Recombinant microRNAs in xenobiotic and nutrient disposition

重组 microRNA 在异生素和营养物质配置中的作用

基本信息

批准号：
10576880
负责人：
Aiming Yu
金额：
$ 36.89万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10576880
关键词：
Amino Acid Transporter Amino Acids Anabolism Animal Disease Models Basic Science Bioenergetics Biological Biological Assay Biomedical Engineering Biotechnology Cell Survival Cells Cellular Metabolic Process Chemicals Collection Coupled Data Development Disease Drug Exposure Drug Modulation Effectiveness Endotoxins Engineering Enzymes Family Fermentation Fluorouracil Gene Expression Genome Glucose Glucose Transporter Glycolysis Homeostasis Human Human Cell Line In Vitro Knowledge Laboratories Liposomes Malignant Epithelial Cell Mediating Metabolic Metabolism MicroRNAs Modeling Modification Nutrient Oncolytic Oxidases Pharmaceutical Preparations Pharmacodynamics Post-Transcriptional Regulation Primary carcinoma of the liver cells Production Proliferating Proteins Pyridoxal Phosphate RNA Recombinants Regulation Research Role SLC2A1 gene Safety Seminal Techniques Technology Testing Therapeutic Transfer RNA Untranslated RNA Vitamin B6 Vitamin B6 Metabolism Pathway Vitamins Xenobiotic Metabolism Xenobiotics biomaterial compatibility clinically relevant cofactor design drug action drug development drug disposition drug efficacy improved in vivo in vivo Model innovation inorganic phosphate insight inter-individual variation invention large scale production macromolecule metabolome nanocomplexes new therapeutic target novel novel therapeutic intervention nutrient metabolism peptide chemical synthesis polypeptide posttranscriptional pre-miRNA protein expression research and development response solute success sugar synergism technology platform therapy outcome tool tumor xenograft tumorigenesis uptake

项目摘要

PROJECT SUMMARY MicroRNAs (miRNAs or miRs) are genome-derived, functional noncoding RNA (ncRNA) molecules that govern posttranscriptional regulation of target gene expression in cells. Our long-term objective is to gain a mechanistic understanding of miRNA-controlled regulation of xenobiotic & nutrient disposition and drug actions towards the development of new therapeutic strategies. My laboratory pioneered the research on miRNA pharmacoepigenetics that has offered new insights into inter-individual variability in drug disposition and response. Since the diseased carcinoma cells are addicted to continuous supply and metabolism of key xenobiotic nutrients (e.g., amino acids or AAs, vitamins, sugars, etc.) for uncontrolled proliferation and tumorigenesis, understanding the roles of metabolic enzymes (e.g., pyridoxine-5-prime-phosphate oxidase or PNPO) and transporters (e.g., AA transporter SLC7A5/LAT1, glucose transporter SLC2A1/GLUT1, etc.) in nutrient metabolism and transport may help to identify new therapeutic targets. Yet, there is a critical gap in the understanding of important regulatory factors and mechanisms underlying key nutrients’ disposition and homeostasis in carcinoma cells. In addition, current research on miRNA functions and therapeutics are limited to the use of chemo-engineered miRNA “mimics” made in vitro and comprised of extensive and various types of chemical modifications, which are completely different from natural RNA molecules produced and folded in living cells without any or just carrying a limited number of posttranscriptional modifications. This is also in sharp contrast to protein research and therapy that have found ultimate success by using recombinant or bioengineered proteins produced and folded in living cells, rather than polypeptides or proteins synthesized chemically in vitro. Very recently, we have established a novel tRNA/pre-miRNA-based RNA bioengineering technology that permits high-yield and large-scale production of recombinant miRNA agents through in vivo bacterial fermentation. Our studies have showed that recombinant miRNAs are biologically active in regulating target gene expression in human cells, and subsequently modulate drug disposition and response. Furthermore, our preliminary studies have revealed that particular human tRNA (htRNA) can be coupled with human pre-miRNA (hsa-pre-miR) as novel carriers for the production of fully-humanized recombinant or bioengineered miRNA agents, namely hBERA/miRNA. Therefore, in this application, we proposed to (1) establish and utilize novel ncRNA carriers to produce a collection of recombinant hBERA/miRNA molecules (Aim 1), (2) delineate the mechanistic actions of recombinant miRNAs in the control of cellular vitamin B6 metabolism and AA metabolome/homeostasis (Aim 2), and (3) define the effectiveness of recombinant miRNAs in the modulation of pharmacodynamics in disease animal models in vivo (Aim 3). The proposed research will establish a one-of-a-kind RNA biotechnology, and define the mechanistic actions of several human miRNAs in the control of xenobiotic/nutrient metabolism as well as their applications to improving drug efficacy.

项目概要 MicroRNA（miRNA 或 miR）是基因组衍生的功能性非编码 RNA (ncRNA) 分子，控制细胞中靶基因表达的转录后调控。外源物质、营养物质配置和药物的 miRNA 控制调节的机制理解我的实验室率先开展了以下研究： miRNA 药物表观遗传学为药物处置的个体间差异提供了新的见解由于患病癌细胞对关键物质的持续供应和代谢成瘾。外源营养素（例如氨基酸或 AA、维生素、糖等）可导致不受控制的增殖和肿瘤发生，了解代谢酶（例如吡哆醇-5-磷酸氧化酶）的作用或 PNPO）和转运蛋白（例如 AA 转运蛋白 SLC7A5/LAT1、葡萄糖转运蛋白 SLC2A1/GLUT1 等）营养代谢和运输方面的研究可能有助于确定新的治疗靶点，但仍存在重大差距。了解关键营养素配置背后的重要调节因素和机制此外，目前关于 miRNA 功能和治疗的研究正在进行中。仅限于使用体外制造的化学工程 miRNA“模拟物”，并由广泛和多种化学修饰，与天然RNA分子完全不同在活细胞中产生和折叠，没有任何或仅携带有限数量的转录后这也与找到终极的蛋白质研究和疗法形成鲜明对比。成功是通过使用在活细胞中产生和折叠的重组或生物工程蛋白质，而不是最近，我们建立了一种新的体外化学合成的多肽或蛋白质。基于tRNA/pre-miRNA的RNA生物工程技术，可实现高产、大规模生产我们的研究表明，通过体内细菌发酵重组 miRNA 制剂。重组 miRNA 在调节人类细胞中的靶基因表达方面具有生物活性，并且随后调节药物处置和反应。特定的人类 tRNA (htRNA) 可以与人类 pre-miRNA (hsa-pre-miR) 偶联作为新型载体用于生产完全人源化重组或生物工程 miRNA 制剂，即因此，在本申请中，我们建议（1）建立和利用新型ncRNA载体。产生重组 hBERA/miRNA 分子的集合（目标 1），（2）描述机制重组 miRNA 在控制细胞维生素 B6 代谢和 AA 中的作用代谢组/稳态（目标 2）和（3）定义重组 miRNA 在体内疾病动物模型的药效学调节（目标 3）。建立独一无二的 RNA 生物技术，并定义多种人类 miRNA 的机制作用控制异生物质/营养素代谢及其在提高药物功效方面的应用。