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

描述（由申请人提供）：胰腺癌是第十大最常见的癌症诊断，但也是第四大最常见的估计死亡原因。胰腺癌的唯一潜在治愈性治疗是手术切除;然而，很少有患者具有可以考虑的肿瘤。胰腺导管腺癌占胰腺癌的90%，特别具有侵袭性，因为它迅速转移，并且经常表达允许快速生长的几种生长因子和信号成分。替代疗法是迫切需要的，因为最近没有治疗胰腺癌的医学进展。通过过继转移工程化T细胞的免疫疗法可以介导癌症消退并克服肿瘤逃避免疫应答的逃避机制。嵌合抗原受体（CAR）是工程化的分子，其是抗体衍生的抗原结合基序和细胞内信号传导结构域的融合物，并且可以独立于主要组织相容性复合物识别肿瘤抗原，其表达通常被肿瘤细胞丢失。在Discovery Genomics，Inc. (DGI)我们正在开发睡美人转座子系统，以产生用于自体过继性T细胞治疗的T细胞。在这里，我们建议使用SB系统工程T细胞表达CAR识别？小维6，一种在胰腺癌细胞上高度表达的整合素。此外，我们将稳定共表达嵌合受体（4？？），由IL-4受体的融合体组成细胞外结构域和共同的内结构域？受体，其是IL-2和IL-15受体两者的组分。4的表达方式？受体抗？？小维6 T细胞将允许CAR阳性细胞的选择性扩增。白细胞介素-4与4？受体将激活增殖信号，从而刺激T细胞分裂。工程化T细胞将通过流式细胞术进行免疫表型表征。抗肿瘤细胞毒活性将评估在体外通过测量细胞因子分泌和脱粒后，工程化的T细胞暴露于？小维6阳性胰腺癌细胞和表达CAR的T细胞裂解？V？6例胰腺癌细胞阳性。来自这些实验的结果将提供SB工程化T细胞在杀死胰腺癌细胞中的有效性的评估。在胰腺癌动物模型中的后续研究将导致通过施用使用睡美人转座子系统离体遗传工程化的人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.