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Principal Investigator: Chuanju Liu
Organization: YALE UNIVERSITY
Fiscal Year: 2024
Award: $388,041
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Project Summary
Pro-inflammatory cytokine TNFα is believed to be responsible for the delayed fracture healing observed in diabetes. However, there is no consensus on the effect of TNFα inhibition on the bone formation, indicating the unmet need in searching for new regents with unique features other than pure TNF inhibitors for diabetic fracture healing. Our genetic screen led to the identification of TNFR as the novel receptor of progranulin (PGRN) (Tang, et al, Science, 2011), a chondrogenic factor that has been shown to be therapeutic against autoimmune inflammatory arthritis. Type 1 diabetes is the most common autoimmune disease, characterized by chronic inflammation and elevated TNFα activity. Although TNFα activity is mediated primarily through TNFR1, we were excited to find that PGRN-stimulated bone regeneration largely depends on TNFR2. These paradoxical findings suggest that the regenerative PGRN/TNFR2 pathway plays a major role in PGRN-stimulated fracture healing. In addition, 14-3-3ε was identified as a component of TNFR2 pathway in response to PGRN stimulation. Further, we have developed an engineered protein named Atsttrin which is composed of three TNFR-binding domains of PGRN, and Atsttrin is more effective than PGRN in inflammatory arthritis. Given that elevated TNFα is believed to be responsible for delayed diabetic fracture healing, we hypothesize that PGRN and Atsttrin stimulate diabetic fracture healing through a) inhibition of TNFα/TNFR1 inflammatory and bone resorption pathway; and primarily b) recruitment of 14-3-3ε to TNFR2, followed by activation of bone regeneration pathway. The Specific Aims are: (1) To determine the role of PGRN, especially its derivative Atsttrin, in diabetic fracture healing. We will use both systemic and inducible PGRN knockout mice to determine whether knockout of PGRN delays diabetic fracture healing, and whether recombinant PGRN and Atsttrin can reverse it (SA#1A); which stage of fracture healing requires PGRN for successful completion of diabetic fracture healing (SA#1B); and whether PGRN, especially Atsttrin, has therapeutic efficacy in treating diabetic fracture (SA#1C). We will use an appropriate injectable hydrogel to locally deliver various dosages of PGRN or Atsttrin. (2) To elucidate the molecular mechanisms by which PGRN and Atsttrin stimulate diabetic fracture healing. We will determine the effects of PGRN, Atsttrin, and TNFα on chondrogenesis of diabetic bone marrow stem cells, signaling pathways, interplays and dependence on TNFR and 14-3-3ε (SA#2A); whether both TNFRs are important for mediating PGRN's role in diabetic bone healing (SA#2B); and whether the protective effects of PGRN and Atsttrin depend on 14-3-3ε by establishing diabetic fracture models with inducible 14-3-3ε[-/-] mice (SA#2C). Proposed studies will not only advance our understanding of the molecular events underlying diabetic fracture healing, but could also lead to novel therapeutic interventions for diabetic fracture healing and other conditions in which fracture healing is impaired.
Terms: <(TNF)-α><3-D print><3-D printer><3D Print><3D printer><3D printing><4H3 protein><Affinity><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Autoimmune><Autoimmune Diseases><Binding><Biochemical><Bone Formation><Bone Marrow Stem Cell><Bone Regeneration><Bone Resorption><Brittle Diabetes Mellitus><CD 120a Antigen><CD 120b Antigen><CD120a Antigens><CD120b Antigens><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cachectin><Cachectin Receptors><Cas nuclease technology><Cell Communication and Signaling><Cell Isolation><Cell Segregation><Cell Separation><Cell Separation Technology><Cell Signaling><Chondrogenesis><Chronic><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Complex><Compound Fractures><Consensus><Data><Dependence><Development><Diabetes Mellitus><Disease><Disorder><Effectiveness><Event><Exhibits><Force of Gravity><Fracture><Fracture Healing><Gene set enrichment analysis><Genes><Genetic><Genetic Screening><Gravities><Growth Agents><Growth Factor><Growth Substances><Healing abnormal><Healing delayed><Hydrogels><IDDM><Impaired healing><Impairment><In Vitro><Inflammation><Inflammatory><Inflammatory Arthritis><Inflammatory Response><Injectable><Injections><Insulin-Dependent Diabetes Mellitus><Intracellular Communication and Signaling><Juvenile-Onset Diabetes Mellitus><KO mice><Ketosis-Prone Diabetes Mellitus><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Laboratories><Ligands><Macrogols><Macrophage-Derived TNF><Maleimides><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Mice><Mice Mammals><Modeling><Molecular><Molecular Interaction><Monocyte-Derived TNF><Murine><Mus><Names><Null Mouse><Open Fractures><Osteoclastic Bone Loss><Osteogenesis><PC cell-derived growth factor><PCDGF><PGRN gene><PGRN protein><Pathway interactions><Pilot Projects><Play><Polyethylene Glycols><Polyethylene Oxide><Polyethyleneoxide><Polyoxyethylenes><Progranulin><Protein Engineering><Proteins Growth Factors><Receptor Protein><Receptors, Tumor Necrosis Factor, Type II><Recombinants><Regenerative pathway><Risk><Role><STZ><Science><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Molecule><Site><Streptozocin><Streptozotocin><Sudden-Onset Diabetes Mellitus><T1 DM><T1 diabetes><T1D><T1DM><TNF><TNF A><TNF Alpha><TNF Receptor Family Protein><TNF Receptor Superfamily><TNF Receptor p55><TNF Receptors><TNF gene><TNF-R2><TNF-RII><TNF-sR55><TNF-α><TNF-α receptor><TNFA><TNFAR><TNFBR><TNFR><TNFR p60><TNFR p75><TNFR, 55-kD><TNFR, 60-kD><TNFR-I><TNFR1><TNFR2><TNFR55><TNFR60><TNFR80><TNFRSF1A><TNFRSF1A Receptor><TNFRSF1A gene><TNFRSF1B><TNFRSF1B Receptor><TNFRSF1B gene><TNFalpha receptor><TNFα><TNFα receptor><Testing><Therapeutic><Treatment Efficacy><Tumor Necrosis Factor><Tumor Necrosis Factor Beta Receptor><Tumor Necrosis Factor Receptor><Tumor Necrosis Factor Receptor 1><Tumor Necrosis Factor Receptor 2><Tumor Necrosis Factor Receptor 55><Tumor Necrosis Factor Receptor 75><Tumor Necrosis Factor Receptor Family><Tumor Necrosis Factor Receptor Superfamily><Tumor Necrosis Factor Receptor Type 2><Tumor Necrosis Factor-alpha><Type 1 Diabetes Mellitus><Type 1 diabetes><Type I Diabetes Mellitus><Zanosar><autoimmune condition><autoimmune disorder><autoimmunity disease><biological signal transduction><bone fracture><bone fracture healing><bone fracture repair><bone healing><bone marrow stromal stem cell><bone tissue formation><bone wound healing><cell sorting><cytokine><developmental><diabetes><diabetic><diabetic patient><dosage><fracture repair><genetic protein engineering><granulin precursor><in vivo><inhibitor><insulin dependent diabetes><insulin dependent type 1><interest><intervention efficacy><juvenile diabetes><juvenile diabetes mellitus><ketosis prone diabetes><mimecan><name><named><naming><new drug target><new druggable target><new pharmacotherapy target><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutic target><new therapy approaches><new therapy target><new treatment approach><new treatment strategy><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutic target><novel therapy approach><novel therapy target><osseous wound healing><osteoglycin><osteoinductive factor><pathway><pilot study><prevent><preventing><progranulin gene><progranulin protein><protective effect><protein design><receptor><recruit><regenerate bone><regeneration pathway><regenerative><response><scaffold><scaffolding><social role><therapeutic efficacy><therapy efficacy><three dimensional printing><treatment effect><tumor necrosis factor alpha receptor><tumor necrosis factor receptor 1A><tumor necrosis factor receptor superfamily, member 1B><tumor necrosis factor α receptor><type I diabetes><type one diabetes>