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Principal Investigator: Daniel John Siegwart
Organization: UT SOUTHWESTERN MEDICAL CENTER
Fiscal Year: 2023
Award: $383,887
Funding agency: National Cancer Institute
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.
Terms: <3D cell culture><3D culture><Affect><Antibodies><Applications Grants><B7-H1><B7H1><Biological Markers><Body Tissues><Brain><Brain Nervous System><CD274><CRISPR><CRISPR/Cas system><Cancer Model><Cancer Treatment><CancerModel><Cancers><Cell Body><Cell-Extracellular Matrix><Cells><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats><Coupled><Data><ECM><Encapsulated><Encephalon><Endocytosis><Exhibits><Extracellular Matrix><Focal Adhesion Kinase 1><Formulation><Future><GEM model><GEMM model><Gene Inactivation><Gene Proteins><Gene Silencing><Generalized Growth><Genes><Genetic Enhancement><Genetic Suppression><Genetically Engineered Mouse><Grant Proposals><Growth><Hepatic Cancer><Hepatocarcinoma><Hepatocarcinoma model><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatoma><Immune infiltrates><Intravenous><Kinetics><Lipids><Liver><Liver Cells Carcinoma><Lung><Lung Respiratory System><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Lung><Malignant neoplasm of liver><Malignant neoplasm of lung><Measures><Mechanics><Mediating><Messenger RNA><Mice><Mice Mammals><Modeling><Modification><Murine><Mus><Muscle><Muscle Tissue><NSCLC><NSCLC - Non-Small Cell Lung Cancer><Non-Small Cell Lung Cancer><Non-Small-Cell Lung Carcinoma><Nonsmall Cell Lung Carcinoma><Nucleic Acids><Organ><Outcome><PD-L1><PDL-1><PDL1><PTK2 Protein Tyrosine Kinase 2><Pathway interactions><Penetration><Primary carcinoma of the liver cells><Programmed Cell Death 1 Ligand 1><Programmed Death Ligand 1><Protein Gene Products><Proteins><Pulmonary Cancer><Pulmonary malignant Neoplasm><Series><Short interfering RNA><Single-Stranded DNA><Small Interfering RNA><Solid Neoplasm><Solid Tumor><Spleen><Spleen Reticuloendothelial System><System><Therapeutic><Tissue Growth><Tissues><Toxic effect><Toxicities><Treatment Efficacy><Tropism><Tumor Burden><Tumor Cell><Tumor Load><Tumor Tissue><Work><anti-cancer immunotherapy><anti-cancer therapy><anticancer immunotherapy><anticancer therapy><bio-markers><biologic marker><biomarker><cancer immunotherapy><cancer microenvironment><cancer progression><cancer therapy><cancer-directed therapy><design><designing><endogenous substrate pp120><focal adhesion kinase><focal adhesion protein tyrosine kinase><focal adhesion-associated protein tyrosine kinase pp125FAK><gene corrected><gene correction><gene repair><gene-editing therapy><genetically engineered mouse model><genetically engineered murine model><genome editing><genome editing based therapy><genome editing therapy><genome editing treatment><genome editing-based therapeutics><genomic editing><hepatic body system><hepatic organ system><hepatocellular carcinoma cancer model><hepatocellular carcinoma model><immune cell infiltrate><immune-based cancer therapies><immunotherapy for cancer><immunotherapy of cancer><improved><in vivo><innovate><innovation><innovative><intervention efficacy><intravenous administration><knock-down><knockdown><lipid based nanoparticle><lipid nanoparticle><liver cancer><liver cancer model><liver carcinoma><liver malignancy><lung cancer><lung cancer cell><mRNA><mRNA delivery><malignancy><malignant liver tumor><mechanic><mechanical><mechanical force><mechanical properties><muscular><nano particle><nano particle delivery><nano-sized particle><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanosized particle><neoplasm progression><neoplasm/cancer><neoplastic cell><neoplastic progression><nucleic acid delivery><ontogeny><pathway><programmed cell death ligand 1><programmed cell death protein ligand 1><protein death-ligand 1><pulmonary><repair><repaired><siRNA><ssDNA><synergism><therapeutic editing><therapeutic efficacy><therapeutic genome editing><therapy efficacy><three dimensional cell culture><transcriptional silencing><tumor><tumor microenvironment><tumor progression><uptake>