Cancer cell-specific RNA interference in vivo

体内癌细胞特异性RNA干扰

• 批准号: 405833972
• 负责人: Professor Dr. Andriy Mokhir
• 金额: --
• 依托单位: Skolkovo Institute of Science and Technology (Skoltech)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/405833972?language=en
• 关键词: Cancer; cell; specific; RNA; interference

RNA interference (RNAi) caused by small interfering RNAs (siRNAs) is a general method of sequence-specific control of gene expression in cells, both in vitro and in vivo. A problem of design of siRNAs for every gene of interest can be solved in silico by bioinformatics and validated in a limited number of experiments in vitro and in vivo. Therefore, development of siRNA-based drugs can potentially be less time-consuming and costly than that of classical drugs and can further promote development of personalized medicine. Recent breakthrough in delivery of siRNAs in vivo enabled the initiation of a number of clinical trials with siRNA-based drugs (treatment of hyperlipidemia, age related macular degeneration, etc.), some of which reached advanced stages (e.g. TTR amyloidosis). However, despite these important successes in the field, cancer-cell specific delivery/activation of siRNAs still remains a problem, for which no simple and general solution is known. Therefore, the number of targets for siRNA drugs is limited to genes, which are either strongly overexpressed or present exclusively in cancer cells. Application of cancer-specific prodrugs is expected to not only extend the number of possible targets for siRNA drug, but also reduce side effects of the treatment. In this project we will work on a simple solution for designing cancer specific siRNA-prodrugs, which are activated in the presence of reactive oxygen species (ROS). The cell specificity will be based on the experimental fact that cancer cells overproduce ROS, whereas their concentration in normal cells is negligible. ROS-induced activation is a potential general approach for the design of cancer specific siRNAs, since elevated ROS is believed to be crucial for cancer phenotypes. Therefore, the proposed siRNA-prodrugs should be active against many different cancer types. As a proof-of-principle, ROS-responsive siRNA-prodrugs targeting Ubr ligases will be developed and tested in the treatment of hepatocellular carcinoma (HCC) in mice models. This model was selected since no satisfactory treatment for HCC is currently known. Moreover, methods of liver specific delivery of siRNAs are well developed. The principles, worked out in this project, will be potentially applicable towards the design of siRNA-prodrugs for the treatment of cancers in organs other than the liver, providing that organ-specific delivery will become available in the future. This study capitalizes on the extensive experience of (a) the group of Mokhir in synthesis and cellular applications of “caged” siRNAs and (b) the group of Zatsepin on optimization of siRNA design and formulation as well synthesis and application of siRNA-based conjugates for inhibition of gene expression in vivo in mice models.

由小干扰RNA（siRNA）引起的RNA干扰（RNAi）是体外和体内序列特异性控制细胞中基因表达的一般方法。针对每个感兴趣的基因的siRNA的设计问题可以通过生物信息学在计算机中解决，并且在有限数量的体外和体内实验中验证。因此，基于siRNA的药物的开发可能比经典药物更耗时和成本更低，并且可以进一步促进个性化医疗的发展。最近在体内递送siRNA方面的突破使得能够开始使用基于siRNA的药物（治疗高脂血症、年龄相关性黄斑变性等）的许多临床试验，其中一些达到晚期（例如TTR淀粉样变性）。然而，尽管在该领域取得了这些重要的成功，siRNA的癌细胞特异性递送/活化仍然是一个问题，对此没有简单和通用的解决方案。因此，siRNA药物的靶点数量仅限于基因，这些基因要么强烈过表达，要么只存在于癌细胞中。癌症特异性前药的应用不仅有望扩大siRNA药物的可能靶点，而且还有望减少治疗的副作用。在这个项目中，我们将致力于设计癌症特异性siRNA前药的简单解决方案，这些前药在活性氧（ROS）的存在下被激活。细胞特异性将基于癌细胞过度产生ROS的实验事实，而它们在正常细胞中的浓度可以忽略不计。ROS诱导的活化是设计癌症特异性siRNA的潜在通用方法，因为升高的ROS被认为对癌症表型至关重要。因此，所提出的siRNA前药应该对许多不同的癌症类型具有活性。作为原理验证，将开发靶向Ubr连接酶的ROS响应性siRNA前药，并在小鼠模型中治疗肝细胞癌（HCC）。选择该模型是因为目前尚无满意的HCC治疗方法。此外，肝特异性递送siRNA的方法已得到很好的开发。在该项目中制定的原则将可能适用于设计用于治疗肝脏以外器官癌症的siRNA前药，前提是器官特异性递送将在未来可用。本研究利用了（a）Mokhir小组在“笼状”siRNA的合成和细胞应用方面的丰富经验，以及（B）Zatsepin小组在siRNA设计和制剂优化以及基于siRNA的缀合物的合成和应用方面的丰富经验，以抑制小鼠模型体内的基因表达。