In vivo fermentation production of humanized noncoding RNAs carrying payload miRNAs for targeted anticancer therapy.
In vivo fermentation production of humanized noncoding RNAs carrying payload miRNAs for targeted anticancer therapy.
复制标题
作者：
Li PC;Tu MJ;Ho PY;Batra N;Tran MML;Qiu JX;Wun T;Lara PN;Hu X;Yu AX;Yu AM
原理：非编码rna （ncRNAs）如microRNAs （miRs或miRNAs）通过转录后基因调控在细胞过程的控制中发挥重要作用。然而，ncRNA的研究仅限于利用体外合成的RNA试剂。在活细胞中产生和折叠的重组rna能够更好地概括生物rna。方法：在此，我们开发了一个新的平台，用于在体内发酵生产人源化重组ncRNA分子，即hBERAs，携带有效载荷miRNAs或sirna。用阴离子交换FPLC法纯化目标hBERAs。研究了hBERA/ mirna在人癌细胞中的功能，并在小鼠原位骨肉瘤异种移植自发肺转移模型中测定了其抗肿瘤活性。结果：鉴定出合适的人类trna与最佳的hsa-pre-miR-34a偶联，作为新的完全人源化的ncRNA载体，以容纳弹头mirna或sirna。一组30个目标hBERAs均异质过表达（每个占细菌总RNA的40%），这有利于大规模生产（从1L细菌培养中获得8-31 mg单个hBERAs）。模型hBERA/miR-34a-5p和miR-124-3p在人癌细胞中选择性加工成弹头mirna，调节靶基因表达，促进细胞凋亡，抑制侵袭性。此外，生物工程miR-34a-5p和miR-124-3p制剂均可显著降低原位骨肉瘤异种移植物肿瘤生长和自发性肺转移。结论：这种新型的ncRNA生物工程技术和由此产生的重组ncRNA是传统技术和基础研究和药物开发工具的独特补充。
Rationale: Noncoding RNAs (ncRNAs) such as microRNAs (miRs or miRNAs) play important roles in the control of cellular processes through posttranscriptional gene regulation. However, ncRNA research is limited to utilizing RNA agents synthesized in vitro. Recombinant RNAs produced and folded in living cells shall better recapitulate biologic RNAs. Methods: Herein, we developed a novel platform for in vivo fermentation production of humanized recombinant ncRNA molecules, namely hBERAs, carrying payload miRNAs or siRNAs. Target hBERAs were purified by anion exchange FPLC method. Functions of hBERA/miRNAs were investigated in human carcinoma cells and antitumor activities were determined in orthotopic osteosarcoma xenograft spontaneous lung metastasis mouse models. Results: Proper human tRNAs were identified to couple with optimal hsa-pre-miR-34a as new fully-humanized ncRNA carriers to accommodate warhead miRNAs or siRNAs. A group of 30 target hBERAs were all heterogeneously overexpressed (each accounting for >40% of total bacterial RNA), which facilitated large-scale production (8-31 mg of individual hBERAs from 1L bacterial culture). Model hBERA/miR-34a-5p and miR-124-3p were selectively processed to warhead miRNAs in human carcinoma cells to modulate target gene expression, enhance apoptosis and inhibit invasiveness. In addition, bioengineered miR-34a-5p and miR-124-3p agents both reduced orthotopic osteosarcoma xenograft tumor growth and spontaneous pulmonary metastases significantly. Conclusion: This novel ncRNA bioengineering technology and resulting recombinant ncRNAs are unique additions to conventional technologies and tools for basic research and drug development.
登录
查看更多内容
影响因子：
11.2
作者：
Bader AG;Brown D;Winkler M
通讯作者：
Winkler M
DOI：
10.1007/s40272-015-0134-4
发表时间：
2015-08
期刊：
Paediatric drugs
影响因子：
--
作者：
Rivera-Valentin RK;Zhu L;Hughes DP
通讯作者：
Hughes DP
影响因子：
64.5
作者：
Hatziapostolou M;Polytarchou C;Aggelidou E;Drakaki A;Poultsides GA;Jaeger SA;Ogata H;Karin M;Struhl K;Hadzopoulou-Cladaras M;Iliopoulos D
通讯作者：
Iliopoulos D
影响因子：
3.3
作者：
Li, Kay Ka Wai;Pang, Jesse Chung-sean;Ng, Ho-keung
通讯作者：
Ng, Ho-keung
miR-34a 的纳米颗粒递送通过直接靶向 C22ORF28 消除长期培养的乳腺癌干细胞
影响因子：
12.4
作者：
Lin, Xiaoti;Chen, Weiyu;Xie, Xiaoming
通讯作者：
Xie, Xiaoming