Department of Biochemistry and Molecular Medicine, UC Davis School of Medicine, Sacramento, CA 95817, USA.
Department of Pathology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Biochem Pharmacol. 2021 Jul;189:114392. doi: 10.1016/j.bcp.2020.114392. Epub 2021 Jan 3.
Lung cancer remains the leading cause of cancer deaths worldwide and accounts for more than 22% of all cancer-related deaths in the US. Developing new therapies is essential to combat against deadly lung cancer, especially the most common type, non-small cell lung cancer (NSCLC). With the discovery of genome-derived functional small noncoding RNA (ncRNA), namely microRNAs (miRNA or miR), restoration of oncolytic miRNAs lost or downregulated in NSCLC cells represents a new therapeutic strategy. Very recently, we have developed a novel technology that achieves in vivo fermentation production of bioengineered miRNA agents (BERA) for research and development. In this study, we aimed at simultaneously introducing two miRNAs into NSCLC cells by using single recombinant "combinatorial BERA" (CO-BERA) molecule. Our studies show that single CO-BERA molecule (e.g., let-7c/miR-124) was successfully processed to two miRNAs (e.g., let-7c-5p and miR-124-3p) to combinatorially regulate the expression of multiple targets (e.g., RAS, VAMP3 and CDK6) in human NSCLC cells, exhibiting greater efficacy than respective BERA miRNAs in the inhibition of cell viability and colony formation. Furthermore, we demonstrate that CO-BERA let-7c/miR-124-loaded lipopolyplex nanomedicine was the most effective among tested RNAs in the control of tumor growth in NSCLC patient-derived xenograft mouse models. The anti-tumor activity of CO-BERA let-7c/miR-124 was associated with the suppression of RAS and CDK6 expression, and enhancement of apoptosis. These results support the concept to use single ncRNA agent for dual-targeting and offer insight into developing new RNA therapeutics for the treatment of lethal NSCLC.
肺癌仍然是全球癌症死亡的主要原因，占美国所有癌症相关死亡的 22%以上。开发新的治疗方法对于对抗致命的肺癌至关重要，尤其是最常见的非小细胞肺癌（NSCLC）。随着基因组衍生的功能性小非编码 RNA（ncRNA），即 microRNAs（miRNA 或 miR）的发现，恢复 NSCLC 细胞中丢失或下调的肿瘤溶瘤 miRNA 代表了一种新的治疗策略。最近，我们开发了一种新的技术，能够在体内发酵生产用于研究和开发的生物工程 miRNA 剂（BERA）。在这项研究中，我们旨在通过使用单个重组“组合 BERA”（CO-BERA）分子同时将两种 miRNA 引入 NSCLC 细胞。我们的研究表明，单个 CO-BERA 分子（例如，let-7c/miR-124）成功地被加工成两种 miRNA（例如，let-7c-5p 和 miR-124-3p），从而组合调控多个靶标（例如，RAS、VAMP3 和 CDK6）在人类 NSCLC 细胞中的表达，其抑制细胞活力和集落形成的效果优于各自的 BERA miRNAs。此外，我们证明 CO-BERA let-7c/miR-124 负载的脂质多聚体纳米药物在控制 NSCLC 患者来源异种移植小鼠模型中的肿瘤生长方面是所有测试 RNA 中最有效的。CO-BERA let-7c/miR-124 的抗肿瘤活性与抑制 RAS 和 CDK6 表达以及增强细胞凋亡有关。这些结果支持使用单个 ncRNA 剂进行双重靶向的概念，并为开发用于治疗致命性 NSCLC 的新型 RNA 疗法提供了思路。
