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Principal Investigator: Shuying Yang
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2024
Award: $553,248
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Abstract. Rheumatoid arthritis (RA) is a debilitating chronic joint disease, causing functional disability, severe
pain, joint deterioration, and substantial socioeconomic burdens. Presently, there is no cure for RA. While some
patients manage RA with methotrexate and other disease-modifying anti-rheumatic drugs (DMARDs), around
40% do not respond. Even with biologic treatments, 6-21% RA patients are unresponsive. Thus, a deeper
understanding of the mechanisms of RA pathogenesis is crucial for developing effective treatments for RA
patients. In RA, synovial macrophages (MΦs) are abundant and major sources of pro-inflammatory cytokines
like TNFα, IL1, and IL6, contributing to RA pathogenesis. By analyzing single-cell RNA-seq databases of human
synovial tissues from RA patients and mouse synovial MΦs in CAIA model, we discovered a significant reduction
in INPP5E expression. INPP5E, a 5-phosphatase, dephosphorylates PI(4,5)P2, PI(3,5)P2, PI(3,4,5)P3, known
for regulating the immune response and MΦ phagocytosis via PIP3/PI3K/AKT signaling. However, the role of
INPP5E in regulating MΦs, RA pathogenesis, and its potential as a drug candidate remain poorly understood.
Our preliminary data showed INPP5E deletion in MΦs worsened inflammation and bone destruction in a mouse
CAIA model, while overexpression of INPP5E inhibited inflammation in IL1 induced MΦs. Moreover, we found
that INPP5E associates with Aurora kinase A (AURKA), and overexpression of INPP5E suppressed NF-κB and
DNA damage markers like H2AX and ATM/ATR phosphorylation. Proteomics results indicated INPP5E affects
oxidative reduction, inflammation, NF-κB signaling, and DNA repair, suggesting anti-RA potential. Moreover, to
overexpress INPP5E in RA MΦs, we developed novel F4/80 Ab Fab'-functionalized LNPs (Ab-LNPs) for INPP5E
mRNA delivery. Our preliminary data showed that these LNPs can accumulate and target synovial MΦs and can
effectively express GFP and INPP5E mRNA. Our preliminary data support two key hypotheses: 1) Macrophagic
INPP5E inhibits RA pathogenesis by regulating AURKA/AKT/NF-κB and DNA damage/ATM/ATR signaling
pathways; 2) Introduction of INPP5E through a novel targeted LNPs carrying INPP5E mRNA mitigates RA
pathogenesis. To test hypothesis, in Aim 1, we will test if deletion or overexpression of INPP5E in MΦs
significantly alter inflammation and bone erosion in CAIA model. We will explore pathomechanisms by
characterizing AURKA/AKT/NF-κB, DNA damage/ATM/ATR and other INPP5E downstream signaling pathways.
We will further characterize INPP5E interacting proteins and their function by performing mass spectrometry
and RNA interference analysis. In Aim 2, we will demonstrate that targeted delivery of INPP5E mRNA to MΦ
using a novel F4/80 Ab conjugated mRNA LNPs platform can effectively attenuate RA in acute and chronic RA
mouse models. The completion of the proposed project is expected to reveal the novel role and mechanism of
INPP5E in RA pathogenesis and provide a novel targeted nanoparticle mRNA therapy for RA and other
inflammatory diseases.
Terms: <(TNF)-α><AIK gene><AKT><AKT Signaling Pathway><ARK1><AURKA><AURORA2><Ablation><Acute><Affect><Aik protein><Akt protein><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Antibodies><Atrophic Arthritis><Attenuated><Aurora-Related Kinase 1><Aurora/IPL1-Like Kinase><Autoimmune Diseases><B-Cell Differentiation Factor Gene><B-Cell Stimulatory Factor 2 Gene><BSF-2 Gene><BSF2 Gene><BTAK><Basal Transcription Factor><Basal transcription factor genes><Behavior><Beta-2 Gene Interferon><Biological Response Modifier Therapy><Biological Therapy><Body Tissues><Cachectin><Cell Communication and Signaling><Cell Function><Cell Growth in Number><Cell Multiplication><Cell Physiology><Cell Process><Cell Proliferation><Cell Signaling><Cellular Function><Cellular Physiology><Cellular Process><Cellular Proliferation><Chronic><Clinic><Clinical Treatment Moab><Co(beta)-cyano-7''-(2-methyl)adeninylcobamide><Collagen Type II><Culturing, in vitro Vertebrate, Primary><DMARD><DMARDs><DNA Damage><DNA Damage Repair><DNA Injury><DNA Repair><Data><Data Bases><Databases><Dephosphorylation><Deterioration><Disease><Disease Management><Disease-Modifying Antirheumatic Drugs><Disease-Modifying Second-Line Drugs><Disorder><Disorder Management><Economic Burden><Embryo Development><Embryogenesis><Embryonic Development><General Transcription Factor Gene><General Transcription Factors><Genetic Alteration><Genetic Change><Genetic defect><HSF Gene><Hepatocyte Stimulatory Factor Gene><Human><Hybridoma Growth Factor Gene><IFNB2 Gene><IL-1><IL-6 Gene><IL1><IL6><IL6 gene><IP3-5-phosphatase><Immune response><Immune system><Immunological response><Inflammation><Inflammatory><Inflammatory Response><Interleukin 6 (Interferon, Beta 2) Gene><Interleukin I><Interleukin-1><Interleukin-6 Gene><Intracellular Communication and Signaling><Joint Diseases><Joints><Joubert syndrome><Joubert-Boltshauser syndrome><Lymphocyte-Stimulating Hormone><Macrophage><Macrophage Cell Factor><Macrophage-Derived TNF><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Meckel syndrome><Meckel-Gruber syndrome><Mediating><Messenger RNA><Methotrexate><Methotrexate Methylaminopterin><Methotrexatum><Metotrexato><Mice><Mice Mammals><Microphthalmos><Modeling><Modern Man><Monoclonal Antibodies><Monocyte-Derived TNF><Murine><Mus><Mutation><Mφ><Nervous System Physiology><Neurologic function><Neurological function><Osteoclasts><Oxidation-Reduction><PI(3,4,5)P3><PI-3K/AKT><PI3K/AKT><Pain><Painful><Pathogenesis><Patients><Phagocytosis><Phenotype><Phosphatases><Phosphohydrolases><Phosphomonoesterases><Phosphoric Monoester Hydrolases><Phosphorylation><Phosphorylation Inhibition><Physiologic><Physiological><Play><Population><Post-Transcriptional Gene Silencing><Posttranscriptional Gene Silencing><Primary Cell Cultures><Protein Dephosphorylation><Protein Kinase B><Protein Phosphorylation><Proteins><Proteomics><Proto-Oncogene Proteins c-akt><RAC-PK protein><RNA Interference><RNA Silencing><RNAi><Redox><Refractory><Rheumatoid Arthritis><Role><STK15><STK6><STK6 gene><STK6, Mouse, Homolog of><Safety><Sequence-Specific Posttranscriptional Gene Silencing><Serine/Threonine Protein Kinase 15><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Situs Inversus><Source><Specificity><Subcellular Process><T Helper Factor><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFα><Technology><Testing><Tissues><Transcription Factor Proto-Oncogene><Transcription factor genes><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><Unscheduled DNA Synthesis><antibody conjugate><arthropathic><arthropathies><arthropathy><attenuate><attenuates><aurora kinas><aurora kinase A><aurora-kinase A><autoimmune condition><autoimmune disorder><autoimmunity disease><biological signal transduction><biological therapeutic><biological treatment><biologically based therapeutics><biotherapeutics><biotherapy><bone><bone erosion><brain deformity><brain malformation><c-akt protein><cell type><cerebellar vermis agenesis><cerebelloparenchymal disorder IV><collagen antibody induced arthritis><congenital brain anomaly><congenital brain deformity><cytokine><data base><deliver mRNA><deliver messenger RNA><delivery system for mRNA><drug candidate><effective therapy><effective treatment><factor A><functional disability><genome mutation><heterotaxia syndrome><heterotaxy><host response><immune system response><immunoresponse><inositol 5-phosphatase><inositol polyphosphate 5-phosphatase><inositol triphosphatase><inositol triphosphate 5-phosphatase><inositol-1,4,5-trisphosphate 5-phosphatase><insight><intense pain><inversion of viscera><joint disorder><laterality sequence><lipid based nanoparticle><lipid nanoparticle><lymphocyte activating factor><mAbs><mRNA><mRNA delivery><messenger RNA delivery><microphthalmia><monoclonal Abs><mouse model><murine model><myoinositol trisphosphatase><nano particle><nano particle delivery><nano-sized particle><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanophthalmos><nanosized particle><nervous system function><novel><overexpress><overexpression><oxidation reduction reaction><phosphoinositide-3,4,5-triphosphate><proto-oncogene protein RAC><proto-oncogene protein akt><rac protein kinase><related to A and C-protein><rheumatic arthritis><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><site targeted delivery><situs abnormality><situs inversus viscerum><skeletal abnormality><social role><socio-economic><socio-economically><socioeconomically><socioeconomics><targeted delivery><transcription factor><transposition of viscera><visceral heterotaxy><visceral transposition>