Ovarian cancer using novel nanoparticle formulations

描述(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，即使在手术和化疗之后，癌症也会在相对较短的时间内复发。因此，免疫治疗策略可能成为控制卵巢癌复发或进展的另一种选择。口服疫苗相对容易接种。然而，疫苗的抗原组分在胃肠道中的降解是一个主要问题。在本研究中，我们将制定和评价口服卵巢癌纳米颗粒疫苗在小鼠体内预防/延缓卵巢癌生长的效果。疫苗的抗原性材料将从ID8小鼠卵巢癌细胞中制备，因为该细胞系在各种相似标记方面与人卵巢癌细胞系具有很好的相关性，并为研究卵巢癌进展和免疫系统完整的小鼠的治疗前试验提供了独特的模型。在这项建议中，卵巢癌抗原、M细胞靶向配体、免疫刺激细胞因子和Toll样受体(Toll-like Receptor，TLR9)配体，如能够诱导T细胞反应的CpG寡核苷酸，将被包裹在纳米颗粒中，该纳米颗粒由可生物降解和生物相容的聚合物基质组成，其中含有防止它们在胃内酸性条件下降解的肠道聚合物的混合物，以及能够以受控方式释放抗原的缓释聚合物。这些疫苗纳米粒将通过口服途径注射给带有卵巢肿瘤的雌性C57BL/6小鼠。这些颗粒针对存在于肠道Peyer氏斑中的M细胞，这些细胞接受封装的疫苗，通过将抗原递送给树突状细胞和巨噬细胞来产生免疫。采集血清标本，测定抗原特异性免疫球蛋白水平，以评估系统免疫功能。我们还将开展机制研究，以探讨CD+T细胞、CD8+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。我们还将评估疫苗微粒的有效性，包括a)M细胞诱导RANKL，b)小剂量环磷酰胺或抗CD25抗体的Treg耗竭研究，以评估在现有肿瘤模型中的免疫耐受性，以及c)过继转移T细胞。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，纳米技术实验室在配方方法方面取得了重大进展，使我们能够在一步过程中使用喷雾干燥方法生产纳米颗粒。从将疫苗配方从试验台推进到临床的观点来看，这是一个重大优势，因为这一过程无需进一步修改就可以实现扩大规模。

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.