Therapy of Melanoma with Bortezomib and Interferon-alpha

描述（由申请人提供）：用硼替佐米和干扰素- α免疫治疗黑色素瘤对细胞毒性治疗的耐药性是恶性黑色素瘤的标志。目前治疗转移性疾病的标准是单药达卡巴嗪，总有效率仅为15%。因此，必须探索具有新的作用机制的新药剂。Bortezomib （Velcade，原PS-341）是一种新型抗肿瘤化合物，是26S蛋白酶体的特异性和选择性抑制剂，26S蛋白酶体是细胞蛋白降解的泛素-蛋白酶体途径的核心成分。用硼替佐米治疗恶性细胞通过多种机制导致生长停滞和凋亡细胞死亡，包括细胞周期蛋白的改变和促生存信号的阻断。我们注意到硼替佐米诱导的黑色素瘤细胞凋亡在干扰素- α (IFN-a)存在下协同增强，干扰素- α是一种具有独特促凋亡作用的药物。进一步分析表明，细胞凋亡与bcl-2水平的降低和caspase蛋白的激活有关。使用小鼠恶性黑色素瘤模型，我们还证明，与单独使用任何一种药物相比，硼替佐米和IFN-a联合治疗后，荷瘤小鼠的存活率显著提高（p = 0.02）。我们也有初步的数据表明，用硼替佐米预处理外周血单个核细胞会导致响应IFN-a的Jak-STAT信号转导通路的延长激活。这是第一次研究蛋白酶体抑制剂与细胞因子联合抗肿瘤作用的实验。我们现在建议在转移性恶性黑色素瘤患者中进行一项由研究者发起的硼替佐米和IFN-a2b的临床试验。我们假设IFN-a会增强晚期恶性黑色素瘤患者硼替佐米诱导的肿瘤细胞凋亡。在治疗前后，肿瘤将被活检，以便进行相关的免疫组织化学染色。对患者免疫细胞中ifn - A诱导的信号通路的仔细分析也将使用一种新的流式细胞分析方法进行。这些研究将有助于确定由硼替佐米和IFN-a2b联合调节的特定凋亡和免疫刺激途径。

DESCRIPTION (provided by applicant): Immunotherapy of Melanoma with Bortezomib and Interferon-Alpha Resistance to cytotoxic therapy is a hallmark of malignant melanoma. The current standard of care for metastatic disease is single-agent dacarbazine which has an overall response rate of just 15%. New agents with new mechanisms of action must therefore be explored. Bortezomib (Velcade, formerly PS-341) is a novel anti-tumor compound that is a specific and selective inhibitor of the 26S proteasome, a central component of the ubiquitin-proteasome pathway for the degradation of cellular proteins. Treatment of malignant cells with bortezomib leads to growth arrest and apoptotic cell death via multiple mechanisms including altered turnover of cell cycle proteins and blockade of pro-survival signals. We noted that bortezomib-induced apoptosis of melanoma cells was synergistically enhanced in the presence of interferon-alpha (IFN-a), an agent that has unique pro-apoptotic effects of its own. Further analysis revealed that apoptosis was associated with reduced levels of bcl-2 and activation of caspase proteins. Using a murine model of malignant melanoma, we also demonstrated that the survival of tumor-bearing mice was significantly enhanced following treatment with the combination of bortezomib and IFN-a as compared to either agent alone (p = 0.02). We also have preliminary data to suggest that pre-treatment of peripheral blood mononuclear cells with bortezomib leads to prolonged activation of the Jak-STAT signal transduction pathway in response to IFN-a. These are the first experiments to examine the anti-tumor effects of a proteasome inhibitor when combined with a cytokine. We now propose to conduct an investigator-initiated clinical trial of bortezomib and IFN-a2b in patients with metastatic malignant melanoma. We hypothesize that IFN-a will enhance bortezomib-induced tumor cell apoptosis in patients with advanced malignant melanoma. Tumors will be biopsied before and after therapy in order that correlative immunohistochemical stains can be performed. A careful analysis of IFN-a-induced signaling pathways in patient immune cells will also be conducted using a novel flow cytometric assay. These studies will help to define the specific apoptotic and immunostimulatory pathways that are being modulated by the combination of bortezomib and IFN-a2b.