Mutagenic chain reaction-facilitated immunotherapy

诱变链式反应促进的免疫疗法

基本信息

批准号：
9163059
负责人：
ETHAN BIER
金额：
$ 65.1万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-09 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9163059
关键词：
Alleles Allergic Anopheles Genus Autoimmunity Biomedical Research Cancerous Cell Surface Receptors Cell division Cells Chromosomes Chronic Communicable Diseases DNA Sequence Alteration Disease Drosophila melanogaster Ecosystem Elements Equilibrium Event Feedback Frequencies Gene Conversion Generations Genes Genetic Genetic Heterogeneity Genome Goals Guide RNA Human Hypersensitivity Immune Immune system Immunity Immunotherapeutic agent Immunotherapy Individual Inflammatory Insertion Mutation Instruction Ligands Lymphocyte Malignant Neoplasms Mediating Mediator of activation protein Metabolic Metastatic Melanoma Methods Mission Modeling Monoclonal Antibodies Motion Mutation Parasites Pathway interactions Patients Plasmodium Play Point Mutation Population Pre-Clinical Model Radiation therapy Reaction Research Resistance Retrieval Ribonucleoproteins Series Signal Transduction Solid Suicide System T-Lymphocyte Techniques Time Tissues Transgenes Translating United States National Institutes of Health abstracting base cancer cell cancer immunotherapy design fighting genetic approach genetic element genetic manipulation immunopathology innovation killings mouse model neoplastic cell novel novel therapeutics parasitism pressure repaired research study tool tumor tumor microenvironment vector

项目摘要

Project Summary/Abstract The objectives of this project are to reprogram primary T cells as a means of eliminating cancer cells—a major goal of biomedical research as mandated by the National Institutes of Health. Using new genetic tools applied in novel ways, we will overcome present limitations to cancer immunotherapy by produc- ing T cells that have been relieved of multiple inhibitory feedback mechanisms. However, the more ef- fective the T cell population in eliminating cancerous cells, the more likely they are to cause inflammato- ry immunopathology. Thus, an important requirement for this reprogramming is to provide a provision for their elimination. The means for producing such T cells relies on a recently developed autocatalytic gene conversion method based on the Crispr/Cas9 system and known as the mutagenic chain reaction (MCR). There are two primary advantages to our integrated system for reprogramming T-cells. First, our meth- od should be substantially more efficient in generating biallelic insertions that can render cells defective for multiple genes. Second, because MCR vectors integrate larger inserts into the genome with high effi- ciency and fidelity, we will be able to introduce a multifunctional cassette of genetic elements (e.g., sgR- NAs targeting multiple inhibitory pathways and an inducible suicide module). Combined, these two fea- tures will make it possible to reprogram effector T cells in a single round of ex vivo treatment reducing the number of cell divisions that take place in culture and minimizing the time from cell retrieval to rein- fusion of tumor-fighting T cells. The anti-tumor efficacy of reprogrammed T cells will be studied in models of solid and metastatic melanoma to identify the optimal combination of targeted inhibitory pathways. Initial experiments will take advantage of mouse models, with experiments progressing to the reprogramming of cultured human T cells. These techniques will enable a generation of novel therapies to treat cancer, but also chronic infectious diseases, autoimmunity, and allergic hypersensitivity diseases.

项目摘要/摘要该项目的目标是将原代T细胞重新编程为消除癌细胞的一种手段，A 国家卫生研究院规定的生物医学研究的主要目标。使用新通用以新颖的方式应用的工具，我们将通过生产克服对癌症免疫疗法的表现限制 - 已减轻多种抑制反馈机制的T细胞。但是，越多人体T细胞种群消除了取消细胞，引起炎症的可能性越大 RY免疫病理学。这是此重编程的重要要求是提供一项规定因为他们的进化。产生此类T细胞的手段取决于最近开发的自催化基于CRISPR/CAS9系统的基因转换方法，称为诱变链反应（MCR）。我们的集成系统用于重编程T细胞有两个主要优点。首先，我们的大都会 OD在产生可能使细胞有缺陷的生物插入时应该更有效对于多个基因。其次，因为MCR载体将较大的插入物集成到基因组中，并具有高效率 - 效率和保真度，我们将能够引入遗传元素的多功能盒（例如，Sgr- NA靶向多个抑制途径和诱导自杀模块）。合并，这两个fea- 在一轮体内治疗中，可以重新编程效应子T细胞减少在培养中发生的细胞分裂数量，并最大程度地减少从细胞检索到恢复的时间抗肿瘤T细胞的融合。重编程T细胞的抗肿瘤效率将在固体和转移性黑色素瘤的模型，以确定靶向抑制的最佳组合途径。最初的实验将利用鼠标模型，实验发展为重编程培养的人T细胞。这些技术将使一代新颖的疗法能够治疗癌症，以及慢性传染病，自身免疫性和过敏性超敏反应。