Humanizing the Mouse Immune System using BAC

使用 BAC 人性化小鼠免疫系统

• 批准号: 6882152
• 负责人: HIROAKI SHIZUYA
• 金额: $ 12.9万
• 依托单位: ALIVA BIOPHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-15 至 2006-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882152
• 关键词: Escherichia coli; animal genetic material tag; animal tissue; artificial chromosomes; bacterial DNA; biotechnology; embryonic stem cell; genetic library; genetically modified animals; human genetic material tag; immunoglobulin genes; laboratory mouse; polymerase chain reaction; recombinant DNA; southern blotting; technology /technique development

DESCRIPTION (provided by applicant): Antibodies have become the foundation of the new field of protein therapeutics. They are used to treat immune related diseases and proliferative disorders such as cancer. Six different human antibody therapeutics have already been approved by the FDA for treatment of cancer, autoimmune diseases and to prevent transplant rejection and many more of these protein therapeutics are in clinical development. The generation of completely human antibodies in animals provides for the development of molecules that will avoid the problems of rejection and immunogenicity .inherent in the development of "humanized" or "chimeric" animal antibodies. However, the generation and development of human antibodies in animals is has been challenging The process involves insertion of near-mega base length human sequences encoding portions of one or more immunoglobulin (Ig) loci into embryonic stem (ES) cells and then transfer to animals whose host immune system have been knocked out. Transgenic animals are then challenged with antigen to produce human antibodies and B-cells isolated to manufacture the desired monoclonal antibody via hybridoma technology. While many methods have been used to generate animals that express various human proteins, most approaches are not capable of reliable and accurately transferring the large amounts of genetic material needed for antibody production. In additional, the initial approaches used to transfer the human Ig loci into immune deficient mice, YAC and transchromosomal technology, have a number of significant drawbacks. Aliva is a new start-up biotechnology company that has developed a technology that can provide important advantages over present approaches to generating human antibodies in animals. Aliva is focused on developing bacterial artificial chromosome (BAG) technology, first invented by Aliva's founder and Chief Scientific Officer, Dr.Hiroaki Shizuya, as a new generation system that will for the first time allow both targeted and sequential transfer of large stretches of genetic material to a host animal to produce a diverse repertoire of fully human therapeutic antibodies. We propose in this phase I SBIR grant, proof of principle studies .to demonstrate the sequential and targeting capabilities of the Aliva BAG technology, and to begin to generate a mouse that will produce true human antibodies. For this, we will create and use BAG to replace a major portion of the mouse heavy chain constant region with a 600-700Kb region of the human heavy chain in an ES cell line. This will provide the foundation for future phase II SBIR studies to generate Fl mice from the ES cells and replace the remaining heavy and light chain regions to generate mice capable of produce a full repertoire of human antibodies

描述（由申请人提供）：抗体已成为蛋白质治疗新领域的基础。它们用于治疗免疫相关疾病和增殖性疾病，例如癌症。 FDA 已批准六种不同的人类抗体疗法用于治疗癌症、自身免疫性疾病和预防移植排斥，还有更多的蛋白质疗法正在临床开发中。 在动物中产生完全人类抗体提供了分子的开发，该分子将避免“人源化”或“嵌合”动物抗体开发中固有的排斥和免疫原性问题。然而，在动物体内产生和开发人类抗体一直具有挑战性。该过程涉及将编码一个或多个免疫球蛋白（Ig）基因座部分的近兆碱基长度的人类序列插入胚胎干（ES）细胞，然后转移到动物体内其宿主免疫系统已被敲除。然后用抗原激发转基因动物，产生人类抗体，并分离 B 细胞，通过杂交瘤技术制造所需的单克隆抗体。尽管已使用许多方法来产生表达各种人类蛋白质的动物，但大多数方法无法可靠且准确地转移抗体生产所需的大量遗传物质。此外，最初用于将人类 Ig 基因座转移到免疫缺陷小鼠体内的方法（YAC 和转染色体技术）具有许多显着的缺点。 Aliva 是一家新成立的生物技术公司，该公司开发了一种技术，与现有的在动物体内产生人类抗体的方法相比，该技术具有重要优势。 Aliva 专注于开发细菌人工染色体 (BAG) 技术，该技术由 Aliva 创始人兼首席科学官 Hiroaki Shizuya 博士首次发明，作为新一代系统，该系统将首次允许大段遗传基因的定向和顺序转移材料注入宿主动物体内，产生多种全人类治疗性抗体。我们建议在第一阶段 SBIR 拨款中进行原理研究证明，以证明 Aliva BAG 技术的顺序和靶向能力，并开始培育能够产生真正人类抗体的小鼠。为此，我们将在 ES 细胞系中创建并使用 BAG 将小鼠重链恒定区的主要部分替换为人重链的 600-700Kb 区域。这将为未来 II 期 SBIR 研究奠定基础，以从 ES 细胞产生 F1 小鼠，并替换剩余的重链和轻链区域，以产生能够产生完整人类抗体库的小鼠