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

描述(由申请人提供)：本建议书描述了一项为期5年的皮肤病研究学术生涯发展培训计划。这位首席研究员已经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，并将在人类临床翻译方法、临床试验设计和免疫监测分析方面扩展她的科学技能。在已故的拉尔夫·斯坦曼博士获得201年诺贝尔医学奖之前，她曾接受过他的指导，她最初是与他一起形成了这个申请和项目。她现在将得到Michel Nussenzweig博士的密切指导--他是一位杰出的HHMI适应性免疫研究员。Flt3L是一种有效的DC造血素，能在小鼠体内扩增CD8交叉呈递的树突状细胞(DC)。研究将集中在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靶向免疫结合时在小鼠的临床前疗效。Flt3L已经在300多名患者身上证明了安全性，但由于商业化问题，这种药物的开发停止了。FDA最近解除了对Flt3L的临床控制，因此重新评估该制剂的研究可以继续进行。中心假设是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 Treatment(一种针对前列腺癌的DC治疗性疫苗)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，以在体外产生DC用于疫苗接种。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和更具生理性的DC前体，作为当前白细胞分离步骤的替代。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，并开发一种创新的方法来产生佐剂、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提高皮肤免疫力的机制，重点是生物方法在这一应用中增强特定的DC。假设Flt3L扩展了跨物种的交叉递呈DC等价物，并可用作佐剂。这一假说将通过追求两个特定目标来验证：(1)建立和描述人类CD8Flt3L的�等价物，比较人类DC亚群功能，评估Flt3L治疗后从头和召回免疫反应的变化；(2)评估Flt3L与人用佐剂和小鼠DC靶向免疫时是否提高了免疫力，从而允许体内靶向而无需白细胞渗透。这是创新的，因为使用Flt3L来增强以DC亚群靶向为目标的人类交叉呈递DC是新颖的-它使用先进的免疫方法来靶向DC，该疫苗是由申请人的实验室开发的基于蛋白质的疫苗，目前正在人体上进行测试。该培训计划旨在确保掌握树突状细胞生物学，将其应用于免疫治疗和辅助设计，以改善健康和免疫力。米歇尔·努森茨韦格教授将指导首席研究员的科学发展。Nussenzweig博士是DC开发、靶向的关键领导者，并领导适应性免疫领域。他与斯坦曼博士的合作使人们对耐受性和免疫力的控制有了新的理解。努森茨韦格博士曾指导过许多初级和高级教职员工。培训活动将通过在一个正式的临床翻译计划中集中讲授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板凳强化培训和翻译科学技能的理想环境。这种环境将为首席研究员在皮肤病学研究领域的学术生涯做好准备，在那里她将阐述免疫佐剂在提高皮肤免疫力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.