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: 8840196
• 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/8840196
• 关键词: Adverse effects; Affinity; Antibodies; Bilateral; Binding; 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; Protein Binding; 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; female reproductive system; gene therapy; immunogenic; improved; in vivo; influenzavirus; innovation; killings; macrophage; mortality; neoplastic cell; neutralizing antibody; novel; novel strategies; 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）：晚期卵巢癌是美国女性生殖系统癌症患者中死亡率最高的。现有的卵巢癌治疗方法，例如手术和化疗，具有显着的副作用，并且很少能对局部晚期或转移性疾病患者产生长期治愈。缺乏治愈性治疗方法和高比例的诊断为晚期卵巢癌的患者强调迫切需要开发创新的靶向疗法来控制晚期卵巢癌。在当前的提案中，我们的目标是通过使用人乳头瘤病毒（HPV）假病毒粒子（psV）将治疗性DNA构建体传递到卵巢肿瘤细胞，从而通过细胞介导和控制来控制感染和未感染的卵巢肿瘤细胞，从而克服当前疗法的主要局限性。体液免疫反应。最近，我们证明HPV psV能够在体内优先感染卵巢肿瘤。此外，我们还发现，含有肿瘤归巢分子 NKG2D 的嵌合蛋白与免疫球蛋白 G (IgG2a) 的 Fc 片段融合，能够包裹肿瘤细胞并产生抗肿瘤作用（参见初步数据）。肿瘤归巢分子 NKG2D 以高亲和力与 NKG2D 配体结合，与正常细胞相比，NKG2D 配体在大多数卵巢肿瘤细胞中过度表达。 NKG2D 与 Fc 片段融合 (NKG2D-Fc) 使具有 Fc 受体的效应细胞（例如巨噬细胞和自然杀伤细胞）与肿瘤细胞结合，从而诱导抗体依赖性和补体依赖性细胞毒性。我们建议将 NKG2D-Fc 与流感病毒的 MHC I 类限制性免疫原性 CTL 表位进一步融合，并由弗林蛋白酶切割位点分隔。这将通过利用预先存在的细胞毒性 T 淋巴细胞 (CTL) 来对抗常见的外来病毒表位（绝大多数个体中都存在这种表位），从而绕过免疫耐受。我们之前已经证明，腹膜内注射含有模型免疫原性 CTL 抗原肽 OVA 的类似嵌合蛋白（两侧带有弗林蛋白酶切割位点）导致嵌合蛋白的靶向递送以及 OVA CTL 肽在 MHC I 类分子上的呈递。荷瘤小鼠体内的肿瘤细胞。因此，我们从嵌合蛋白（NKG2D-Fc-RM）中推断，NKG2D部分将使嵌合蛋白特异性结合HPV psV感染和未感染的卵巢癌细胞，其中免疫原性CTL表位将通过弗林蛋白酶切割而释放出来。包被肿瘤细胞以供预先存在的 CTL 识别。此外，Fc的存在使得结合的肿瘤细胞容易受到体液免疫的攻击。总体而言，我们提出的由 HPV psV 递送的治疗性嵌合蛋白具有靶向肿瘤递送、肿瘤位点特异性弗林蛋白酶切割以及通过现有 CTL 免疫和 Fc 介导的杀伤作用实现有效的肿瘤靶向杀伤的优点，代表了一种具有高转化率的新策略。价值。拟议研究的成功实施将作为一种创新策略，可应用于其他类型晚期癌症的治疗。