Oral immune modulatory therapy using antigens bioencapsulated in plant cells

使用生物封装在植物细胞中的抗原进行口服免疫调节疗法

基本信息

批准号：
8299074
负责人：
HENRY DANIELL
金额：
$ 44.61万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-15 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8299074
关键词：
A-factor (Streptomyces)Acids Address Allergic Allergic Reaction Anaphylaxis Animal Model Antibodies Antibody Formation Antigen Presentation Antigen-Presenting Cells Antigens Binding Bioreactors Blood Coagulation Factor Bypass Canis familiaris Cell Wall Cells Chloroplasts Clinical Cold Chains Complication Development Disease Dose Effectiveness Encapsulated Enzymes Epithelial Factor IX Fermentation Freeze Drying Funding Future Gene Deletion Generations Genes Genetically Modified Plants Glycogen storage disease type II Goals Grant Gut associated lymphoid tissue Hemophilia A Hemophilia B Hemorrhage Hereditary Disease Human IgE Immune Immune Tolerance Immune system Immunosuppression Incidence Inherited Injection of therapeutic agent Intravenous infusion procedures Lettuce - dietary Life Ligands Link Lysosomal Storage Diseases Mediating Methods Morbidity - disease rate Mus Nephrotic Syndrome Nonsense Mutation Oral Orphan Disease Patients Pharmaceutical Preparations Plants Prevention Production Protein Deficiency Proteins Protocols documentation Reagent Recombinant Proteins Regulation Regulatory T-Lymphocyte Replacement Therapy Risk Role Sterility Stomach System Terminator Codon Testing Therapeutic Time Tobacco Transgenic Organisms Transgenic Plants Translations United States National Institutes of Health base clinical application commensal microbes cost cytokine enzyme replacement therapy experience genetic manipulation immunotoxicity inhibitor/antagonist microbial mortality novel novel strategies oral tolerance prevent prophylactic receptor response standard care therapeutic protein tissue culture translational study

项目摘要

DESCRIPTION (provided by applicant): Current treatment for many inherited protein deficiencies is based on intravenous infusion of recombinant proteins. The infused protein is expensive and is often encountered by antibody responses that neutralize therapy, thereby complicating treatment and further increasing costs. They also cause severe allergic or even life-threatening anaphylactic reactions. Oral immune modulatory therapy represents an ideal antigen-specific approach to prevent such responses because it is non-invasive and does not require immune suppression or genetic manipulation of the patient's cells. This proposal takes advantage of a unique concept based on the use of plant chloroplasts as highly efficient bioreactors for production and oral delivery of therapeutic protein antigens without a need for fermentation, purification, cold chain, or sterile injections. Bioencapsulated antigens are protected in the stomach from acids or enzymes, but are released in the gut when plant cell walls are digested by commensal bacteria. We propose to further advance this method for effective delivery of antigen-ligand fusions to the gut-associated lymphoid tissue. We have also made substantial progress towards development of an oral delivery system (based on the edible crop lettuce) suitable for clinical translation. In treatment of the X-linked bleeding disorder hemophilia, formation of inhibitory antibodies against the therapeutic clotting factor represents a serious complication that substantially increases morbidity and mortality. For example, patients with hemophilia B due to deletion or early stop codon in the factor IX (F.IX) gene are at increased risk for inhibitor formation. Not only are most F.IX inhibitors high-titer and difficult to eliminate, but these antibodies are often associated with potentially life-threatening anaphylactic reactions. Recently, we demonstrated that oral delivery of F.IX, produced by tobacco chloroplasts and bioencapsulated in plant cells, prevents formation of inhibitors and anaphylaxis in protein replacement therapy for hemophilia B (featured in PNAS 107:7101). These results illustrate the effectiveness of our approach towards immune modulatory therapy for inherited protein deficiencies. The goals of this proposal are to develop an oral immune modulatory protocol for hemophilia B that prevents or mitigates pathogenic antibody responses (including anaphylaxis); to dissect the underlying mechanisms of antigen presentation and immune regulation; to find alternative transmucosal carriers for effective antigen delivery; to test the approach for a different disease with similar antibody/anaphylactic response (the lysosomal storage disorder Pompe disease); to develop transplastomic lettuce to express F.IX for future oral delivery in humans; to investigate stability and microbial contamination of the freeze-dried transgenic material; and to perform translational studies in a large animal model of hemophilia B. We hypothesize that our low cost approach can be utilized in a prophylactic fashion in several diseases, such as hemophilia and lysosomal storage disorders, thereby eliminating the need for immune suppression.

描述（由申请人提供）：当前对许多遗传蛋白缺陷的治疗方法基于重组蛋白的静脉注射。注入的蛋白质很昂贵，经常会遇到中和治疗的抗体反应，从而使治疗复杂化并进一步增加成本。它们还会引起严重的过敏性甚至威胁生命的过敏反应。口服免疫调节疗法代表了一种理想的抗原特异性方法，可以防止这种反应，因为它是无创的，并且不需要对患者细胞的免疫抑制或遗传操纵。该建议利用了一个独特的概念，基于将植物叶绿体用作高效生物反应器的生产和口服治疗性蛋白质抗原而无需发酵，纯化，冷链或无菌注射的高效生物反应器。生物封装的抗原受到胃或酶的保护，但在植物细胞壁被共生细菌消化时释放在肠道中。我们建议进一步促进这种方法，以有效地递送抗原 - 配体融合到肠道相关的淋巴组织。我们还取得了巨大的进展，以开发适合临床翻译的口服输送系统（基于可食用的作物生菜）。在治疗X连锁出血障碍血友病时，对治疗性凝血因子的抑制性抗体形成代表了严重的并发症，大大增加了发病率和死亡率。例如，由于缺失或因素IX（f.ix）基因的早期停止密码子引起的血友病患者的抑制剂形成风险增加。不仅大多数f.ix抑制剂高光剂且难以消除，而且这些抗体通常与潜在的威胁生命的过敏反应有关。最近，我们证明了F.IX的口服递送，该F.IX是由植物细胞中烟草叶绿体产生的，可防止抑制剂和蛋白质替代疗法中的抑制剂和过敏反应的形成（PNAS 107：7101）。这些结果说明了我们针对遗传蛋白缺乏症的免疫调节治疗方法的有效性。该提案的目标是为血友病B开发一种口服免疫调节方案，以防止或减轻致病性抗体反应（包括过敏反应）；剖析抗原表现和免疫调节的潜在机制；找到有效抗原递送的替代透射载体；测试具有类似抗体/过敏反应的不同疾病的方法（溶酶体储存障碍疾病）；开发移植生菜来表达f.ix，以供人类未来的口服分娩；研究冷冻干燥的转基因材料的稳定性和微生物污染；并在大型血友病动物模型中进行翻译研究。我们假设可以在多种疾病中以预防性的方式使用低成本方法，例如血友病和溶酶体储存障碍，从而消除了对免疫抑制的需求。