Core B: Genome Engineering Core

核心 B：基因组工程核心

• 批准号: 9891734
• 负责人: Frederic D Bushman
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9891734
• 关键词: Adverse event; Aliquot; Behavior; Categories; Cell physiology; Cells; Chromosomal translocation; Cleaved cell; Clinical; Clonal Expansion; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA; Data; Event; Evolution; Frequencies; Gene therapy trial; Gene-Modified; Genes; Genetic; Genetic Transcription; Genome; Genome engineering; Genomics; Growth; HIV; HIV-1; HMGA2 gene; Human; Human Genome; Immunotherapy; Infusion procedures; Insertional Mutagenesis; Lentivirus Vector; Methods; Modification; Molecular; Monitor; Nature; Outcome; Pathway interactions; Patients; Procedures; Protocols documentation; Reagent; Recurrence; Safety; Site; Specificity; T-Cell Proliferation; T-Lymphocyte; Technology; Therapeutic Effect; Time; base; beta Thalassemia; cell growth; cellular transduction; clinical efficacy; design; engineered T cells; experience; experimental study; gene therapy; genotoxicity; human subject; improved; integration site; lentiviral integration; nuclease; safety assessment; targeted nucleases; therapeutic cloning; vector

Abstract Core B. Quantifying outcomes of genome modification Core B will track the nature and consequences of genome modifications carried out in Projects 1, 3 and 4, and use these data to design improved reagents for CART genome modification. Core B will analyze several types of genome modification as dictated by the needs of each project. Integration of lentiviral vectors necessarily disrupts the host cell locus at the site of integration. We and others have found that insertional mutagenesis by vectors used to deliver CARs can be associated with altered cell growth, which has resulted in clinical adverse events, but also clinically favorable clonal expansions that have bolstered therapeutic effects. The core will use these data to identify genes and pathways to manipulate to optimize CART expansion and persistence. Vector integration also marks each transduced cell uniquely, allowing tracing of the partitioning of descendant cells, information useful in tracking outcomes and understanding mechanisms of possible adverse events. Cleavage of the host genome using targeted nucleases is also an effective means of achieving genome modification; however, off-target cleavage and associated deletions are safety concerns. The core will monitor off-target cleavage using our iGUIDE technology. Lastly, in protocols where multiple sgRNA/CAS9 complexes cleave multiple targets in a single cell, nuclease cleavage at multiple sites can result in rejoining of DNA ends in a “swapped” fashion, causing chromosomal translocations. Core B will monitor molecular outcomes in each of these categories to allow optimization of reagents, assessment of safety, and scoring of cell function.

抽象核心B.量化基因组修饰的结果 核心B将跟踪基因组修饰的性质和后果 项目1、3和4，并使用这些数据设计改进的CART基因组试剂 修改。核心B将分析几种类型的基因组修饰，如 每个项目的需求。慢病毒载体的整合必然会破坏宿主 整合位点的细胞基因座。我们和其他人发现插入 用于输送汽车的向量的诱变可能与细胞改变有关 增长，导致临床不良事件，但在临床上也有利 增强治疗作用的扩张。核心将使用这些数据来 确定操纵基因和途径以优化购物车扩展和 持久性。向量集成还标记每个翻译的单元格，允许 跟踪后代单元的分区，可用于跟踪结果的信息 并了解可能不利事件的机制。主持人的裂开 使用靶向核的基因组也是实现基因组的有效手段 修改;但是，脱靶裂解和相关的删除是安全的 关注。核心将使用我们的Iguide技术监视脱靶裂解。 最后，在协议中，多个sgrna/cas9复合物清除了多个目标 单细胞，多个位点的核酸酶切割可能导致DNA在A中的末端重新加入 “交换”时尚，导致染色体易位。核B将监测分子 这些类别中的每个类别的结果以允许优化试剂，评估 安全性和细胞功能的评分。