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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/摘要人类乳头瘤病毒(HPV)是一种引起多种疾病的病原体，尤其是宫颈癌，导致了HPV靶向治疗的发展，包括治疗性HPV 用于治疗HPV相关恶性肿瘤的疫苗。然而，通过以下方式证明的有效方法临床前研究中的治疗性HPV候选疫苗通常没有在临床环境中得到反映。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。我们假设一个理想的临床前HPV肿瘤模型应该是具有以下特点：1)形成自发的、定位的、表达HPV致癌蛋白的肿瘤；2)显示肿瘤形态；3)具有局部免疫抑制肿瘤与临床HPV阳性肿瘤相似的微环境(TME)；4)肿瘤的形成应遵循临床从癌前病变到侵袭和转移状态的进展；5)适用于不同的MHC I类背景；6)荷瘤小鼠对免疫治疗应有适当的反应策略和产生抗肿瘤免疫。初步数据：我们制定了一项战略，以产生基于原位注射致癌基因的临床前自发性HPV宫颈阴道癌编码HPV16-E6、HPV16-E7、组成活性Akt、荧光素酶报告基因和睡美人利用电穿孔技术将转座酶(SB)导入小鼠宫颈阴道部，以提高转基因效率。 SB的随后表达诱导了质粒DNA整合到转基因细胞的基因组中，导致癌基因的持续表达和转基因细胞的自发转化。在一个阴道内短期应用抗CD3抗体引起的全身性免疫抑制致癌基因载体介导的人乳头瘤病毒阳性肿瘤的自发形成特点。我们建议通过加入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度表达到我们的自发性HPV宫颈阴道肿瘤模型中消除短期CD3耗尽的需要。此外，我们还将进一步利用遗传异种小鼠和HPV16 假病毒载体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感染诱导的癌基因导入。此外，我们还将检查各种治疗策略，例如将治疗性HPV疫苗接种与抑制剂相结合免疫抑制分子，在克服免疫抑制TME的基础上产生改良治疗抗肿瘤反应。Impact：一种新的临床前HPV宫颈阴道癌模型概述临床情况将加强关键的HPV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了更好的预测，以及允许测试针对免疫抑制基因的新型分子干预。