MOLECULAR PROFILE OF MELANOMA AND NEUROBLASTOMA ANTIGENS

主要目标是确定实验方法， 降低自发转移的肿瘤细胞播散率 神经外胚层肿瘤的模型。 待检验的主要假设为 无论是旨在激活免疫系统的免疫疗法， 效应细胞和/或干扰细胞外 基质（ECM）的金属蛋白酶（MMP）降解足以 干扰多步转移过程，足以减少 体外和体内肿瘤细胞侵袭和转移率。 抗GD 2单克隆抗体的初步临床结果令人鼓舞 (mAb)在神经母细胞瘤中的应用提供了关注设计和 评价这种恶性肿瘤的新型免疫治疗方式， 包括与细胞因子IL-2，TNF-α， α，TNF-β，GM-CSF。 这些模式的潜力将首先 在鼠的同基因自发转移模型中进行测试 在A/J小鼠中的神经母细胞瘤，然后在这样的人模型中， 移植了人免疫效应细胞的scid小鼠中的神经母细胞瘤。 将比较各自的杀灭效果 转移性黑色素瘤细胞在体外和体内与HL-A I类- 限制性黑色素瘤特异性同种异体细胞毒性T细胞，或 抗肿瘤mAb和mAb之间的重组双特异性构建体 针对CD 3或Fc γ III受体。 试图降低 黑色素瘤细胞的侵袭/转移潜力，通过操纵MMP- 衍生的ECM降解将包括：1）转染黑素瘤细胞 具有编码MMP的天然组织抑制剂-2的cDNA，和2） 用含有抗肿瘤抗体的mAb-融合蛋白靶向黑色素瘤转移 在体外抑制肿瘤细胞侵袭的前胶原酶肽。 努力确定哪些MMPs可以使非转移性或转移性差 黑色素瘤更具侵袭性的转移包括：1）选择性 通过反义RNA中和单个MMP基因产物，和2） 通过转染肿瘤细胞上调单个MMP 编码它们的cDNA。 这些研究将通过确定 选择整合素作为MMP表达的潜在调节元件 并选择那些可以调节转移性差的肿瘤， 转移得更厉害 预计其中一些 我们的研究结果可能最终有助于改善 转移性癌症的治疗。

The major objective is to identify experimental approaches that will reduce the rate of tumor cell dissemination in spontaneous metastasis models of neuroectodermal tumors. Major hypotheses to be tested are that either immunotherapy modalities designed to activate immune effector cells and/or manipulations that interfere with extracellular matrix (ECM) degradation by metalloproteinases (MMPs) are adequate to disturb the multi-step metastasis process sufficiently to decrease the rate of tumor cell invasion and metastasis in vitro and in vivo. Encouraging initial clinical results with anti-GD2 monoclonal antibody (mAb) in neuroblastoma provided the rationale to focus on the design and evaluation of novel immunotherapy modalities for this malignancy, including recombinant mAb fusion proteins with cytokines IL-2, TNF- alpha, TNF-beta, GM-CSF. The potential of these modalities will first be tested in a syngeneic spontaneous metastasis model for murine neuroblastoma in A/J mice and then in such a model for human neuroblastoma in scid mice engrafted with human immune effector cells. Comparisons will be made between the respective efficacy of killing metastatic melanoma cells in vitro and in vivo with either HL-A Class I- restricted, melanoma-specific allogeneic cytotoxic T cells or recombinant bispecific constructs between anti-tumor mAbs and mAbs directed to either CD3 or FcgammaIII receptor. Attempts to decrease the invasive/metastatic potential of melanoma cells by manipulating MMP- derived ECM degradation will include: 1) transfection of melanoma cells with cDNA encoding the natural tissue inhibitor-2 of MMP, and 2) targeting of melanoma metastasis with a mAb-fusion protein containing a procollagenase peptide that inhibits tumor cell invasion in vitro. Efforts to establish which MMPs can render non- or poorly metastatic melanoma more aggressively metastatic include: 1) selective neutralization of individual MMP gene products by anti-sense RNA, and 2) up-regulation of individual MMPs by transfection of tumor cells with cDNAs encoding them. These studies will be rounded out by identifying selected integrins as potential regulatory elements of MMP expression and selecting those that can modulate poorly metastatic tumors to metastasize more aggressively. It is anticipated that some of the results from our studies may ultimately contribute to improve the treatment of metastatic cancer.