SBIR-STTR Award

The objective of this project is to develop a therapeutic strategy for H|V-associated malignancy based on the role of HPV in the pathogenesis. HPV related cancers express the E6/E7 oncoproteins of HPV that are ideal targets for immune inducing vaccines. We will develop a vaccine based upon a novel adenovirus serotype-S vector (Ads) platform with unique and additional deletions of the viral DNA polymerase and the pre- terminal protein in the early gene 2b (E2b) region (Ads [E1-, E2b-]). In studies employing HIV antigens, we reported that the new Ad5 [E1-, E2b-]-HlV vector vaccine was superior to a current Ads [E1-]-H|V vector vaccine (containing deletion in the early gene 1 (E1) region) when used to induce CMI responses in a multiple immunization regimen. Significant antigen specific CMI responses were induced in mice and monkeys despite the presence of pre-existing AdS immunity. We will construct and produce an AdS [E1-, E2b-] vector vaccine that contains the HPV oncoproteins E6/E7 with non-oncogenic function. This new recombinant adenovirus will induce immune responses by expressing the modified HPV-E6/E7 antigens after direct transfection of antigen presenting cells. We will evaluate this in combination with a toll-like receptor agonist (TLRa) designed to enhance immune responses induced by the vaccine. Our pre-clinical data indicate that the AdS [E1-, E2b-] vec*or induces robust CMI responses against tumor associated antigen [fAA), even in the presence of pre- existing AdS immunity. In a murine cancer model employing the carcinoembryonic antigen (CEA) gene insert, tl.g -CEA immunogenicity and in vivo anti-tumor effects of repeated immunizations with i new AdS [E1-, E2b-l- GEA were compared to those observed with a current generation Ads tE1-l-CEA. These AdS vectors were tested in a clinically relevant AdS immune setting. We observed that AdS immune mice immunized multiple times with AdS [E1-, E2b-]-CEA induced CEA-specific CMI responses that were significantly increased over those detected in AdS immune mice immunized multiple times with a current generation AdS tE1-l-CEA. Ads immune mice bearing CEA expressing tumors that were treated with AdS [E1-, E2b-J-CEA had an increased anti-tumor response as compared to AdS [E1-I-CEA treated mice. These results demonstrate that AdS [E1-, Ezb-l-CEA can induce CMI immune responses that result in tumor growth inhibition despite the presence of pre-existing AdS immunity. We have also utilized the Ads [E1-, Ezb-] vector platform expressing the TAA HER2lneu as a breast cancer immunotherapeutic agent. Ads [E1-, E2b-l-HERZlneu induced potent CMI against HER2/neu and significantly inhibited progression of established tumors in Ad5 immune mice. These data demonstrate that in vivo delivery of Ad5 [E1-, E2b-] vectors expressing TM can induce anti-TAA immunity and inhibit progression of tumors in AdS immune animals.

Public Health Relevance:
In this study, we will develop a new adenoviral based drug (AdS [E1-, E2b-I-HPV-E6/7) to treat HtV+ patients with HPv-associated oropharyngeal and tonsillar ,ncers. The treatment platform is needed to overcome pre-existing AdS immunity that has prevented the widespread use of this type of technology.

Thesaurus Terms:
Aids Antigens;Aids Virus;Aids Therapy;Atgn;Acquired Immune Deficiency Syndrome Virus;Acquired Immunodeficiency Syndrome Virus;Adenoviridae;Adenoviruses;Adverse Effects;Agonist;Animals;Anti-Retroviral Agents;Antigen Targeting;Antigen-Presenting Cells;Antigens;Antiretroviral Agents;Breast Cancer;Cd66e Antigen;Cancer Model;Cancermodel;Cancers;Carcinoembryonic Antigen;Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5;Cells;Clinical Data;Combined Modality Therapy;Dna Polymerases;Dna-Dependent Dna Polymerases;Dna-Directed Dna Polymerase;Data;Disease;Disorder;Drugs;Ec 2.7.7.7;Erbb2;Erbb2 Gene;Fap-1 Protein;Fas-Associated Phosphatase-1;Generations;Genes;Her -2;Her-2;Her-2 Genes;Her2;Her2 Genes;Her2/Neu;Hiv;Hiv Antigens;Hiv-Associated Antigens;Hnscc;Hpv;Htlv-Iii;Htlv-Iii Antigens;Htlv-Iii-Lav Antigens;Head And Neck Carcinoma;Head And Neck Squamous Cell Carcinoma;Human Egf Receptor 2 Gene;Human Immunodeficiency Viruses;Human Papilloma Virus;Human Papillomavirus;Human T-Cell Leukemia Virus Type Iii;Human T-Cell Lymphotropic Virus Type Iii;Human T-Lymphotropic Virus Type Iii;Human T-Lymphotropic Virus Type Iii Antigens;Human Papilloma Virus Infection;Human Papillomavirus Infection;Immune;Immune Targeting;Immune Response;Immunity;Immunization;Immunologic Accessory Cells;Immunologic Sensitization;Immunologic Stimulation;Immunological Sensitization;Immunological Stimulation;Immunologically Directed Therapy;Immunostimulation;Immunotherapeutic Agent;Immunotherapy;In Vitro;Incidence;Individual;Infectious Human Wart Virus;Lav Antigens;Lav-Htlv-Iii;Lymphadenopathy-Associated Antigens;Lymphadenopathy-Associated Virus;Malignant Neoplasms;Malignant Oropharyngeal Neoplasm;Malignant Oropharyngeal Tumor;Malignant Tonsillar Neoplasm;Malignant Tonsillar Tumor;Malignant Tumor;Meconium Antigen 100;Mediating;Medication;Mice;Mice Mammals;Monitor;Monkeys;Monocytes / Macrophages / Apc;Morbidity;Morbidity - Disease Rate;Mortality;Mortality Vital Statistics;Multimodal Therapy;Multimodal Treatment;Multimodality Treatment;Murine;Mus;Oncogene Products;Oncogene Proteins;Oncoproteins;Oral;Oropharnyx Cancers;Oropharyngeal;Oropharyngeal Cancer;Oropharyngeal Carcinoma;Oropharynx;Oropharynx Cancer;Oropharynx Carcinoma;Oropharynxs;Ptpn13;Pathogenesis;Patients;Pharmaceutic Preparations;Pharmaceutical Preparations;Plaque Assay;Prevalence;Proteins;Recombinants;Regimen;Reporting;Risk;Role;Scchn;Serotyping;Tkr1;Tlr Protein;Technology;Telomerase;Testing;Therapeutic;Time;Timeline;Toll-Like Receptors;Tonsil;Tonsil Cancer;Tonsillar Cancer;Transfection;Treatment Side Effects;Tumor Antigens;Tumor-Associated Antigen;United States;Vaccines;Viral Vector;Virus-Hiv;Virus-Related Malignancy;Virus-Related Malignant Neoplasm;Western Blotting;Western Immunoblotting;Accessory Cell;Anti-Retroviral;Antiretroviral;Base;C-Erbb-2;C-Erbb-2 Genes;C-Erbb-2 Proto-Oncogenes;Cancer Progression;Carcinoembryonal Antigen;Clinical Relevance;Clinically Relevant;Combination Therapy;Combined Modality Treatment;Combined Treatment;Design;Designing;Develop A Vaccine;Development Of A Vaccine;Disease/Disorder;Drug/Agent;Erbb-2 Genes;Erbb2 [{C0242957}];Experiment;Experimental Research;Experimental Study;Gene Product;Host Response;Immune Therapy;Immunogen;Immunogenicity;Immunologic Preparation;Immunoresponse;Immunotherapeutics;In Vivo;Malignancy;Malignant Breast Neoplasm;Malignant Breast Tumor;Malignant Oropharynx Neoplasm;Malignant Oropharynx Tumor;Malignant Tonsil Neoplasm;Malignant Tonsil Tumor;Multimodality Therapy;Neoplasm Progression;Neoplasm/Cancer;Neoplastic Progression;Neu Genes;New Vaccines;Next Generation Vaccines;Non-Oncogenic;Novel;Novel Vaccines;Oral Pharyngeal;Pandemic;Pandemic Disease;Plasmid Vaccine;Pre-Clinical;Preclinical;Prevent;Preventing;Protein Blotting;Research Study;Response;Side Effect;Social Role;Therapeutic Vaccine;Therapy Adverse Effect;Tonsillar;Treatment Adverse Effect;Tumor;Tumor Growth;Tumor Progression;Tumor-Specific Antigen;Vaccine Development;Vector;Vector Vaccine;Vector-Induced;Viral Dna;Virus Dna;Virus Related Cancer;Wart Virus

The objective of this project is to further develop an immunotherapeutic strategy for HIV-associated malignancy based on the role of HPV in the pathogenesis and perform the pre-clinical experiments with a new drug which allow us to enter clinical trials to test its safety and effectiveness. HPV related cancers express the E6/E7 oncoproteins of HPV that are ideal targets for immune inducing vaccines. We developed a vaccine based upon a novel adenovirus serotype-5 vector (Ad5) platform with unique and additional deletions of the viral DNA polymerase and the pre-terminal protein in the early gene 2b (E2b) region (Ad5 [E1-, E2b-]). In studies employing infectious disease and cancer antigens, we reported that new Ad5 [E1-, E2b-] vector vaccines are superior to current Ad5 [E1-] vector vaccines (containing deletion in the early gene 1 (E1) region) when used to induce CMI responses in a multiple homologous immunization regimen. We have demonstrated that significant antigen specific CMI responses are induced in mice and monkeys despite the presence of pre- existing Ad5 immunity. Our data indicates that the new Ad5 [E1-, E2b-] vectors induces robust CMI responses against tumor associated antigens (TAA) resulting in anti-tumor activity, even in the presence of pre-existing Ad5 immunity. We have constructed and produced a modified HPV-E6/E7 gene that expresses non-oncogenic early gene 6 (E6) and early gene 7 (E7) HPV proteins. We have incorporated this modified HPV-E6/E7 gene into the new recombinant Ad5 [E1-, E2b-] vector platform (Ad5 [E1-, E2b-]-HPV-E6/E7) to be used as an immunotherapeutic for the treatment of patients with HPV-E6/E7 expressing cancers. This recombinant platform induces immune responses by expressing the modified HPV-E6/E7 antigens after direct transfection of antigen presenting cells via injection. We evaluated this dru combination with a toll-like receptor agonist (TLRa) platform designed to enhance immune responses induced by the vector. In an Ad5 immune murine cancer model employing the modified HPV-E6/E7 gene insert, the immunogenicity and in vivo anti-tumor effects of repeated immunizations with the Ad5 [E1-, E2b-]-HPV-E6/E7 with or without the addition of Ad5 [E1-, E2b-]-TLRa were compared. Significantly higher levels of HPV-E6/E7 directed CMI activity was induced in mice immunized with Ad5 [E1-, E2b-]-HPV-E6/E7 plus Ad5 [E1-, E2b-]-TLRa as compared with mice immunized with Ad5 [E1-, E2b-]-HPV-E6/E7 alone. Importantly, mice treated by immunotherapy with Ad5 [E1-, E2b-]-HPV-E6/E7 with or without the addition of Ad5 [E1-, E2b-]-TLRa experienced large reductions in tumor growth as compared with control mice injected with Ad5 [E1-, E2b-]-null (empty vector). These results indicate that immunotherapy of HPV-E6/E7 expressing tumors using the new Ad5 [E1-, E2b-]-HPV-E6/E7 induces potent anti-tumor activity. Our previous research data also indicates that this new immunotherapeutic drug may be used in conjunction with chemotherapy/irradiation treatment to induce effective anti-tumor activity. The studies in the present pre-clinical program will allow us to advance manufacturing approaches and acquire data that will allow us to advance to FDA approved clinical trials

Public Health Relevance Statement:


Public Health Relevance:
In this study, we will develop a new immunotherapy to treat HPV induced cancers such as oropharyngeal and tonsillar cancers as well as certain cervical cancers. The new approach is to use an adenoviral based drug (Ad5 [E1-, E2b-]-HPV-E6/7) to treat HIV+ patients with HPV-associated Head and Neck cancers. Our published research data indicates that this novel immunotherapeutic drug induces anti-tumor activity and may be used in conjunction with chemotherapy/irradiation treatment for patients. These studies set the stage for a first in man clinical trial to test this approach in HIV+ Head and Neck cancer.

Project Terms:
Adenoviruses; Adjuvant; Agonist; AIDS therapy; Animals; Anti-Retroviral Agents; Antibodies; Antigen Targeting; Antigen-Presenting Cells; Antigens; base; Biological Assay; Cancer Model; cell mediated immune response; Cells; chemotherapy; Clinical Trials; Combined Modality Therapy; Communicable Diseases; Data; design; Disease; DNA-Directed DNA Polymerase; Dose; Effectiveness; Enzyme-Linked Immunosorbent Assay; experience; FDA approved; Flow Cytometry; Gene Delivery; Genes; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Health; HIV; Human; Human papilloma virus infection; Human Papillomavirus; Immune; Immune response; Immune Targeting; Immunity; Immunization; immunogenicity; Immunotherapeutic agent; Immunotherapy; in vivo; Incidence; Individual; Injection of therapeutic agent; irradiation; Malignant neoplasm of cervix uteri; Malignant Neoplasms; malignant oropharynx neoplasm; malignant tonsil neoplasm; man; Mediating; Monkeys; Morbidity - disease rate; Mortality Vital Statistics; Mus; non-oncogenic; novel; novel strategies; novel vaccines; Oncogene Proteins; Oropharyngeal; pandemic disease; Pathogenesis; Patients; Pharmaceutical Preparations; Phase; phase 1 study; pre-clinical; preclinical study; Prevalence; programs; Proteins; Publishing; Recombinants; Regimen; Reporting; Research; research study; Risk; Role; Safety; Serotyping; Small Business Innovation Research Grant; Staging; Testing; Therapeutic; therapeutic vaccine; TimeLine; Toll-like receptors; Tonsil; Transfection; tumor; Tumor Antigens; tumor growth; United States; vaccine development; Vaccines; vector; vector vaccine; vector-induced; viral DNA; Viral Proteins; virus related cancer