An in vivo screen for biological and chemical regulators of mammalial PDEs

哺乳动物 PDE 生物和化学调节剂的体内筛选

基本信息

批准号：
7337165
负责人：
CHARLES S. HOFFMAN
金额：
$ 19.43万
依托单位：
BOSTON COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7337165
关键词：
Adenylate Cyclase Adverse effects Allergic rhinitis Alzheimer&apos s Disease Behavior Biological Biological Assay Biological Process Candidate Disease Gene Cardiac Cells Chemicals Condition Cyclic AMP Cyclic GMP Cystic Fibrosis Detection Development Disease Dot Immunoblotting Engineering Environment Enzymes Facility Construction Funding Category Family Fission Yeast Fluorescence Future Genes Genetic Transcription Glucose Goals Growth Hematologic Neoplasms Huntington Disease Individual Isoenzymes Kidney Diseases Lead Libraries Lung Malignant Neoplasms Measurement Measures Mental Depression Multiple Sclerosis Mus Mutation PDE4A4 PDE4B Pathway interactions Pharmacotherapy Phenotype Phosphodiesterase Inhibitors Pilot Projects Preclinical Drug Evaluation Process Psoriasis RNA Splicing Range Rate Regulation Relative (related person)Reporter Repression Research Personnel Resistance Rheumatoid Arthritis Role Schizophrenia Screening procedure Second Messenger Systems Signal Pathway Signal Transduction Specificity System Therapeutic Therapeutic Index Thinking Tissues Treatment Efficacy Variant Work Yeasts base cDNA Library cell growth diethylstilbestrol monophosphate expression vector high throughput screening human tissue in vitro Assay in vivo inhibitor/antagonist member next generation novel phosphoric diester hydrolase programs research study response second messenger small molecule libraries

项目摘要

Many biological processes are regulated by a cell's ability to sense molecules in its environment and create an intracellular signal to effect an appropriate biological response. One major signaling pathway involves the regulation of cAMP levels, which is a function of cAMP synthesis by adenylate cyclases and cAMP destruction by cAMP phosphodiesterases (PDE). In the fission yeast, Schizosaccharomyces pombe, cAMP levels are regulated by a glucose signaling pathway that includes a single PDE gene. We have developed reporter constructs,which confer growth phenotypes that reflect the cell's intracellular cAMP level. We propose to introduce mammalian PDE genes into our strains, such that the growth behavior will be a function of PDE activity. We will use such strains to carry out the following two aims. 1) We will conduct high throughput screening for chemical inhibitors of specific PDEs. Utilizing strains expressing various murine PDEs (4A, 4B, 8A, 8B), we expect to identify both nonspecific and specific inhibitors. Of note, there are no known PDE8-specific inhibitors, thus making it difficult to determine the relative role of PDES enzymes in various biological processes. 2) We will use these strains to screen a cDNA library for biological activatorsof the target PDE and identify the tissues in which these activatorsare expressed. Strains expressing both the activator and the target PDE will be subjected to chemical library screens for compounds that inhibit the activator, rather than the PDE itself. As these activators may be expressed in a subset of tissues in which the PDE is found, compounds that target the activator may provide a more tissue-specific effect on PDE activity, and thus provide a therapeutic benefit with less of a side-effect than would be possible for compounds that target the PDE directly. There is a broad range of diseases that are currently being treated with PDE inhibitors, or are thought to be amenable to treatment with PDE inhibitors. Therefore, the development of this in vivo platform to identify chemical and biological regulators of PDEs has the potential of identifying the next generation of PDE-related Pharmaceuticalsfor the treatment of cardiac, pulmonary, and renal diseases, as well as certain cancers, cystic fibrosis, multiple sclerosis, rheumatoid arthritis, Huntington's Disease, allergic rhinitis, psoriasis, schizophrenia, Alzheimer's disease and depression.

许多生物过程是由细胞感知其环境中的分子并产生化学反应的能力来调节的。产生适当的生物反应的细胞内信号。一个主要的信号通路涉及 cAMP 水平的调节，这是腺苷酸环化酶和 cAMP 合成 cAMP 的功能 cAMP 磷酸二酯酶 (PDE) 破坏。在裂殖酵母中，裂殖酵母 (Schizosaccharomyces pombe) 中，cAMP 水平由包含单个 PDE 基因的葡萄糖信号传导途径调节。我们开发了报告基因构建体，赋予反映细胞内 cAMP 水平的生长表型。我们建议将哺乳动物 PDE 基因引入我们的品系中，使生长行为成为函数 PDE 活性。我们将利用这些菌株来实现以下两个目标。 1）我们将进行高特定 PDE 化学抑制剂的通量筛选。利用表达各种小鼠的菌株 PDE（4A、4B、8A、8B），我们期望识别非特异性和特异性抑制剂。值得注意的是，没有已知的 PDE8 特异性抑制剂，因此很难确定 PDES 酶在各种生物过程。 2) 我们将使用这些菌株来筛选 cDNA 文库的生物激活剂目标 PDE 并识别表达这些激活剂的组织。表达两者的菌株激活剂和目标 PDE 将接受化学库筛选，寻找抑制 PDE 的化合物激活器，而不是偏微分方程本身。由于这些激活剂可能在一部分组织中表达，其中发现 PDE，靶向激活剂的化合物可能对 PDE 活性提供更具组织特异性的作用，从而提供比以下化合物可能具有的治疗益处和更少的副作用直接针对 PDE。目前正在使用 PDE 治疗多种疾病抑制剂，或被认为适合用 PDE 抑制剂治疗。因此，发展这个用于识别 PDE 的化学和生物调节剂的体内平台有可能识别下一代 PDE 相关药物用于治疗心脏、肺和肾脏疾病，以及某些癌症、囊性纤维化、多发性硬化症、类风湿性关节炎、亨廷顿病、过敏性鼻炎、牛皮癣、精神分裂症、阿尔茨海默病和抑郁症。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Use of a Fission Yeast Platform to Identify and Characterize Small Molecule PDE Inhibitors.

DOI：
10.3389/fphar.2021.833156
发表时间：
2021
期刊：
Frontiers in pharmacology
影响因子：
5.6
作者：
Hoffman CS
通讯作者：
Hoffman CS

A yeast-based chemical screen identifies a PDE inhibitor that elevates steroidogenesis in mouse Leydig cells via PDE8 and PDE4 inhibition.

DOI：
10.1371/journal.pone.0071279
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Demirbas D;Wyman AR;Shimizu-Albergine M;Cakici O;Beavo JA;Hoffman CS
通讯作者：
Hoffman CS

Fission yeast-based high-throughput screens for PKA pathway inhibitors and activators.

基于裂变酵母的 PKA 途径抑制剂和激活剂高通量筛选。

DOI：
10.1007/978-1-4939-2269-7_6
发表时间：
2015
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
deMedeiros,AnaSantos;Kwak,Grace;Vanderhooft,Jordan;Rivera,Sam;Gottlieb,Rachel;Hoffman,CharlesS
通讯作者：
Hoffman,CharlesS

Methods to Assess Phosphodiesterase and/or Adenylyl Cyclase Activity Via Heterologous Expression in Fission Yeast.

通过裂殖酵母中的异源表达评估磷酸二酯酶和/或腺苷酸环化酶活性的方法。