Genetic information transfer to hematopoietic cells: Role of microvesicles

遗传信息传递至造血细胞：微泡的作用

• 批准号: 8064149
• 负责人: PETER J. QUESENBERRY
• 金额: $ 38.63万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8064149
• 关键词: Address; Alveolar Cell Type I; Antibodies; B-Lymphocytes; Behavior; Blood Vessels; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Brain; Cancerous; Candidate Disease Gene; Cell Culture Techniques; Cell Lineage; Cell Separation; Cells; Cellular biology; Clara cell-specific protein; Clinical; Coculture Techniques; Disease; Electrophoresis; Endothelial Cells; Epithelial; Epithelial Cells; Genes; Genetic; Genetic Transcription; Genome; Heart; Hematopoietic; Hematopoietic stem cells; Immunohistochemistry; In Vitro; Individual; Label; Lead; Liver; Lung; Lung diseases; Malignant - descriptor; Marrow; Mediating; Membrane; Messenger RNA; MicroRNAs; Molecular; Mus; Mutation; PKH 26; Phenotype; Population; Proteins; Proteome; Publishing; Relative (related person); Reverse Transcriptase Polymerase Chain Reaction; Role; Smooth Muscle Myocytes; T-Lymphocyte; Thinking; Time; Tissues; Transplantation; Two-Dimensional Gel Electrophoresis; Vascular Smooth Muscle; Vesicle; Viral Vector; Work; Wound Healing; alpha Actin; alveolar type II cell; aquaporin 5; base; cancer therapy; cellular transduction; comparative; cytokine; granulocyte; in vivo; mRNA Expression; macrophage; monocyte; novel; novel strategies; prevent; programs; research study; surfactant; tissue regeneration; transcription factor; uptake; von Willebrand Factor; water channel

DESCRIPTION (provided by applicant): We have previously demonstrated that membrane enclosed microvesicles derived from murine lung cells contain lung-specific mRNA, micro RNA and protein and that these microvesicles enter murine marrow cells and effect a change in genetic phenotype manifest by expression of the lung-specific mRNAs; surfactants A-D, aquaporins and clara cell specific protein and in function manifest by increased conversion to epithelial lung cells after transplantation into lethally irradiated mice. The observed mRNA changes appear to be due to transfer of both mRNA and of a tissue specific transcriptional protein. Murine heart, liver and brain also effect tissue specific genetic changes in target marrow cells. The present proposal is a natural extension of these now published studies. We plan to elucidate the specific target cells in marrow for microvesicle genetic changes and will also study individual lung cells for their specific capacity to induce genetic change in marrow; previous studies were with whole lung. We plan to evaluate the stability of the observed genetic changes in marrow both in vitro and in vivo. Lastly, we will extend our characterization of the potential for reprogramming and attempt to identify specific transcriptional proteins or microRNA which may mediate the observed phenotype changes. To carry out these studies we will employ fluorescent activated cell sorting of cell populations, Real-time RT-PCR, immunohistochemistry, cell culture, murine marrow transplantation, SILAC to study protein transfer, lente viral vector transduction of cells and comparative 2-D gel electrophoresis and difference electrophoresis (DIGE). These studies should establish a more precise platform for studies of microvesicles, establish the stability of microvesicle induced genetic changes and begin to unravel the mechanisms behind such changes. This work could alter our thinking toward cell biology in general and open new strategies for approaching a number of diseases characterized by tissue degeneration or damage. PUBLIC HEALTH RELEVANCE: The focus of this project is to determine if vesicles shed by lung cells are able to influence the identity of bone marrow cells that consume them to the point that they behave like lung cells. Such an approach could lead to new approaches for tissue regeneration and repair for a variety of lung diseases. Alternatively, by preventing the transfer of malignant behavior from cancerous cells to non-cancerous cells, novel therapies for cancer could be developed.

描述(申请人提供)：我们先前已经证明来自小鼠肺细胞的膜包被微囊含有肺特异的mRNA、Micro RNA和蛋白质，并且这些微囊进入小鼠骨髓细胞后，通过表达肺特异的mRNAs、表面活性物质A-D、水通道蛋白和Clara细胞特异的蛋白来改变遗传表型，并且在移植到致死性照射的小鼠体内后，通过增加向上皮细胞的转化来显示其功能。观察到的信使核糖核酸的变化似乎是由于信使核糖核酸和一种组织特异性转录蛋白的转移。小鼠的心脏、肝脏和大脑也会影响目标骨髓细胞的组织特异性基因变化。目前的建议是这些现已发表的研究的自然延伸。我们计划阐明骨髓中微泡基因变化的特定靶细胞，并将研究单个肺细胞诱导骨髓基因变化的特定能力；以前的研究是针对整个肺的。我们计划评估在体外和体内观察到的骨髓基因变化的稳定性。最后，我们将扩展我们对重新编程潜力的描述，并试图识别可能介导观察到的表型变化的特定转录蛋白或microRNA。为了进行这些研究，我们将使用荧光激活的细胞分类，实时定量RT-PCR，免疫组织化学，细胞培养，小鼠骨髓移植，SILAC来研究蛋白质转移，Lente病毒载体转导细胞，以及比较的双向凝胶和差异电泳(DGE)。这些研究应该为微囊泡的研究建立一个更精确的平台，建立微囊泡诱导的遗传变化的稳定性，并开始揭开这些变化背后的机制。这项工作可能会改变我们对细胞生物学的总体看法，并为解决一些以组织退化或损伤为特征的疾病开辟新的策略。 与公共卫生相关：该项目的重点是确定肺细胞释放的小泡是否能够影响消耗它们的骨髓细胞的身份，以至于它们的行为与肺细胞相似。这种方法可能会为各种肺部疾病的组织再生和修复带来新的方法。或者，通过防止恶性行为从癌细胞转移到非癌细胞，可以开发癌症的新疗法。