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Principal Investigator: JOSHUA ALAN CHOE
Organization: UNIVERSITY OF WISCONSIN-MADISON
Fiscal Year: 2022
Award: $37,443
Funding agency: National Institute on Aging
PROJECT SUMMARY / ABSTRACT
Age related traumatic and pathologic fractures are expected to increase greatly in incidence in the coming
decades with aging populations. Older patients are at higher risk for non-healing, ‘critical-size’, bone defects
following fracture. This is exacerbated in composite injuries with concomitant soft tissue defects. These require
multiple surgical interventions and still a majority of patients remain significantly disabled. Therapies, like
bone marrow aspirate concentrate (BMAC), or recombinant bone morphogenetic protein-2 are inconsistent
and ineffective or limited by dose related side effects, such as inflammation, respectively. These drawbacks
may be overcome by osteogenic stimulation with biomaterial platforms for non-viral gene delivery.
The Murphy laboratory has developed nanostructured mineral coated microparticles (MCM) that improve
and sustain delivery of therapeutic nucleic acid complexes. These biomimetic mineral coatings were inspired
by the structure and characteristics of fossils. Ancient DNA has been recovered from fossils which implicates
a role in mineralized surfaces preserving nucleic acids. New stabilization strategies are essential as current
mRNA products, like COVID-19 vaccines, require extensive cold-chain infrastructure limiting their
application. Biomimetic mineralized materials may provide a platform for the binding, long term stabilization,
and delivery of mRNA therapeutics.
This fellowship proposal will develop a biomaterials-based method for storage and delivery of mRNA
encoding for osteogenic growth factors and train the applicant for a career as a physician scientist. We
hypothesize that MCM delivery of mRNA can 1) improve mRNA transfection after lyophilization, 2) promote
an osteogenic phenotype in BMAC and 3) mRNA for osteogenic growth factors will reduce inflammation and
improve bone defect healing. Aim 1 will optimize and characterize how MCMs stabilize mRNA complexes
after lyophilization. Aim 2 will assess the impact of MCM delivery of mRNA for bone morphogenetic protein
-2/-7 heterodimers in a rat model of a critical size bone defect. Aim 3 will investigate the efficacy of R-
Spondin-2 mRNA on a rat model of composite muscle and bone injury. Outcomes will focus on 1) transfection
after simulated accelerated degradation studies, 2/3) gene expression as well as radiographic,
biomechanical and histologic bone healing. The goal of this work is to improve storage and application of
materials guided mRNA therapeutics for bone regeneration. The proposal will combine the biomaterials and
orthopedic expertise of the proposal sponsor, with the vast medical, scientific, graduate training and
translational resources available at the University of Wisconsin – Madison. This pre-doctoral fellowship will
provide for the development of research, clinical, mentorship, innovation, and communication skills
necessary for a career as an independent physician scientist.
Terms: <2019-nCoV vaccine><Adjuvant Therapy><Affinity><Aging><Animals><Archaeology><Aspirate><Aspirate substance><Autograft><Autologous><Autologous Transplantation><Autotransplant><BMP-2><BMP-2A><BMP2><BMP2 gene><BMP2A Gene><Binding><Biocompatible Materials><Biological Mimetics><Biomaterials><Biomechanics><Biomimetics><Body Tissues><Bone Formation><Bone Injury><Bone Marrow><Bone Marrow Reticuloendothelial System><Bone Morphogenetic Protein 2 Gene><Bone Morphogenetic Protein 2A Gene><Bone Regeneration><COVID-19 vaccine><COVID19 vaccine><Cellular Expansion><Cellular Growth><Characteristics><Clinical><Cold Chains><Common Rat Strains><Communication><Complex><Cryofixation><Cryopreservation><Cues><DNA><DNA Therapy><Defect><Deoxyribonucleic Acid><Development and Research><Developmental Biology><Disabled Persons><Disabled Population><Dose><Economics><Elderly><Embryonic Muscle Cells><Evaluation><Fellowship><Fossils><Fracture><Freeze Drying><Freeze Dryings><Freezing><Gene Expression><Gene Transfer Clinical><Genetic Intervention><Goals><Gold><Growth Agents><Growth Factor><Growth Substances><Handicapped><Harvest><Health Care Systems><Healthcare Systems><Histologic><Histologically><Human><In Vitro><Incidence><Inflammation><Inflammatory><Infrastructure><Injury><Isoforms><Laboratories><Lyophilization><Lytotoxicity><Measures><Mediating><Medical><Mentorship><Messenger RNA><Methods><Minerals><Modeling><Modern Man><Molecular Interaction><Muscle><Muscle Tissue><Musculoskeletal><Myoblasts><Nanostructures><Natural regeneration><Nucleic Acids><Operative Procedures><Operative Surgical Procedures><Orthopedic><Orthopedic Surgical Profession><Orthopedics><Osteogenesis><Outcome><Pathologic Fracture><Pathological fracture><Patients><People with Disabilities><Persons with Disabilities><Phenotype><Physicians><Play><Population><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Precursor Muscle Cells><Procedures><Production><Protein Isoforms><Protein Modification><Proteins><Proteins Growth Factors><R & D><R&D><Radiography><Rat><Rats Mammals><Rattus><Recombinants><Recovery><Regeneration><Regenerative Medicine><Research Resources><Resources><Roentgenography><Role><SARS-CoV-2 vaccine><SARS-CoV2 vaccine><SARS-coronavirus-2 vaccine><Scientist><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vaccine><Source><Spontaneous Fractures><Structure><Surface><Surgical><Surgical Interventions><Surgical Procedure><Temperature><Testing><Therapeutic><Tissues><Training><Transfection><Universities><WNT Signaling Pathway><WNT signaling><Wisconsin><Work><advanced age><age dependent><age related><autologous graft><autotransplantation><base><biological material><biomechanical><bone><bone fracture><bone healing><bone morphogenetic protein 2><bone tissue formation><bone wound healing><calcium phosphate><career><cell growth><cold preservation><cold storage><corona virus disease 2019 vaccine><coronavirus disease 2019 vaccine><coronavirus disease-19 vaccine><cytotoxicity><disabled><disabled individual><disabled people><effective therapy><effective treatment><elderly patient><elders><gene repair therapy><gene therapy><gene-based therapy><genetic therapy><genomic therapy><geriatric><healing><high risk><improved><individuals with disabilities><ineffective therapies><ineffective treatment><injuries><innovate><innovation><innovative><interest><late life><later life><long bone><mRNA><mRNA Stability><mRNA delivery><mesenchymal stromal cell><minimally invasive><muscular><myogenesis><nano-structures><neuromuscular><new drug treatments><new drugs><new therapeutics><new therapy><next generation therapeutics><non-viral gene delivery><nonviral gene delivery><novel drug treatments><novel drugs><novel therapeutics><novel therapy><nucleic acid therapy><nucleic acid-based therapeutics><older adult><older patient><older person><osseous wound healing><osteogenic><patient variability><patient variation><pre-doc><pre-doctoral><predoc><predoctoral><preservation><protective effect><protein folding><radiologic imaging><radiological imaging><recombinant human bone morphogenetic protein-2><regenerate><regenerate bone><regeneration biology><regenerative approach><regenerative biology><regenerative strategy><regenerative technique><repair><repaired><research and development><response><rhBMP-2><senior citizen><side effect><skills><social role><soft tissue><surgery><therapeutic nucleic acids><vaccine against 2019-nCov><vaccine against SARS-CoV-2><vaccine against SARS-CoV2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine for novel coronavirus><variability between patients><variation between patients>