Document text
Principal Investigator: BIN GUO
Organization: OHIO STATE UNIVERSITY
Fiscal Year: 2024
Award: $497,002
Funding agency: National Cancer Institute
PROJECT SUMMARY
Targeted nanotechnologies have shown great promise in overcoming roadblocks in cancer therapeutics.
Tumor metastasis has a limited response or resistance to chemotherapeutics and a moderate response to new
antibody therapies. Our RNA nanotechnology has shown great promise in targeting metastatic disease to deliver
both gene silencing and chemical drug therapies.
Our long goal is to overcome colon cancer, the second most common cause of cancer death primarily due
to the mutations to KRAS and subsequent lung metastasis with expected survival of months. KRAS, the gene
that codes for the K-Ras protein, is considered “undruggable”. KRAS mutations are found in up to 45% of
colorectal cancers. We have designed an epigenetic repressor to silence mutant K-Ras through epigenome
editing. We created a fusion protein consisting of nuclease-inactive dCas9 and the histone deacetylase HDAC1
and targeted dCas9-HDAC1 to the promoter of mutant KRAS. We can load the recombinant dCas9-HDAC1-
gRNA ribonucleoprotein (RNP) complex into exosomes and silence K-Ras; we designed RNA nanoparticles
carrying mutant K-Ras siRNA to inhibit KRAS mutant lung cancer; we also successfully constructed RNA 4WJ
carrying SN-38 to inhibit colon cancer lung metastasis.
The goal of this proposal is to identify mechanisms that govern the high delivery platform to silence K-Ras in
colon cancer. We intend to deactivate mutant KRAS via RNA-ligand displaying exosomes loaded with dCas9-
HDAC1-gRNA ribonucleoprotein, siRNAs, and chemical drugs individually or in combination in colorectal cancer
primary and metastasis tumors, using orthotopic and PDX xenograft models. We will investigate the mechanism
of action in K-Ras inhibition, including the conditions for the integration and assembly of dCas9-HDAC1 and
gRNA or crRNA such as sequence and length requirement for silencing mutant KRAS and suppressing colon
cancer cells. We will apply RNA nanoparticle orientation to display targeting ligands on the surface of exosomes.
Instead of delivering dCas9 plasmids, we will deliver a ribonucleoprotein complex of dCas9-recombinant protein
and gRNA. Exosomes will display RNA nanoparticles with an aptamer to bind colon cancer cells specifically. We
will engineer RNA nanoparticles and increase the surface display density of the negatively charged RNA ligands
to enhance the negative zeta potential of exosomes for preventing binding to the vital organs and healthy cells
that normally have negatively charged lipid membranes. We will try to enhance therapeutic efficacy and reduce
toxicity by overcoming endosome trapping and non-specific cell entry through RNA ligand manipulation. Zonal
and density gradient ultracentrifugation or size exclusion columns will select exosomes smaller than 100 nm to
escape macrophage engulfment and improve biodistribution. This project with a multidisciplinary approach will
build a strong foundation from which researchers can deploy large protein complexes to treat cancer by tumor-
specific delivery and effectively targeting those previously difficult targets like K-Ras.
Terms: <Antibody Therapy><Antigen Defined by Monoclonal Antibody AUAI><Binding><Biodistribution><C-K-RAS><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cancer Cause><Cancer Etiology><Cancer Treatment><Cancers><Cas nuclease technology><Cell Body><Cell Membrane Lipids><Cells><Cessation of life><Charge><Chemicals><Chimera Protein><Chimeric Proteins><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Code><Coding System><Colon><Colon Cancer><Colon Carcinoma><Colorectal Cancer><Complex><Cytosol><Data><Death><Density Gradient Centrifugation><Density Gradient Fractionation><Disease><Disorder><Drug Delivery><Drug Delivery Systems><Drug Kinetics><Drug Therapy><Drug resistance><Drugs><EGF Receptor><EGFR><ERBB Protein><Endosomes><Engineering><EpCAM><Epidermal Growth Factor Receptor><Epidermal Growth Factor Receptor Kinase><Epidermal Growth Factor Receptor Protein-Tyrosine Kinase><Epidermal Growth Factor-Urogastrone Receptors><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Epithelial Cellular Adhesion Molecule><Exclusion><Foundations><Fusion Protein><GA733-2><Gastrointestinal Tumor-Associated Antigen 2, 35-KD Glycoprotein><Gene Inactivation><Gene Silencing><Genes><Genetic Alteration><Genetic Change><Genetic defect><Goals><Guide RNA><HD1><HDAC><HDAC Proteins><HDAC1><HDAC1 gene><HER1><Histone Deacetylase><Histone Deacetylase 1><Human><Individual><Investigators><K-RAS2A><K-RAS2B><K-Ras><K-Ras 2A><K-Ras-2 Oncogene><KRAS><KRAS2><KRAS2 gene><Ki-RAS><Length><Ligands><Liver><Lung><Lung Respiratory System><M4S1><MIC18><Macrophage><Malignant Cell><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Lung><Malignant neoplasm of lung><Medication><Membrane Component, Chromosome 4, Surface Marker 1><Membrane Lipids><Metastasis><Metastasis to the Lung><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Neoplasm to the Lung><Metastatic Tumor><Metastatic Tumor to the Lung><Micro RNA><MicroRNAs><Modeling><Modern Man><Molecular Interaction><Molecular Sieve Chromatography><Molecular Tumor Suppression><Mutation><Mφ><Nanotechnology><Neoplasm Metastasis><Non-Polyadenylated RNA><Nucleoproteins><Oncogene K-Ras><Organ><PDX model><Pathway interactions><Patient derived xenograft><Pharmaceutical Preparations><Pharmacokinetics><Pharmacotherapy><Plasmids><Post-Transcriptional Gene Silencing><Posttranscriptional Gene Silencing><Proteins><Pulmonary Cancer><Pulmonary malignant Neoplasm><RAS inhibition><RASK2><RNA><RNA Gene Products><RNA Interference><RNA Silencing><RNA Stability><RNAi><RPD3-Like 1><RPD3L1><Receptor Protein><Receptosomes><Recombinant Proteins><Recombinants><Recurrence><Recurrent><Reduced Potassium Dependency 3, Yeast, Homolog-Like 1><Research Personnel><Researchers><Resistance><Ribonucleic Acid><Ribonucleoproteins><SN-38><Secondary Neoplasm><Secondary Tumor><Sequence-Specific Posttranscriptional Gene Silencing><Short interfering RNA><Size Exclusion Chromatography><Small Interfering RNA><Specificity><Surface><System><TACSTD1><TACSTD1 gene><TGF-alpha Receptor><Therapeutic><Thermodynamic><Thermodynamics><Toxic effect><Toxicities><Transforming Growth Factor alpha Receptor><Treatment Efficacy><Tumor Suppression><Tumor-Associated Calcium Signal Transducer 1><Ultracentrifugation><Urogastrone Receptor><Xenograft Model><anti-cancer therapy><antibody based therapies><antibody treatment><antibody-based therapeutics><antibody-based treatment><aptamer><c-erbB-1><c-erbB-1 Protein><cancer cell><cancer in the colon><cancer metastasis><cancer therapy><cancer-directed therapy><chemical stability><density><density gradient ultracentrifugation><design><designing><drug resistant><drug treatment><drug/agent><epigenetically><epigenome editing><epigenomic editing><erbB-1><erbB-1 Proto-Oncogene Protein><erbBl><exosome><gRNA><genome mutation><hepatic body system><hepatic organ system><improved><inhibitor><interdisciplinary approach><intervention efficacy><lung cancer><lung metastasis><lung metastatic><malignancy><metastasize to the lung><metastatic colo-rectal><metastatic colo-rectal cancer><metastatic colo-rectal carcinoma><metastatic colon cancer><metastatic colorectal><metastatic colorectal cancer><metastatic colorectal carcinoma><miRNA><miRNAs><multidisciplinary approach><mutant><nano particle><nano sized><nano tech><nano technology><nano-sized particle><nano-technological><nanoparticle><nanosized><nanosized particle><nanotech><nanotechnological><neoplasm/cancer><nuclease><pathway><patient derived xenograft model><prevent><preventing><promoter><promotor><protein complex><proto-oncogene protein c-erbB-1><pulmonary><pulmonary metastasis><pulmonary metastatic><ras Gene Products><ras Proteins><receptor><resistance to Drug><resistant><resistant to Drug><response><siRNA><therapeutic efficacy><therapy efficacy><transcriptional silencing><tumor><tumor cell metastasis><v-Ki-RAS2 Kirsten Rat Sarcoma 2 Viral Oncogene Homolog><xenograft transplant model><xenotransplant model><zeta potential>