The clinical use of Flt3L - an immune adjuvant to potentiate Dendritic Cells

描述（由申请人提供）：该提案描述了一个为期5年的研究皮肤病学学术生涯发展培训计划。主要研究者完成了纽约大学皮肤科住院医师学位和斯坦福大学医学博士学位。她在基础科学研究方面有着良好的记录，并将扩展她在人类临床转化方法、临床试验设计和免疫监测分析方面的科学技能。她之前受到已故的拉尔夫·斯坦曼博士的指导，在他收到201年诺贝尔医学奖之前，她最初与他一起形成了这个应用程序和项目。她现在将由米歇尔·努森茨威格博士密切指导，他是HHMI在适应性免疫方面的杰出研究者。Flt3L是一种有效的DC造血素，可扩大小鼠CD8交叉呈递树突状细胞（DC）。研究将侧重于Flt3L作为一种免疫佐剂的临床开发，以扩大人类DC亚群的数量，测试其对人类免疫的影响，并测试其与基于蛋白质的DC靶向免疫联合使用时在小鼠中的临床前疗效。Flt3L已经在300多名患者中证明了安全性，但由于商业化问题，该药物的开发停止了。FDA最近解除了对Flt3L的临床限制，因此重新评估该药物的研究可以继续进行。核心假设是Flt3L将动员和扩大人类的交叉呈递dc，促进优越的T细胞反应。在Provenge疗法（一种针对前列腺癌的DC治疗疫苗）中，患者首先要经历昂贵而复杂的白细胞分离程序，以获得足够的自体细胞来体外产生用于疫苗接种的DC。使用Flt3L提供了一种替代方法，可以在简单注射后在体内产生充足且更具生理性的DC前体，替代目前的白细胞分离步骤。因此，这些实验可能会为这种基于dc的抗癌策略提供实质性的改进，并开发出一种创新的方法来生产佐剂、治疗性dc疫苗和其他组合免疫疗法。长期目标是研究克服耐受性和改善皮肤免疫的机制，重点是生物方法来增强这种应用中的特定DC。假设是Flt3L在物种间扩展交叉呈现的DC等价物，可以用作佐剂。这一假设将通过追求两个特定目标来验证：1)建立和分析Flt3L患者的CD8 -等量物，比较人类DC亚群功能，评估Flt3L治疗后新生和回忆免疫反应的变化；2)评估Flt3L与佐剂偶联后是否能改善免疫，用于人类和DC靶向免疫小鼠，允许体内靶向而不需要白细胞沉淀。它是创新的，因为使用Flt3L增强人类交叉呈递DC，以DC亚群靶向为目标是新颖的，它使用先进的免疫方法，用申请人实验室开发的基于蛋白质的疫苗靶向DC，目前正在人体中进行测试。该培训计划旨在确保树突细胞生物学应用于免疫治疗和辅助设计，以改善健康和免疫力。Michel Nussenzweig教授将指导主要研究者的科学发展。Nussenzweig博士是DC开发，靶向和领导适应性免疫领域的关键领导者。他和斯坦曼博士的合作让我们对控制耐受性和免疫力有了新的认识。努森茨威格博士指导过许多初级和高级教员。培训活动将通过在正式的临床转化项目中重点指导高级免疫学和转化生物学来加强。一个由杰出的内科科学家组成的科学咨询委员会将提供科学和职业建议。洛克菲勒大学的细胞生理学、免疫学和分子免疫学实验室是解决临床免疫学复杂领域所需的实验室和转化科学技能强化培训的理想场所。这个环境将为研究皮肤病学的学术生涯做好准备，在那里她将解决免疫佐剂在改善皮肤免疫和预防和治疗皮肤癌方面的作用。

DESCRIPTION (provided by applicant): This proposal describes a 5-year training program for the development of an academic career in Investigative Dermatology. The principle investigator has completed a residency in Dermatology from New York University and her MD/PhD from Stanford University. She has a strong track record of basic science research and will expand her scientific skills in human clinical translational methods, clinical trial design, and immuno-monitoring assays. She was previously mentored by the late Dr. Ralph Steinman, prior to his receipt of the 201 Nobel Prize in Medicine and with whom she initially formed this application and project. She will now be mentored closely by Dr. Michel Nussenzweig now- an outstanding HHMI investigator in adaptive immunity. Flt3L, a potent DC hematopoietin, expands CD8 cross-presenting dendritic cells (DC) in mice. Research will focus on the clinical development of Flt3L as an immune adjuvant to expand DC subset numbers in humans, testing its impact on immunity in humans and testing its preclinical efficacy in mice, when combined with protein-based DC targeted immunization. Flt3L has demonstrated safety in over 300 patients, but development of this agent was ceased due to commercialization issues. The FDA has recently lifted a clinical hold on Flt3L so studies to re-evaluate this agent can continue. The central hypothesis is that Flt3L will mobilize and expand cross-presenting DCs in humans, promoting superior T cell responses. In Provenge therapy (a DC therapeutic-vaccine targeted against prostate cancer), patients first undergo an expensive and complex leukapheresis procedure to obtain sufficient autologous cells to generate the DCs ex vivo to use for the vaccination. Using Flt3L offers an alternative approach to generate ample and more physiological DC precursors in vivo after simple injection, as a substitute for the current leukapheresis step. Thus these experiments may offer substantial improvement in such a DC-based anti-cancer strategy, and also develop an innovative approach to generating adjuvants, therapeutic DC-vaccines, and other combinatorial immunotherapy. The long-term goal is to investigate mechanisms to overcome tolerance and improve cutaneous immunity, focusing on biologic methods to potentiate particular DC in this application. The hypothesis is that Flt3L expands cross-presenting DC equivalents across species and can be used as an adjuvant. This hypothesis will be tested by pursuing 2 specific aims to (1) establish and profile CD8� equivalents in humans with Flt3L and compare human DC subset functions and assess alterations in de novo and recall immune responses after Flt3L treatment (2) assess whether Flt3L improved immunity when coupled to adjuvants for human use and DC targeted immunization in mice, that allow in vivo targeting without leukophoresis. It is innovative because the use of Flt3L to potentiate human cross-presenting DC with the goal of DC subset targeting is novel~ it uses advanced immunization methods to target DC with protein-based vaccines developed in the applicant's laboratory, and currently being tested in humans. The training program has been designed to ensure a command of dendritic cell biology as applied to immunotherapy and adjuvant design towards improved health and immunity. Prof. Michel Nussenzweig will mentor the principle investigator's scientific development. Dr. Nussenzweig is a key leader of the DC development, targeting, and leads the adaptive immune field. His work with Dr. Steinman has led to a new understanding of the control of tolerance and immunity. Dr. Nussenzweig has mentored numerous junior and senior faculty members. Training activities will be enhanced through focused instruction in advanced immunology and translational biology in a formal clinical translational program. A scientific advisory committee, composed of exceptional physician scientists, will provide scientific and career advice. The Laboratories of Cellular Physiology and Immunology and Molecular Immunology at Rockefeller University are the ideal setting for intensive training in bench and translational science skills required to address a complex area of clinical immunology. This environment will prepare the principle investigator for an academic career in investigative dermatology where she will address a role for immune adjuvants in improving cutaneous immunity and the prevention and treatment of skin cancers.