Development of Novel Spontaneous HPV Cervicovaginal Carcinoma Models for Cancer Immunotherapy

用于癌症免疫治疗的新型自发性 HPV 宫颈阴道癌模型的开发

基本信息

批准号：
10374864
负责人：
TZYY-CHOOU WU
金额：
$ 57.12万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10374864
关键词：
Antigens Antitumor Response Automobile Driving Biology C57BL/6 Mouse CD3 Antigens CD8-Positive T-Lymphocytes Cancer Model Carcinoma Cells Cervical Squamous Cell Carcinoma Cervix carcinoma Characteristics Clinic Clinical DNA Data Development Electroporation Gene Expression Generations Genes Genetic Genome Human Human Papilloma Virus Vaccine Human Papilloma Virus-Related Malignant Neoplasm Human Papillomavirus Human papillomavirus 16 Immune Immune Evasion Immune Targeting Immune checkpoint inhibitor Immune response Immunity Immunocompetent Immunosuppression Immunotherapeutic agent Immunotherapy Infection Injections Intervention Kinetics Lesion Luciferases MHC Class I Genes Malignant Neoplasms Malignant neoplasm of cervix uteri Mediating Methodology Modeling Molecular Monitor Monoclonal Antibody HuM291 Morphology Mus Oncogenes Oncogenic Oncoproteins Performance Plasmids Population Process Proteins Reporter Genes Research Role Shapes Sleeping Beauty System T-Cell Depletion Testing Therapeutic Therapeutic Effect Time Transfection Transposase Treatment Efficacy Tumor Immunity Tumor Markers Vaccinated Vaccines anti-tumor immune response base biological research cancer imaging cancer immunotherapy cervicovaginal clinically relevant experience immunosuppressed improved inhibitor luminescence neoplastic cell novel overexpression patient population plasmid DNA pre-clinical preclinical study predict clinical outcome premalignant success targeted treatment therapeutic HPV vaccinations therapeutic vaccine treatment strategy tumor tumor growth tumor microenvironment tumor-immune system interactions tumorigenesis vaccination strategy vaccine candidate

项目摘要

PROJECT SUMMARY / ABSTRACT The identification of human papillomavirus (HPV) as a causative agent for a host of conditions, particularly cervical cancer, has led to the development of HPV-targeting therapeutics, including therapeutic HPV vaccines, for the treatment of HPV-associated malignancies. However, the potent efficacies demonstrated by the therapeutic HPV vaccine candidates in preclinical studies are often not reflected in clinical settings. This discrepancy is potentially due to the inability of existing preclinical HPV tumor models to fully replicate the biology of clinical HPV-associated cancers. We hypothesize that an ideal preclinical HPV tumor model should possess the following characteristics: 1) forms spontaneous, localized, HPV oncogenic proteins-expressing tumors; 2) displays carcinoma morphology; 3) possesses a locally immunosuppressive tumor microenvironment (TME) resembling that of clinical HPV+ tumors; 4) tumor formation should follow clinical progression starting from a precancerous to an invasive and metastatic state; 5) be applicable to different MHC class I backgrounds; and 6) the tumor-bearing mice should respond appropriately to immunotherapeutic strategies and generate anti-tumor immunity. Preliminary data: We developed a strategy for the generation of preclinical spontaneous HPV cervicovaginal carcinoma based on orthotopic injection of oncogenic plasmids encoding HPV16-E6, HPV16-E7, constitutively active Akt, luciferase reporter gene, and Sleeping Beauty Transposase (SB) into the cervicovaginal tract of mice with electroporation to enhance transfection efficiency. Subsequent expression of SB induces the integration of plasmid DNA into the genome of transfected cells, resulting in persistent oncogenes expression and spontaneous transformation of transfected cells. In a systemic immunosuppressed setting induced by short-term anti-CD3 administration, intracervicovaginal oncogenic plasmid transfection led to the spontaneous formation of HPV+ tumors with carcinoma characteristics. We propose to further optimize our model by incorporating immunosuppressive molecules that are often overexpressed in clinical cervical cancers into our spontaneous HPV cervicovaginal tumor model and eliminate the need of short-term CD3 depletion. Also, we will further utilize genetic outbred mice and HPV16 pseudovirion delivery of oncogenes for the generation of spontaneous tumors, thereby recapitulating the genetic diverse patient population and HPV16 infection-induced oncogene introduction. Furthermore, we will examine various treatment strategies, such as the combination of therapeutic HPV vaccination with inhibitors of immunosuppressive molecules, in overcoming the immunosuppressive TME for the generation of improved therapeutic antitumor responses. Impact: A novel preclinical HPV cervicovaginal cancer model that faithfully recapitulates the clinical situation would potentiate crucial immunotherapeutic and biological research for HPV- associated cancers, provide better predictions for clinical outcomes of HPV-specific immunotherapies, and permit testing of novel molecular interventions targeting immune suppressive genes.

项目概要/摘要人乳头瘤病毒 (HPV) 被鉴定为多种疾病的病原体，特别是宫颈癌，导致了 HPV 靶向疗法的发展，包括治疗性 HPV 疫苗，用于治疗 HPV 相关恶性肿瘤。然而，所证明的强大功效临床前研究中的治疗性 HPV 候选疫苗通常不会反映在临床环境中。这差异可能是由于现有的临床前 HPV 肿瘤模型无法完全复制临床 HPV 相关癌症的生物学。我们假设理想的临床前 HPV 肿瘤模型应该具有以下特征：1）形成自发的、局部的、表达HPV致癌蛋白的肿瘤； 2) 显示癌形态； 3) 患有局部免疫抑制肿瘤类似于临床 HPV+ 肿瘤的微环境 (TME)； 4）肿瘤的形成应遵循临床从癌前状态进展到侵袭性和转移性状态； 5) 适用于不同的MHC I 类背景； 6) 荷瘤小鼠应对免疫治疗产生适当反应策略并产生抗肿瘤免疫力。初步数据：我们制定了生成的策略基于致癌质粒原位注射的临床前自发性 HPV 宫颈阴道癌编码 HPV16-E6、HPV16-E7、组成型活性 Akt、荧光素酶报告基因和睡美人通过电穿孔将转座酶 (SB) 导入小鼠子宫颈阴道以提高转染效率。随后 SB 的表达诱导质粒 DNA 整合到转染细胞的基因组中，导致癌基因持续表达和转染细胞自发转化。在一个短期抗 CD3 给药诱导的全身免疫抑制环境，宫颈阴道内致癌质粒转染导致 HPV+ 肿瘤自发形成特征。我们建议通过结合免疫抑制分子来进一步优化我们的模型在我们的自发性 HPV 宫颈阴道肿瘤模型中，临床宫颈癌中经常过度表达，并且消除短期 CD3 耗尽的需要。此外，我们将进一步利用基因远交小鼠和HPV16 假病毒粒子传递癌基因以产生自发性肿瘤，从而概括了遗传多样性的患者群体和 HPV16 感染诱发癌基因的引入。此外，我们将检查各种治疗策略，例如治疗性 HPV 疫苗接种与抑制剂的组合免疫抑制分子，克服免疫抑制 TME，产生改进的治疗性抗肿瘤反应。影响：一种新型临床前 HPV 宫颈阴道癌模型，忠实地概括临床情况将加强 HPV 的重要免疫治疗和生物学研究相关癌症，为 HPV 特异性免疫疗法的临床结果提供更好的预测，以及允许测试针对免疫抑制基因的新型分子干预措施。