Preclinical Testing of Galectin-3C for Multiple Myeloma

描述(申请人提供)：总体目标是开发一种新的人类蛋白质Galectin-3C，作为多发性骨髓瘤(MM)的治疗方法，可以与其他药物一起使用，以提高对治疗有反应的患者的百分比和存活率。尽管引入了新的治疗方案，但MM很少被治愈，迫切需要更有效的治疗方法。10年生存率最近有所上升，但仍只有17.4%。目前治疗多发性骨髓瘤的药物包括地塞米松、沙利度胺和硼替佐米，被认为部分是通过诱导细胞凋亡发挥作用的。Galectin-3C含有Galectin-3的碳水化合物识别结构域，但缺乏参与多聚化的N-末端结构域。Galectin-3C通过与相同的碳水化合物配体结合而不是多聚化，成为Galectin-3的主要负抑制因子。此前，我们在人类乳腺癌的小鼠模型中显示Galectin-3C减少了肿瘤的生长和转移。我们的初步数据表明，Galectin-3C可以诱导或促进地塞米松等抗癌药物介导的多发性骨髓瘤和其他类型癌细胞的凋亡。第一阶段研究的具体目标如下：目的#1.确定Galectin-3C抑制患者和人骨髓瘤细胞系生长的能力，并为其作用机制和潜在的生物标志物提供证据。GAL3C对多发性骨髓瘤细胞系和患者细胞增殖和凋亡的影响将被评估。CD138阴性的MM细胞被认为具有肿瘤干细胞的启动和维持特性，将被分离出来，并用Galectin-3C进行测试。将在粘连于永生化的人类基质细胞上的MM细胞上测试其疗效，条件是产生保护MM细胞免受凋亡的“细胞外基质耐药”。检测Galectin-3和Bcl2家族成员Mcl-1、Bax、Bad和Bcl2的表达。目的#2.评估Galectin-3C促进患者和人类多发性骨髓瘤细胞对地塞米松、沙利度胺和硼替佐米的反应的能力。将确定Galectin-3C和每种药物对细胞系的增殖，并将用患者细胞测试最佳组合。目的#3.为了确定Galectin-3C单独和联合治疗人MM的NOD/SCID小鼠模型的效果，将根据AIM#2的结果选择一种药物，并与Galectin-3C一起试验在播散性和皮下NOD/SCID小鼠MM模型中。在播散性模型中，Galectin-3C将通过肌肉注射和微量泵持续静脉注射的方式给予。产生最佳反应的方法将用于皮下模型的给药。第二阶段将专注于完成临床前开发，以启动第一阶段的人类临床试验。公共卫生相关性：尽管引入了新的治疗方案，但多发性骨髓瘤很少能治愈，需要更有效的治疗方法。预计2007年美国多发性骨髓瘤新增病例将超过19,900例，死亡10,700例。虽然10年存活率最近有所上升，但只有17.4%。这项研究的总体目标是开发一种新的人类蛋白Galectin-3C，作为治疗多发性骨髓瘤的药物，它可以与其他药物一起使用，以促进细胞凋亡的诱导，提高对治疗有反应的患者的比例和存活率。

DESCRIPTION (provided by applicant): The overall goal is development of a novel human protein, Galectin-3C, as a treatment for multiple myeloma (MM) that can be given with other agents to increase the percentage of patients who are responsive to therapy, and the survival rate. MM is rarely curable despite the introduction of new therapeutic regimens and more efficacious treatments are sorely needed. The 10-year survival rate has recently increased, but is still just 17.4%. Current therapeutic agents for MM including dexamethasone, thalidomide, and bortezomib are thought to act partially through induction of apoptosis. Galectin-3C contains the carbohydrate recognition domain of Galectin-3 but lacks its N-terminal domain that functions in multimerization. Galectin-3C acts as a dominant negative inhibitor of Galectin-3 by binding to the same carbohydrate ligands but not multimerizing. Previously, we showed that Galectin-3C reduced the growth and metastasis of tumors in a mouse model of human breast cancer. Our preliminary data indicate that Galectin-3C can induce or facilitate apoptosis mediated by anticancer agents such as dexamethasone in multiple myeloma and other types of cancer cells. The Specific Aims of the Phase I research are as follows: Aim #1. To determine the ability of Galectin-3C to inhibit growth of MM cells from patients and human cell lines and to provide evidence of the mechanism of action and potential biomarkers. The effect of GAL3C on proliferation and apoptosis of MM cell lines, and patient cells will be assessed. The CD138negative MM cells that are postulated to possess the tumor-initiating and maintaining characteristics of cancer stem cells will be isolated and tested with Galectin-3C. Efficacy will be tested on MM cells adherent to immortalized human stromal cells in conditions that give rise to "extracellular matrix drug resistance" that protects MM cells from apoptosis. We will determine the expression of Galectin-3 and Bcl-2 family members, Mcl-1, Bax, Bad, and Bcl-2. Aim #2. To assess the ability of Galectin-3C to facilitate response to dexamethasone, thalidomide, and bortezomib in MM cells from patients and human cell lines. Proliferation of cell lines with Galectin- 3C and each agent will be determined, and the best combination will be tested with patient cells. Aim #3. To determine the effect of Galectin-3C alone and in combination therapy of NOD/SCID mouse models of disseminated and subcutaneous human MM. One agent will be selected based on results in Aim #2 and tested with Galectin-3C in disseminated and subcutaneous NOD/SCID mouse models of MM. Galectin-3C will be administered by intramuscular injection and continuous intravenous delivery by mini-pump in the disseminated model. The methodology producing the best response will be used for administration in the subcutaneous model. Phase II would focus on completing preclinical development towards initiation of a Phase 1 human clinical trial. PUBLIC HEALTH RELEVANCE: Multiple myeloma is rarely curable despite the introduction of new therapeutic regimens and more efficacious treatments are needed. More than 19,900 new cases and 10,700 deaths from multiple myeloma are expected in the U.S. in 2007. Although the 10-year survival rate has recently increased it is only 17.4%. The overall goal of this research is the development of a novel human protein, Galectin-3C, as a treatment for multiple myeloma that can be given with other agents to facilitate the induction of apoptosis, to increase the percentage of patients who are responsive to therapy, and the survival rate.