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

睡美人介导治疗表达 Alpha V Beta 6 的胰腺癌

• 批准号: 8523478
• 负责人: Kendra A. Hyland
• 金额: $ 20.02万
• 依托单位: DISCOVERY GENOMICS, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-12 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8523478
• 关键词: Adoptive Transfer; Adult; Affinity; Alternative Therapies; Animal Model; Antibodies; Antigen Receptors; Applications Grants; Autologous; Binding; Biological Assay; CD28 gene; CD3 Antigens; CD8B1 gene; Cancer Etiology; Cause of Death; Cell Degranulation; Cell Line; Cell Therapy; Cells; Cessation of life; Chronic Lymphocytic Leukemia; Clinical Treatment; Clinical Trials; Coculture Techniques; Complex; Cytolysis; DNA; Data; Detection; Development; Diagnosis; Disease; Disease remission; Drug Formulations; Early Diagnosis; Effectiveness; Elements; Engineering; Enzyme-Linked Immunosorbent Assay; Epithelial; Excision; Exposure to; Extracellular Domain; Fc domain; Flow Cytometry; Genetic Engineering; Genomics; Growth Factor; Human; Human Engineering; IgG4; Immune response; Immunotherapy; In Vitro; Integrins; Interleukin-15; Interleukin-2; Interleukin-4; Lead; Ligands; Lymphocyte; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Mediating; Medical; Neoplasm Metastasis; Neuroblastoma; Operative Surgical Procedures; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Patients; Peptides; Phase I Clinical Trials; Plasmids; Preclinical Testing; Proteins; Resectable; Signal Transduction; Sleeping Beauty; Survival Rate; Symptoms; System; Systems Integration; T cell therapy; T-Cell Receptor; T-Lymphocyte; Testing; Time; Tissues; Transmembrane Domain; Transposase; Tumor Antigens; Viral Vector; advanced disease; antigen binding; base; cancer diagnosis; cancer regression; cancer therapy; cellular engineering; cytokine; cytotoxic; cytotoxicity; flexibility; gene therapy; improved; in vitro activity; killings; neoplastic cell; non-viral gene therapy; novel strategies; outcome forecast; pancreatic cancer cells; pancreatic neoplasm; peripheral blood; pre-clinical; public health relevance; rapid growth; receptor; receptor binding; research study; tumor

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.

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死亡原因。唯一可能治愈胰腺癌的治疗方法是手术切除；然而，很少有患者有可以被尊重的肿瘤。胰腺导管腺癌占胰腺癌的90%，其侵袭性特别强，因为它转移迅速，并且通常表达几种允许快速生长的生长因子和信号成分。替代疗法是迫切需要的，因为没有最近的医学进展治疗胰腺腺癌。通过过继性转移工程T细胞的免疫治疗可以介导癌症消退和克服逃避机制，通过肿瘤避免免疫反应。嵌合抗原受体（CAR）是一种工程分子，是抗体衍生的抗原结合基序和细胞内信号域的融合，可以独立于主要的组织相容性复合体识别肿瘤抗原，其表达通常在肿瘤细胞中丢失。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T细胞治疗的T细胞。在这里，我们建议使用SB系统来改造T细胞来表达CAR识别V？一种在胰腺癌细胞上高度表达的整合素。此外，我们将稳定地共表达一个嵌合受体（4??），由IL-4受体的融合组成？胞外结构域和胞内结构域的共同之处？-受体，它是IL-2和IL-15受体的组成部分。4?？抗- V受体T细胞允许CAR阳性细胞选择性扩增。白细胞介素-4与4?？受体会激活增殖信号，从而刺激T细胞分裂。工程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通过测量工程T细胞暴露于V? V?后的细胞因子分泌和脱颗粒来评估。6个阳性胰腺癌细胞和表达car的T细胞裂解V？6个阳性胰腺癌细胞。这些实验的结果将为sb工程T细胞杀死胰腺癌细胞的有效性提供评估。胰腺癌动物模型的后续研究将导致胰腺癌治疗的临床试验，通过使用睡美人转座子系统进行体外基因工程改造的人类t细胞。