Development of an in vivo screening technology for cancer vaccine immunogens

癌症疫苗免疫原体内筛选技术的开发

• 批准号: 8508685
• 负责人: Steven L. Zeichner
• 金额: $ 23.02万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-19 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8508685
• 关键词: Address; Animals; Antibody Formation; Antigens; Bacteria; Bar Codes; Cancer Vaccines; Cell Extracts; Cells; Chimeric Proteins; Cloning; Computer software; DNA; DNA biosynthesis; Development; Devices; Discipline; Doctor of Philosophy; Ecology; Escherichia coli; Feces; Gastrointestinal tract structure; Genes; Genetic Engineering; Genomics; Gram-Negative Bacteria; Immune; Immune response; Immune system; Institution; Knowledge; Libraries; Life; Malignant Neoplasms; Microarray Analysis; Modeling; Monitor; Mucosal Immune Responses; Mus; Peptide Library; Peptide Vaccines; Peptides; Plasmids; Principal Investigator; Procedures; Production; Proteins; Recombinants; Relative (related person); Research Personnel; Salmonella; Salmonella Vaccines; Serum; Site; Study of serum; Surface; System; Techniques; Technology; Testing; Theoretical model; Therapeutic; Time; Tumor Antigens; Tumor Cell Derivative Vaccine; Vaccines; base; cancer immunotherapy; cancer therapy; comparative efficacy; design; experience; expression vector; feeding; gastrointestinal; genetic analysis; high risk; high throughput screening; immunogenic; immunogenicity; improved; in vivo; innovation; insight; member; microbial; microbiome; mouse model; new technology; novel; novel vaccines; oncology; programs; rRNA Genes; research study; response; screening; survivin; synthetic biology; technology development; tumor; vaccine candidate

DESCRIPTION (provided by applicant): This application addresses the mandates of the RFA by proposing to develop a highly novel and previously untested technology to identify, assess, and compare immunogens for cancer vaccines, employing an in vivo immune system in a high throughput screening mode. The development of the new technology will bring together multiple investigators experienced in a wide variety of complementary techniques, from different institutions and disciplines. If successful, the new technology will have great impact on oncology by providing investigators with an important new technology to evaluate and compare immunogens for cancer vaccines. We will develop and then employ this new technology in a test case to identify immunogens that elicit immune responses to the survivin tumor-associated antigen, an attractive and widely studied tumor vaccine candidate useful for this developmental proof-of-principle study due to its demonstrated activity in cancer vaccine models and relatively small size. The technology exploits an intact GI mucosal immune system as a massively parallel in vivo screening device. To develop this new screening technology we will: 1) Create a library of bacteria transformed with DNA bar-coded plasmids that include expression cassettes that place large amounts of chimeric protein, which will include sequences encoding an exhaustive, overlapping library of peptides derived from survivin, on the Gram-negative bacterial surface, 2) Feed the library to mice, 3) Use PhyloChip microarray technologies (or high throughput sequencing) to identify, via the barcodes, members of the library showing decreased relative abundance in the mouse feces over time, which we would take as evidence of the induction of a mucosal immune response against the chimeric proteins encoded by plasmids showing decreased abundance. We will rescreen the clones to confirm their ability to induce an immune response and evaluate sera and cells from animals inoculated with bacteria expressing the proteins and animals immunized with the proteins identified in the screen to determine if they induce anti-survivin humoral and cellular immune responses. Clones and the peptides encoded by the clones able to induce an anti-survivin response will be studied in an in vivo tumor vaccine model and compared to peptides already known to induce an anti-tumor immune response in the model. The study proposes to develop an innovative, rapid, high-throughput approach to the identification of potential immunogens useful as cancer vaccine employing the survivin tumor associated antigen as a model. If successfully developed, the technology also would be useful in the search for other immunogens for cancer vaccines and for comparing the immunogenicity of different candidate proteins/peptides. Since the proteins are expressed in Gram-negative bacteria, including vaccine strains of Salmonella, the technology could be used to rapidly and cheaply produce a candidate vaccine. Although the project is risky, the project could therefore potentially have a very large impact on cancer immunotherapy.

描述（由申请人提供）：本申请通过建议开发一种高度新颖且未经测试的技术来识别，评估和比较癌症疫苗的免疫原子，以高吞吐量筛选模式采用体内免疫系统，以探讨RFA的任务。新技术的发展将汇集到来自不同机构和学科的各种补充技术中经验丰富的多个研究人员。如果成功，新技术将通过为研究人员提供一项重要的新技术来评估和比较癌症疫苗的免疫原子，从而对肿瘤学产生重大影响。我们将在测试案例中开发并采用这项新技术，以鉴定引起对Survivin肿瘤相关抗原的免疫反应的免疫原子，这是一种有吸引力且经过广泛研究的肿瘤疫苗候选者，可用于此发育证明基本研究，因为它在癌症疫苗中证明了活性，并且相对较小。该技术利用完整的GI粘膜免疫系统作为体内筛查装置的大规模平行。为了开发这种新的筛查技术，我们将：1）创建一个细菌库，该细菌库由DNA棒编码的质粒转化，其中包括表达盒式盒，这些盒式磁带上放置了大量的嵌合蛋白，其中包括编码详尽的，详尽的，重叠的肽库的序列，从gram-negrip swimential criperies criperiation criber phy phy for phy，3），3）吞吐量测序）为了通过条形码识别文库的成员，随着时间的推移，小鼠粪便中的相对丰度降低，我们将其作为诱导粘膜免疫反应的证据，该粘膜免疫反应针对质粒蛋白质，该嵌合蛋白被质粒所编码的嵌合蛋白显示出降低的丰度。我们将重新分组克隆，以确认其诱导免疫反应的能力，并评估用表达蛋白质和动物接种的动物的血清和细胞，这些蛋白质和动物在筛选中鉴定出的蛋白质免疫，以确定它们是否诱导抗细胞体验的体液和细胞免疫反应。将在体内肿瘤疫苗模型中研究克隆和能够诱导抗阴性反应的克隆编码的肽，并与已知在模型中诱导抗肿瘤免疫反应的肽进行了比较。该研究建议开发一种创新，快速，高通量的方法，以鉴定使用与Survivin肿瘤相关的抗原作为模型的癌症疫苗的潜在免疫原子。如果成功开发，该技术也将在寻找其他免疫原癌疫苗以及比较不同候选蛋白/肽的免疫原性的情况下很有用。由于蛋白质在革兰氏阴性细菌（包括沙门氏菌的疫苗菌株）中表达，因此该技术可用于快速，便宜地生产候选疫苗。尽管该项目有风险，因此该项目可能会对癌症免疫疗法产生很大的影响。