Role of Nitric Oxide and Cyclic GMP in Stem Cells

一氧化氮和环鸟苷酸在干细胞中的作用

基本信息

批准号：
7483772
负责人：
FERID MURAD
金额：
$ 28.22万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
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：
2007-08-10 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7483772
关键词：
Address Affect Animal Disease Models Animal Model Basic Science Biology Cardiac Cardiac Myocytes Cardiovascular Models Cell Lineage Cell Proliferation Cell Separation Cell Transplantation Cells Cellular biology Clinical Research Code Condition Coronary Cyclic GMP Cyclic GMP-Dependent Protein Kinases Cyclic Nucleotides Data Disease Embryo Embryonic Stem Cell Transplantation Engineering Engraftment Experimental Designs Future Gene Expression Profile Goals Growth Guanylate Cyclase Homing Human Immunohistochemistry In Vitro Injury Investigation Ligation Measures Mediating Messenger RNA Modeling Mus Myocardial Neurons Nitric Oxide Nitric Oxide Donors Nitric Oxide Signaling Pathway Nitric Oxide Synthase Outcome Parkinson Disease Pharmaceutical Preparations Play Process Property Proteins Protocols documentation Publishing Purpose Rate Research Reverse Transcriptase Polymerase Chain Reaction Risk Role Signal Pathway Signal Transduction Soluble Guanylate Cyclase Stem cells Structure of beta Cell of islet System Teratoma Tissues Transplantation Undifferentiated United States National Institutes of Health WA09 Cell Line Work cell motility cell type clinical application embryonic stem cell gene delivery system genetic manipulation human embryonic stem cell human stem cells improved in vivo inhibitor/antagonist interest migration nucleotide analog precursor cell progenitor repaired research study self-renewal stem tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): In this revised application, we will primarily focus on studying the role of NO and cyclic GMP signaling in proliferation and differentiation of human and mouse embryonic stem cells into cardiomyocytes. Our preliminary studies strongly suggest a role for nitric oxide and cyclic GMP in mouse and human embryonic stem cell proliferation and differentiation. The mRNA (RT-PCR and Northerns) and protein (Western and activity) in mouse and human stem cells (WA09) will be examined during stem cell proliferation and differentiation for nitric oxide synthases (NOS 1,2,3) and soluble guanylyl cyclase, the receptor for NO. The cellular localization will be confirmed with immunohistochemistry for these proteins and cellular markers and cell sorting. The effects of activators and inhibitors of NOS, soluble guanylyl cyclase, protein kinase G, and cyclic nucleotide phosphodieserases will also be examined to confirm the role of NO and cGMP in stem cell proliferation and differentiation into myocardial cells. With an understanding of the presence or absence of the proteins participating in nitric oxide and cyclic GMP synthesis and function in stem cells and differentiated cells with their in vitro culture, we expect to be able to design experimental protocols with various pharmacologic agents such as NO donor drugs, cyclic nucleotide analogues, phosphodieterase inhibitors, and guanylyl cyclase activators that should modify stem cell proliferation and differentiation into various cellular lineages. The function of these stem cells and differentiated cells will also be examined in several cardiovascular models. This should be extremely valuable information for future clinical studies by others and us for the purpose of tissue replacement and perhaps drug and gene delivery systems. Several colleagues with expertise in stem cell biology and/or in vivo animal models will serve as advisors, consultants, and collaborators. Our preliminary experiments and the work of others indicate that our studies proposed are feasible and are likely to be successful to determine the roles of nitric oxide and cyclic GMP in stem cell proliferation, differentiation, and function.

描述（由申请人提供）：在此修订后的应用中，我们将主要集中于研究NO和环状GMP信号传导在人类和小鼠胚胎干细胞扩散和分化中的作用。我们的初步研究强烈表明一氧化氮和环状GMP在小鼠和人类胚胎干细胞增殖和分化中的作用。在干细胞增殖和一氧化氮合酶（NOS 1,2,3）和可溶性胰鸟叶尼尔环酶的过程中，将检查小鼠和人类干细胞中小鼠和人类干细胞中的mRNA（RT-PCR）和蛋白质（WA09）（WA09）（WA09）的mRNA（WA09）和蛋白质（WA09）。细胞定位将通过免疫组织化学确认这些蛋白质，细胞标记和细胞分类。还将检查NOS，可溶性鸟叶基环酶，蛋白激酶G和环状核苷酸磷酸二酶的活化剂和抑制剂的影响，以确认NO和CGMP在干细胞增殖中的作用以及分化为心肌细胞。了解参与一氧化氧化物和环状GMP合成的蛋白质的存在或不存在，并在干细胞中的功能以及与其体外培养的分化细胞中的分化细胞，我们希望能够使用各种药理剂（如无供体药物）（例如均可构成核苷酸，磷酸固定剂，造成的，造成的，造成的，造成的，造成的，造成的，造成的，造成的，造成的，造成刺激剂，以及造成的，造成的构造剂，造成刺激性的构造，并将其磷酸化型构造构成构造，并构成了造成的构造，并将其磷酸化构造构成构造剂，并构成了造成的构造。分化为各种细胞谱系。这些干细胞和分化细胞的功能也将在几种心血管模型中进行检查。对于组织置换以及药物和基因递送系统的目的，这对于其他人和我们的未来临床研究应该是极其有价值的信息。几位具有干细胞生物学专业知识和/或体内动物模型的专业知识的同事将担任顾问，顾问和合作者。我们的初步实验和其他人的工作表明，我们提出的研究是可行的，很可能成功地确定一氧化氮和环状GMP在干细胞增殖，分化和功能中的作用。