Bioengineered Oral Mucosa from Embryonic Stem Cells for Regenerative Medicine

用于再生医学的胚胎干细胞生物工程口腔粘膜

• 批准号: 7755039
• 负责人: JONATHAN A GARLICK
• 金额: $ 31.06万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-05 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7755039
• 关键词: Basement membrane; Behavior; Biological; Biological Process; Biology; Biomedical Engineering; Cells; Commit; Complex; Cues; Dentistry; Development; Differentiation and Growth; Engineering; Epithelial; Epithelial Cells; Epithelium; Event; Extracellular Matrix Proteins; Face; Fibroblasts; Future; Generations; Goals; Growth; Human; Link; Mediating; Membrane Proteins; Mesenchymal; Molecular; Mouth Diseases; Mucous Membrane; Oral; Oral cavity; Oral mucous membrane structure; Organ; Pattern; Phenotype; Process; Property; Regenerative Medicine; Regulation; Research; Research Personnel; Research Proposals; Role; Signal Pathway; Signal Transduction; Source; Specific qualifier value; Staging; Stem cells; Structure; Technology; Testing; Therapeutic; Tissue Transplantation; Tissues; Transplantation; base; clinical application; embryonic stem cell; human embryonic stem cell; in vivo; insight; keratinocyte; keratinocyte differentiation; new technology; novel; novel strategies; oral cavity epithelium; prevent; programs; regenerative; regenerative therapy; self-renewal; stem cell biology; stem cell niche; success; tissue regeneration

DESCRIPTION (provided by applicant): Human embryonic stem cells provide a potentially unlimited source of oral mucosal tissues that can revolutionize the treatment of oral diseases. These stem cells are essential to the success of novel regenerative therapies, due to their plasticity and to their tremendous regenerative potential that can overcome the shortcomings and currently limited supply of post-natal, oral epithelial stem cells. However, many obstacles still limit their therapeutic application, as it is not known if human embryonic stem cells can 1- form 3D epithelial tissues with hierarchical structure, 2 - give rise to cells similar to post-natal, epithelial stem cells or 3 - maintain their differentiated state in 3D tissues. To help overcome these barriers, the immediate goal of this application is to use our novel approaches in 3D tissue biology to provide key insights into the biological properties of human embryonic stem cells to generate 3D oral epithelial tissues that will demonstrate the feasibility of using these new technologies for future oral therapies. We hypothesize that the tissue microenvironment, instructed by basement membrane proteins and fibroblast-derived soluble factors, regulates the self-renewal and differentiation of keratinocytes derived from human embryonic stem cells to generate specialized tissues that mimic oral epithelium. To test this hypothesis, our AIMS are to: 1- Determine if keratinocytes derived from human embryonic stem cells give rise to progenitor cells with properties of post-natal stem cells that can undergo long-term, self-renewal and can differentiate to form 3D epithelia and 2- Begin to explore the cellular- and tissue-based cues, mediated by the stem cell "niche" at the basement membrane interface and by fibroblast-derived soluble factors, that direct these events. We expect to set the stage for future studies that will dissect signaling pathways that direct these cues and optimize the growth, differentiation and survival of 3D epithelia. This research is made possible by two major advances in our lab: 1 - Development of engineered, 3D tissues that mimic the oral mucosa to a significant degree and have established a link between the tissue microenvironment, self- renewal of post-natal stem cells and predictable tissue regeneration after in vivo transplantation. 2 - Propagation of H9, human embryonic stem cells (NIH-WA09) committed to an epithelial lineage and generation of immature but multilayer 3D epithelial tissues from their progeny. Our long-term goal is to harness the developmental potential and growth capacity of pluripotent, human embryonic stem cells by gaining convincing evidence that oral epithelial tissues derived from them can serve as long-term, functional replacement tissues to treat oral diseases for which there are no current practical therapies. By doing so, we will advance our understanding of stem cell biology in a fundamental manner to enable future efforts to tailor and engineer lineage-specific tissues of therapeutic importance in humans.

描述（由申请人提供）：人类胚胎干细胞提供了潜在无限的口腔粘膜组织来源，可以彻底改变口腔疾病的治疗。这些干细胞对于新型再生疗法的成功至关重要，因为它们具有可塑性和巨大的再生潜力，可以克服出生后口腔上皮干细胞的缺点和目前供应有限的问题。然而，许多障碍仍然限制了它们的治疗应用，因为尚不清楚人类胚胎干细胞是否能够 1- 形成具有分层结构的 3D 上皮组织，2- 产生类似于出生后上皮干细胞的细胞或 3- 维持它们在 3D 组织中的分化状态。为了帮助克服这些障碍，该应用的直接目标是利用我们在 3D 组织生物学方面的新方法，为人类胚胎干细胞的生物学特性提供关键见解，以生成 3D 口腔上皮组织，从而证明使用这些新方法的可行性未来口腔治疗的技术。我们假设，组织微环境在基底膜蛋白和成纤维细胞衍生的可溶性因子的指导下，调节源自人类胚胎干细胞的角质形成细胞的自我更新和分化，以产生模仿口腔上皮的特殊组织。为了检验这一假设，我们的目标是： 1- 确定源自人类胚胎干细胞的角质形成细胞是否产生具有产后干细胞特性的祖细胞，这些祖细胞可以进行长期、自我更新并可以分化形成 3D上皮和 2- 开始探索基于细胞和组织的线索，由基底膜界面处的干细胞“生态位”和成纤维细胞衍生的可溶性因子介导，指导这些事件。我们期望为未来的研究奠定基础，这些研究将剖析指导这些线索并优化 3D 上皮细胞的生长、分化和存活的信号通路。我们实验室的两项重大进展使这项研究成为可能： 1 - 开发出工程化的 3D 组织，该组织在很大程度上模仿口腔粘膜，并在组织微环境、产后干细胞的自我更新和体内移植后可预测的组织再生。图 2 - H9 人类胚胎干细胞 (NIH-WA09) 的增殖，致力于形成上皮谱系，并从其后代中生成未成熟但多层的 3D 上皮组织。我们的长期目标是通过获得令人信服的证据来利用多能人类胚胎干细胞的发育潜力和生长能力，证明源自多能人类胚胎干细胞的口腔上皮组织可以作为长期的功能性替代组织来治疗口腔疾病。目前尚无实用疗法。通过这样做，我们将从根本上增进对干细胞生物学的理解，以便未来能够努力定制和设计对人类具有治疗重要性的谱系特异性组织。