Specialized Proresolving Lipid Mediator-Enhanced Stem Cell Therapy and Tissue Regeneration

项目摘要/摘要 牙周病仍然是一个重大的公共卫生问题。致炎的口腔微生物群失调导致宿主介导的牙周组织破坏，导致慢性未解炎症。由于无法消除炎症，牙周再生的努力显示出有限的成功。炎症的消退是由专门的促分解脂质介体(SPM)启动的。SPM包括脂氧素、溶血素、保护素、树脂素及其蛋白结合物，它们表现出多管齐下的作用，在实验模型和临床试验中改善炎症相关病理的结果。人牙周膜干细胞(PDLSC)释放SPM来调节免疫调节和促进愈合的特性。在SPM中，Maresin 1(MaR1)在调节炎症消退和疼痛、伤口修复和组织再生方面显示出强大的作用。然而，MaR1诱导组织再生(包括骨)的机制尚不明确。我们建议的研究将通过使用系统生物学方法结合LM-SPM代谢脂组学和PDLSC的RNA测序来发现在PDLSC组织再生中受MaR1调控的可能途径。通过利用美国约克郡猪作为大型动物再生模型，我们将测试MaR1增强的PDLSC疗法对移植到人类的牙周组织再生的治疗潜力。提出了三个具体的目标：1)在PDLSC中建立MaR1生物合成途径。2)明确MaR1调控组织再生的机制。3a)研究MaR1增强PDLSC介导的猪牙周组织再生。3b)阐明MaR1在BMP2-PDLSC联合治疗中的作用。这些研究的结果将解决NIDCR的一个使命：“支持开发与人类疾病和伤害相关的动物模型，用于口腔和颅面复合体的组织再生，优先考虑大型动物模型。” 候选人是一名牙医兼科学家，目前是德克萨斯大学圣安东尼奥健康分校的博士后研究员。这项建议包括一个全面的指导和培训计划，以提高候选人在免疫学、代谢性蛋白质组学、RNA测序和翻译科学方面的技能和知识，并在NIH资助的强大团队的指导下，这些科学家包括Kenneth Hargreaves博士、Stephen Harris博士、Charles Serhan博士和George Huang博士。该提案建立在候选人之前在干细胞、BMP2和牙周组织形成方面的研究基础上，通过整合新的专业领域。该计划包括一个旨在优化学习和获得新技能的指导阶段(K99)，随后是R00阶段，该阶段专门设计用于利用申请者的优势开发一条新的研究途径，以确定炎症消退和干细胞在BMP2介导的再生中的作用。R00阶段还包括派驻培训。完成这一全面的培训计划将使应聘者具备一套独特的跨学科技能，使她能够过渡到专门从事基于干细胞的牙周组织再生治疗的独立专业。

PROJECT SUMMARY/ABSTRACT Periodontal disease remains a significant public health problem. Chronic unresolved inflammation in response to proinflammatory oral microbiome dysbiosis induces host-mediated destruction of the periodontal tissues. Periodontal regeneration efforts have shown limited success due to the inability to resolve inflammation. Resolution of inflammation is initiated by Specialized Proresolving lipid Mediators (SPMs). SPMs include the lipoxins, resolvins, protectins, maresins, as well as their protein conjugates, which exhibit multipronged actions that improve the outcome of inflammation-related pathologies in experimental models and clinical trials. Human periodontal ligament stem cells human (PDLSC) release SPMs to regulate immunomodulatory and pro-healing properties. Among SPMs, Maresin 1 (MaR1) displays potent actions in regulating inflammation resolution and pain, wound repair and tissue regeneration. However, there is no defined mechanism by which MaR1 induces tissue regeneration, including bone. Our proposed research will address this gap by using a systems biology approach with LM-SPM metabolipidomics, and RNA-sequencing of PDLSC to discover possible pathways modulated by MaR1 in PDLSC tissue regeneration. By utilizing the American Yorkshire Pig as a large animal regeneration model, we will test the therapeutic potential of MaR1-enhanced PDLSC therapy for regeneration of periodontal tissues for translation to humans. Three specific aims are proposed: 1) To establish MaR1 biosynthetic pathway in PDLSC. 2) To define the mechanism by which MaR1 controls tissue regeneration. 3a) To demonstrate MaR1-enhanced PDLSC-mediated periodontal regeneration in the Pig. 3b) To elucidate the impact of MaR1 on combined BMP2-PDLSC treatment in the Pig. Results from these studies will address a mission of NIDCR: ”to support the development of human disease- and injury-relevant animal models for tissue regeneration of the oral and craniofacial complex, with preference given to large animal models.” The candidate is a dentist-scientist and is currently a postdoctoral research fellow at the University of Texas Health San Antonio. This proposal includes a comprehensive mentorship and training plan to advance the candidate’s skills and knowledge in immunology, metabolipidomics, RNA-sequencing and translational science under the guidance of a strong team of NIH-funded scientists; Dr. Kenneth Hargreaves, Dr. Stephen Harris, Dr. Charles Serhan and Dr. George Huang. The proposal builds on the candidate’s previous research in stem cells, BMP2 and periodontium formation by integrating new areas of expertise. The plan includes a mentored phase designed to optimize learning and acquisition of new skills (K99) followed by the R00 phase, which is specifically designed to capitalize on the strengths of the applicant to develop a new path of research that will determine the role of resolution of inflammation and stem cells in BMP2 mediated regeneration. The R00 phase also includes residency training. Completion of this comprehensive training plan will position the candidate with a unique set of cross disciplinary skills that will enable her to transition to independence specializing in stem cell-based periodontal tissue regenerative therapies.