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

 描述（由申请人提供）：本申请要求为眼科培训的眼科病理学家提供指导性职业发展奖，该病理学家对理解可用于治疗性治疗的关键细胞通路具有科学兴趣。候选人最初将专注于Src家族激酶（SFKs）和Src激活和信号分子（Srcasm）通路在眼表伤口愈合和肿瘤发生，因为先前的研究已经显示其在皮肤角质形成细胞伤口和肿瘤局部治疗策略的重要性和顺从性。候选人将在奖励期间有以下目标：1）通过指导，教学法，相关研讨会和负责任的研究行为指导个人研究计划的发展; 2）通过指导和相关活动的方式完善研究和管理技能; 3）通过演讲，手稿和赠款准备提高沟通和写作技能; 4）通过收集初步结果和设计实验目标来获得未来的资金。在奖励期结束时，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项目，以新颖，合理的方法战略性地解决眼科研究领域的重要，关键问题。 该项目将在眼科和皮肤科的首席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部门之间的这种合作努力。此外，宾夕法尼亚大学有一个强大的遗产，即促进和发展初级教师到领导职位。导师们在个人和各自的部门都坚持这一传统。眼科和皮肤科的部门都是领先的机构，在美国国立卫生研究院赠款资金，在高影响力期刊上的出版物，以及杰出的医生和科学家的培训中排名第一。为候选人的目标和职业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安排的会议;教学法;研讨会;以及提高候选人研究，管理，沟通和写作技能的活动。除了他们的指导，导师将提供必要的资源，设备和专业知识，以执行提案的具体目标。整个校园有多个核心和机构，以及最先进的设备，这将有助于促进执行 提出的目标。宾夕法尼亚大学的互动和充满活力的科学社区将为候选人提供众多机会，以促进K 08奖的结果并推进候选人的职业生涯。 眼表疾病引起的视力丧失是全球范围内第二大致盲原因。虽然原因多种多样，但导致视力丧失的最常见途径是眼表瘢痕。在伤口愈合过程的早期进行治疗干预对于解决这一全球负担至关重要。SFK/Srcasm途径似乎调节伤口愈合中的细胞增殖、迁移和分化，使其适合于治疗方法。然而，该途径在肿瘤发生中也起着重要作用，这是预期的，因为伤口和肿瘤之间存在共性。因此，从研究眼表伤口愈合和肿瘤发生中的这一途径获得的结果将导致这两种疾病的潜在治疗靶点。这一目标将通过以下3个具体目标来实现。本提案中概述的机构，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。这一结构化的职业发展计划将使候选人在授标期结束时能够独立工作。目的1）观察SFKs对角膜上皮损伤愈合的影响。我们将确定Fyn，SFK成员，在体内角膜上皮伤口愈合的基因修饰小鼠的影响，并在体外人类器官型角膜伤口模型，使用慢病毒敲低和Fyn的过表达。目的2）观察Srcasm对角膜上皮损伤愈合的影响。我们将使用基因修饰的小鼠来定义Sracsm在体内角膜上皮伤口愈合中的作用;以及使用慢病毒敲低Srcasm在体外人器官型角膜伤口模型中的作用。目的3）研究眼表鳞状细胞瘤（OSSN）的分子和转录水平，以及SFKs和Srcasm在OSSN发生发展中的作用。我们将用免疫组织化学和RNA测序分析结膜OSSN;并表征慢病毒介导的Fyn过表达和Srcasm敲低在体外结膜肿瘤模型中的作用。

 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.