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Principal Investigator: CYRUS R SAFINYA
Organization: UNIVERSITY OF CALIFORNIA SANTA BARBARA
Fiscal Year: 2020
Award: $295,627
Funding agency: National Institute of General Medical Sciences
Project Summary/Abstract
The current level of research activity involving gene therapy with either synthetic vectors (carriers) or engineered viruses
is unprecedented. Liposomes are the most widely studied nonviral carriers worldwide for nucleic acid (NA) and drug
delivery applications. Cationic liposomes (CLs) are relatively safe nonviral vectors used in ongoing clinical trials. CLs
may either be complexed via electrostatic interactions with therapeutic NAs (anionic DNA or short interfering RNA) for
gene delivery and silencing, or used as vectors of potent cytotoxic hydrophobic drugs, encapsulated within their lipid
bilayer, in cancer therapeutics. Among the biggest advantages of nonviral vectors (over viral vectors which are currently
more efficient in in vivo settings) are their safety, their low immunogenicity and their ability to transfer entire genes
(containing coding and noncoding sequences) and regulatory sequences into cells (currently not feasible with engineered
viruses because of capsid size limitations). The development of nonviral lipid-based vectors with efficacy competitive
with viral vectors in vivo will require a mechanistic understanding of how synthetic vectors may be functionalized to
overcome the major intracellular hurdle of endosomal escape. Successful endosomal escape is required for release of
therapeutic nucleic acid within the cell cytosol and therefore maximum efficacy. The first aim of this research application
is to employ modern biophysical and synthetic approaches to the rational design of functionalized CL–NA nanoparticles
(NPs) with synergistic, complementary dual-function PEG-lipid and fusogenic components for optimized endosomal
escape. Modern methods of organic and solid phase chemistry will be employed to synthesize dual-function PEG-lipids
with cell targeting and endosome escaping properties. The second aim of this research application is to optimize efficacy
of a new class of CL-based carriers of the hydrophobic drug paclitaxel (PTXL) for cancer therapeutics. This will be
achieved by developing a mechanistic understanding of the relation between physical and chemical properties of the
carrier (i.e. size of the functionalized CL carrier, membrane spontaneous curvature, and lipid tail structure) and functional
efficacy (i.e. PTXL membrane solubility, cell uptake of vector and PTXL delivery leading to cytotoxicity against human
cancer cells). The structures of CL-based vectors of NAs and hydrophobic drugs will be characterized using cryogenic
electron microscopy and synchrotron x-ray diffraction techniques. The interactions between CL vectors and cell
organelles will be directly visualized with spinning disk confocal fluorescence microscopy. Their structures will be
correlated to their biological activity in human cancer cells. The broad, long-term objective of our research is to develop a
fundamental science base through mechanistic studies that will lead to the design and synthesis of nonviral vectors of
nucleic acids and hydrophobic drugs for gene and cancer therapeutics.
Terms: <1,2-Ethanediol><2-Hydroxyethanol><Abscission><Achievement><Achievement Attainment><Acids><Adhesions><Albumin-Stabilized Nano particle Paclitaxel><Albumin-Stabilized Nanoparticle Paclitaxel><Allergy><Anzatax><Asotax><Assay><Bioassay><Biologic Assays><Biological><Biological Assay><Biophysics><Bristaxol><Cancer cell line><Cancers><Capsid><Cations><Cell Body><Cell Survival><Cell Viability><Cells><Charge><Chemicals><Chemistry><Chemotherapy Protocol><Chemotherapy Regimen><Chemotherapy, Cancer, General><Chemotherapy-Oncologic Procedure><Clinical Trials><Code><Coding System><Combination Chemotherapy Regimen><Communities><Complex><Cone><Custom><Cyclic Peptides><Cytosol><DNA><DNA Therapy><Data><Deoxyribonucleic Acid><Dependence><Development><Dihydroxyethanes><Distal><Drug Delivery><Drug Delivery Systems><Drugs><Electron Microscopy><Electrostatics><Encapsulated><Endosomes><Engineering><Ethanediols><Ethylene Glycols><Excision><Exons><Extirpation><Fluorescence Light Microscopy><Fluorescence Microscopy><Functional RNA><Gastric Body Cancer><Gastric Cancer><Gastric Cardia Cancer><Gastric Fundus Cancer><Gastric Pylorus Cancer><Gene Delivery><Gene Inactivation><Gene Silencing><Gene Transfer Clinical><Genes><Genetic Intervention><Goals><Homing><Hospitals><Human><Hydrophobicity><Hypersensitivity><In Vitro><Intervening Sequences><Introns><Knowledge><Laboratories><Ligands><Lipid Bilayers><Lipids><Liposomal><Liposomes><Lytotoxicity><Malignant Cell><Malignant Gastric Neoplasm><Malignant Gastric Tumor><Malignant Neoplasms><Malignant Tumor><Mediating><Medication><Membrane><Methods><Micelles><Modern Man><Modernization><Monoethylene Glycol><Nanoparticle Paclitaxel><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-Viral Vector><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nucleic Acids><Organelles><Organic Chemistry><Paclitaxel><Paclitaxel (Taxol)><Penetration><Peptides><Pharmaceutic Preparations><Pharmaceutical Preparations><Phase><Praxel><Probability><Property><Proteins><Quimioterapia><RNA><RNA Gene Products><Reaction><Receptosomes><Removal><Research><Research Activity><Ribonucleic Acid><Roentgen Rays><Safety><Science><Shapes><Short interfering RNA><Small Interfering RNA><Solid><Solubility><Stomach Cancer><Structure><Surgical Removal><Synchrotrons><Tail><Taxol><Taxol A><Taxol Konzentrat><Techniques><Testing><Therapeutic><Transfection><Tumor Tissue><Untranslated RNA><Vesicle><Viral Vector><Virus><X ray diffraction><X ray diffraction analysis><X-Radiation><X-Ray Radiation><X-ray><Xray><Xray diffraction><base><biophysical analysis><biophysical approaches><biophysical characteristics><biophysical characterization><biophysical foundation><biophysical measurement><biophysical methodology><biophysical methods><biophysical parameters><biophysical principles><biophysical properties><biophysical sciences><biophysical studies><biophysical techniques><cancer cell><cancer chemotherapy><chemical property><cryogenics><cytotoxic><cytotoxicity><design><designing><developmental><disease control><disorder control><drug/agent><efficacy testing><ethylene glycol><gastric malignancy><gene therapy><gene-based therapy><genetic therapy><genomic therapy><human model><immunogenic><immunogenicity><in vivo><lipid bilayer membrane><liposomal carrier><liposomal nanocarrier><liposomal vehicle><liposome vector><malignancy><malignant stomach neoplasm><malignant stomach tumor><membrane structure><model of human><mouse model><murine model><nano meter scale><nano meter sized><nano particle><nano particle Paclitaxel><nano scale><nano-sized particle><nanometer scale><nanometer sized><nanoparticle><nanoscale><nanosized particle><neoplasm/cancer><noncoding><nonviral vector><novel><nucleic acid delivery><nucleic acid-based therapeutics><physical property><resection><siRNA><stomach fundus cancer><stomach pylorus cancer><therapeutic nucleic acids><tool><transcriptional silencing><tumor><uptake><vector>