Recombinant microRNAs in xenobiotic metabolism and disposition

重组 microRNA 在外源代谢和处置中的作用

• 批准号: 10165376
• 负责人: Aiming Yu
• 金额: $ 38.5万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10165376
• 关键词: Address; Animal Model; Area; Biomedical Engineering; Cells; Chemicals; Engineering; Enzymes; Fermentation; Gene Expression; Goals; Hepatocyte; Human; Hybrids; In Vitro; Laboratories; Medical; MicroRNAs; Modification; Molecular; Pharmaceutical Preparations; Pharmacotherapy; Post-Transcriptional Regulation; Process; Prodrugs; Production; Proteins; RNA; Recombinants; Research; Research Proposals; Science; Small Interfering RNA; Technology; Untranslated RNA; Xenobiotic Metabolism; base; body system; cost effective; drug development; drug disposition; drug metabolism; improved; in vivo Model; innovation; large scale production; milligram; novel; pharmacokinetics and pharmacodynamics; polypeptide; recombinant RNA; research and development; success; tRNA Precursor; therapy outcome; tool

PROJECT SUMMARY Understanding the molecular mechanisms behind variable drug metabolism and disposition (DMD) is critical to improve pharmacotherapy and drug development. The ultimate goal of my research is to decipher novel posttranscriptional regulation mechanisms controlled by noncoding microRNAs (miRNAs) and establish their importance in DMD. My laboratory pioneered miRNA research in the fields of DMD that is expected to fill the critical gaps in understanding new posttranscriptional regulation mechanisms. Nevertheless, current studies on miRNA functions in DMD, as well as in the broad areas of general medical sciences, are limited to the use of chemo-engineered RNA “mimics” made in vitro, which, comprised of extensive and various types of chemical modifications, are fundamentally different from natural RNA molecules that are produced and folded in cells and do not carry or just have minimal degree of posttranscriptional modifications. This is also in sharp contrast to protein research that has found the ultimate success by using recombinant or bioengineered proteins produced and folded in living cells, instead of synthetic polypeptides or proteins made in vitro. Very recently, we have developed a novel recombinant RNA technology, based upon hybrid tRNA/pre-miRNA carriers, which offers high-yield and large-scale production of recombinant miRNA agents (& small interfering RNAs or siRNAs) through cost-effective bacterial fermentation. Our studies have also demonstrated that bioengineered miRNA agent (BERA) acts as a “prodrug”, which is specifically processed to target miRNA molecule in human cells to selectively regulate target gene expression. Further, the access to large quantity (tens milligrams from 1 L bacterial culture) of high-purity (>98%) recombinant miRNA molecules allows us to readily investigate the impact of miRNAs on pharmacokinetics and pharmacodynamics (PK/PD) in animal models in vivo. These highly innovative approaches and one-of-its-kind recombinant miRNA tools shall open up new avenues for miRNA research and development, as well as broadly general medical sciences. In this MIRA application, we propose to produce and utilize one-of-a-kind recombinant miRNAs to dissect new regulatory mechanisms in DMD by addressing the following key questions: 1) Can the tRNA/pre-miRNA-based technology be re-innovated for the production of fully-humanized recombinant miRNAs (hBERA/miRNAs)? 2) Will hBERA/miRNAs be processed to target miRNAs in human hepatocytes and liver cells to selectively modulate drug-metabolizing enzyme or transporter expression, and alter cellular DMD capacity? 3) To what degrees will miRNA alter PK and PD in the whole-body system?

项目摘要 了解可变药物代谢和性格（DMD）背后的分子机制是关键的 改善药物治疗和药物开发。我的研究的最终目标是破译小说 通过非编码microRNA（miRNA）控制的转录后调节机制并建立 它们在DMD中的重要性。我的实验室在DMD领域进行了MiRNA研究，这预计将 填补了解新的转录后调节机制的关键空白。然而，目前 DMD中的miRNA功能以及一般医学科学的广泛领域的研究仅限于 在体外制造的化学工程RNA“模仿”的使用，由广泛和各种类型组成 化学修饰的基本上与产生的天然RNA分子根本不同，并且 折叠在细胞中，不携带或仅具有最低程度的转录后修饰。这也是 与蛋白质研究形成鲜明对比的是，蛋白质研究通过使用重组或使用重组或 在活细胞中生产和折叠的生物工程蛋白，而不是合成多肽或蛋白质 在体外制造。最近，我们开发了一种基于混合动力的新型重组RNA技术 tRNA/pre-miRNA载体，可提供高产和大规模生产重组miRNA剂 （＆小型干扰RNA或siRNA）通过具有成本效益的细菌发酵。我们的研究也 证明生物工程miRNA剂（BERA）充当“前药”，专门处理 靶向人类细胞中的miRNA分子以选择性调节靶基因表达。此外，访问 大量（来自1 L细菌培养的毫克数十毫克）高纯度（> 98％）重组miRNA 分子使我们能够轻松研究miRNA对药代动力学和 在体内动物模型中的药效学（PK/PD）。这些高度创新的方法 独一无二的重组miRNA工具应为miRNA研发开辟新的途径， 以及广泛的医学科学。在此Mira应用中，我们建议生产和利用 通过解决该dmd中的新调节机制，通过解决该机制的一种重组miRNA 以下是关键问题：1）可以重新开入基于tRNA/基于miRNA的技术以进行生产 完全人性化的重组miRNA（hbera/miRNA）？ 2）将处理hbera/miRNA以靶向 人肝细胞和肝细胞中的miRNA，以选择性调节药物代谢酶或 转运蛋白表达并改变细胞DMD的能力？ 3）在什么程度上会改变PK和PD 全身系统？