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

• 批准号: 10618787
• 负责人: TZYY-CHOOU WU
• 金额: $ 57.08万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 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/10618787
• 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-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; 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 HPV vaccine; therapeutic vaccine; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions; tumorigenesis; vaccination strategy; vaccine candidate

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.

项目总结/摘要 人乳头瘤病毒（HPV）作为许多病症的病原体的鉴定， 宫颈癌，导致HPV靶向治疗的发展，包括治疗性HPV 疫苗，用于治疗HPV相关的恶性肿瘤。然而，通过以下方法证明的有效性 在临床前研究中的治疗性HPV疫苗候选物通常不反映在临床环境中。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 具有以下特点：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）具有局部免疫抑制肿瘤 与临床HPV+肿瘤相似的微环境（TME）; 4）肿瘤形成应遵循临床 从癌前病变到浸润和转移状态的进展; 5）适用于不同的MHC I类背景;和6）荷瘤小鼠应适当地对免疫应答 策略并产生抗肿瘤免疫。初步数据：我们开发了一种策略， 原位注射致癌质粒治疗临床前自发性HPV宫颈阴道癌 编码HPV 16-E6、HPV 16-E7、组成型活性Akt、荧光素酶报告基因和睡美人 转座酶（SB）电穿孔进入小鼠宫颈阴道，以提高转染效率。 SB的后续表达诱导质粒DNA整合到转染细胞的基因组中， 导致持续的癌基因表达和转染细胞的自发转化。中 宫颈阴道内短期抗CD 3给药诱导的全身性免疫抑制 致癌质粒转染导致HPV+肿瘤与癌的自发形成 特色我们建议通过引入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， 消除短期CD 3耗竭的需要。此外，我们将进一步利用遗传远交小鼠和HPV 16 假病毒体递送癌基因以产生自发性肿瘤，从而重述了 遗传多样的患者群体和HPV 16感染诱导的癌基因引入。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结合 免疫抑制分子，在克服免疫抑制性TME的产生改善， 治疗性抗肿瘤反应。影响：一种新的临床前HPV宫颈阴道癌模型， 概述了临床情况将加强对HPV的重要免疫学和生物学研究- 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， 允许测试靶向免疫抑制基因的新型分子干预。