Anti-Inflammatory Mesenchymal Stem Cell Therapy for Dental Applications

 描述（由申请人提供）：用于牙科应用的抗炎间充质干细胞疗法Vining博士是一名普通牙医和博士。学生，在哈佛工程与应用科学学院接受大卫穆尼教授的培训。Vining博士的长期研究目标是解决细胞外微环境的生物物理参数如何用于开发口腔和颅面健康的新型抗炎疗法。具体而言，拟议的研究将检查微环境的机械刚度是否可用于控制MSC的抗炎功能，以用于口腔和颅面健康的治疗用途。Vining博士致力于成为一名独立的临床科学家，因此他寻求这个指导临床科学家奖，以促进他作为口腔健康科学和工程临床科学家的成长和发展。拟议的研究计划将调查的假设，水凝胶刚度可用于控制抗炎旁分泌功能的封装骨髓来源的骨髓间充质干细胞在体外，并提高其在啮齿动物牙髓损伤模型中的体内功能。在具体目标1中，Vining博士将研究机械刚度水平如何影响胶原-藻酸盐互穿网络水凝胶中成人骨髓来源的MSC的免疫调节分泌和转录行为。MSC的行为将通过测量促炎因子和抗炎因子的表达和分泌来表征不同的基质硬度。基于目标1中散装水凝胶的最佳基质刚度，在特定目标2中，Vining博士将研究人骨髓源性MSC对人巨噬细胞M1与M2表面标志物的体外功能效应。在具体目标3中，Vining博士将研究MSC的体内抗炎功能，这些功能通过微环境的刚度来调节，以增加抗炎因子的分泌，如目标1 - 2所示。他将通过将目标1 - 2中的水凝胶包裹的MSC局部注射到啮齿动物牙髓损伤模型中，在体内测试机械调节的人类MSC。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测量MSC对宿主炎症反应的功能作用。尽管在医学上得到了广泛的研究，但MSC免疫疗法在牙科领域却被忽视了。Vining博士建议将这些发现应用于开发一种用于口腔和颅面应用的新型MSC免疫疗法。这些目标将通过探索生物物理线索如何调节MSC的免疫调节旁分泌信号传导对基础科学知识产生影响。作为Vining博士职业发展计划的一部分，他将学习工程和应用科学以及细胞生物学和免疫学课程，以帮助他实现研究目标。Vining博士将通过参加研究研讨会和研究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。Vining博士将接受其博士学位的指导穆尼教授在他的博士学位期间担任顾问。通过正式的论文，双周一次的个人会议。Kai Wucherpfennig博士将为该项目的免疫学方面提供共同指导。Vining博士培训的一个关键组成部分将包括通过与Mooney实验室的各种成员以及哈佛大学的合作实验室进行互动，在实验室进行非正式培训。Vining博士将通过哈佛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优秀的教师和导师。该计划为Vining博士从辅导博士成功晋升奠定了框架学生，独立博士后研究员，最后独立首席研究员在一个终身制的学术职位。

 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.