Cellular Immunity Targeting Epithelial Ovarian Cancer

在美国，卵巢癌是妇科恶性肿瘤死亡的主要原因，估计每年造成14，000人死亡。尽管治疗方法有所改进，但晚期卵巢癌的5年生存率仍约为20- 30%。卵巢癌差异表达某些蛋白质，这些蛋白质通常只在胚胎发育期间存在于其他组织上。在这些癌胚蛋白中有WT 1蛋白、糖基化粘蛋白Ca 125（MUC 16）和HER 2/neu。最近，我们和其他人已经鉴定出来源于WT 1和MUC 16的潜在免疫原性肽，其结合常见的HLA等位基因HLA A0201。我们对免疫原性WT 1肽和候选MUC 16肽的初步研究表明，这些肽可用于敏化来自正常个体和卵巢癌患者的T细胞，其可在体外扩增。用WT 1和MUC 16致敏的T细胞响应于肽产生IFN γ和其他细胞因子，并特异性裂解原发性HLA A2+ WT 1+或MUC 16+白血病和选定的癌，包括体外卵巢癌细胞。在SCID小鼠、人肿瘤异种移植物模型中，我们发现过继转移的WT 1特异性HLA A2+ T细胞归巢并特异性诱导某些HLA A2+ WT 1+白血病和选定的癌包括卵巢癌的消退。在这项授权中，我们首先提出鉴定和表征WT-1和MUC 16的肽，它们对HLAA 2具有中度至高度的亲和力，然后对其他HLA等位基因具有中度至高度的亲和力。然后，我们将使用负载有WT 1、MUC 16或HER 2/neu肽或转导以表达这些蛋白的EBV BLCL，因为它们能够敏化和刺激能够在体外裂解原代卵巢癌细胞和建立的细胞系的肽特异性T细胞的扩增。我们还将探索一种新的十五肽方法来鉴定WT 1和MUC 16中的新抗原。然后，我们将确定这些肿瘤的特征，这些特征可以增强或抑制这些T细胞在体外的杀肿瘤活性。我们还将比较评估从患者和正常个体产生的T细胞群与这些T细胞群之间的差异。 肽，以确定这些T细胞的表型或功能特征的差异是否影响体外肿瘤细胞相互作用。第三，我们将测试这些肽特异性细胞的能力，回家，使用一种新的体内成像方法，并在SCID小鼠模型中诱导人卵巢癌异种移植物的消退。最后，我们建议进行WT 1或MUC 16肽特异性T细胞治疗晚期卵巢癌妇女的I/II期试验。这些研究可能会提供重要的新信息的潜力，离体产生的T细胞对这些免疫原性肽过继细胞治疗卵巢癌。

Ovarian carcinoma is the leading cause of death from gynecologic malignancies in the United States, responsible for an estimated 14,000 deaths per year. Despite improvements in therapy, 5-year survival for advanced ovarian cancer remains approximately 20-30%. Ovarian cancers differentially express certain proteins normally present on other tissues only during embryonal development. Among these onco fetal proteins are the WT1 protein, the glycosylated mucin Ca125 (MUC16) and HER2/neu. Recently, we and others have identified potentially immunogenic peptides derived from WT1 and MUC16 which bind the common HLA allele, HLA A0201. Our initial studies with immunogenic WT1 peptides and with candidate MUC16 peptides suggest that these peptides can be used to sensitize T cells from normal individuals and from ovarian cancer patients, which can be expanded in vitro. The T cells sensitized with WT1 and MUC16 generate IFNy and other cytokines in response to peptide and specifically lyse primary HLA A2+ WT1+ or MUC16+ leukemias and selected carcinomas, including ovarian cancer cells in vitro. In a SCID mouse, human tumor xenograft model, we have found that adoptively transferred WT1 specific HLA A2+ T cells home to and specifically induce regressions of certain HLA A2+ WT1+ leukemia and selected carcinomas including ovarian cancer. In this grant, we propose first, to identify and characterize peptides of WT-1 and MUC16 which have moderate to high affinity for HLA A2, and thereafter, other HLA alleles. We will then use EBV BLCL either loaded with WT1, MUC16 or HER2/neu peptides or transduced to express these proteins, for their capacity to sensitize and stimulate the expansion of peptide-specific T cells capable of lysing primary ovarian cancer cells and established cell lines in vitro. We will also explore a novel pentadecapeptide approach to identify novel antigens in WT1 and MUC16. We will then identify features of these tumors which can potentiate or inhibit the tumoricidal activity of these T cells in vitro. We will also comparatively evaluate T cell populations generated from patients and from normal individuals against these peptides to determine whether disparities in phenotypic or functional features of those T cells affect tumor cell interactions in vitro. Third, we will test these peptide-specific cells for their capacity to home to, using a novel in vivo imaging approach, and induce regressions of human ovarian carcinoma xenografts in a SCID mouse model. Lastly, we propose to conduct a phase I/II trial of WT1 or MUC16 peptide-specific T cells in the treatment of women with advanced ovarian carcinoma. These studies may provide important new information regarding the potential of T cells generated ex vivo against these immunogenic peptides for adoptive cell therapy of ovarian cancer.