Sleeping Beauty Mediated Therapy for Alpha V Beta 6-Expressing Pancreatic Cancer

描述(申请人提供)：胰腺癌在最常见的癌症诊断中排名第十，但在最常见的估计死亡原因中排名第四。胰腺癌唯一可能的根治方法是手术切除；然而，很少有患者有可以尊重的肿瘤。胰腺导管腺癌占胰腺癌的90%，它的侵袭性特别强，因为它转移得很快，通常表达几种允许快速生长的生长因子和信号成分。由于胰腺癌的治疗最近没有医学上的进展，因此迫切需要替代疗法。通过过继转移工程T细胞进行的免疫治疗可以介导肿瘤的消退，并克服肿瘤逃避免疫反应的逃避机制。嵌合抗原受体(CAR)是一种融合了抗体衍生的抗原结合基序和细胞内信号域的工程分子，可以独立于主要组织相容复合体识别肿瘤抗原，而主要组织相容复合体的表达经常被肿瘤细胞丢失。在发现基因组公司(DGI)，我们正在开发睡美人转座子系统，以产生用于自体过继T细胞治疗的T细胞。在这里，我们建议使用SB工程T细胞系统来表达CAR识别？V？6，这是一种在胰腺癌细胞上高度表达的整合素。此外，我们还将稳定地共表达一个嵌合受体(4？？)，它由IL-4受体？共同受体的胞外区和内区，后者是IL-2和IL-15受体的组成部分。4的表达？？受体通过抗V6 T细胞选择性扩增CAR阳性细胞。白介素4与4？？受体会激活增殖信号，从而刺激T细胞分裂。基因工程的T细胞将通过流式细胞仪进行免疫表型鉴定。体外实验将通过检测基因工程T细胞与V6阳性胰腺癌细胞接触后细胞因子的分泌和脱颗粒以及表达CAR的T细胞对V6阳性胰腺癌细胞的杀伤能力来评价其抗肿瘤细胞毒作用。这些实验的结果将为SB工程T细胞在杀死胰腺癌细胞方面的有效性提供评估。随后对胰腺癌动物模型的研究将导致通过使用睡美人转座子系统在体外使用基因工程的人类T细胞来治疗胰腺癌的临床试验。

DESCRIPTION (provided by applicant): Pancreatic cancer represents the 10th most common cancer diagnosis, yet the 4th most common estimated cause of death. The only potential curative therapy for pancreatic cancer is surgical resection; however, few patients have tumors which can be respected. Pancreatic ductal adenocarcinoma, which represents 90% of pancreatic cancers, is particularly aggressive, since it rapidly metastasizes and often expresses several growth factors and signaling components that permit rapid growth. Alternative therapies are desperately needed, as there have been no recent medical advances for treatment of pancreatic adenocarcinoma. Immunotherapy by adoptive transfer of engineered T cells can mediate cancer regression and overcome evasive mechanisms by which tumors avoid immune responses. Chimeric antigen receptors (CAR) are engineered molecules that are a fusion of an antibody-derived antigen-binding motif and intracellular signaling domains, and can recognize tumor antigens independently of the major histocompatiblity complex, expression of which is often lost by tumor cells. At Discovery Genomics, Inc. (DGI), we are developing the Sleeping Beauty transposon system to generate T cells for autologous adoptive T cell therapy. Here we propose to use the SB system for engineering T cells to express CAR recognizing ?V?6, an integrin that is highly expressed on pancreatic cancer cells. In addition, we will stably co-express a chimeric receptor (4??), consisting of a fusion of the IL-4 receptor ? extracellular domain and the endodomain of the common ?- receptor, which is a component of both IL-2 and IL-15 receptors. Expression of the 4?? receptor by anti-??V?6 T cells will allow selective expansion of CAR positive cells. Binding of interleukin-4 to the 4?? receptor will activate a proliferative signal, thus stimulating the T cells to divide. The engineered T cells will be immunophenotypically characterized by flow cytometry. Antitumor cytotoxic activity will be assessed in vitro by measuring cytokine secretion and degranulation after exposure of engineered T cells to ?V?6 positive pancreatic cancer cells and the ability of CAR-expressing T cells to lyse ?V?6 positive pancreatic cancer cells. Results from these experiments will provide an assessment of the effectiveness of SB-engineered T cells in killing pancreatic cancer cells. Subsequent studies in animal models of pancreatic cancer will lead to a clinical trial for the treatment of pancreatic adenocarcinoma by administering human T-cells genetically engineered ex vivo using the Sleeping Beauty transposon system.