Ovarian cancer gene therapy using HPV pseudovirion

使用 HPV 假病毒颗粒进行卵巢癌基因治疗

• 批准号: 9026581
• 负责人: TZYY-CHOOU WU
• 金额: $ 33.62万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-18 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026581
• 关键词: Adverse effects; Affinity; Antibodies; Bilateral; Binding; Binding Proteins; Bypass; CD8B1 gene; Cancer Control; Cells; Chimeric Proteins; Complement-Dependent Cytotoxicity; Cytotoxic T-Lymphocytes; DNA; DNA Vaccines; Data; Diagnosis; Diagnostic Neoplasm Staging; Disease; Distant Metastasis; Effector Cell; Enzymes; Epitopes; Exhibits; Face; Fc Immunoglobulins; Fc Receptor; Generations; Greater sac of peritoneum; Gynecologic; HLA-A2 Antigen; Health; Histocompatibility Antigens Class I; Homing; Human; Human Papillomavirus; Human papillomavirus 16; Humoral Immunities; Immune Tolerance; Immune response; Immunity; Immunoglobulin G; Indium; Individual; Influenza; Innovative Therapy; Intraperitoneal Injections; Ligands; Link; MHC Class I Genes; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Membrane Proteins; Modeling; Mus; Natural Immunity; Natural Killer Cells; Normal Cell; Oncogenes; Operative Surgical Procedures; Papillomavirus; Patients; Peptides; Proteins; Role; Site; Staging; Stromal Cells; Surface; T-Lymphocyte; T-Lymphocyte Epitopes; Therapeutic; Transgenic Mice; Transplantation; United States; Vaccination; Viral; Viral Vector; Virus; advanced disease; antitumor effect; cancer cell; cell killing; chemotherapy; curative treatments; female reproductive system; gene therapy; immunogenic; improved; in vivo; influenzavirus; innovation; killings; macrophage; mortality; neoplastic cell; neutralizing antibody; novel strategies; novel therapeutics; ovarian neoplasm; overexpression; retinal S antigen peptide M; targeted delivery; targeted treatment; therapeutic DNA; therapeutic gene; therapeutic protein; tumor; tumor microenvironment

DESCRIPTION (provided by applicant): Advanced ovarian cancer has the highest mortality rate among patients with cancers of the female reproductive system in the United States. Existing therapies for ovarian cancer, such as surgery and chemotherapy, have significant side effects and rarely result in long-term cures for patients with locally advanced or metastatic disease. The lack of curative treatments and high proportion of patients diagnosed with advanced disease underscores the urgent need to develop innovative and targeted therapies to control advanced stage ovarian cancer. In the current proposal, we aim to overcome major limitations of current therapies by employing human papillomavirus (HPV) pseudovirions (psVs) to deliver a therapeutic DNA construct to ovarian tumor cells in order to control infected and uninfected ovarian tumor cells through cell-mediated and humoral immune responses. Recently, we demonstrated that HPV psV is able to preferentially infect ovarian tumors in vivo. Furthermore, we have shown that a chimeric protein containing a tumor-homing molecule, NKG2D, fused to the Fc fragment of immunoglobulin G (IgG2a) was capable of coating tumor cells and generating antitumor effects (see preliminary data). The tumor-homing molecule, NKG2D, binds with high affinity to NKG2D ligand, which is overexpressed on the majority of ovarian tumor cells, compared to normal cells. NKG2D fused to the Fc fragment (NKG2D-Fc) enables the binding of effector cells exhibiting Fc receptor, such as macrophages and natural killer cells, to the tumor cells for the induction of antibody-dependent and complement-dependent cytotoxicity. We propose to further fuse NKG2D-Fc, with an MHC class I-restricted immunogenic CTL epitope for influenza virus, separated by a furin cleavage site. This will bypass immune tolerance by exploiting pre-existing cytotoxic T lymphocytes (CTLs) against a common foreign viral epitope, which is found in the vast majority of individuals. We have previously demonstrated that intraperitoneal injection of a similar chimeric protein containing model immunogenic CTL antigenic peptide, OVA, flanked with a furin cleavage site, led to the targeted delivery of the chimeric protein and the presentation of OVA CTL peptide on MHC class I molecules of tumor cells in tumor-bearing mice. Therefore, we reason that from the chimeric protein (NKG2D-Fc-RM), the NKG2D portion will make the chimeric protein bind specifically to HPV psV infected and uninfected ovarian cancer cells, where the immunogenic CTL epitopes will be released through cleavage by furin and coat the tumor cells for recognition by pre-existing CTLs. In addition, the presence of Fc renders the bound tumor cell susceptible to attack by humoral immunity. Overall, our proposed therapeutic chimeric protein delivered by HPV psV has the advantages of targeted tumor delivery, specific furin cleavage at the tumor site, and potent tumor-targeted killing through existing CTL immunity and Fc-mediated killing, representing a novel strategy with high translational value. The successful implementation of the proposed study would serve as an innovative strategy that may be applied to the treatment of other types of late stage cancers.

描述（由申请人提供）：在美国，女性生殖系统癌症患者中，晚期卵巢癌的死亡率最高。现有的卵巢癌疗法，例如手术和化学疗法，对局部晚期或转移性疾病患者的长期治愈产生显着副作用。缺乏治疗性疗法和高比例的诊断患有晚期疾病的患者强调了开发创新和有针对性的疗法以控制晚期卵巢癌的需求。在目前的提案中，我们旨在通过采用人类乳头瘤病毒（HPV）假病毒（PSV）来克服当前疗法的主要局限性，以通过细胞介导的和体形免疫反应来控制感染和未感染的卵巢肿瘤细胞，以控制卵巢肿瘤细胞的治疗性DNA构建体。最近，我们证明了HPV PSV能够在体内优先感染卵巢肿瘤。此外，我们已经表明，含有肿瘤分子NKG2D的嵌合蛋白与免疫球蛋白G（IgG2a）的FC片段融合在一起，能够涂有肿瘤细胞并产生抗肿瘤效应（请参阅初步数据）。与正常细胞相比，与NKG2D配体的高亲和力相比，肿瘤分子NKG2D与NKG2D配体的高亲和力与NKG2D配体结合，该配体在大多数卵巢肿瘤细胞上过表达。 NKG2D与FC片段（NKG2D-FC）融合，使表现出表现FC受体的效应细胞（例如巨噬细胞和天然杀伤细胞）的结合到肿瘤细胞中，以诱导抗体依赖性和补体依赖性的细胞毒性。我们建议将NKG2D-FC与MHC I限制性的免疫原性CTL表位进行进一步融合，以用于流感病毒，并由Furin裂解位点隔开。这将通过利用预先存在的细胞毒性T淋巴细胞（CTL）来绕过免疫耐受性，以针对常见的外源病毒表位，这在绝大多数个体中都发现。我们先前已经证明，腹膜内注射类似的嵌合蛋白含有模型的免疫原性CTL抗原肽，OVA，侧面有脂蛋白裂解位点，导致嵌合蛋白的靶向递送，并在MHC类I型MHC I-Molecules of Tumor-bire tumor-beare的OVA CTL肽上的呈现。 Therefore, we reason that from the chimeric protein (NKG2D-Fc-RM), the NKG2D portion will make the chimeric protein bind specifically to HPV psV infected and uninfected ovarian cancer cells, where the immunogenic CTL epitopes will be released through cleavage by furin and coat the tumor cells for recognition by pre-existing CTLs.另外，FC的存在使结合的肿瘤细胞容易受到体液免疫的攻击。总体而言，我们提出的HPV PSV递送的治疗性嵌合蛋白具有靶向肿瘤递送，肿瘤部位的特异性脂蛋白裂解以及通过现有的CTL免疫力和FC介导的杀伤性杀死的有效杀害的优点，代表了具有高转化价值的新型策略。拟议研究的成功实施将作为一种创新策略，可以应用于其他类型的晚期癌症的治疗。