Oral Tolerance for Hemophilia

血友病的口服耐受性

• 批准号: 9297901
• 负责人: HENRY DANIELL
• 金额: $ 66.84万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-10 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9297901
• 关键词: Address; Affect; Anaphylaxis; Animal Model; Animals; Antibodies; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Biomedical Engineering; Birth; Blood; Blood Coagulation Disorders; Bypass; CD3 Antigens; CD4 Positive T Lymphocytes; Canis familiaris; Cells; Cellular Metabolic Process; Chloroplasts; Clinical; Clinical Trials; Complication; Data; Dendritic Cells; Disease; Dose; Epithelium; F8 gene; FOXP3 gene; Factor IX; Factor VIII; Factor VIIa; Funding; Future; Future Generations; Genetic Engineering; Grant; Health Care Costs; Helper-Inducer T-Lymphocyte; Hemophilia A; Hemophilia B; Immune; Immune Tolerance; Immune response; Immune system; Immunosuppression; Injection of therapeutic agent; Interdisciplinary Study; Intravenous; Intravenous infusion procedures; Laboratories; Lamina Propria; Lettuce - dietary; Life; Light; Link; Liver; Methods; Modeling; Morbidity - disease rate; Mus; Nature; Nephrotic Syndrome; Oral; Ovalbumin; Pain; Patients; Pharmaceutical Preparations; Plants; Production; Proteins; Protocols documentation; Quality of life; Reagent; Recombinants; Regulatory T-Lymphocyte; Risk; Safety; Science; Site; Small Intestines; Structure of aggregated lymphoid follicle of small intestine; System; T-Lymphocyte; Temperature; Time; TimeLine; Transgenic Organisms; Transgenic Plants; Translations; Work; antibody inhibitor; base; cost; cytokine; editorial; genomic tools; immunoregulation; immunotoxicity; improved; inhibitor/antagonist; male; mortality; mouse model; next generation; novel; oral tolerance; overexpression; pediatric patients; prevent; prophylactic; public health relevance; recombinant antihemophilic factor VIII; response; scale up; single molecule; standard of care; success; therapeutic protein; tool; translational study; uptake

PROJECT SUMMARY/ABSTRACT. The current standard of care for the X-linked bleeding disorder hemophilia is intravenous (IV) infusion of recombinant factor VIII (FVIII, for hemophilia A) or factor IX (FIX for hemophilia B). These protein products are expensive, require frequent repeated IV injections (which is painful and inconvenient), and are often targeted by antibody (“inhibitor”) responses; thereby complicating/neutralizing therapy, creating immunotoxicities, and further increasing costs. In fact, inhibitor formation, an antigen-specific CD4+ T helper cell-driven B cell response, is widely considered the most serious complication of current therapy for hemophilia. Forming an interdisciplinary research team to address this problem, our laboratories have been closely collaborating since 2007 to develop a bioengineering-based approach. We conceived and now validated the concept that oral delivery of bioencapsulated FVIII and FIX antigens produced by the chloroplasts of transgenic plants could suppress inhibitor formation in animal models of hemophilia. FVIII and FIX antigens were effectively delivered to the epithelium of the small intestine when expressed as fusions to a transmucosal carrier, and subsequently taken up by dendritic cells (DCs) in the lamina propria (LP) and Peyer's patches (PP). Tolerance was established (over a wide range, including very low antigen doses) by induction of multiple subsets of regulatory T cells (Treg) in an IL-10 dependent manner. In addition, this approach prevented and also reversed anaphylaxis against FIX. Importantly, we were able to generate chloroplast transgenic edible crop plants (lettuce), and scale up of production was successful to a commercial system that is used to cost- effectively generate GMP material. To further advance this approach, we propose the following specific aims: 1. Generate the next generation of transplastomic edible crop plants expressing FVIII antigen using cutting-edge chloroplast genomics tools and alternative transmucosal carriers. 2. Continue to define the mechanism of oral tolerance induction/immune regulation, in part through use of a model antigen. 3. Identify optimal plants for oral tolerance induction in hemophilia A mice; perform translational studies in hemophilia A dogs; and further strengthen oral tolerance by enhancing immune regulation or manipulating T cell metabolism. This work will facilitate translation of oral tolerance for hemophilia into clinical trials, define the mechanism of how plant cell-based oral tolerance is accomplished, pave the way for future combination protocols for optimal oral tolerance induction, and further advance genetic engineering of plants for biomedical applications.

项目摘要/摘要。 X连锁出血障碍血友病的当前护理标准是静脉内（IV）输注 重组因子VIII（FVIII，用于血友病A）或第IX因子（血友病B）。这些蛋白质产物 价格昂贵，需要经常重复的IV注射（这很痛苦且不便），并且经常是 由抗体（“抑制剂”）反应靶向；从而使疗法复杂化/中和，创造 免疫毒性，并进一步增加成本。实际上，抑制剂形成，抗原特异性CD4+ T助手 细胞驱动的B细胞反应被广泛认为是当前治疗中最严重的并发症 血友病。组建一个跨学科研究团队来解决这个问题，我们的实验室已经 自2007年以来，紧密合作，开发一种基于生物工程的方法。我们构思并现在验证 叶绿体产生的生物封装的FVIII和固定抗原的口服的概念 转基因植物可以抑制血友病动物模型中的抑制剂形成。 FVIII并修复抗原 当表达为透射蛋白的融合时，有效地输送到小肠的上皮 载体，随后被树突状细胞（DC）在固有层（LP）和Peyer的斑块（PP）中占用。 通过诱导多种 调节T细胞（TREG）的子集以IL-10依赖性方式。此外，这种方法阻止了 还逆转过敏反应。重要的是，我们能够生成叶绿体转基因食用作物 植物（生菜）和生产规模取得了成功，用于商业系统，用于成本 - 有效生成GMP材料。为了进一步推进这种方法，我们提出以下特定目的：1。 生成下一代的移植作物植物，使用尖端表达FVIII抗原 叶绿体基因组学工具和替代性跨粘膜载体。 2。继续定义口服的机制 耐受性诱导/免疫调节，部分通过使用模型抗原。 3。确定最佳植物 血友病A小鼠的口服耐受性诱导；在血友病A狗中进行翻译研究；还有 通过增强免疫调节或操纵T细胞代谢来增强口服耐受性。这项工作将 促进将血友病的口服耐受性转化为临床试验，定义了植物的机制 实现了基于细胞的口服耐受性，为未来的组合方案铺平了道路，以获得最佳的口服 耐受性诱导，并进一步推进生物医学应用的植物基因工程。