘤并递送细胞毒性药物的个性化诱导神经干细胞（iNSC），是治疗各种类型癌症的一种极具前景的方法。然而，尚未探索从癌症患者的皮肤中生成 iNSC 的能力。在这里，我们通过从接受侵袭性脑癌胶质母细胞瘤（GBM）治疗的人类患者的皮肤活检中生成 iNSC，朝着临床应用迈出了重要的一步。然后，我们利用一系列功能和基因组研究来研究这些患者来源细胞的疗效和肿瘤归巢能力，以及基因组分析，以表征这种个性化细胞疗法中个体间变异性的影响。从皮肤组织活检中，我们建立了成纤维细胞，并将这些细胞转分化为 iNSC。基因组和功能测试显示，在患者系中，成纤维细胞向 iNSC 转化过程中的细胞生长速度、治疗剂产生和基因表达存在显著差异。体内测试表明，患者来源的 iNSC 归巢至肿瘤，但归巢相关途径的比率和表达在患者间存在差异。使用侵袭性人 CD133 胶质母细胞瘤细胞的手术切除小鼠模型和连续动态成像，我们发现“高表现”患者来源的 iNSC 系将 GBM 细胞的体积减少了 60 倍，并将生存期从 28 天延长至 45 天。用“低表现”患者系进行治疗对肿瘤生长的影响很小，但通过增加腔内剂量可以挽救抗肿瘤作用。这些数据首次表明，可以从癌症患者的皮肤中生成归巢肿瘤的 iNSC，并有效地抑制肿瘤生长。我们还开始定义遗传标记，这些标记可用于鉴定包含对人类患者肿瘤归巢和杀伤最有效属性的细胞，包括已知参与神经元细胞迁移的神经丝蛋白-3B（SEMA3B）的高基因表达。这些研究应该为胶质母细胞瘤的临床治疗提供重要的指导，其中优化的 iNSC 治疗的个性化特性有可能避免移植排斥并最大限度地延长治疗效果的持久性。
