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

项目概要/摘要 牙周病仍然是一个重大的公共卫生问题。慢性未解决的炎症反应促炎性口腔微生物菌群失调诱导宿主介导的牙周组织破坏。由于无法解决炎症，牙周膜再生的努力已经显示出有限的成功。炎症的缓解是由专门的促溶解脂质介体（SPMs）启动的。SPM包括脂氧素、消退素、保护素、maresins以及它们的蛋白质缀合物，它们在实验模型和临床试验中表现出改善炎症相关病理结果的多管齐下的作用。人牙周膜干细胞（PDLSC）释放SPM来调节免疫调节和促愈合特性。在SPM中，Maresin 1（MaR 1）在调节炎症消退和疼痛、伤口修复和组织再生中显示出有效的作用。然而，MaR 1诱导组织再生（包括骨）的机制尚不明确。我们提出的研究将通过使用LM-SPM代谢脂质组学的系统生物学方法和PDLSC的RNA测序来解决这一差距，以发现MaR 1在PDLSC组织再生中调节的可能途径。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MaR 1增强的PDLSC疗法用于牙周组织再生的治疗潜力，以转化为人类。具体目标有三：1）建立牙周膜干细胞MaR 1生物合成途径。2)明确MaR 1控制组织再生的机制。3a）证明猪中MaR 1增强的PDLSC介导的牙周再生。3b）阐明MaRl对猪中BMP 2-PDLSC组合治疗的影响。这些研究的结果将解决NIDCR的使命：“支持开发人类疾病和损伤相关的动物模型，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。” 候选人是一名牙医科学家，目前是德克萨斯大学健康圣安东尼奥的博士后研究员。该提案包括一个全面的指导和培训计划，以提高候选人在免疫学，代谢脂质组学，RNA测序和转化科学方面的技能和知识，由NIH资助的科学家组成的强大团队指导; Kenneth Hargreaves博士，Stephen Harris博士，Charles Serhan博士和乔治Huang博士。该提案建立在候选人之前在干细胞，BMP 2和牙周组织形成方面的研究基础上，整合了新的专业领域。该计划包括一个指导阶段，旨在优化学习和获得新技能（K99），然后是R 00阶段，该阶段专门旨在利用申请人的优势开发一条新的研究路径，以确定炎症和干细胞在BMP 2介导的再生中的作用。R 00阶段还包括住院医师培训。完成这一全面的培训计划将使候选人具备一套独特的跨学科技能，使她能够过渡到独立从事基于干细胞的牙周组织再生疗法。

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.