Optimizing Cancer Immunotherapy Safety and Efficacy using Genome Editing

使用基因组编辑优化癌症免疫治疗的安全性和有效性

基本信息

批准号：
9980799
负责人：
Benjamin Peter Kleinstiver
金额：
$ 24.9万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9980799
关键词：
Achyrocline Address Adopted Adoptive Cell Transfers Adoptive Transfer Advisory Committees Affinity Allogenic Antigens Area Autologous Award B lymphoid malignancy B-Lymphocytes Biology Boston CRISPR/Cas technology Cancer Biology Cancer Model Cell physiology Cellular biology Clinic Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Communities Conflict of Interest DNA DNA cassette Development Discipline Disease Engineering Environment Faculty Future Gene Expression Gene Targeting General Hospitals Genes Genetic Screening Genome Genomics Goals Health Hematologic Neoplasms Human Human Resources Immunology In Vitro International Journals Knock-out Knowledge Lymphocyte Malignant Neoplasms Massachusetts Mediating Mentors Mentorship Methods Monoclonal Antibodies Mus Names Oncogenes Patients Pattern Performance Phase Positioning Attribute Probability Process Property Reagent Receptor Cell Refractory Relapse Research Research Personnel Safety Science Site Specificity T cell therapy T-Cell Receptor T-Lymphocyte Techniques Technology Therapeutic Training Tumor Antigens Virus Integration Work anti-cancer therapeutic base cancer cell cancer genome cancer immunotherapy cancer therapy cancer type career career development chimeric antigen receptor clinically relevant data framework design engineered T cells engineered nucleases experience gene therapy genome editing genome wide screen genome-wide graft vs host disease improved in vivo innovation insight integration site knockout gene medical schools meetings mouse model neoplastic cell next generation novel nuclease off-target site receptor response responsible research conduct skills technology development tumor tumor eradication

项目摘要

The adoptive transfer of lymphocytes engineered to recognize tumor cells has shown tremendous promise in patients with relapsed or refractory B cell malignancies. In this approach, patient derived T cells are programmed in vitro with engineered T cell receptors (TCRs) or chimeric antigen receptors (CARs) that have affinity for cancer- or lineage-specific antigens. Subsequent autologous reinfusion of the engineered T cells enables tumor targeting and eradication. More recently, genome editing approaches have been proposed to improve engineered T cell performance by knocking out genes that mediate graft-versus-host disease (for allogeneic transfer), that are involved in tumor-mediated suppression of T cell efficacy, or that are recognized by potent clinically relevant monoclonal antibodies. However, to this point no robust characterization of genome editing efficiency or specificity has been performed in T cells. Much of my postdoctoral work has focused on developing methods to characterize and improve the utility and genome-wide specificity of CRISPR-Cas nucleases, making me uniquely suited to address this largely outstanding question. The primary aims of this proposal are therefore: 1) to characterize and optimize the efficiency and specificity of genome editing in T cells, 2) utilize genome editing to improve processes involved in T cell engineering, and 3) leverage CRISPR-Cas screens to enhance the overall efficacy of engineered T cells. The proposed research will provide considerable insight into the feasibility of implementing genome editing strategies in T cells as a means to improve tumor killing efficacy, persistence, or manufacturing. Significant findings relevant to the fields of cancer immunotherapy, cancer biology, genome editing, and gene therapy are expected. Areas of additional scientific training that will enable successful completion of this proposal are knowledge of T cell biology and immunology, mentorship on in vivo cancer modeling in mice, and experience implementing genome-wide CRISPR-Cas screens. The mentored phase of the award will be supported by Dr. Keith Joung, a world-leader in genome editing technology development, and by Dr. Marcela Maus, an expert in cancer immunotherapy. Dr. Joung, Dr. Maus, and nearly all other senior/key personnel of this project are located at the Massachusetts General Hospital or in the greater Boston scientific community. Professional and career development activities will include training in mentorship, responsible conduct of research, grantsmanship, finance, and conflict of interest among other topics. National and international meetings will continue to be attended to disseminate findings from the proposed research. The academic and professional development environment at the Massachusetts General Hospital and Harvard Medical School, combined with a top level mentorship team and scientific advisory committee, will offer the best opportunity for further training as I transition to independence.

旨在识别肿瘤细胞的淋巴细胞的过继转移在以下领域显示出巨大的前景：患有复发性或难治性 B 细胞恶性肿瘤的患者。在这种方法中，患者来源的 T 细胞被编程在体外使用对癌症具有亲和力的工程化 T 细胞受体 (TCR) 或嵌合抗原受体 (CAR) 或谱系特异性抗原。随后自体回输工程化 T 细胞可实现肿瘤靶向和消灭。最近，基因组编辑方法被提出来改善工程 T 细胞通过敲除介导移植物抗宿主病（用于同种异体转移）的基因来实现性能，这些基因是参与肿瘤介导的 T 细胞功效抑制，或被有效的临床相关性所认可单克隆抗体。然而，到目前为止，还没有对基因组编辑效率或基因组编辑效率的可靠表征。特异性已在 T 细胞中进行。我的大部分博士后工作都集中在开发方法表征并提高 CRISPR-Cas 核酸酶的实用性和全基因组特异性，使我独一无二适合解决这个很大程度上悬而未决的问题。因此，本提案的主要目标是： 1) 表征和优化 T 细胞基因组编辑的效率和特异性，2) 利用基因组编辑改进 T 细胞工程的流程，以及 3) 利用 CRISPR-Cas 筛选来增强整体工程化 T 细胞的功效。拟议的研究将为我们的可行性提供深入的见解在 T 细胞中实施基因组编辑策略作为提高肿瘤杀伤功效、持久性或制造业。与癌症免疫治疗、癌症生物学、基因组领域相关的重大发现编辑和基因治疗有望实现。有助于成功的额外科学培训领域该提案的完成包括 T 细胞生物学和免疫学知识以及体内癌症指导小鼠建模，并拥有实施全基因组 CRISPR-Cas 筛选的经验。指导阶段该奖项将得到基因组编辑技术开发领域的世界领先者 Keith Joung 博士的支持，以及由癌症免疫治疗专家 Marcela Maus 博士撰写。 Joung 博士、Maus 博士以及几乎所有其他高级/关键人员该项目的人员位于马萨诸塞州总医院或大波士顿科学中心社区。专业和职业发展活动将包括指导培训、责任培训研究行为、资助、财务和其他主题中的利益冲突。国家和将继续出席国际会议，传播拟议研究的结果。这马萨诸塞州总医院和哈佛大学的学术和专业发展环境医学院与顶级导师团队和科学顾问委员会相结合，将提供最好的当我过渡到独立时，有机会接受进一步的培训。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Prediction of off-target activities for the end-to-end design of CRISPR guide RNAs.

DOI：
10.1038/s41551-017-0178-6
发表时间：
2018-01
期刊：
Nature biomedical engineering
影响因子：
28.1
作者：
Listgarten J;Weinstein M;Kleinstiver BP;Sousa AA;Joung JK;Crawford J;Gao K;Hoang L;Elibol M;Doench JG;Fusi N
通讯作者：
Fusi N

Activities and specificities of CRISPR/Cas9 and Cas12a nucleases for targeted mutagenesis in maize.

DOI：
10.1111/pbi.12982
发表时间：
2019-03
期刊：
Plant biotechnology journal
影响因子：
13.8
作者：
Lee K;Zhang Y;Kleinstiver BP;Guo JA;Aryee MJ;Miller J;Malzahn A;Zarecor S;Lawrence-Dill CJ;Joung JK;Qi Y;Wang K
通讯作者：
Wang K

Plant genome editing branches out.