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Principal Investigator: Paul Joshua Hurst
Organization: STANFORD UNIVERSITY
Fiscal Year: 2024
Award: $76,408
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract
The proposed research will improve the understanding of structure-property-function
relationships in formulations of charge-altering releasable transporter (CART) gene delivery vectors.
These materials demonstrate remarkable efficiency for mRNA transfection with low cytotoxicity due
to charge-neutralizing degradative chemistry and have shown remarkable selectivity towards
different organs in vivo depending on the chemical structure of the CART. A predictive understanding
of how chemical variation affects nanostructure and biological activity will be established. If
successful, a set of design rules for CARTs will guide the optimization of mRNA/CART complexes
for improved cellular transfection and catered cellular/organ selectivity enabling the development of
site-specific CART / RNA therapeutics.
The fellow will be the first to perform detailed structural and dynamic studies using advanced
cryogenic transmission electron microscopy (cryoEM) and in situ X-ray scattering, illuminating the
morphological properties of mRNA/CART nanoparticles. Additionally, this proposal will establish if
nanostructure variation arises from changes in formulation technique and CART chemical structures
(cation, lipid) and relate these nano-structural variation to differences in biological function. The
fellow will develop microfluidic mixing protocols (t-mixer) and screen mRNA/CARTs with differing
sizes/morphologies in vitro with cell cultures to establish correlations between functional outcome
and nanostructure. In addition to cryoEM and scattering, supplementary measurements of zeta
potential, size, phase, thermal transitions, supramolecular interactions, etc. will be carried out to
determine the effect physico-chemical properties has on nanostructure in different CART
formulations. In separate future research not covered by this proposal, the most distinct
mRNA/CARTs will be further studied with in vivo experimentation in mouse models with our
collaborators. The fellow will also receive formal and informal training in the responsible conduct of
research, teaching, career development skills applicable to their future career goals of becoming a
research professor and participate in outreach and mentoring. These studies will take place in a
highly interdisciplinary training environment at Stanford University in the lab of Prof. Robert
Waymouth, in close collaboration with Dr. Christopher Tassone at SSRL (SLAC national laboratory).
Terms: <2,5-Diaminopentanoic Acid><2019-nCoV vaccine><Address><Affect><Benchmarking><Best Practice Analysis><Biological><Biological Function><Biological Process><COVID-19 vaccine><Cancer Treatment><Cations><Cell Body><Cell Culture Techniques><Cells><Cellular Assay><Charge><Chemical Structure><Chemicals><Chemistry><Collaborations><Complex><Cryo-electron Microscopy><Cryo-electron tomography><Cryoelectron Microscopy><DNA Therapy><Data><Development><Educational process of instructing><Electron Cryomicroscopy><Environment><Exhibits><Formulation><Freeze Drying><Freeze Dryings><Future><Gene Delivery><Gene Transfer Clinical><Genetic Intervention><Goals><Immune mediated therapy><Immunologically Directed Therapy><Immunotherapy><In Situ><In Vitro><Laboratories><Lipids><Liver><Lung><Lung Respiratory System><Lyophilization><Lytotoxicity><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Measurement><Medicine><Mentors><Messenger RNA><Methods><Microfluidics><Microscopy><Modeling><Morphology><Nanostructures><Nature><Non-Polyadenylated RNA><Organ><Ornithine><Phase><Property><Protocol><Protocols documentation><RNA><RNA Gene Products><RNA based therapeutics><RNA based therapy><RNA therapy><Research><Ribonucleic Acid><Roentgen Rays><SARS-CoV-2 vaccine><SARS-coronavirus-2 vaccine><Sampling><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vaccine><Site><Spleen><Spleen Reticuloendothelial System><Structure><System><Teaching><Techniques><Training><Transfection><Transmission Electron Microscopy><Universities><Unsaturated Fats><Vaccination><Variant><Variation><X-Radiation><X-Ray Radiation><X-ray><Xray><anti-cancer therapy><benchmark><biologic><cancer therapy><cancer-directed therapy><career><career development><cell assay><cell culture><cell cultures><coronavirus disease 2019 vaccine><coronavirus disease-19 vaccine><cryo-EM><cryo-EM tomography><cryoEM><cryoEM tomography><cryoelectron tomography><cryogenic electron microscopy><cryogenics><cytotoxicity><deliver mRNA><deliver messenger RNA><delivery system for mRNA><delivery vector><delivery vehicle><design><designing><developmental><electron cryo-tomography><electron tomography><functional outcomes><gene repair therapy><gene therapy><gene-based therapy><genetic therapy><genomic therapy><hepatic body system><hepatic organ system><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><improved><in vivo><insight><lipid based nanoparticle><lipid nanoparticle><mRNA><mRNA Translation><mRNA delivery><messenger RNA delivery><mouse model><murine model><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><nano><nano particle><nano-sized particle><nano-sized structures><nano-structures><nanoparticle><nanosized particle><next generation><outreach><particle><physical property><professor><pulmonary><responsible research conduct><skill acquisition><skill development><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic RNA><therapeutically effective><unsaturated dietary fat><unsaturated dietary lipid><unsaturated lipid><vaccine against 2019-nCov><vaccine against COVID-19><vaccine against SARS-CoV-2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine candidates against SARS-CoV-2><vaccine for novel coronavirus><vaccines preventing COVID><vaccines to prevent COVID><zeta potential><µfluidic>