Preclinical Testing of Galectin-3C for Multiple Myeloma

描述（由申请人提供）：总体目标是开发一种新型人类蛋白质半乳糖凝集素-3C，作为多发性骨髓瘤（MM）的治疗药物，可与其他药物联合使用，以增加对治疗有反应的患者百分比和生存率。尽管引入了新的治疗方案，但MM很少是可治愈的，并且迫切需要更有效的治疗。10年生存率最近有所上升，但仍然只有17.4%。目前用于MM的治疗剂包括地塞米松、沙利度胺和硼替佐米被认为部分通过诱导细胞凋亡起作用。半乳糖凝集素-3C含有半乳糖凝集素-3的碳水化合物识别结构域，但缺乏其在多聚化中起作用的N-末端结构域。半乳糖凝集素-3C通过与相同的碳水化合物配体结合但不多聚化而作为半乳糖凝集素-3的显性负性抑制剂。之前，我们在人乳腺癌小鼠模型中证明了半乳糖凝集素-3C减少了肿瘤的生长和转移。我们的初步数据表明，半乳糖凝集素-3C可以诱导或促进多发性骨髓瘤和其他类型的癌细胞中由抗癌药物如地塞米松介导的细胞凋亡。第一阶段研究的具体目标如下：目标1。确定半乳糖凝集素-3C抑制患者和人细胞系MM细胞生长的能力，并提供作用机制和潜在生物标志物的证据。将评估GAL 3C对MM细胞系和患者细胞的增殖和凋亡的影响。将分离假定具有癌症干细胞的肿瘤起始和维持特征的CD 138阴性MM细胞，并使用半乳糖凝集素-3C进行检测。将在产生“细胞外基质耐药性”（保护MM细胞免于凋亡）的条件下，对粘附于永生化人基质细胞的MM细胞进行疗效检测。我们将确定Galectin-3和Bcl-2家族成员Mcl-1、Bax、Bad和Bcl-2的表达。目标2。评估半乳糖凝集素-3C促进患者MM细胞和人细胞系对地塞米松、沙利度胺和硼替佐米反应的能力。将测定具有半乳糖凝集素-3C和每种药剂的细胞系的增殖，并且将用患者细胞测试最佳组合。目标3。为了确定半乳糖凝集素-3C单独和联合治疗播散性和皮下人MM的NOD/SCID小鼠模型的效果。将基于目标#2中的结果选择一种药剂，并在MM的播散性和皮下NOD/SCID小鼠模型中用半乳糖凝集素-3C进行测试。在播散性模型中，半乳糖凝集素-3C将通过肌内注射和通过微型泵连续静脉内递送进行给药。皮下给药模型将采用产生最佳缓解的方法。第二阶段的重点是完成临床前开发，以启动第一阶段的人体临床试验。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年美国将有19,900多例多发性骨髓瘤新发病例和10,700例死亡病例。虽然10年生存率最近有所增加，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质Galectin-3C，作为多发性骨髓瘤的治疗方法，可以与其他药物一起使用，以促进诱导细胞凋亡，增加对治疗有反应的患者的百分比和生存率。

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.