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Principal Investigator: Georgios Theocharidis
Organization: BETH ISRAEL DEACONESS MEDICAL CENTER
Fiscal Year: 2024
Award: $152,236
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
ABSTRACT
The career development activities and studies delineated in this K01 application are designed to equip Dr.
Theocharidis, the Principal Investigator (PI), with expertise in biomaterials and multi-omics in order to become
an independent investigator. Non-healing diabetic foot ulcers (DFUs) affect millions of Americans and lead to
devastating consequences. Existing treatments are inefficient in promoting wound closure. The ongoing steep
surge in diabetic population necessitates new strategies to accelerate healing of diabetic wounds. Unlike acute
uncomplicated wounds, the linear progression from one phase of wound healing to the next is impaired in DFU,
which is characterized by a chronic low-grade inflammation. This could be the primary reason that growth factor
treatments that act during the proliferative phase have been unsuccessful. Single-cell RNA-sequencing (sc-
RNAseq) offers extensive insights into cell function and disease pathophysiology by allowing the mapping of the
transcriptomic landscape of individual cells in heterogeneous tissues like DFUs. A recently completed study led
by the PI focused on differences between DFU patients who healed their ulcers and those who failed to heal
them and investigated molecular changes via sc-RNAseq of surgically removed DFUs. Comparative analysis
unveiled genes and pathways significantly associated with successful wound repair.
Delivery of messenger RNA (mRNA) into recipient cells has the potential to enable functional protein
expression with tremendous therapeutic implications. Ongoing work by the PI has revealed that alginate hydrogel
dressings that facilitate lipid nanoparticle (LNPs)-mediated delivery of healing associated modified CHI3L1
mRNA and IL-2 mRNA to the injury site, markedly improve wound healing in diabetic mice. Based on these
findings, I hypothesize that (i) topical wound treatment with advanced biomaterials capable of tailorable delivery
of mRNAs, corresponding to genes shown to promote DFU healing can lead to novel treatments. Further, I
hypothesize that (ii) altering the LNPs’ surface by conjugating antibodies targeting specific cell type receptors
can streamline their application and maximize their pro-reparative impact.
These hypotheses will be explored across three aims. The first aim will focus on fabricating alginate hydrogels
for effective topical delivery of mRNA loaded LNPs. During the second aim, I will employ multi-omics approaches
to establish LNP cell internalization and intracellular mRNA translation processes, together with particle
biodistribution, as well as to identify mechanisms of action. In the third aim, LNPs will be endowed with cell
specificity by conjugation of antibodies on their surface to maximize the platform’s efficiency and also evaluated
in a mouse model of increased clinical relevance. Successful completion of this multidisciplinary proposal will
result in highly novel data that will substantially expand our knowledge on DFU pathophysiology and advance
the fields of biomaterials, RNA therapeutics and cutaneous wound repair. This can lead to the development of
innovative much-needed interventions for the management of DFU.
Terms: <Acceleration><Acute><Advanced Development><Affect><Alginates><American><Amputation><Antibodies><Award><B-Cell Differentiation Factor Gene><B-Cell Stimulatory Factor 2 Gene><BSF-2 Gene><BSF2 Gene><Bandage><Beta-2 Gene Interferon><Biocompatible Materials><Biodistribution><Biomaterials><Body Tissues><CHI3L1><CHI3L1 gene><Cartilage Glycoprotein 39><Cell Body><Cell Communication and Signaling><Cell Function><Cell Physiology><Cell Process><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chitinase 3-Like 1><Chondrocyte Protein YKL40><Chronic><Co-Stimulator><Coloring Agents><Costimulator><Data><Data Set><Development><Diabetes Mellitus><Diabetic Foot Ulcer><Diabetic mouse><Disease><Disorder><Dose><Dyes><Dysfunction><Encapsulated><Endowment><Engraftment><Epidermal Thymocyte Activating Factor><Fibroblasts><Functional disorder><Genes><Growth Agents><Growth Factor><Growth Substances><HIF 1 alpha><HIF-1alpha><HIF1-Alpha><HIF1A><HIF1A gene><HIF1α><HSF Gene><Hepatocyte Stimulatory Factor Gene><Human><Hybridoma Growth Factor Gene><Hydrogel Bandage><Hydrogels><IFNB2 Gene><IL-2><IL-6 Gene><IL2 Protein><IL6><IL6 gene><Immune><Immunes><Impairment><In Vitro><Individual><Inflammation><Inflammatory><Injury><Interleukin 2><Interleukin 2 Precursor><Interleukin 6 (Interferon, Beta 2) Gene><Interleukin II><Interleukin-2><Interleukin-6 Gene><Interleukine 2><Interleukine 2 Precursor><Interleukine II><Interstitial Collagenase><Intervention><Intervention Strategies><Intracellular Communication and Signaling><Investigation><Investigators><Knowledge><Lower Extremity><Lower Limb><Lymphocyte Mitogenic Factor><MMP-1><MMP-1Fibroblast Collagenase><MMP1><MMP11><MMP11 gene><MMP3><MMP3 gene><MOP1><Macrophage><Maps><Matrix Metalloproteinase 11><Matrix Metalloproteinase-1><Mediating><Membrum inferius><Mentors><Messenger RNA><Mitogenic Factor><Modern Man><Molecular><Molecular Biology Techniques><Mφ><Names><Network Analysis><Operative Procedures><Operative Surgical Procedures><Outcome><Pathway Analysis><Pathway interactions><Patients><Phase><Physiologic><Physiological><Physiopathology><Population><Principal Investigator><Proteins><Proteins Growth Factors><Proteomics><QOL><Quality of life><RNA based therapeutics><RNA based therapy><RNA therapy><Receptor Protein><Reporter Genes><Research><Research Personnel><Researchers><SL-1><SL-3><ST-3 Protein><ST3><ST3 Matrix Metalloproteinase><STMY><STMY1><STMY3><STR1><Series><Shapes><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skin><Specificity><Subcellular Process><Surface><Surgical><Surgical Interventions><Surgical Procedure><System><T cell growth factor><T-Cell Growth Factor><T-Cell Stimulating Factor><Therapeutic><Therapeutic Intervention><Thymocyte Stimulating Factor><Time><Tissues><Topical Drug Administration><Topical application><Translation Process><Treatment Factor><Ulcer><Ulceration><Work><Wound Repair><administer topically><antibody conjugate><apply topically><biological material><biological signal transduction><career development><cell type><clinical relevance><clinically relevant><comparative><cutaneous wound><db/db mouse><deliver mRNA><deliver messenger RNA><deliver topically><delivery system for mRNA><dermal wound><design><designing><developmental><diabetes><diabetes management><diabetes mellitus management><diabetes mouse model><diabetic><diabetic foot wound><diabetic management><diabetic wound healing><gp39><healing><hydrogel dressing><improved><in vitro Assay><in vivo><injuries><innovate><innovation><innovative><insight><intervention therapy><interventional strategy><lipid based nanoparticle><lipid nanoparticle><mRNA><mRNA delivery><messenger RNA delivery><mmp-11><mouse model><multidisciplinary><multiomics><multiple omics><murine model><name><named><naming><novel><overexpress><overexpression><panomics><particle><pathophysiology><pathway><protein expression><proteogenomics><receptor><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><skin wound><stromelysin 3><surgery><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic RNA><tissue wound><topical administration><topical delivery><topical drug application><topical treatment><topically administered><topically applied><topically delivered><topically treated><transcriptomics><translational opportunities><translational potential><treat topically><treat wound><uptake><wound><wound closure><wound healing><wound management><wound recovery><wound resolution><wound therapeutics><wound therapy><wound treatment><wounding><wounds>