Mouse modeling of HPV infection

总结 我们的总体目标是建立一个基于实验室小鼠的人乳头瘤病毒（HPV）感染模型， 支持新型HPV疫苗的开发。转化研究需要动物模型 这是人类病理学的强大代表，用于测试临床重要性的问题， 为开发新的干预措施和患者获益提供可靠的信息。HPV是主要的 全球所有癌症中至少5%的病原体，主要是宫颈癌和其他肛门和生殖系统癌症的子集， 头颈癌以及潜在的非黑色素瘤皮肤癌。不幸的是，没有具体的 抗病毒治疗，但基于疫苗的方法是非常有前途的。许多团体带来了候选人 治疗性HPV疫苗，包括我们的pNGLV 4aCRTE 6 E7 L2 DNA，但迄今为止， 尽管在目前的标准动物中疫苗的数据很有希望，但在治疗患者方面的成功有限 模型这表明需要更多的预测动物模型。HPV不会完成其生命周期 并在小鼠或细胞培养单层中产生病毒体， 结构经常使用。该系统不能完全模拟病毒衣壳的组装和成熟， 表达E4的分化上皮或提供疾病终点。一种从 需要在乳头状瘤中产生并表达临床相关HPV序列的病毒。所以我们 建议通过整合关键的HPV来改变小家鼠乳头瘤病毒1型（MmuPV 1）的效用 序列，并证明其可用作测试新型治疗性和保护性HPV疫苗的模型。 具体目的1：开发掺入HPV序列的MmuPV 1病毒。器官移植受者 （OTR）和HIV+患者表现出更严重和进行性的HPV疾病， HPV相关恶性肿瘤。免疫受损患者的非黑色素瘤皮肤癌（NMSC）是 与最初在疣状表皮发育不良（EDV）中描述的β HPV过多相关 患者具体目的2：开发一种基于MmuPV 1的人类队列小鼠攻击模型， HPV相关癌症发展的正常和高风险。为了验证这些新的小鼠 HPV感染和疾病的模型，我们建议检查许可的HPV疫苗和两种 我们的临床级实验疫苗，将很快进入早期阶段的测试：RG 1-VLP，一个单一的病毒样 颗粒抗原，旨在提供针对不同HPV类型的广泛免疫力， DNA疫苗pNGLV 4aCRTE 6 E7 L2通过电穿孔施用。具体目标3：比较 Gardasil 9、pNGLV 4aCRTE 6 E7 L2 DNA和RG 1-VLP疫苗对疾病和病毒的效力 在健康受试者和HPV相关癌症高风险受试者的鼠模型中的终点。

SUMMARY Our overall goal is to create a laboratory mouse-based model of human papillomavirus (HPV) infection and disease to support the development of novel HPV vaccines. Translational research requires animal models that are robust representations of human pathology in which to test questions of clinical importance, and provide reliable information for the development of novel interventions and patient benefit. HPV is the primary etiologic agent of at least 5% of all cancers worldwide, mostly cervical and subsets of other anogential and head and neck cancers and potentially also non-melanoma skin cancers. Unfortunately, there is no specific antiviral therapy, but vaccine-based approaches are very promising. Many groups are bringing candidate therapeutic HPV vaccines to the clinic, including our pNGLV4aCRTE6E7L2 DNA, but to date there has been limited success in treating patients despite promising data with the vaccines in the current standard animal models. This demonstrates the need for more predictive animal models. HPV does not complete its life cycle and produce virions in mice or in cell culture monolayers and so HPV pseudovirions delivering a reporter construct are often used. This system does not fully mimic the assembly and maturation of the viral capsid in E4-expressing differentiating epithelium or provide a disease endpoint. A model that produces disease from virus produced in a papilloma and expressing clinically-relevant HPV sequences is required. Therefore, we propose to transform the utility of Mus musculus papillomavirus type 1 (MmuPV1) by incorporating key HPV sequences and credential it for use as a model for testing novel therapeutic and protective HPV vaccines. SPECIFIC AIM 1: To develop MmuPV1 viruses incorporating HPV sequences. Organ transplant recipients (OTRs) and HIV+ patients exhibit more severe and progressive HPV disease, and dramatically higher rates of HPV-associated malignancies. Non-melanoma skin cancers (NMSC) in immune-compromised patients are associated with a plethora of βHPVs that were initially described in epidermodysplasia verruciformis (EDV) patients. SPECIFIC AIM 2: To develop an MmuPV1-based mouse challenge model of human cohorts at normal and high risk for the development of HPV-associated cancer. To validate these new murine models of HPV infection and disease we propose to examine the efficacy of a licensed HPV vaccine and two of our clinical grade experimental vaccines that will shortly enter early phase testing: RG1-VLP, a single virus-like particle antigen intended to provide broad immunity against diverse HPV types, and the candidate therapeutic DNA vaccine pNGLV4aCRTE6E7L2 administered via electroporation. SPECIFIC AIM 3: To compare the efficacy of the Gardasil 9, pNGLV4aCRTE6E7L2 DNA and RG1-VLP vaccines against disease and viral endpoints in murine models of healthy subjects and those at high risk for HPV-related cancer.