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慢病毒技术

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发表于 2015-10-30 22:48:31 | 显示全部楼层 |阅读模式
Keywords: folate receptor alpha, chimeric antigen receptor, adoptive immunotherapy, ovarian cancer, T cells
Received: May 12, 2015  Accepted: August 20, 2015  Published:September 02, 2015
Rigorous optimization and validation of potent RNA CAR T cell therapy for the treatment of common epithelial cancers expressing folate receptor
方案的优化和确定:使用 RNA 嵌合抗原受体修饰的T细胞治疗表达叶酸受体的常见上皮癌
Using lentiviral technology, we recently demonstrated that incorporation of CD27 costimulation into CARs greatly improves antitumor activity and T cell persistence.Still, virus-mediated gene transfer is expensive, laborious and enables long-term persistence, creating therapies which cannot be easily discontinued if toxic.To address these concerns, we utilized a non-integrating RNA platform to engineer human T cells to express FRα-specific, CD27 CARs and tested their capacity to eliminate human FRα+ cancer. Novel CARs comprised of human components were constructed, C4-27z and C4opt-27z, a codon-optimized variant created for efficient expression.
Following RNA electroporation, C4-27z and C4opt-27z CAR expression is initially ubiquitous but progressively declines across T cell populations. In addition, C4-27z and C4opt-27z RNA CAR T cells secrete high levels of Th-1 cytokines and display strong cytolytic function against human FRα+ cancers in a time- and antigen-dependent manner.Further, C4-27z and C4opt-27z CAR T cells exhibit significant proliferation in vivo, facilitate the complete regression of fully disseminated human ovarian cancer xenografts in mice and reduce the progression of solid ovarian cancer. These results advocate for rapid progression of C4opt-27z RNA CAR to the clinic and establish a new paradigm for preclinical optimization and validation of RNA CAR candidates destined for clinical translation
使用慢病毒技术,我们最近发现,合并CD27渗入CARs,会大大提高T细胞的抗肿瘤活性和持久性。然而, 病毒介导的基因转移非常昂贵,费力并且会长期存在,如果有毒性,不容易停止,需要进行新的治疗。为了解决这些问题,我们构建了一个非集成的RNA平台来改造人T细胞,使其表达特异的叶酸受体α和CD27嵌合抗原受体,来检测它们消灭人表达叶酸受体α类癌症的能力。我们构建了新型的CARs,C4-27zC4opt-27z,该变异体密码子得到了优化,表达效率高。随着RNA电穿孔,C4-27z 和C4opt-27z CAR的表达刚开始非常容易,但是当通过T细胞群落时,很快减少。此外,C4-27z 和 C4opt-27z RNA CAR T 细胞分泌大量的TH-1细胞因子,而且对于FRα+癌症显示出强大的溶细胞功能,并且是时间-抗原依赖性的方式。此外,C4-27z 和 C4opt-27z CAR T细胞在体内能够有显著的增值,帮助异种移植人卵巢癌小鼠完全消除扩散的肿瘤。这些结果倡导了C4opt-27z RNA CAR用于临床,而且对于临床治疗的候选RNA CAR的优化和验证,建立一个新的范式。
关键词:叶酸受体α,嵌合抗原受体,免疫疗法,卵巢癌,T细胞http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=5029&pubmed-linkout=1

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