Packaging Cell Lines for Lentiviral Vector Products

描述（由申请人提供）：基于HIV-1的慢病毒载体正成为一种越来越有吸引力的将转基因整合到靶细胞中的方法。病媒正在进行或进入针对多种疾病的临床试验，包括遗传病、艾滋病和癌症。为了将靶细胞的范围扩大到HIV-1 （CD4+细胞）之外，载体用各种病毒包膜进行假型。Rimedion已经确定罗斯河病毒（RRV）和猫RD114包膜糖蛋白是生产稳定包装细胞系的有希望的候选者。在这个STTR申请中，Rimedion和它的研究合作伙伴，印第安纳大学医学院提出利用这些包膜开发新的、稳定的慢病毒包装细胞系。这些细胞系将有助于进一步的临床前开发，也将适用于未来的临床应用。具体目标1:Rimedion将生成表达RRV和RD114信封的稳定包装线。特异性目标2：利用RRV和RD114的包装细胞系，产生稳定的慢病毒产生细胞系，表达绿色荧光蛋白，用于载体滴度、细胞系长期稳定性和慢病毒复制能力检测的研究。这些细胞系将取代目前慢病毒载体的生产方法（瞬时转染），这种方法不适合大规模生产，并且限制了慢病毒载体获得FDA许可的潜力。如果成功，这些细胞系将为一大批新的生物制剂提供技术平台。第二阶段将包括符合FDA指南的认证，以便这些生产线可以用于良好生产规范。作为II期的一部分，RD114细胞系将用于在I期临床试验中创建用于治疗腺苷脱氨酶缺乏症的生产细胞系。通过适当的材料协议，这些线路也将通过NIH赞助的项目（如国家基因载体生物资源库）提供给学术研究人员用于研究，非商业用途。

DESCRIPTION (provided by applicant): HIV-1 based lentiviral vectors are becoming an increasingly attractive means of integrating transgenes into target cells. Vectors are in or enterning clinical trials for a diverse group of ailments, including genetic disease, AIDS and cancer. To expand the range of target cells beyond that of HIV-1 (CD4+ cells), vectors are pseudotyped with various viral envelopes. Rimedion has identified the Ross River Virus (RRV) and the feline RD114 envelope glycoproteins to be promising candidates for production of stable packaging cell lines. In this STTR application, Rimedion and its research partner, Indiana University School of Medicine proposes to develop novel, stable lentiviral packaging cell lines using these envelope. These lines will be useful for further preclinical development and will also be suitable for future clinical use. Specific Aim 1: Rimedion will generate stable packaging lines expressing the RRV and RD114 envelopes. Specific Aim 2: Using the packaging cell lines for RRV and RD114, generate stable lentiviral producer cell lines expressing the Green Fluorescent Protein to be used for studies of vector titer, long-term cell line stability and replication competent lentiviral testing. These cell lines will replace the current manufacutring methodolgy (transient transfection) for lentiviral vectors which is not suitable for large scale productions and limits the potential of lentiviral vectors from achieving licensure by the FDA. If successful, these cell lines will provide the platform technology for a large group of new biologic agents. Phase II will include the certification to meet FDA Guidances so the lines may be used in Good Manufacturing Practices. As part of Phase II, the RD114 cell line would be used to create a producer cell line for treatment of adenosine deaminase deficieny in a Phase I clinical trial. Through appropriate material agreements, the lines will also be made available to academic investigators for investigational, non-commerical use through NIH sponsored programs such as the National Gene Vector Biorepostory. PUBLIC HEALTH RELEVANCE: Gene therapy mediated by lentiviral vectors is now in clinical trials and holds promise for a wide variety of genetic diseases, AIDS and cancer. A major limitation to commercialization of these biologic products is large- scale production technology. Rimedion proposes to generate a series of lentiviral packaging cell lines that will allow manufacturing of lentiviral products suitable for FDA licensure specifications.