Anti-Inflammatory Mesenchymal Stem Cell Therapy for Dental Applications

 描述(申请人提供)：用于牙科应用的抗炎间充质干细胞疗法维宁博士是一名普通牙医和博士生，在哈佛大学工程和应用科学学院接受大卫·穆尼教授的培训。维宁博士的长期研究目标是解决如何利用细胞外微环境的生物物理参数来开发口腔和颅面健康的新型抗炎疗法。具体地说，这项拟议的研究将检查微环境的机械硬度是否可以用于控制MSCs的抗炎功能，以用于口腔和颅面健康的治疗。维宁博士致力于成为一名独立的临床科学家，因此他寻求获得这一临床科学家导师奖，以促进他作为口腔健康科学和工程领域的临床科学家的成长和发展。在啮齿动物牙髓损伤模型中，该研究计划将探讨水凝胶硬度可用于体外控制包裹的骨髓间充质干细胞的抗炎旁分泌功能和增强其体内功能的假说。在具体目标1中，维宁博士将研究机械硬度水平如何影响成年人骨髓来源的间充质干细胞在胶原-海藻酸盐互穿网络水凝胶中的免疫调节分泌和转录行为。通过测量促炎和抗炎因子的表达和分泌，MSCs的行为将表现为不同的基质硬度。基于目标1的块状水凝胶的最佳基质硬度，在特定的目标2中，Vning博士将研究人骨髓来源的MSCs对人类巨噬细胞M1和M2表面标志的体外功能影响。在具体的目标3中，Vning博士将研究MSCs的体内抗炎功能，如AIMS 1-2所示，MSCs通过其微环境的刚性来调节以增加抗炎因子的分泌。他将通过将AIMS 1-2的水凝胶中包裹的MSCs局部注射到啮齿动物牙髓损伤模型中，在体内测试经过机械调整的人MSCs。骨髓间充质干细胞对宿主炎症反应的功能效应将通过髓过氧化物酶探针、组织学和免疫组织化学以及基因表达来衡量。尽管在医学上进行了广泛的研究，但在牙科领域，MSC免疫疗法在很大程度上被忽视了。维宁博士建议将这些发现应用于开发一种新的MSC免疫疗法，用于口腔和颅面应用。这些目标将通过探索生物物理线索如何调节MSCs的免疫调节旁分泌信号来影响基础科学知识。作为维宁博士职业发展计划的一部分，他将参加工程学和应用科学以及细胞生物学和免疫学的课程，以帮助他实现研究目标。威宁博士将通过参加研究研讨会和学习小组以及在年度重大科学会议上发表演讲来发展他的专业技能。维宁博士将在他的博士导师穆尼教授的指导下，通过每两周一次的正式个人会议，在他的整个博士论文中进行指导。Kai Wucherpfennig博士将为该项目的免疫学方面提供共同指导。维宁博士培训的一个关键部分是，通过与穆尼实验室和哈佛大学合作实验室的不同成员进行互动，在实验室进行非正式培训。维宁博士将通过哈佛大学文理学院和工程与应用科学学院的教学活动学习如何成为一名好的教师和导师。这项计划为维宁博士成功地从有指导的博士后到独立的博士后研究人员，最后成为终身教职学术职位的独立首席研究员奠定了基础。

 DESCRIPTION (provided by applicant): Anti-Inflammatory Mesenchymal Stem Cell Therapy for Dental Applications Dr. Vining is a general dentist and Ph.D. student, training with Professor David Mooney at Harvard School of Engineering and Applied Sciences. Dr. Vining's long-term research goal is to address how biophysical parameters of the extracellular microenvironment can be used to develop novel anti-inflammatory therapies in oral and craniofacial health. Specifically, the proposed research will examine whether mechanical stiffness of the microenvironment can be used to control the anti-inflammatory function of MSCs for therapeutic use in oral and craniofacial health. Dr. Vining is committed to become an independent clinician-scientist, so he seeks this mentored clinical scientist award to further his growth and development as a clinician-scientist in oral health sciences and engineering. The proposed research plan will investigate the hypothesis that hydrogel stiffness can be used to control the anti-inflammatory paracrine function of encapsulated bone marrow-derived MSCs in vitro and to enhance their in vivo function in a rodent dental pulp injury model. In Specific Aim 1, Dr. Vining will examine how the level of mechanical stiffness affects the immunomodulatory secretory and transcriptional behavior of adult human bone marrow-derived MSCs in a collagen-alginate interpenetrating network hydrogel. The MSCs' behavior will be characterized in varying matrix stiffness by measuring expression and secretion of pro- and anti-inflammatory factors. Based on the optimal matrix stiffness in bulk hydrogels from Aim 1, in Specific Aim 2 Dr. Vining will investigate the in vitro functional effects of human bone marrow-derived MSCs on human macrophages M1 versus M2 surface markers. In Specific Aim 3, Dr. Vining will investigate the in vivo anti-inflammatory function of MSCs, which are tuned by the stiffness of their microenvironment to increase secretion of anti-inflammatory factors as shown in Aims 1 - 2. He will test the mechanically-tuned human MSCs in vivo by locally injecting MSCs encapsulated in the hydrogel from Aims 1 - 2 into a rodent dental pulp injury model. MSCs' functional effects on the host inflammatory response will be measured by a myeloperoxidase probe, histology and immunohistochemistry, and gene expression. Despite being widely investigated in medicine, MSC immunotherapies have largely been overlooked in dentistry. Dr. Vining proposes to apply these findings to develop a novel MSC-immunotherapy for oral and craniofacial applications. These aims will have an impact on basic science knowledge by exploring how biophysical cues modulate MSCs' immunomodulatory paracrine signaling. As part of Dr. Vining's career development plan, he will take courses in engineering and applied sciences, as well as cell biology and immunology, to help him achieve his research objectives. Dr. Vining will develop his professional skills through participating in research seminars and study groups and presenting at annual major scientific conferences. Dr. Vining will be mentored by his Ph.D. advisor, Professor Mooney, throughout his Ph.D. thesis via formal, bi-weekly individual meetings. Dr. Kai Wucherpfennig will provide co-mentoring for the immunology aspects of the project. A key component of Dr. Vining's training will involve informal training in the lab via interaction with various members of the Mooney laboratory, and collaborating laboratories at Harvard University. Dr. Vining will learn how to be a good teacher and mentor through teaching activities through Harvard Faculty of Arts and Sciences and School of Engineering and Applied Sciences. This plan lays a framework for Dr. Vining's successful progression from mentored Ph.D. student, to independent post- doctoral researcher, and finally independent principal investigator in a tenure-track academic position.