The Role of Src-Family Tyrosine Kinases and Srcasm in Ocular Surface Epithelial Wound Repair and Neoplasia

 描述(由申请者提供)：本申请申请获得指导职业发展奖，授予受过眼科训练的眼科病理学家，该医生对了解可作为治疗目标的关键细胞通路具有科学兴趣。候选人首先将重点放在眼表创伤愈合和肿瘤发生中的Src家族激酶(SFKs)和Src激活和信号分子(SRCasm)途径，因为先前的研究已经表明其在皮肤角质形成细胞创伤和肿瘤的局部治疗策略中的重要性和适用性。候选人在获奖期间将有以下目标：1)通过导师、教学、相关研讨会和负责任的研究行为指导，发展个人研究计划；2)通过导师和相关活动，完善研究和管理技能；3)通过演讲、手稿和拨款准备，增强沟通和写作技能；4)通过收集初步结果和设计实验目标，获得未来的资金。到获奖期结束时，候选人将获得设计、提出和实施视觉研究实验所需的指导、知识和宝贵的实践经验。候选人的长期目标是领导一个研究计划，以新的、合理的方法战略性地解决眼科研究领域的重要、关键问题。该项目将在来自眼科和皮肤科的高级研究员的监督下进行，他们将共同为拟议的研究和候选人的职业发展提供指导。宾夕法尼亚大学特殊的科学环境促进了各系之间的合作努力。此外，宾夕法尼亚大学还有一个强大的遗产，就是提拔和发展初级教职员工担任领导职务。导师个人和他们各自的部门都坚持这一传统。眼科和皮肤科都是领先的机构，在NIH拨款、在高影响力期刊上发表文章以及培训杰出的医生和科学家方面都名列前茅。已经为候选人的目标和职业生涯的逐步成长和发展制定了全面的指导计划。该计划包括导师和候选人之间定期的预定会议；教学；研讨会；以及提高候选人的研究、管理、沟通和写作技能的活动。除了他们的指导外，导师还将提供必要的资源、设备和专业知识来执行提案的具体目标。整个园区有多个核心和机构，配备了最先进的设备，将有助于促进执行 建议的目标。宾夕法尼亚大学现有的互动和充满活力的科学社区将为候选人提供无数机会来宣传K08奖的结果，并推动候选人的职业生涯。眼表疾病导致的视力丧失是全球第二大致盲原因。虽然原因多种多样，但导致视力丧失的最常见途径是眼表疤痕。在伤口愈合过程中的早期治疗干预对于解决这一全球负担至关重要。SFK/srCasm通路似乎在伤口愈合过程中调节细胞的增殖、迁移和分化，使其适用于治疗方法。然而，鉴于创伤和肿瘤之间的共性，这一途径在肿瘤的发生中也起着重要的作用。因此，通过研究这一途径在眼表创伤愈合和肿瘤发生中的作用，将为这两种疾病提供潜在的治疗靶点。这一目标将以下面列出的三个具体目标实现。该提案中概述的机构、导师和合作者致力于研究和培养早期研究人员的职业生涯。这一有组织的职业发展计划将使候选人在获奖期结束时获得独立资格。目的1)研究SFKs对角膜上皮损伤愈合的影响。我们将使用转基因小鼠确定SFK成员Fyn在体内角膜上皮伤口愈合中的作用；并在体外使用慢病毒敲除和过表达Fyn来确定人类器官型角膜创伤模型的效果。目的2)明确sRCasm对角膜上皮损伤愈合的影响。我们将使用转基因小鼠在体内确定SRACSM在角膜上皮伤口愈合中的作用；在体外使用慢病毒敲除SRCasm来确定SRACSM在人类器官型角膜创伤模型中的作用。目的3)研究眼表鳞状上皮性肿瘤(OSSN)的分子和转录水平，以及sFKs和sRCasm在OSSN发生发展中的作用。我们将通过免疫组织化学和RNA测序来描述结膜OSSN；并在体外结膜肿瘤模型中表征慢病毒介导的Fyn过表达和srCasm基因敲除的效果。

 DESCRIPTION (provided by applicant): This application requests a mentored career development award for an ophthalmology trained ocular pathologist with a scientific interest in understanding key cellular pathways that can be targeted for therapeutic treatment. The candidate will focus initially on the Src-family kinases (SFKs) and Src-activating and signaling molecule (Srcasm) pathway in ocular surface wound healing and tumorigenesis because of prior studies that have shown its significance and amenability for topical therapeutic strategies in skin keratinocyte wounds and tumors. The candidate will have the following objectives during the award period: 1) development of an individual research program through mentorship, didactics, related workshops, and instruction in responsible conduct of research; 2) refinement of research and management skills by way of mentorship and related activities; 3) enhancement of communication and writing skills by means of presentations, manuscript and grant preparations; 4) and acquisition of future funding by assembling preliminary results and designing experimental aims. By the end of the award period, the candidate will have gained the mentorship, knowledge, and valuable hands-on experience required to design, propose and implement experiments in vision research. The candidate's long-term goal is to lead a research program that strategically addresses significant, critical questions in the field of ophthalmic research with novel, rational approaches. This project will be under the supervision of premier, senior investigators from the Department of Ophthalmology and Dermatology, who will jointly provide guidance in the proposed research and candidate's career development. The exceptional scientific environment at the University of Pennsylvania promotes such collaborative efforts between departments. In addition, there is a strong legacy at the University of Pennsylvania of promoting and developing junior faculty members to positions of leadership. The mentors uphold this tradition personally and in their respective departments. Both the Department of Ophthalmology and Dermatology are leading institutions, ranking in the top tier in NIH grant funding, publications in high impact journals, and training of outstanding physicians and scientists. A comprehensive mentoring plan has been created for the progressive growth and development of the candidate's goals and career. The plan includes regular, scheduled meetings between the mentors and candidate; didactics; workshops; and activities to enhance the candidate's research, management, communication, and writing skills. In addition to their guidance, the mentors will provide the necessary resources, equipment, and expertise to execute the specific aims of the proposal. There are multiple cores and institutions throughout the campus, as well, with state-of-the-art equipment that will help facilitate the execution of the proposed aims. The interactive and spirited scientific community that exists at the University of Pennsylvania will provide numerous opportunities for the candidate to promote the results from the K08 award and advance the candidate's career. Vision loss due to ocular surface disease is the second major cause of blindness worldwide. Although the causes are varied, the most common pathway leading to the loss of vision is scarring of the ocular surface. Therapeutic interventions early in the process of wound healing are critical to addressing this global burden. The SFK/Srcasm pathway appears to regulate cellular proliferation, migration, and differentiation in wound healing, making it amenable to therapeutic approaches. This pathway, however, also plays a significant role in tumorigenesis as well, which is expected given the commonalities between wounds and tumors. The results obtained from studying this pathway in both ocular surface wound healing and tumorigenesis, therefore, will lead to potential therapeutic targets in both disorders. This goal will be accomplished with 3 specific aims as listed below. The institution, mentors, and collaborators outlined in this proposal are dedicated to research and cultivating the careers of early investigators. This structured career development plan will position the candidate for independence at the end of the award period. AIM 1) Determine the effect of SFKs on corneal epithelial wound healing. We will determine the effect of Fyn, an SFK member, in in vivo corneal epithelial wound healing using genetically-modified mice; and in in vitro human organotypic corneal wound model using lentiviral knockdown and overexpression of Fyn. AIM 2) Define the effect of Srcasm on corneal epithelial wound healing. We will define the effect of Sracsm in in vivo corneal epithelial wound healing using genetically-modified mice; and in in vitro human organotypic corneal wound model using lentiviral knockdown of Srcasm. AIM 3) Determine the molecular and transcriptional profile of ocular surface squamous neoplasias (OSSN, which are the epithelial tumors of the ocular surface), as well as the effect of SFKs and Srcasm in the development of OSSNs. We will profile conjunctival OSSNs with immunohistochemistry and RNA sequencing; and characterize the effect of lentiviral-mediated Fyn overexpression and Srcasm knockdown in an in vitro conjunctival tumor model.