Biologic Activities of IL-36 Cytokines in Psoriasis

描述（由申请人提供）：完成我的博士学位后。在英国沃里克大学生物科学系，我加入了冰岛国立大学医院免疫学系Helgi Valdimarsson教授的研究团队，在那里我获得了银屑病免疫学和遗传学方面的广泛培训，发表了6篇出版物和2篇综述文章。后来我加入了J. T博士。在密歇根大学皮肤病学系的Elder团队中，经过18个月的博士后指导，作为国家皮肤病基金会和皮肤病学基金会研究员，我被任命为研究助理教授，并作为初级教师开始了我在皮肤免疫学方面的研究生涯。我在银屑病免疫学方面有良好的基础，本计划书中概述的培训将使我能够实现这些五年的职业目标：（1）成为IL-1/IL-36生物学和银屑病发病机制方面的专家;（2）成为一家大型研究机构的知名、资金充足的首席研究员;（3）成为本科生、研究生和博士后水平培训生的优秀导师，成功培养他们的职业道路。我的长期职业目标是成为一名独立的学术教员，与一个跨学科的研究团队合作，设计协作方法来解决复杂的生物医学问题。更具体地说，我预计我的知识和技能将直接转化为对银屑病发病机制的新见解，并确定新的治疗靶点和治疗这种疾病的方法。这个K 01奖将让我沉浸在一个丰富的环境中，一个多学科的研究团队将教我实现职业目标所需的生物统计学，免疫学和分子生物学技能。研究环境：我有在我的处置所需的所有资源，成功完成这个项目，包括400平方米。英尺实验室空间配备设备， 细胞培养、RNA分离、PCR、流式细胞术。我得到了Elder博士的指导，并有一个由生物统计学（Goncalo Abecasis博士）、T细胞生物学（Weiping Zou博士）、IL-1/炎性小体生物学（Gabriel Nunez博士）和皮肤病学（John Voorhees博士）的著名专家组成的咨询委员会。在导师的指导下，为我的职业发展制定了全面的培训计划：（1）培养统计和系统生物学分析技能-在Abecasis博士和Elder博士的指导下，有针对性地举办了一些正式课程/研讨会。(2)发展角质形成细胞生物学和功能方面的知识和技能-来自埃尔德博士的指导和正式课程，并与西北大学皮肤病资源中心角质形成细胞核心设施合作。(3)发展研究皮肤免疫系统和T细胞生物学的知识和技能-由邹博士指导，并在密歇根大学流式细胞术核心设施接受正式培训。(4)为了磨练赠款写作技能，并学习编写有效的内部审查委员会（IRB）的建议，对人类受试者进行道德研究，包括负责任的研究行为（RCR）-我将使用密歇根大学的一些资源来开发赠款，IRB建议和RCR问题培训的技能。(5)为了培养领导力，指导和团队建设技能-这将通过与密歇根大学本科生研究机会计划中的轮换本科生和研究生以及学生密切合作来实现，在那里我将能够招募并与本科生一起工作多年，以促进他们的科学发展和对医学研究的兴趣。(6)提高书面和口头沟通技巧-通过参加皮肤病学和免疫学部门的研究会议，以及参加国家免疫学和皮肤病学会议。研究项目：银屑病是一种由T细胞、抗原递呈细胞（APC）、角质形成细胞和细胞因子相互作用驱动的炎症性和过度增生性皮肤疾病。IL-36细胞因子（IL-36，？和IL-36 Ra）是最近确定的IL-1家族成员，并且在损伤性银屑病组织中过表达。该提案的广泛，长期目标是了解IL-10的表达，加工和功能， 36种细胞因子对银屑病中观察到的表皮增生的影响并确定其机会 开发创新的治疗策略。我们的中心假设是IL-36细胞因子通过（a）直接驱动角质形成细胞的先天宿主防御， 自分泌方式，和（B）激活APC以引发Th 17 T细胞应答，然后（c）与IL-36协同放大角质形成细胞应答。有许多未表征的步骤调节角质形成细胞分泌活性IL-36，IL-36对人APC和KC的作用仍有待阐明。为了验证这一假设，我们提出了以下具体目标：（1）确定KC中是否发生IL-36蛋白水解，并阐明IL-36分泌的机制;（2）确定IL-36的活性差异，？（3）阐明IL-36对银屑病患者APC功能和Th 1/Th 17 T细胞偏倚的影响。

DESCRIPTION (provided by applicant): After completing my Ph.D. in the Department of Biological Sciences, University of Warwick, UK, I joined the research team of Prof. Helgi Valdimarsson in the Department of Immunology at the National University Hospital, Iceland, where I gained extensive training in psoriasis immunology and genetics, generating 6 publications and 2 review articles. I subsequently joined Dr. J.T. Elder's team in the Department of Dermatology at the University of Michigan, and after 18 months' post-doctoral mentoring as a National Psoriasis Foundation and Dermatology Foundation Fellow, I was appointed Research Assistant Professor, and as junior faculty have set about establishing my research career in skin immunology. I have a sound foundation in psoriasis immunology and the training outlined in this proposal will allow me to achieve these five-year career goals: (1) To become an expert in IL-1/IL-36 biology and psoriasis pathogenesis; (2) To become an established, well-funded principal investigator at a major research institution; (3) Be an excellent mentor to undergraduate, graduate and post-doctoral level trainees and successfully foster their career paths. My long- term career goal is to be an independent academic faculty member, working with an interdisciplinary research team to devise collaborative approaches to solve complex biomedical problems. More specifically, I anticipate that my broadened knowledge and skill sets will directly translate into new insights into the pathogenesis of psoriasis and the identification of novel therapeutic targets and approaches to treating this disease. This K01 award will allow me to become immersed in a rich environment where a multidisciplinary team of investigators will teach me the biostatistics, immunology and molecular biological skills needed to achieve my career goals. Research Environment: I have at my disposal all of the resources required to successfully complete this project, including 400 sq. ft. of lab space furnished with equipment for cell culture, RNA isolation, PCR, flow cytometry. I have the mentorship of Dr. Elder and an Advisory Committee composed of prominent experts in biostatistics (Dr. Goncalo Abecasis), T cell biology (Dr. Weiping Zou), IL-1/inflammasome biology (Dr. Gabriel Nunez) and dermatology (Dr. John Voorhees). Under the supervision of my mentors, a comprehensive training plan has been devised for my career development: (1) To develop skills in statistics and systems biology analysis - a number of formal courses/workshops have been targeted together with the guidance of Drs. Abecasis and Elder. (2) To develop knowledge and skills in keratinocyte biology and function - mentoring from Dr. Elder and a formal course and collaboration with the Skin Disease Resource Center Keratinocyte Core facility, Northwestern University. (3) To develop knowledge and skills to study the skin immune system and T cell biology - mentoring by Dr. Zou and formal training with the Flow Cytometry Core Facility, University of Michigan. (4) To hone grant writing skills and learn to write effective internal review board (IRB) proposals for ethically conducted research on human subjects including the responsible conduct of research (RCR) - I will use of a number of resources at University of Michigan to develop skills for grantsmanship, IRB proposals, and training in RCR issues. (5) To foster leadership, mentoring and team-building skills - this will be achieved by working closely with rotating undergraduate and graduate students and students in the University of Michigan Undergraduate Research Opportunity Program where I will be able to recruit and work with an undergraduate student on a multiple year basis to foster their scientific development and interest in medical research. (6) To improve written and oral communication skills - through attending both Dermatology and Immunology departmental research meetings as well as participating in national immunology and dermatology meetings. Research Project: Psoriasis is an inflammatory and hyperproliferative skin disease driven by interactions of T cell, antigen-presenting cells (APC), keratinocytes and cytokines. The IL-36 cytokines (IL-36¿, ¿, ? and IL-36Ra) are recently defined members of the IL-1 family and are overexpressed in lesional psoriasis tissue. The broad, long-term objective of this proposal is to understand how the expression, processing and function of IL- 36 cytokines impact on the epidermal hyperplasia seen in psoriasis and identify opportunities for the development of innovative therapeutic strategies. Our central hypothesis is that IL-36 cytokines contribute to psoriatic inflammation by (a) directly driving keratinocyte innate host defense in an autocrine manner, and (b) activating APC to invoke Th17 T cell responses which then (c) synergize with IL-36 in amplifying the keratinocyte response. There are a number of uncharacterized steps regulating the secretion of active IL-36 from keratinocytes and the effects of IL-36 on human APC and KC remain to be elucidated. To test this hypothesis, we set out the following Specific Aims: (1) Determine whether IL-36 proteolysis occurs in KC and delineate the mechanism of IL-36 secretion; (2) Determine the differences in the activity of IL-36¿, ¿ and ? and their contribution to skin inflammation; (3) Elucidate the role of IL-36 on APC function and the Th1/Th17 T cell bias in psoriasis.