Molecular regulation underlying differentiation of dental pulp stem cells

牙髓干细胞分化的分子调控

基本信息

批准号：
8812561
负责人：
Shaomian Yao
金额：
$ 36.22万
依托单位：
LOUISIANA STATE UNIV A&M COL BATON ROUGE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8812561
关键词：
3&apos Untranslated Regions Adult Binding Bioinformatics Bone Marrow Stem Cell Transplantation Cell Separation Cell Therapy Cells Code Dental Dental Pulp Development Disease Down-Regulation Elements Ethical Issues Fluorescence Future Gene Expression Gene Proteins Genes Global Change Goals Heat shock proteins Immune response In Vitro Injury Late Effects Light Medicine Messenger RNA Methods MicroRNAs Molecular Natural regeneration Northern Blotting Organ Patients Play Polyadenylation Porifera Property Proteins Proteome Rattus Regenerative Medicine Regulation Regulator Genes Regulatory Element Research Response Elements Reverse Transcriptase Polymerase Chain Reaction Role Signal Transduction Small RNA Source Stem cells Teratoma Testing Therapeutic Time Tissue Engineering Tissues Tooth structure Transfection Translations Transplantation Two-Dimensional Gel Electrophoresis Western Blotting adult stem cell design embryonic stem cell expression vector gain of function human disease improved induced pluripotent stem cell leukemia loss of function novel novel strategies overexpression prevent public health relevance repaired stem stem cell differentiation stemness tissue regeneration tumorigenesis two-dimensional vector

项目摘要

DESCRIPTION (provided by applicant): Utilization of stem cells to repair diseased or damaged tissues/organs represents a novel approach for remedies in regenerative medicine. Although embryonic stem cells (ESC) and induced pluripotent stem cells (iPS) have been studied, there are major drawbacks and concerns in using ESC and iPS; for example, tumorigenesis and teratomas formation can occur after their transplantation. It is not clear if the obstacles pertaining to ESC and iPS can be overcome in order to use them for therapeutics; at least there is still long way to go. In contrast, adult stem cells (AdSC) are used for treatment of human diseases. One example is the transplantation of bone marrow stem cells to cure or improve certain disorders, such as leukemia. Such cell- based therapies usually require large numbers of stem cells. Because AdSC lose their stem cell properties (stemness) including differentiation capability when cultured in vitro, primary isolated AdSC can only be used for limited passages, and propagation of a large quantity of high-potential stem cells from primary isolation is difficult. Thus, large amounts of tissues are needed for stem cell isolation, and continuous isolation of AdSC is required for multiple treatments. However, tissues, e.g., dental tissues that can be used for AdSC isolation are often limited. Stem cells isolated from dental pulp of extracted and exfoliated teeth have been shown to be a good source of stem cells for regenerative medicine. However, like other AdSC, these dental pulp stem cells (DPSCs) also lose their differentiation potential when cultured in vitro, which hampers their applications. The long-term goal of this research is to elucidate the genes and molecules regulating differentiation capability in the DPSCs. Preliminary studies showed that heat shock protein B8 (HspB8) was downregulated in cultured DPSCs when the cells lost differentiation capability, and knockdown of HspB8 expression in the early passage DPSCs caused the cells to lose differentiation capability. Further studies suggested that microRNAs (miRNAs) are responsible for downregulation of HspB8 expression in DPSCs. The central hypothesis is that upregulated expression of certain miRNAs in late passage DPSCs downregulates HspB8 and other regulatory genes, and in turn causes DPSCs to lose differentiation capability during in vitro culture. The project Aims: (1) to identify the miRNAs whose expression is substantially increased in late passage DPSCs, designated as late-passage upregulated miRNAs (LPU-miRNAs); (2) to identify the LPU-miRNAs that downregulate HspB8 expression; i.e., HspB8-targeting miRNAs, and (3) to study the effects of LPU-miRNAs and HspB8-targeting miRNAs on DPSC differentiation and on regulating other potential genes. Accomplishment of the Aims would be significant in understanding the roles of miRNAs and HspB8 in regulating the loss of differentiation seen in the long-term cultured DPSCs and will shed light on other AdSC as well. This would facilitate the development of cutting- edge methods for maintaining the stem cell properties in culturing DPSCs and AdSC, such that a large quantity of stem cells with high-differentiation potential can be obtained from expansion of primary isolated stem cells.

描述（通过应用提供）：使用干细胞修复解散或受损的组织/器官代表了再生医学中的一种新方法。尽管胚胎干细胞（ESC）和诱导的多能干细胞（IPS）已经研究了，但使用ESC和IPS存在主要的缺点和关注。例如，肿瘤发生后可能发生在移植后。目前尚不清楚是否是可以克服与ESC和IP有关的障碍，以便将其用于治疗；至少还有很长的路要走。相反，成年干细胞（ADSC）用于治疗人类疾病。一个例子是骨髓干细胞的移植以治愈或改善某些疾病，例如白血病。这种基于细胞的疗法通常需要大量的干细胞。由于ADSC在体外培养时会失去其干细胞特性（Stem），包括分化能力，因此只能用于有限的传递，而将大量高电位干细胞从主要分离中传播。干细胞分离需要大量组织，并且需要连续隔离ADSC来进行多种治疗。但是，例如可用于ADSC隔离的牙科组织通常受到限制。从提取的牙齿和去角质牙齿的牙髓中分离出来的干细胞已被证明是再生医学干细胞的良好来源。但是，与其他ADSC一样，这些牙髓干细胞（DPSC）在体外培养时也会失去其分化潜力，从而阻碍其应用。这项研究的长期目标是阐明基因和分子调节DPSC中的分化能力。初步研究表明，当细胞失去分化能力时，热休克蛋白B8（HSPB8）在培养的DPSC中被下调，并且在早期传递DPSC中敲低HSPB8表达会导致细胞失去分化能力。进一步的研究表明，microRNA（miRNA）负责DPSC中HSPB8表达的下调。中心假设是，在晚期DPSC中，某些miRNA的表达更新了HSPB8和其他调节基因，进而导致DPSC在体外培养过程中失去分化能力。该项目的目的是：（1）确定在晚期DPSC中表达的miRNA，被指定为上调的miRNA（LPU-MIRNA）；（2）识别下调HSPB8表达的LPU-MIRNA；即，靶向Hspb8靶向miRNA，以及（3）研究LPU-MIRNA和HSPB8靶向miRNA对DPSC分化以及调节其他潜在基因的影响。实现目标对于理解miRNA和HSPB8在确定长期培养的DPSC中看到的分化损失方面的作用将是重要的，并且还将阐明其他ADSC。这将支持开发用于维持DPSC和ADSC中干细胞特性的尖端方法，从而可以通过膨胀的主要分离干细胞的扩展获得了大量具有高分化潜力的干细胞。