MOLECULAR PROFILE OF MELANOMA AND NEUROBLASTOMA ANTIGENS

主要目标是确定实验方法， 降低自发转移中肿瘤细胞的转移率 神经外胚层肿瘤模型。需要检验的主要假设是 要么是旨在激活免疫的免疫疗法 干扰细胞外的效应细胞和/或操作 基质(ECM)被金属蛋白酶(MMPs)降解足以 充分干扰多步转移过程，减少肿瘤转移。 体内外肿瘤细胞侵袭转移率。 抗GD2单抗的初步临床结果令人鼓舞 (MAb)在神经母细胞瘤中提供了专注于设计和 对这种恶性肿瘤的新的免疫治疗方法的评价， 包括重组单抗与细胞因子IL-2、TNF-2的融合蛋白 α、肿瘤坏死因子-β、粒细胞集落刺激因子。这些模式的潜力将首先 在小鼠同基因自发性转移模型中进行试验 A/J小鼠和人类神经母细胞瘤模型 植入人免疫效应细胞的SCID小鼠的神经母细胞瘤。 我们将对杀戮的效果进行比较 转移性黑色素瘤细胞的体外和体内实验研究 限制性黑色素瘤特异性同种异体细胞毒性T细胞或 抗肿瘤单抗与单抗之间的重组双特异性构建 定向到CD3或FcGammaIII受体。试图减少 通过调控基质金属蛋白酶-1的表达对黑色素瘤细胞侵袭/转移潜能的影响 衍生的细胞外基质降解将包括：1)转染人黑色素瘤细胞 带有编码基质金属蛋白酶的天然组织抑制因子-2的cDNA，以及2) 单抗融合蛋白靶向黑色素瘤转移的实验研究 抑制肿瘤细胞体外侵袭的胶原酶原肽。 努力确定哪些MMP可以不转移或转移较差 更具侵袭性转移的黑色素瘤包括：1)选择性 反义RNA对单个基质金属蛋白酶基因产物的中和作用 转染人MMPs对肿瘤细胞单个MMPs的上调 编码它们的DNA。这些研究将通过确定 选择整合素作为基质金属蛋白酶表达的潜在调控元件 并选择那些可以调节转移不良的肿瘤的基因 更具侵略性地转移。预计将会有一些 我们的研究结果最终可能有助于改善 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.