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: 7844933
• 负责人: William Robb MacLellan
• 金额: $ 19.25万
• 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/7844933
• 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）：内细胞团的细胞分化为形成构成活生物体的复杂组织所需的特化细胞，传统上被视为单向过程，胚胎中的细胞逐渐致力于特定的细胞类型。然而，体细胞核移植实验表明，卵母细胞可以使成体分化细胞的细胞核恢复为多能胚胎样状态。虽然人们对诱导这一过程的因素知之甚少，但最近的几份报告描述了四种转录因子的能力，这些转录因子的逆转录病毒过度表达能够在小鼠成纤维细胞中诱导多能状态。同时过表达多能性相关 POU 结构域 5 类转录因子 1 (Oct3/4)、含有基因 2 (Sox2)、原癌基因 myc (c-Myc) 和 Kruppel 样因子 4 (Klf4) 的 SRY-box，导致诱导多能干 (iPS) 细胞的产生，其表现出与胚胎干 (ES) 相似的形态和生长特性 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.这些人类 iPS 细胞具有正常的核型，表达端粒酶活性、细胞表面标记和人类 ES 细胞的典型基因，并保持分化为所有三个初级胚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使用人类 ES 细胞的争议性需求，而且还提供了一种可能生成定制的、患者特异性多能细胞的方法，用于包括心血管组织工程在内的再生医学工作。然而，这并不排除需要严格研究 iPS 细胞的分化行为，因为定向分化方案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，小鼠 iPS 细胞可以分化为心血管和造血谱系的细胞，并且有可能从具有分化为心血管谱系所有三种细胞类型的能力的分化 iPS 细胞中分离出 Flk1 阳性祖细胞。直接对体细胞进行重编程以产生与患者匹配的多能干细胞，这些干细胞可以作为自体材料的无限来源，可能会彻底改变心脏病的治疗； however, the differentiation and therapeutic capacity of iPS cells is still unknown.该应用将建立在我们已经取得的进展的基础上，进一步探索 iPS 衍生的心血管祖细胞的生物学和治疗潜力。 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.将分化的人类体细胞重编程为多能状态不仅可以消除有争议的人类 ES 细胞使用的需要，而且还可以提供一种机制来生成定制的、患者特异性的多能细胞，用于包括心血管组织工程在内的再生医学工作。