Defining BMP-responsive lncRNA for bone regeneration

定义用于骨再生的 BMP 响应性 lncRNA

• 批准号: 9278145
• 负责人: Derrick Wan
• 金额: $ 12.65万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-02 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9278145
• 关键词: Bone Morphogenetic Proteins; Bone Regeneration; Calvaria; Cell Fate Control; Cell Line; Cell Maintenance; Cell Therapy; Cell surface; Cells; ChIP-seq; Chromatin; Clinical; Code; Cultured Cells; Data; Defect; Derivation procedure; Development; Developmental Process; Epigenetic Process; Ganciclovir; Gene Silencing; Genes; Genetic Transcription; Growth; Head and neck structure; Implant; Length; Literature; Maintenance; Messenger RNA; Modeling; Nucleotides; Osteogenesis; Pattern; Play; Pluripotent Stem Cells; Post-Translational Protein Processing; Proteins; RNA Interference; Recruitment Activity; Regenerative Medicine; Repair Complex; Risk; Role; Teratoma; Therapeutic; Tissues; Transcript; Transforming Growth Factors; Untranslated RNA; Work; adverse outcome; bone; bone morphogenetic protein 2; cell fate specification; cell type; clinical application; embryonic stem cell; epigenetic regulation; histone methylation; histone methyltransferase; histone modification; human disease; implantation; in vivo; induced pluripotent stem cell; innovation; member; novel; osteogenic; overexpression; pluripotency; public health relevance; regenerative; repaired; response; risk minimization; skeletal; small molecular inhibitor; small molecule; suicide gene; tool; transcriptome; transcriptome sequencing; tumor

DESCRIPTION (provided by applicant): With the ability to differentiate into almost any cell in the body, pluripotent cells represent a powerful tool for regenerative medicine and repair of deficient tissues such as bone. Several challenges to their clinical application remain, however, chief among which is their propensity to form tumors. Spontaneous teratoma formation has been frequently observed following implantation of induced pluripotent cells or embryonic stem cells. While approaches such as immunodepletion of SSEA-5+ cells by FACS, incorporation of suicide genes responsive to ganciclovir, and use of small molecular inhibitors have been described, none have proven completely successful. Recent studies have now shown the epigenetic landscape to play a key role in the specification of cell fate, and understanding how this may be regulated would be critial to facilitate safe and efficient use of pluripotent cells in therapeutic strategies to replace deficient tissues such as bone. Emerging data have described a novel class of noncoding RNAs ranging from 200 nucleotides to over 10 kilobases that are roughly as diverse in a given cell type as protein---coding mRNA. These long noncoding RNAs have been found to interact with small molecule regulators of histone methylation, targeting specific domain for transcriptional activity or gene silencing. How long noncoding RNAs are controlled and their precise role in regulating differentiation remains undefined, but with better understanding, the potential exists to redirect the developmental process in pluripotent cells. This will be the focus of our proposal. We will evaluate how induced pluripotent stem cells and embryonic stem cells respond to BMP-2 with definition of whole transcriptome response and changes to the chromatin landscape. Integrating data from both cell lines, we will then identify novel long noncoding transcripts responsive to BMP-2. We wil look to manipulate these novel long noncoding RNA transcripts and evaluate for changes to the pluripotent state and bone differentiation potential. Finally, we will also evaluae the in vivo response of pluripotent cells following manipulation of novel long noncoding RNAs. Specifically, the ability for both bone regeneration in a critical-sized calvaril defect model and the teratoma formation potential will be evaluated. We expect result of these innovative studies to identify a ground---breaking approach to understand and regulate cell fate determination, allowing for development of safe cell---based therapies employing pluripotent cells.

描述（由申请人提供）：多能细胞具有分化为体内几乎任何细胞的能力，是再生医学和修复骨等缺陷组织的有力工具。然而，它们的临床应用仍然面临着一些挑战，其中最主要的是它们形成肿瘤的倾向。自发性畸胎瘤的形成在诱导多能细胞或胚胎干细胞植入后经常被观察到。虽然已经描述了一些方法，如FACS对SSEA-5+细胞的免疫消耗，对更昔洛韦有反应的自杀基因的结合，以及使用小分子抑制剂，但没有一种被证明是完全成功的。最近的研究表明，表观遗传景观在细胞命运的规范中起着关键作用，了解如何调节这一点对于促进在治疗策略中安全有效地使用多能细胞来替代缺陷组织（如骨）至关重要。新出现的数据描述了一类新的非编码rna，其范围从200个核苷酸到超过10万个碱基，在给定的细胞类型中与蛋白质编码mRNA大致相同。这些长链非编码rna已被发现与组蛋白甲基化的小分子调节因子相互作用，靶向转录活性或基因沉默的特定区域。非编码rna被控制多长时间以及它们在调节分化中的确切作用尚不清楚，但随着对其更好的理解，多能细胞的发育过程存在着重新定向的潜力。这将是我们提案的重点。我们将评估诱导多能干细胞和胚胎干细胞如何通过定义全转录组反应和染色质景观的变化来响应BMP-2。整合来自两种细胞系的数据，我们将鉴定对BMP-2有反应的新型长非编码转录物。我们将寻求操纵这些新的长链非编码RNA转录本，并评估多能状态和骨分化潜力的变化。最后，我们还将评估多能细胞在操纵新型长链非编码rna后的体内反应。具体来说，将评估临界尺寸颅骨缺损模型的骨再生能力和畸胎瘤形成的潜力。我们期望这些创新研究的结果能够确定一种突破性的方法来理解和调节细胞命运的决定，从而允许开发基于多能细胞的安全细胞疗法。