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Controllable In Vivo Genome Editing for Immune-Checkpoint Blockade in Solid Tumors

用于实体瘤免疫检查点封锁的可控体内基因组编辑

基本信息

批准号：
9767834
负责人：
Sheng Tong
金额：
$ 16.94万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-21 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9767834
关键词：
Address Advanced Malignant Neoplasm Adverse effects Agonist Allografting Antibodies Antigen-Antibody Complex Antitumor Response Biodistribution CRISPR/Cas technology Cancer Patient Cell Culture Techniques Clinical Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats Colon Adenocarcinoma Combination immunotherapy Combined Modality Therapy Complex Coupled DNA Double Strand Break DNA Sequence Data Development Disease remission Drug Kinetics Endocytosis Engineering Gene Deletion Genes Genetic Genome Guide RNA Human Hybrids Immune Immune checkpoint inhibitor Immune response Immune system Immunity Immunologics Immunooncology Immunosuppressive Agents Immunotherapy In Vitro Individual Insect Viruses Lead Light MC38 Magnetic nanoparticles Magnetism Malignant Neoplasms Mediating Modeling Modification Mus Natural Immunity Nonhomologous DNA End Joining Normal tissue morphology OX40 Organ PD-1 blockade PDCD1LG1 gene Pancreatic Ductal Adenocarcinoma Pathologic Processes Pathway interactions Phenotype Proteins Signal Transduction Solid Neoplasm Specificity System Testing Therapeutic Therapeutic Effect Tissues Treatment Efficacy Tumor Immunity Tumor Tissue Viral Viral Vector anti-cancer anti-tumor immune response base cancer heterogeneity cancer immunotherapy cancer type clinical application clinical translation design engineered nucleases gene delivery system genome editing genotoxicity human disease image guided immune checkpoint immune checkpoint blockade immunoengineering immunogenic in vivo individual patient insertion/deletion mutation iron oxide nanoparticle magnetic field mouse model multiplexed imaging novel nuclease patient subsets personalized strategies preclinical study response success therapeutic genome editing transduction efficiency tumor tumor microenvironment vector vector-induced viral nanoparticle

项目摘要

Project Summary The blockade of immune-checkpoint pathways has emerged as a promising therapeutic strategy for a variety of cancers. However, the diverse tumor responses to immunotherapy seen in preclinical and clinical studies prompt the development of combination immunotherapies that can be tailored to the complex immune milieu of individual patients. On the other hand, the severe adverse effects associated with the combination therapies with multiple antagonist antibodies address the necessity for alternative safe and effective therapeutic approaches. In light of this, we aim to develop a hybrid nanoparticle-viral vector system for CRISPR/Cas9- based in vivo therapeutic genome editing, which will be used for multiplexed disruption of immune suppressive pathways in the tumor microenvironment. CRISPR/Cas9 systems are very efficient in generating DNA double- strand breaks, thus disrupting genes through the non-homologous end-joining (NHEJ) pathway. However, inducing uncontrolled CRISPR/Cas9 activities in vivo may lead to systemic genotoxicity. We will develop a novel in vivo gene delivery system that integrates a baculoviral vector (BV) with magnetic nanoparticles (MNPs). Our studies have shown that by taking advantage of the interplay between the MNP-mediated BV margination and endocytosis and the innate immunity against insect viruses, this delivery system can provide spatial and temporal control of CRISPR/Cas9 activity. We will use the MNP-BV system to deliver optimized CRISPR/Cas9 for gene disruption of immune suppressive signals PD-L1 and TGF- in the tumor tissue. We will evaluate CRISPR/Cas9 induced anti-tumor immune responses using two well-established mouse models, an immunogenic model (MC38) where monotherapy with PD-1 blockade is sufficient, and non-immunogenic models pancreatic ductal adenocarcinoma (PDAC) which portrays most non-immunogenic human solid tumors that require combination strategies. In aim 1 studies, we will design and optimize CRISPR/Cas9 gRNAs for targeting PD-L1 and TGF-, package Cas9 and gRNAs into a BV vector, and construct the MNP- BV system. In aim 2 studies, we will evaluate MNP-BV-induced multiplexed gene disruption in cell culture, and determine the on-target and off-target indel rates. In aim 3 studies, we will test the controlled in vivo delivery of CRISPR/Cas9, determine the immunological and therapeutic effects of local gene disruption vs. systemic blockade with antagonist antibody in mouse tumor models. The success of the proposed studies will provide a multiplexed intratumoral immunoengineering platform and pave the way for the clinical translation of a highly efficient in vivo genome editing strategy for personalized cancer immunotherapy.

项目总结