Document text
Principal Investigator: David J Mooney
Organization: BETH ISRAEL DEACONESS MEDICAL CENTER
Fiscal Year: 2024
Award: $774,435
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
ABSTRACT
Impaired wound healing is an alarming problem in diabetes, attributed to the development of over 750,000
diabetic foot ulcerations (DFU) and 70,000 lower extremity amputations per year in the USA. Single-cell RNA-
sequencing (scRNASeq) analysis provides valuable insights into transcriptional features and disease
pathophysiology by allowing the profiling of individual cells in heterogeneous tissues. A recently conducted study
in our unit primarily focused on divergent characteristics between DFU patients who healed their ulcers (Healers)
and those who failed to heal them (non-Healers) and investigated molecular changes via scRNASeq analysis of
surgically removed DFUs. Our analysis revealed the enrichment of a unique population of fibroblasts,
overexpressing MMP1, MMP3, MMP11, HIF1A, CHI3L1, and TNFAIP6 genes in the Healers. Ongoing work in
our lab has shown that specifically designed alginate bandages that can provide controlled lipid nanoparticle
(LNPs) delivery of modified CHI3L1mRNA, IL-17AmRNA, FGF-2 and IL-2mRNA considerably improve wound
healing in diabetic db/db mice. Based on these findings, we hypothesize that topical wound treatment with new
biomaterials that can release the above mRNAs can lead to the development of new therapeutic approaches.
Furthermore, we hypothesize that combination of various mRNAs with specific temporal delivery can
physiologically imitate the activation of pathways that are associated with DFU healing.
To fully explore our hypotheses, we propose the following Specific Aims: 1.) Develop a modified-mRNA-LNPs
therapeutics delivery system to target different stages of diabetic wound healing and provide proof of concept
regarding their efficacy in wounds of diabetic mice. Alginate hydrogels fabricated into bandages capable of
sustained release will be employed to deliver mRNA-LNPs in 3D human skin equivalents and wounds of db/db
mice. We will identify the most efficient mRNA or combination and the most appropriate dosing and timing of
delivery in improving diabetic wound repair. 2.) Investigate efficacy in a larger animal model, the diabetic Yucatan
minipig and confirm mechanisms of action in both animal models. We will also examine mechanisms of action
of pro-healing mRNA(s) and will compare them to the results from the rodent studies. 3.) Conduct translational
studies in human diabetic wound healing to verify the therapeutic potential. We will conduct ex vivo wound
healing studies with discarded skin from patients with and without diabetes and in vivo studies with wounding of
human skin engrafted onto diabetic nude mice. We will also employ targeted multi-omics analyses to establish
molecular mechanisms of action and compare them with the ones that have been observed in our previous
extensive human studies. We believe that our proposal involves state-of-the-art “bench to bedside” research that
can culminate in the development of innovative new therapeutic approaches. Successful completion of the
project will facilitate the conduction of Phase I/II clinical trials in DFU patients.
Terms: <3-D><3-Dimensional><3D><Adverse Experience><Adverse event><Aldesleukin Gene><Alginates><Amputation><Animal Model><Animal Models and Related Studies><Athymic Mice><Athymic Nude Mouse><B-Cell Differentiation Factor Gene><B-Cell Stimulatory Factor 2 Gene><BSF-2 Gene><BSF2 Gene><Bandage><Basic Fibroblast Growth Factor><Basic Fibroblast Growth Factor Gene><Beta-2 Gene Interferon><Biocompatible Materials><Biodistribution><Biomaterials><Body Tissues><CHI3L1><CHI3L1 gene><CTLA-8><CTLA-8 Gene><CTLA8><CTLA8 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><Characteristics><Chitinase 3-Like 1><Chondrocyte Protein YKL40><Chronic><Complications of Diabetes Mellitus><Cytotoxic T-Lymphocyte-Associated Antigen 8><Cytotoxic T-Lymphocyte-Associated Antigen 8 Gene><Cytotoxic T-Lymphocyte-Associated Serine Esterase 8><Cytotoxic T-Lymphocyte-Associated Serine Esterase 8 Gene><Data Set><Development><Diabetes Complications><Diabetes Mellitus><Diabetes-Related Complications><Diabetic Complications><Diabetic Foot Ulcer><Diabetic mouse><Diabetic wound><Disease><Disorder><Dysfunction><Engraftment><FGF-2><FGF2><FGF2 gene><FGFB><Family suidae><Fibroblast Growth Factor 2><Fibroblast Growth Factor 2 Gene><Fibroblasts><Functional disorder><Gene Transcription><Genes><Genetic Transcription><Goals><Growth Agents><Growth Factor><Growth Substances><HBGF-2><HIF 1 alpha><HIF-1alpha><HIF1-Alpha><HIF1A><HIF1A gene><HIF1α><HSF Gene><Health system><Heparin-Binding Growth Factor 2><Heparin-Binding Growth Factor Class II><Hepatocyte Stimulatory Factor Gene><Human><Hybridoma Growth Factor Gene><Hydrogels><IFNB2 Gene><IL-17><IL-17 Gene><IL-17A><IL-17A Gene><IL-2 Gene><IL-6 Gene><IL17><IL17 Protein><IL17 gene><IL17A><IL17A Gene><IL2><IL2 gene><IL6><IL6 gene><Immune><Immunes><Immunocompromised><Immunocompromised Host><Immunocompromised Patient><Immunosuppressed Host><Impaired tissue repair><Impaired wound healing><Impairment><Individual><Inflammation><Inflammatory><Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8)><Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8) Gene><Interleukin 17 Precursor><Interleukin 17 Precursor Gene><Interleukin 2 Precursor Gene><Interleukin 6 (Interferon, Beta 2) Gene><Interleukin-17><Interleukin-2 Gene><Interleukin-6 Gene><Interstitial Collagenase><Intracellular Communication and Signaling><Lower Extremity><Lower Limb><MMP-1><MMP-1Fibroblast Collagenase><MMP1><MMP11><MMP11 gene><MMP3><MMP3 gene><MOP1><Matrix Metalloproteinase 11><Matrix Metalloproteinase-1><Mediating><Membrum inferius><Messenger RNA><Methodology><Mice><Mice Mammals><Miniature Swine><Minipigs><Modality><Modern Man><Molecular><Molecular Mechanisms of Action><Murine><Mus><Names><Network Analysis><Nude Mice><Operative Procedures><Operative Surgical Procedures><Pathway Analysis><Pathway interactions><Patients><Phase><Phase 1/2 Clinical Trial><Phase I/II Clinical Trial><Physiologic><Physiological><Physiopathology><Pigs><Population><Prostate Epithelial Cell Growth Factor><Proteins><Proteins Growth Factors><RNA Expression><Research><Rodent><Rodentia><Rodents Mammals><SL-1><SL-3><ST-3 Protein><ST3><ST3 Matrix Metalloproteinase><STMY><STMY1><STMY3><STR1><Severities><Signal Transduction><Signal Transduction Systems><Signaling><Skin><Subcellular Process><Suidae><Surgical><Surgical Interventions><Surgical Procedure><Swine><System><T-Cell Growth Factor Gene><TCGF Gene><Therapeutic><Tissues><Toxic effect><Toxicities><Transcription><Transplantation><Ulcer><Ulceration><Work><Wound Repair><Wound models><abnormal tissue repair><appropriate dose><bFGF><bench bed side><bench bedside><bench to bed side><bench to bedside><bench to clinic><bench to clinical practice><biological material><biological signal transduction><db/db mouse><delayed wound healing><deliver mRNA><deliver messenger RNA><delivery system for mRNA><design><designing><developmental><diabetes><diabetes mouse model><diabetes ulcer><diabetic><diabetic foot wound><diabetic skin wound><diabetic ulcer><diabetic wound healing><effective therapy><effective treatment><efficacy testing><experiment><experimental research><experimental study><experiments><gp39><healing><human data><immunosuppressed patient><improved><in vivo><innovate><innovation><innovative><insight><lipid based nanoparticle><lipid nanoparticle><mRNA><mRNA delivery><messenger RNA delivery><mini pig><mini-swine><miniswine><mmp-11><model of animal><multiomics><multiple omics><name><named><naming><nano particle delivery><nanoparticle delivered><nanoparticle delivery><nanoparticle therapy><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapy approaches><new treatment approach><new treatment strategy><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapy approach><optimal drug dosage><optimal drug dose><overexpress><overexpression><panomics><pathophysiology><pathway><porcine><pre-clinical><preclinical><protein expression><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><stromelysin 3><suid><surgery><therapeutic nanoparticles><three dimensional><tissue wound><transcriptome profiling><transcriptomic profiling><translational study><transplant><treat wound><wound><wound healing><wound healing models><wound management><wound recovery><wound resolution><wound therapeutics><wound therapy><wound treatment><wounding><wounds>