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.

项目摘要 /摘要鉴定人乳头瘤病毒（HPV）是许多疾病的病因，尤其是宫颈癌导致了HPV靶向疗法的发展，包括治疗性HPV 疫苗，用于治疗HPV相关的恶性肿瘤。但是，由临床前研究中的治疗性HPV疫苗候选物通常不会在临床环境中反映出来。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肿瘤模型应具有以下特征：1）形成自发，局部，HPV致癌蛋白表达的特征肿瘤； 2）显示癌形态； 3）拥有局部免疫抑制肿瘤微环境（TME）类似于临床HPV+肿瘤； 4）肿瘤形成应遵循临床从癌前到侵入性和转移状态的发展； 5）适用于不同的MHC I类背景； 6）含有肿瘤的小鼠应适当反应免疫治疗策略并产生抗肿瘤免疫力。初步数据：我们制定了一种生成的策略临床前自发的HPV子宫颈阴道癌基于原位注射致癌质粒的原位注射编码HPV16-E6，HPV16-E7，组成性活跃的AKT，荧光素酶记者基因和睡美人转座酶（SB）带有电穿孔的小鼠的宫颈阴道道，以提高转染效率。随后的SB表达诱导质粒DNA整合到转染细胞的基因组中，导致转染细胞的持续性癌基因表达和自发转化。在由短期抗CD3给药引起的全身免疫抑制设置致癌质粒转染导致HPV+肿瘤的自发形成特征。我们建议通过合并免疫抑制分子来进一步优化我们的模型通常在临床宫颈癌中过表达我们自发的HPV子宫颈肿瘤模型和消除了短期CD3耗竭的需求。此外，我们将进一步利用遗传杂种小鼠和HPV16 拟驱动的癌基因递送以生成自发性肿瘤，从而概括了遗传多样的患者人群和HPV16感染引起的癌基因引入。此外，我们会的检查各种治疗策略，例如治疗性HPV疫苗接种与抑制剂的组合免疫抑制分子，在克服改进的免疫抑制TME中治疗性抗肿瘤反应。影响：一种新型的临床前HPV子宫颈癌模型，忠实地概括临床情况将增强HPV-的关键免疫治疗和生物学研究相关的癌症，为HPV特异性免疫疗法的临床结果提供更好的预测，以及允许对靶向免疫抑制基因的新型分子干预进行测试。