Ovarian cancer using novel nanoparticle formulations

描述（由申请人提供）：卵巢癌是美国女性癌症相关死亡的第五大原因。据观察，即使在手术和化疗后，癌症也会在相对短的时间内复发。因此，免疫抑制策略可能作为控制卵巢癌复发或进展的替代方案。口服疫苗相对容易接种。然而，疫苗的抗原成分在胃肠道中的降解是一个主要问题。在这项研究中，我们将制定和评估口服卵巢癌纳米颗粒疫苗在小鼠中预防/延缓卵巢癌生长的有效性。疫苗的抗原材料将由ID 8鼠卵巢癌细胞制备，因为该细胞系在各种类似标志物方面与人卵巢癌细胞系具有良好的相关性，并提供了一种独特的模型来研究卵巢癌进展和具有完整免疫系统的小鼠中的治疗前试验。在该方案中，卵巢癌抗原、M细胞靶向配体、免疫刺激性细胞因子和toll样受体（TLR 9）配体如可诱导T细胞应答的CpG寡核苷酸，所述纳米颗粒由可生物降解和生物相容的聚合物基质组成，所述聚合物基质含有肠溶聚合物和缓释聚合物的混合物，以受控的方式释放抗原。这些疫苗纳米颗粒将通过口服途径给予患有卵巢肿瘤的雌性C57 BL/6小鼠。这些颗粒靶向肠中派伊尔集合淋巴结中存在的M细胞，其吸收包封的疫苗以通过将抗原呈递给树突细胞和巨噬细胞来产生免疫力。将获得血清样本，以测定抗原特异性IgG水平，从而评估全身免疫力。我们还将进行机制研究，以研究CD+ T细胞、CD 8 + T细胞、NK细胞和B细胞在口服疫苗诱导的抗肿瘤免疫中的作用。我们还将在a）包括M细胞诱导RANKL B）低剂量环磷酰胺或抗CD 25 Ab的Treg耗竭研究后评价疫苗微粒的有效性，以评价既存肿瘤模型中的免疫耐受性和c）T细胞过继转移。还将评估疫苗的长期记忆反应，因为这是成功疫苗的必要组成部分。最近，在纳米技术实验室的配方方法方面取得了重大进展，使我们能够在一个步骤中使用喷雾干燥方法生产纳米颗粒。从将疫苗制剂从实验室推进到临床的观点来看，这是一个主要优点，因为可以在没有进一步修改的情况下实现该过程的规模扩大。

DESCRIPTION (provided by applicant): Ovarian cancer is the fifth most leading cause of cancer related deaths in women in the US. It has been observed that the cancer relapses within relatively short periods of time even after the surgery and chemotherapy. Therefore, immunotherapeutic strategies may serve as an alternative to control the recurrence or progression of ovarian cancer. Oral vaccines are relatively easy to administer. However, the degradation of the antigenic component of the vaccine in the gastro-intestinal tract is a major problem. In this research study, we will formulate and evaluate the efficacy of oral ovarian cancer nanoparticulate vaccine in mice to prevent/retard the ovarian cancer growth. The antigenic material for the vaccine will be prepared from ID8 murine ovarian cancer cells as this cell line correlates well with human ovarian cancer cell lines in terms of various similar markers and provides a unique model to study ovarian cancer progression and pre-therapeutic trials in mice with intact immune systems. In this proposal, ovarian cancer antigens, M-cell targeting ligands, immuno-stimulatory cytokines and toll-like receptor (TLR9) ligands such as CpG oligonucleotide that can induce T-cell responses, will be encapsulated into nanoparticles made up of a biodegradable and biocompatible polymer matrix containing a mixture of an enteric polymer to prevent their degradation under acidic conditions in the stomach and also a sustained release polymer enabling the release of the antigen in a controlled manner. These vaccine nanoparticles will be administered to female C57BL/6 mice with ovarian tumors by the oral route. These particles are targeted to M-cells present in Peyer's patches in the intestine, which take up the encapsulated vaccine to generate immunity by presenting the antigen to dendritic cells and macrophages. Serum samples will be obtained to determine the antigen specific IgG levels to assess the systemic immunity. We will also carry out mechanistic studies to investigate the role of CD+ T-cells, CD8+ T-cells, NK cells, and B cells in anti-tumor immunity induced by the oral vaccines. We will also evaluate the efficacy of the vaccine microparticles after a) including M-cell inducing RANKL b) Treg depletion studies with low dose of cyclophosphamide or anti-CD25 Ab in order to evaluate immunotolerance in pre-existing tumor models and c) adoptive transfer of T-cells. Long-term memory responses of the vaccine will also be assessed since this is a necessary component of a successful vaccine. Recently, major advances have been made in the formulation methodology at the Nanotechnology Laboratory allowing us to produce nanoparticles using the spray drying methodology in a single step process. This is a major advantage from the standpoint of advancing the vaccine formulation from bench to clinic as scale-up of this process can be achieved with no further modifications.