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.

许多生物过程受细胞在环境中感知分子的能力并创建的能力来调节细胞内信号可实现适当的生物学反应。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的cAMP合成的函数营地磷酸二酯酶（PDE）的破坏。在裂变酵母中，schizosaccharomyces pombe，camp 水平由包括单个PDE基因的葡萄糖信号通路调节。我们已经发展了记者构造，它赋予反映细胞细胞内营地水平的生长表型。我们建议将哺乳动物PDE基因引入我们的菌株，以使生长行为将是一种功能 PDE活性。我们将使用此类菌株来执行以下两个目标。 1）我们将高涨特定PDE的化学抑制剂的吞吐量筛选。利用表达各种鼠的菌株 PDE（4A，4B，8A，8B），我们希望识别非特异性和特定抑制剂。值得注意的是，没有已知的PDE8特异性抑制剂，因此很难确定PDE酶在各种生物过程。 2）我们将使用这些菌株筛选cDNA库的生物激活剂靶PDE并识别这些激活剂表达的组织。表达的菌株激活剂和靶PDE将受到化学库筛选的抑制化合物激活器，而不是PDE本身。由于这些激活剂可以在组织的一部分中表达发现PDE，靶向激活剂的化合物可能会对PDE活性产生更大的特异性影响，因此，提供的治疗益处的副作用较小，而不是直接靶向PDE。当前正在用PDE治疗的广泛疾病抑制剂，或被认为可以接受PDE抑制剂治疗。因此，发展这个识别PDE的化学和生物调节剂的体内平台具有识别的潜力下一代与PDE相关的药物治疗心脏，肺部和肾脏疾病，以及某些癌症，囊性纤维化，多发性硬化症，类风湿关节炎，亨廷顿氏病，过敏性鼻炎，牛皮癣，精神分裂症，阿尔茨海默氏病和抑郁症。

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

New classes of PDE7 inhibitors identified by a fission yeast-based HTS.