1 Center for Translational Medicine, the First Affiliated Hospital of Xi'an Jiaotong University , Xi'an, P.R. China.
2 Key Laboratory for Tumor Precision Medicine of Shaanxi Province, the First Affiliated Hospital of Xi'an Jiaotong University , Xi'an, P.R. China.
Hum Gene Ther. 2018 Dec;29(12):1352-1363. doi: 10.1089/hum.2018.048. Epub 2018 Nov 19.
Targeted therapy produces objective responses in bladder cancer patients, although the responses can be short. Meanwhile, response rates to immune therapy are lower, but the effects are more durable. Based on these findings, it was hypothesized that urothelial carcinoma associated 1 (UCA1)-targeted therapy could synergize with programmed cell death 1 (PD-1) blockade to enhance antitumor activity. To test this hypothesis, the effects of CRISPR-Cas9 targeting of UCA1 and PD-1 were assessed in vitro and in vivo. It was found that gRNA/cas9-targeted UCA1 induced apoptosis of 5637 bladder cancer cells, whereas PD-1 gene knockout could be achieved by electroporation of gRNA/cas9 targeting PD-1, as detected by polymerase chain reaction. In 5637 cell-xenografted humanized SCID mice, stimulation with CRISPR-Cas9 systems, immune phenotypes, and cytokine expression of human dendritic cells (DCs) was detected by flow cytometry, and polymerase chain reaction, respectively. The results of these assays suggested that the gRNA/cas9 treatment upregulated expression of CD80, CD83, and CD86 and significantly increased interleukin (IL)-6, IL-12, and IL-23 and tumor necrosis factor alpha mRNA levels. Co-administration of anti-PD-1 and anti-UCA1 treatment suppressed tumor growth and markedly improved survival of 5637 xenografted mice. Additionally, the combination treatment increased interferon gamma production by T cells that subsequently enhanced the expression of Th1-associated immune-stimulating genes to reduce transcription of regulatory/suppressive immune genes and reshape the tumor microenvironment from an immunosuppressive to a stimulatory state. Finally, anti-UCA1 treatment was shown to induce interferon gamma-dependent programmed cell death ligand 1 expression within 5637 xenograft tumors in vivo. Together, these results demonstrate potent synergistic effects of a combination therapy using LncRNA UCA1-targeted therapy and immune checkpoint blockade of PD-1, thus supporting the translational potential of this combination strategy for clinical treatment of bladder cancer.
靶向治疗可使膀胱癌患者产生客观反应，尽管反应可能短暂。同时，免疫治疗的反应率较低，但效果更持久。基于这些发现，有人假设尿路上皮癌相关 1（UCA1）靶向治疗可以与程序性细胞死亡 1（PD-1）阻断协同作用，增强抗肿瘤活性。为了验证这一假设，评估了 CRISPR-Cas9 靶向 UCA1 和 PD-1 在体外和体内的作用。结果发现，gRNA/cas9 靶向 UCA1 诱导 5637 膀胱癌细胞凋亡，而通过 gRNA/cas9 靶向 PD-1 的电穿孔可以实现 PD-1 基因敲除，这可以通过聚合酶链反应检测到。在 5637 细胞异种移植的人源化 SCID 小鼠中，通过流式细胞术和聚合酶链反应分别检测到 CRISPR-Cas9 系统刺激后人类树突状细胞（DC）的免疫表型和细胞因子表达。这些测定的结果表明，gRNA/cas9 处理上调了 CD80、CD83 和 CD86 的表达，并显著增加了白细胞介素（IL）-6、IL-12 和 IL-23 和肿瘤坏死因子α mRNA 水平。抗 PD-1 和抗 UCA1 联合治疗抑制肿瘤生长，显著改善 5637 异种移植小鼠的生存。此外，联合治疗增加了 T 细胞产生干扰素γ，从而增强了 Th1 相关免疫刺激基因的表达，降低了调节/抑制性免疫基因的转录，并重塑了肿瘤微环境，使其从免疫抑制状态转变为刺激状态。最后，体内实验表明抗 UCA1 治疗可诱导 5637 异种移植肿瘤中干扰素γ依赖性程序性细胞死亡配体 1 的表达。综上所述，这些结果表明，LncRNA UCA1 靶向治疗联合免疫检查点阻断 PD-1 的联合治疗具有强大的协同作用，从而支持这种联合策略用于膀胱癌临床治疗的转化潜力。
