Cardiovascular and Therapeutic Potential of Reprogrammed Human Fibroblasts

重编程人类成纤维细胞的心血管和治疗潜力

• 批准号: 7572264
• 负责人: William Robb MacLellan
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7572264
• 关键词: Adult; Autologous; Behavior; Biology; Boxing; Cardiac; Cardiovascular Diseases; Cardiovascular system; Cell Differentiation process; Cell Line; Cell Nucleus; Cell Therapy; Cell surface; Cells; Chimera organism; Clinical; Clinical Research; Collagen Type IV; Commit; Complex; Derivation procedure; Development; Embryo; Endothelial Cells; Epigenetic Process; Exhibits; Fibroblasts; GKLF protein; Gene Expression; Generations; Genes; Germ Layers; Growth; Heart Diseases; Hematopoietic; Human; Infarction; Inner Cell Mass; Left Ventricular Function; Life; Medical; Methods; Morbidity - disease rate; Morphology; Mus; Mutation; Myocardial Infarction; Myocardial Ischemia; Myocardium; Octamer Transcription Factor-3; Oocytes; Organism; Patients; Pattern; Phosphotransferases; Pluripotent Stem Cells; Process; Property; Proto-Oncogenes; Protocols documentation; Public Health; Regenerative Medicine; Reporting; Research; Research Personnel; Retroviridae; Smooth Muscle; Somatic Cell; Source; Stem cells; System; Telomerase; Tertiary Protein Structure; Testing; Therapeutic; Tissue Engineering; Tissues; Viral; c-myc Genes; cell type; chromatin remodeling; embryonic stem cell; human embryonic stem cell; improved; induced pluripotent stem cell; mortality; overexpression; pluripotency; progenitor; public health relevance; receptor; regenerative; research study; somatic cell nuclear transfer; stem; stem cell technology; transcription factor; tumor

DESCRIPTION (provided by applicant): Differentiation of the cells of the inner cell mass into the specialized cells required for forming the complex tissues that comprise living organisms has traditionally been viewed as a unidirectional process, with cells in the embryo becoming gradually committed to a specific cell type. However, somatic cell nuclear transfer experiments have demonstrated that the oocyte can return the nucleus of an adult differentiated cell into a pluripotent embryonic-like state. While little is known about the factors that induce this process, several recent reports have described the ability of four transcription factors whose retroviral overexpression enabled the induction of a pluripotent state in murine fibroblasts. Simultaneous overexpression of the pluripotency-associated POU domain class 5 transcription factor 1 (Oct3/4), SRY-box containing gene 2 (Sox2), proto-oncogene myc (c-Myc), and Kruppel-like factor 4 (Klf4) led to the generation of induced pluripotent stem (iPS) cells that exhibited morphology and growth properties similar to embryonic stem (ES) cells that were competent for formation of germline chimera. Rrecently investigators have created iPS cells from adult human cells using either a combination of factors similar to the mouse system. These human iPS cells had normal karyotypes, expressed telomerase activity, cell surface markers and genes that typify human ES cells, and maintained the developmental potential to differentiate into advanced derivatives of all three primary germ layers. The successful reprogramming of differentiated human somatic cells into a pluripotent state may not only eliminate the need of controversial use of human ES cells in research applications, it also provides a method to potentially generate customized, patient- specific pluripotent cells for regenerative medicine efforts including cardiovascular tissue engineering. However, this does not obviate the need to critically study the differentiation behavior of iPS cells as directed differentiation protocols will be essential for these stem cell-based therapies to become clinical reality. Our preliminary results suggest that murine iPS cells can be differentiated into cells of the cardiovascular and hematopoietic lineage, and that it is possible to isolate a Flk1-positive progenitor cell from differentiating iPS cells that possesses the ability to differentiate into all three cell types of the cardiovascular lineage. Direct reprogramming of somatic cells to generate patient-matched pluripotent stem cells that could serve as unlimited source of autologous material could revolutionize the treatment of heart disease; however, the differentiation and therapeutic capacity of iPS cells is still unknown. This application will build on the progress we have already made and further explore the biology and therapeutic potential of iPS-derived cardiovascular progenitor cells. PUBLIC HEALTH RELEVANCE: Despite medical advances, cardiovascular disease remains a leading cause of mortality and morbidity. Thus, regenerative cardiovascular therapies that restore normal function would have an enormous societal and financial impact. Reprogramming of differentiated human somatic cells into a pluripotent state may not only eliminate the need of controversial use of human ES cells but it would also provide a mechanism to generate customized, patient-specific pluripotent cells for regenerative medicine efforts including cardiovascular tissue engineering.

描述（由申请人提供）：将内部细胞质量的细胞分化为形成包含活生物体的复杂组织所需的专门细胞，传统上被视为一个单向过程，胚胎中的细胞逐渐致力于特定的细胞类型。然而，体细胞核转移实验表明，卵母细胞可以将成年分化细胞的核返回到多能的胚胎样状态。尽管对引起该过程的因素知之甚少，但最近的几份报告描述了四个转录因子的能力，这些因子逆转录病毒过表达使得能够诱导鼠鼠成纤维细胞中的多能状态。多能相关的POU结构域5类转录因子1（Oct3/4），含有基因2（SOX2），原始癌基因MYC（C-MYC）（C-MYC）和Kruppel因子4（KLF4）的同时过表达，导致了诱导的Pluripotent sys sypriese sypriese sypriese sypriese syply syply syply syply syply syply syply syply syply syply prolipe sys sypory syply。有能力形成种系嵌合体的细胞。研究人员使用类似于小鼠系统的因素组合从成年人类细胞中创建了IPS细胞。这些人IPS细胞具有正常的核型，表达了端粒酶活性，特征的人ES细胞的细胞表面标记和基因，并保持了分化为所有三个主要种植层的高级衍生物的发育潜力。将分化的人类体细胞成功重编程为多能状态不仅可以消除在研究应用中对人ES细胞进行有争议的使用的需求，而且还提供了一种方法，可以使潜在的定制，患者特定的多能细胞用于再生医学努力，包括心血管组织工程。但是，这并不能消除需要批判性地研究IPS细胞的分化行为的必要性对于这些基于干细胞的疗法成为临床现实至关重要。我们的初步结果表明，鼠IPS细胞可以分化为心血管和造血谱系的细胞，并且可以将FLK1阳性祖细胞与分化具有分化为心血管谱系所有三个细胞类型的能力分离的IPS细胞分离。直接重新编程体细胞以产生可以用作无限材料来源的患者匹配的多能干细胞，这可能会彻底改变心脏病的治疗。但是，IPS细胞的分化和治疗能力仍然未知。该应用将建立在我们已经取得的进展的基础上，并进一步探索IPS来源的心血管祖细胞的生物学和治疗潜力。公共卫生相关性：尽管医疗进展，心血管疾病仍然是死亡率和发病率的主要原因。因此，恢复正常功能的再生心血管疗法将产生巨大的社会和财务影响。将分化的人类体细胞重新编程为多能状态，不仅可以消除有争议的人类ES细胞的需求，而且还将提供一种机制，以生成定制的，患者特异性的多能细胞进行再生医学，以进行心血管组织工程（包括心血管组织工程）。