SBIR-STTR Award

Methicillin resistant Staphylococcus aureus (MRSA) and methicillin sensitive S. aureus (MSSA) are highly significant human health threats that are responsible for a range of diseases. Recently, the CDC and colleagues reported that MRSA are the most significant causes of serious infections and infectious disease deaths in the United States. A major factor involved in the lethality of MRSA and MSSA are secreted toxins, known as superantigens. The lethality associated with these superantigens has been appreciated for decades, as the U.S. military stockpiled large amounts of staphylococcal enterotoxin B (SEB) under its old bioweapons program that was terminated in 1969. The mechanism of action of the toxins is well known - they act extracellularly by binding to the T cell receptor on T cells and stimulating massive release of inflammatory cytokines. The objective of the proposed work is to test the feasibility of using a novel potential drug, a soluble T cell receptor protein (G5-8) that has been engineered for very high-affinity binding to SEB, to prevent lethality. Two rabbit models of SEB-induced lethality will be examined, pulmonary exposure to the toxin alone (as would occur in a bioterrorism incident) or pulmonary infection with a strain of MRSA that produces SEB. The hypotheses of the Phase I work are that yields of the engineered V2 protein G5-8 can be improved, and that the doses and timing of administration that will reduce the severity of pulmonary diseases associated with SEB exposure can be determined. The Specific Aims are: 1) To determine the best conditions for optimizing the yield and long-term storage of the V2-TCR G5-8 protein;2) To determine the efficacy of using G5-8 to protect rabbits from death due to intrapulmonary challenge with SEB;and 3) To determine the efficacy of using G5-8 to protect rabbits from death due to intrapulmonary challenge with an SEB+ CA-MRSA USA400 strain. In Phase II work, ImmuVen would test whether long-term stored G5-8 prevents lethality in rabbits exposed to additional SEB+ MRSA strains, and whether the G5-8 can be used in nebulized form. ImmuVen will also begin development of a cGMP process for additional pre-clinical and clinical studies. G5-8 and similar proteins in development for other S. aureus exotoxins (SEC, TSST-1) represent clinical and commercial opportunities to treat a host of diseases caused or worsened by MRSA and MSSA.

Public Health Relevance:
Antibiotic-resistant Staphylococcus aureus isolates frequently initiate serious human diseases through the bacterium's colonization of respiratory mucosa and skin, followed by secretion of potent toxins referred to as superantigens. Illnesses include serious post- viral pneumonias, allergic skin diseases, and delayed wound healing that affect thousands of individuals each year in the United States. The secreted toxins are directly involved in the disease effects, including death. This application studies a novel protein- based drug that acts to block the disease-causing effects of bronchially-administered superantigens (as would be the case in a bioterrorism agent) and of infections with antibiotic-resistant S. aureus.

Thesaurus Terms:
2,6 Dimethoxyphenylpenicillin;4-Thia-1-Azabicyclo(3.2.0)Heptane-2-Carboxylic Acid, 6-((2,6-Dimethoxybenzoyl)Amino)-3,3-Dimethyl-7-Oxo-, (2s-(2alpha,5alpha,6beta))-;Accounting;Aerosols;Affect;Affinity;Allergic;Animals;Antibiotic Resistance;Armed Forces Personnel;Atopic Dermatitis;Bacteria;Binding;Binding (Molecular Function);Biological Terrorism;Bioterrorism;Cdc;Categories;Centers For Disease Control;Centers For Disease Control (U.S.);Centers For Disease Control And Prevention;Centers For Disease Control And Prevention (U.S.);Cessation Of Life;Clinical;Clinical Research;Clinical Study;Communicable Diseases;Cutaneous Disorder;Cyclic Gmp;Death;Dermatitis, Atopic;Dermatoses;Development;Diagnosis;Disease;Disease Management;Disorder;Disorder Management;Dose;Drugs;E Coli Proteins;Eczema, Atopic;Endocarditis;Engineering;Engineerings;Escherichia Coli Proteins;Exotoxins;Exposure To;Foundations;Genetic Engineering Of Proteins;Government;Guanosine Cyclic 3',5'-Monophosphate;Guanosine Cyclic Monophosphate;Guanosine, Cyclic 3',5'-(Hydrogen Phosphate);Healing Abnormal;Healing Delayed;Health;Hour;Human;Human, General;Illinois;Immunoglobulin V;Immunoglobulin Variable Region;Impaired Healing;Impaired Tissue Repair;Impaired Wound Healing;Individual;Infection;Infectious Disease Pathway;Infectious Diseases;Infectious Diseases And Manifestations;Infectious Disorder;Inflammatory;Lead;Legal Patent;Licensing;Lung;Lung Diseases;Mhc Receptor;Mrsa;Major Histocompatibility Complex Receptor;Mammals, Rabbits;Man (Taxonomy);Man, Modern;Marketing;Mediation;Medication;Methicillin;Methicillin Resistant S. Aureus;Methicillin Resistant Staphylococcus Aureus;Military;Military Personnel;Modeling;Molecular Interaction;Negotiating;Negotiation;Neurodermatitis, Atopic;Neurodermatitis, Disseminated;Oryctolagus Cuniculus;Patents;Pathogenicity Factors;Patients;Pb Element;Penicillin, Dimethoxyphenyl;Pharmaceutic Preparations;Pharmaceutical Agent;Pharmaceutical Preparations;Pharmaceuticals;Pharmacologic Substance;Pharmacological Substance;Phase;Process;Programs (Pt);Programs [publication Type];Protein Engineering;Proteins;Pulmonary Diseases;Pulmonary Disorder;Rabbit, Domestic;Rabbits;Receptor Protein;Receptors, Antigen, T-Cell;Reporting;Resistance To Antibiotics;Resistance, Antibiotic;Resistant To Antibiotics;Respiratory Disease;Respiratory Disorder;Respiratory Mucosa;Respiratory System Disease;Respiratory System Disorder;Respiratory System, Lung;Rights;S. Aureus;S.Aureus;Sbir;Sbirs (R43/44);Seb;Severities;Skin;Skin Diseases;Skin Diseases And Manifestations;Small Business Innovation Research;Small Business Innovation Research Grant;Staph Enterotoxin F;Staphylococcal Enterotoxin B;Staphylococcus Aureus;Superantigens;T Cell Receptor;T Lymphocyte;T-Cell Receptor;T-Cells;T-Lymphocyte;Tsst-1;Tst Protein, Staphylococcus Aureus;Technology;Testing;Therapeutic;Thymus-Dependent Lymphocytes;Time;Toxic Shock;Toxic Shock Syndrome;Toxic Shock Syndrome Toxin-1;Toxin;Translating;Translatings;United States;United States Centers For Disease Control;United States Centers For Disease Control And Prevention;Universities;Variable Region;Variable Region, Ig;Viral Pneumonia;Virulence Factors;Work;Yeasts;Abnormal Tissue Repair;Allergic Dermatitis;Allergic Eczema;Antibiotic Resistant;Atopic Dermatitis;Base;Cgmp;Clinical Relevance;Clinically Relevant;Cytokine;Delayed Wound Healing;Disease /Disorder;Disease Prevention;Disease/Disorder;Disorder Prevention;Drug /Agent;Drug/Agent;Enterotoxin F, Staphylococcal;Gene Product;Guanosine 3'5' Monophosphate;Heavy Metal Pb;Heavy Metal Lead;Human Disease;Improved;Innovate;Innovation;Innovative;Interest;Language Translation;Lung Disorder;Methicillin Resistant Staphylococcus Aureus (Organism);Model;New Technology;Novel;Pre-Clinical;Preclinical;Prevent;Preventing;Programs;Protein Folding;Pulmonary;Receptor;Respiratory;Skin Disorder;Staph Enterotoxin B;Technological Innovation;Therapeutic Protein;Thymus Derived Lymphocyte;Toxic-Shock Toxin;University;Viral Pneumonia

Methicillin resistant Staphylococcus aureus (MRSA) and methicillin sensitive S. aureus (MSSA) are highly significant human health threats that are responsible for a range of diseases. Recently, the CDC and colleagues reported that MRSA are the most significant causes of serious infections and infectious disease deaths in the United States. A major factor involved in the lethality of MRSA and MSSA are secreted toxins, known as super antigens. The lethality associated with these super antigens has been appreciated for decades, as the U.S. military stockpiled large amounts of staphylococcal enterotoxin B (SEB) under its old bioweapons program that was terminated in 1969. The mechanism of action of the toxins is well known - they act extracellularly by binding to the T cell receptor on T cells and stimulating massive release of inflammatory cytokines. We have engineered soluble T cell receptors for high-affinity binding to super antigens to prevent toxin-associated lethality. The objective of this proposal is to build on the success of work completed in Phase I where treatment with the novel therapeutic (soluble, high-affinity T cell receptor, called IMV-01) was able to neutralize SEB secreted from MRSA in an intrapulmonary rabbit model. The hypotheses of the Phase II work are that yields of the therapeutics can be improved, that IMV-01 and two soluble T cell receptor leads (IMV-02 and IMV-03) already engineered against the other clinically important super antigens (SEC &TSST-1) can be co-administered as a cocktail to neutralize these toxins, and prevent lethality due to pneumonia caused by MRSA. The Specific Aims are: 1) To determine the best conditions for optimizing the yield and purity of the V?-TCRs IMV-01, IMV-02 and IMV-03;2) To determine the effective dose and timing of IMV-01, IMV-02 or IMV-03 individually to neutralize their respective toxins produced by MRSA strains in an intrapulmonary rabbit model in the presence or absence of vancomycin;and 3) To determine the efficacy of using IMV-01, IMV-02 and IMV- 03 as a cocktail to protect rabbits from death due to intrapulmonary challenge with three different MRSA strains secreting either SEB + SEC, SEB + TSST-1 or SEC + TSST-1, in the presence or absence of vancomycin.

Public Health Relevance:
Antibiotic-resistant Staphylococcus aureus isolates frequently initiate serious human diseases through the bacterium's colonization of respiratory mucosa and skin, followed by secretion of potent toxins referred to as super antigens. Illnesses include serious post viral pneumonias, allergic skin diseases, and delayed wound healing that affect thousands of individuals each year in the United States. The secreted toxins are directly involved in the effects of the, disease and can cause death. In this application, ImmuVen will develop a therapeutic that neutralizes these toxins, and examine its effectiveness in various models of lethal pneumonia caused by the bacterium.