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: 10407500
• 负责人: Aiming Yu
• 金额: $ 38.42万
• 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/10407500
• 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?

项目概要 了解可变药物代谢和处置 (DMD) 背后的分子机制至关重要 我研究的最终目标是破译新的药物治疗和药物开发。 由非编码 microRNA (miRNA) 控制的转录后调控机制并建立 它们在 DMD 中的重要性 我的实验室开创了 DMD 领域的 miRNA 研究，预计将 填补了理解新的转录后调控机制的关键空白。 关于 miRNA 在 DMD 中的功能以及普通医学的广泛领域的研究仅限于 使用体外制造的化学工程 RNA“模拟物”，其中包括广泛且多种类型 化学修饰，与产生和产生的天然RNA分子根本不同 折叠在细胞中并且不携带或仅具有最小程度的转录后修饰。 与通过使用重组或重组获得最终成功的蛋白质研究形成鲜明对比 在活细胞中产生和折叠的生物工程蛋白质，而不是合成的多肽或蛋白质 最近，我们开发了一种基于杂交的新型重组 RNA 技术。 tRNA/pre-miRNA 载体，可高产、大规模生产重组 miRNA 试剂 （以及小干扰 RNA 或 siRNA）通过具有成本效益的细菌发酵。 证明生物工程 miRNA 药物 (BERA) 作为“前药”，经过专门加工 进一步靶向人类细胞中的miRNA分子来选择性调节靶基因的表达。 大量（1 L 细菌培养物中数十毫克）高纯度 (>98%) 重组 miRNA 分子使我们能够轻松研究 miRNA 对药代动力学的影响 这些高度创新的方法和体内动物模型的药效学（PK/PD）。 独一无二的重组 miRNA 工具将为 miRNA 研究和开发开辟新途径， 以及广泛的一般医学科学。在此 MIRA 应用中，我们建议生产和利用。 一种独一无二的重组 miRNA，通过解决 DMD 的新调控机制 以下关键问题：1）基于tRNA/pre-miRNA的技术是否可以重新创新用于生产 完全人源化的重组 miRNA (hBERA/miRNA) 2) hBERA/miRNA 是否会被加工成靶点？ 人类肝细胞和肝细胞中的 miRNA 可选择性调节药物代谢酶或 转运蛋白表达，并改变细胞 DMD 能力？ 3) miRNA 会在多大程度上改变 PK 和 PD 全身系统？