The primary objective of this project is to demonstrate the feasibility of developing the extracellular cold-inducible RNA-binding protein (eCIRP)-neutralizing monoclonal antibody #14 (mAb14) as a novel treatment for septic patients with acute lung injury (ALI). ALI is a critical component of the elevated mortality rate in sepsis no specific treatment has yet been approved to reduce the mortality of such patients. We have discovered that eCIRP is a critical inducer of ALI caused by sepsis and other inflammatory diseases. In our recent studies, we have shown that increased levels of eCIRP aggravated ALI in mice with sepsis induced by cecal ligation and puncture (CLP). Injection of recombinant eCIRP was sufficient to induce ALI in otherwise healthy mice. In our preliminary studies, we have generated a large panel of anti-eCIRP monoclonal antibodies to develop an eCIRP-targeting treatment for ALI, and screened them for their inhibition of eCIRP-induced release of TNF-a by macrophages. We then used the most effective anti-eCIRP monoclonal antibody, mAb14, to treat mice with CLP-induced ALI. Compared with non-immunized IgG (control), CLP mice treated with mAb14 had attenuated lung inflammation as indicated by the decreased lung gene and protein levels of TNF-a, IL-1ß, IL-6, CXCL1, and CXCL2, as well as the decreased neutrophil infiltration of the lungs as indicated by the myeloperoxidase activity. Based on these novel findings, we hypothesize that mAb14 can be developed as a new and effective drug to treat ALI caused by sepsis. In this project, we will further determine mAb14's eCIRP neutralization ability in vitro and in vivo. We will then optimize mAb14's dose to attenuate sepsis-induced ALI and therapeutic window to improve the survival of septic mice. We will also evaluate mAb14's pharmacokinetics (PK) and pharmacotoxicity properties. Our future steps will include developing a humanized form of mAb14 and then conducting its ADME, PK, advanced toxicology, and immunogenicity studies. We will then file with the FDA an investigational new drug (IND) application to initiate clinical trials to treat ALI in patients with sepsis. Our ultimate goal is to obtain commercial utilization of mAb14 as a safe and effective drug to treat patients with ALI in the context of sepsis.
Public Health Relevance Statement: Public Health Relevance Statement: Acute lung injury (ALI) is a common and deadly complication of sepsis. In this project, we will further characterize the anti-inflammatory effects of the new drug mAb14, a monoclonal antibody against extracellular cold-inducible RNA-binding protein (eCIRP), determine its beneficial effects in the preclinical model of sepsis-induced ALI, and establish its pharmacological properties. These proposed studies will provide critical feasibility information towards its clinical trials and eventual approval as a safe and effective treatment for sepsis-induced ALI.
Project Terms: Adult Respiratory Distress Syndrome; Da Nang Lung; Shock Lung; Stiff lung; wet lung; Acute Respiratory Distress Syndrome; Safety; Staining method; Stains; Technology; Testing; Time; Toxicology; Water; Hydrogen Oxide; cytokine; Measures; RNA-Binding Proteins; Alveolar Macrophages; Pulmonary Macrophages; Investigational New Drug Application; Injury; injuries; base; Blood specimen; Blood Sample; improved; Acute; Histologic; Histologically; Lung Alveolar Epithelia; alveolar epithelium; pneumocyte; Blood Serum; Serum; Acute Pulmonary Injury; Acute Lung Injury; Lung damage; pulmonary damage; pulmonary injury; pulmonary tissue damage; pulmonary tissue injury; lung injury; Chemotactic Cytokines; Homologous Chemotactic Cytokines; Intercrines; SIS cytokines; chemoattractant cytokine; chemokine; Therapeutic; fluid; liquid; Liquid substance; Attenuated; Inflammatory; septic; Intravenous; Life; Area Under Curve; Hour; Normal saline; cell type; Pattern; extracellular; American; interstitial; Surface Plasmon Resonance; attenuation; Histopathology; novel; Protein Gene Products; Gene Proteins; Neutrophil Recruitment; Neutrophil Infiltration; (TNF)-a; Cachectin; Macrophage-Derived TNF; Monocyte-Derived TNF; TNF; TNF A; TNF Alpha; TNF-a; TNFA; TNFa; Tumor Necrosis Factor; Tumor Necrosis Factor-alpha; TNF gene; CXCL2; Chemokine, CXC Motif, Ligand 2; GRO Protein, Beta; GRO2; GRO2 Oncogene; GROb; GROß; MIP-2A; MIP2-alpha; MIP2A; MIP2a; SCYB2; Small Inducible Cytokine Subfamily B, Member 2; macrophage inflammatory protein 2; ß-GRO protein; CXCL2 gene; CXCL1; GRO1; GROA; MGSA; SCYB1; CXCL1 gene; Property; Alveolar; Dose; Data; Preclinical Models; Pre-Clinical Model; Recombinants; in vivo; Antiinflammatory Effect; anti-inflammatory effect; Molecular; Development; developmental; immunogenicity; Sepsis; blood infection; bloodstream infection; neutralizing monoclonal antibodies; neutralizing mAb; novel therapeutics; new drug treatments; new drugs; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel therapy; public health relevance; effective therapy; effective treatment; Pulmonary Inflammation; Lung Inflammation; Pneumonitis; Injections; septic patients; sepsis patients; cecal ligation puncture; CLP model; CLP mouse model; Cecal ligation perforation; sepsis induced acute lung injury; ALI caused by sepsis; sepsis associated ALI; sepsis associated acute lung injury; sepsis caused acute lung injury; sepsis induced ALI; sepsis mediated ALI; sepsis mediated acute lung injury; sepsis related acute lung injury; Adult; 21+ years old; Adult Human; adulthood; Animals; Monoclonal Antibodies; Clinical Treatment Moab; mAbs; Bilirubin; Bilirubin IX alpha; Blood Cell Count; Blood Cell Number; Western Blotting; Immunoblotting; Western Immunoblotting; protein blotting; Bone Marrow; Bone Marrow Reticuloendothelial System; Cells; Cell Body; Clinical Trials; Complication; Creatinine; Disease; Disorder; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Dyes; Coloring Agents; Epithelial Cells; Evans blue stain; Azovan Blue; Evans Blue; Future; Goals; Half-Life; Heart; Hemorrhage; Bleeding; blood loss; Immunoglobulin G; 7S Gamma Globulin; IgG; In Vitro; Interleukin-1 beta; Beta Proprotein Interleukin 1; IL-1 beta; IL-1 ß; IL-1-b; IL-1ß; IL1-Beta; IL1-ß; IL1B Protein; IL1F2; IL1ß; Interleukin 1beta; Interleukin-1ß; Preinterleukin 1 Beta; Interleukin-6; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BSF-2; BSF2; HPGF; Hepatocyte-Stimulating Factor; Hybridoma Growth Factor; IFN-beta 2; IFNB2; IL-6; IL6 Protein; MGI-2; Myeloid Differentiation-Inducing Protein; Plasmacytoma Growth Factor; interferon beta 2; Small Intestines; small bowel; Ischemia; Kidney; Kidney Urinary System; renal; Laparotomy; Liver; hepatic body system; hepatic organ system; Lung; Lung Respiratory System; pulmonary; macrophage; MÏ; Mesentery; Mesenteric; mortality; Mus; Mice; Mice Mammals; Murine; Blood Neutrophil; Blood Polymorphonuclear Neutrophil; Marrow Neutrophil; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; neutrophil; Patients; Permeability; Hemi-Myeloperoxidase; Myeloperoxidase; Peroxidases; Pharmacokinetics; Drug Kinetics; Pharmacology; Blood Plasma; Plasma Serum; Reticuloendothelial System, Serum, Plasma; Plasma; lung edema; Pulmonary Edema; Common Rat Strains; Rat; Rats Mammals; Rattus; ARDS; Acute Respiratory Distress; Adult ARDS; Adult RDS
