Multiplexed nanoparticle delivery to increase CRISPR/Cas gene editing for enhanced cancer therapy

多重纳米颗粒递送可增强 CRISPR/Cas 基因编辑，从而增强癌症治疗

基本信息

批准号：
10419618
负责人：
Daniel John Siegwart
金额：
$ 39.17万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10419618
关键词：
Affect Antibodies Applications Grants Biological Markers Brain Cancer Model Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Encapsulated Endocytosis Exhibits Extracellular Matrix Focal Adhesion Kinase 1 Formulation Future Gene Silencing Genes Genetic Enhancement Genetic Suppression Genetically Engineered Mouse Growth Immune Infiltration Intravenous Kinetics Lipids Liver Lung Malignant Neoplasms Malignant neoplasm of liver Measures Mechanics Mediating Messenger RNA Modeling Modification Mus Muscle Non-Small-Cell Lung Carcinoma Nucleic Acids Organ Outcome Pathway interactions Penetration Primary carcinoma of the liver cells Proteins Series Small Interfering RNA Solid Neoplasm Spleen System Therapeutic Tissues Toxic effect Treatment Efficacy Tropism Tumor Burden Tumor Expansion Tumor Tissue Work base cancer immunotherapy cancer therapy design gene correction gene repair genome editing improved in vivo innovation knock-down lipid nanoparticle lung cancer cell mRNA delivery mechanical force mechanical properties nanoparticle nanoparticle delivery neoplastic cell nucleic acid delivery programmed cell death ligand 1 programs repaired therapeutic genome editing three dimensional cell culture tumor tumor microenvironment tumor progression uptake

Affect Antibodies Applications Grants Biological Markers Brain Cancer Model Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Encapsulated Endocytosis Exhibits Extracellular Matrix Focal Adhesion Kinase 1 Formulation Future Gene Silencing Genes Genetic Enhancement Genetic Suppression Genetically Engineered Mouse Growth Immune Infiltration Intravenous Kinetics Lipids Liver Lung Malignant Neoplasms Malignant neoplasm of liver Measures Mechanics Mediating Messenger RNA Modeling Modification Mus Muscle Non-Small-Cell Lung Carcinoma Nucleic Acids Organ Outcome Pathway interactions Penetration Primary carcinoma of the liver cells Proteins Series Small Interfering RNA Solid Neoplasm Spleen System Therapeutic Tissues Toxic effect Treatment Efficacy Tropism Tumor Burden Tumor Expansion Tumor Tissue Work base cancer immunotherapy cancer therapy design gene correction gene repair genome editing improved in vivo innovation knock-down lipid nanoparticle lung cancer cell mRNA delivery mechanical force mechanical properties nanoparticle nanoparticle delivery neoplastic cell nucleic acid delivery programmed cell death ligand 1 programs repaired therapeutic genome editing three dimensional cell culture tumor tumor microenvironment tumor progression uptake

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项目摘要

Project Summary The programmable CRISPR/Cas gene editing system has great potential for cancer treatment due to the ability to precisely inactivate or repair cancer-related genes. However, delivery of CRISPR to solid tumors for efficient cancer therapy remains limited by the uniquely stiff and fibrotic tumor microenvironment that acts as a barrier to nanoparticle uptake. Here, we propose to directly target tumor tissue mechanics via a multiplexed lipid nanoparticle (LNP) approach involving co-delivery of focal adhesion kinase (FAK) siRNA, Cas9 mRNA, and sgRNA (siFAK + CRISPR LNPs) to enable tumor delivery and enhance gene editing efficacy. We will leverage our recently developed non-viral Selective ORgan Targeting (SORT) LNP platform that enables tissue-specific nucleic acid delivery, protein delivery, and genome editing following intravenous (IV) administration. The proposed approach involves a unique combination of siRNA-mediated gene silencing of FAK, a key modulator of tumor ECM, and CRISPR-mediated gene editing (deletion) of tumor-related genes via a single all-in-one nanoparticle approach. We will further leverage Liver and Lung SORT LNPs for to evaluate gene editing as a strategy for permanent inactivation of programmed death-ligand 1 (PD-L1), supported by the clinical limitations of anti-PD-L1 antibody atezolizumab therapy in hepatocellular carcinoma (HCC) and non-small cell lung cancer (NSCLC). In this grant proposal, we Aim to (1) establish how FAK knockdown enhances SORT LNP-mediated mRNA delivery and CRISPR gene editing, (2) optimize Liver and Lung SORT LNP formulations for multiplexed siRNA + Cas9 mRNA + sgRNA delivery, and (3) evaluate the therapeutic efficacy of Liver and Lung SORT siFAK + Cas9 mRNA + sgPD-L1 delivery in orthotopic and genetically engineered mouse models of cancer. Regulating the mechanical properties of tumor cells/ECM for enhancing the genetic suppression in tumor tissues provides an innovative strategy for treating cancer using CRISPR. We anticipate that this general approach could further synergize with additional types of therapeutics in the future.

项目摘要由于能力精确地灭活或修复与癌症相关的基因。但是，将CRISPR递送到实体瘤以提高效率癌症治疗仍然受到独特僵硬和纤维化的肿瘤微环境的限制，该环境充当了障碍纳米颗粒的吸收。在这里，我们建议通过多重脂质直接靶向肿瘤组织力学纳米颗粒（LNP）方法涉及局灶性粘附激酶（FAK）siRNA，Cas9 mRNA和 SGRNA（SIFAK + CRISPR LNP）可实现肿瘤递送并增强基因编辑功效。我们将利用我们最近开发的非病毒选择性器官靶向（排序）LNP平台，可实现组织特异性静脉内（IV）给药后核酸输送，蛋白质递送和基因组编辑。这提出的方法涉及siRNA介导的基因沉默的独特组合，这是一个关键调节剂肿瘤ECM和CRISPR介导的基因编辑（缺失）与肿瘤相关基因的基因纳米颗粒方法。我们将进一步利用肝脏和肺排序LNP，以评估基因编辑在临床局限性的支持下，导致编程死亡配体1（PD-L1）永久失活的策略肝细胞癌（HCC）和非小细胞肺癌的抗PD-L1抗体atezolizumab治疗（NSCLC）。在此赠款建议中，我们的目标是（1）确定FAK敲低如何增强排序LNP介导 mRNA递送和CRISPR基因编辑，（2）优化肝脏和肺排序LNP配方 siRNA + Cas9 mRNA + SGRNA递送，（3）评估肝脏和肺部Sifak的治疗功效 + CAS9 mRNA + SGPD-L1在原位和基因工程的癌症模型中递送。调节肿瘤细胞/ECM用于增强肿瘤组织遗传抑制的机械性能提供了使用CRISPR治疗癌症的创新策略。我们预计这种一般方法可以进一步将来与其他类型的治疗剂协同作用。