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Alternative pre-mRNA splicing of mu opioid receptor gene and mu opioid actions

mu阿片受体基因的选择性前mRNA剪接和mu阿片作用

基本信息

批准号：
10166814
负责人：
YING-XIAN PAN
金额：
$ 36.5万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10166814
关键词：
Absence of pain sensation Affinity Chromatography Agonist Alternative Splicing Analgesics Animal Model Animals Antisense Oligonucleotides Artificial Chromosomes Attenuated Bacteria C-terminal Cell model Cells Clinical Complex Coupled Coupling Dependence Drug abuse Elements Event Exons GTP-Binding Proteins Genes Goals Heroin Human In Vitro Individual Knockout Mice Knowledge Length Mediating Molecular Molecular Chaperones Monitor Morphine Mus Neuroblastoma Oligonucleotides Opioid Opioid Analgesics Pain Pain management Pharmacology Phosphorylation Physical Dependence Play Protein Isoforms Proteins Pruritus RNA RNA Splicing RNA-Protein Interaction Receptor Gene Regulation Reporter Research Proposals Rewards Rodent Role Scientist Small Interfering RNA Sorting - Cell Movement Structure Structure of thyroid parafollicular cell Tail Trans-Activators Transmembrane Domain Variant base cis acting element deletion analysis drug of abuse high throughput screening human disease in vivo insight mRNA Expression mRNA Precursor morphine tolerance mouse model mu opioid receptors novel novel therapeutics receptor side effect

项目摘要

Project Summary/Abstract Mu opioid receptors play a fundamental role in mediating the actions of morphine and most clinical analgesics, as well as drugs of abuse, such as heroin. The single-copy mu opioid receptor gene (OPRM1) creates an array of splicing variants by undergoing extensive alternative pre-mRNA splicing (AS), that is conserved from rodents to humans. These splice variants are categorized into three types based on receptor structure: 1) Full- length carboxyl (C-) terminal variants with 7-transmembrane (TM) domains; 2) Truncated variants containing 6- TM domains; and 3) Truncated variants containing single TM. Increasing evidence suggests that the OPRM1 AS variants are pharmacologically important. Several C-terminal variants display marked differences in region- specific expression, mu agonist-induced G protein coupling, phosphorylation, internalization and post- endocytic sorting. MOR-1D is responsible for morphine-induced itch. Dysregulation of several variant mRNA expressions has been observed in a number of cell and animal models, as well as human diseases. Evidence from our three knockout mouse models suggest that individual C-terminal sequences generated through alternative 3' splicing play distinct roles in various morphine actions, including tolerance, physical dependence and reward. Additionally, intracerebroventricular (i.c.v.) administration of an antisense vivo-morpholino antisense oligo, which blocks 3' splicing from exon 3 to exon 7, significantly attenuates morphine tolerance in mice. Together, these studies not only strongly support our hypothesis that OPRM1 alternative splicing contributes to the regulation of complex opioid actions in animals and humans, but also provide a compelling rationale and scientific premise for further study of the molecular mechanisms controlling OPRM1 alternative splicing and assessing the impact of modulating OPRM1 alternative splicing using antisense vivo-morpholino oligos on morphine actions, as proposed in this application. The primary goal of this application is to further investigate mechanisms and functions of OPRM1 gene alternative splicing by using a variety of in vitro and in vivo approaches. The specific aims include: 1) Decoding molecular mechanisms underlying OPRM1 3' splicing by identifying cis-acting elements and trans-acting factors that regulate the 3' splicing; 2) Investigating the role of the OPRM1 3' splicing in mu opioid actions in both Be(2)C cells and mice using an antisense vivo- morpholino oligo approach. The proposed studies promise to generate significant insights into the mechanism and function of OPRM1 3' splicing, and may have the potential for developing new therapeutics for controlling pain and alleviating the detrimental side-effects of mu opioids.

项目总结/文摘