Vitamin D Analogs as Adjuvants in Chemotherapy

描述（由申请人提供）： 急性和慢性髓性白血病是造成约15，000人死亡的原因， 美国每年这些疾病经常在人们生命的黄金时期折磨他们，并且用当前的疗法，所有人中基本上不到一半实现长期缓解。大多数治疗选择是强烈的和有毒的，需要抗肿瘤的多化疗，并且在某些情况下，治疗与同种异体造血细胞移植一样激进，伴随着风险。该提案的总体目标是开发分化疗法，以补充这组疾病的治疗方案。我们将专注于识别维生素D生理形式的最有效的类似物（deltanoids），1，25-二羟基维生素D3（1，25 D3）以低浓度给予培养中的白血病细胞，以研究基因表达的改变和细胞生理学的其他变化。三角洲类化合物和1，25 D3也将与目前用作食品防腐剂或添加剂的无毒物质或具有这种用途的可能性的无毒物质组合施用，以使三角洲类化合物的分化活性最大化。已建立的白血病细胞系以及从患者新鲜获得的白血病细胞样品将用于细胞分化研究。深入了解分化控制将通过检查信号通路，特别注意MAPK通路和细胞周期蛋白依赖性激酶的表达，以及它们的激活剂和抑制剂的细胞周期调控。这将通过添加药理学试剂、反义寡核苷酸、转录因子诱饵和转染的质粒构建体来研究这些操作的分子后果来实现，这将通过免疫印迹、北方分析、RT-PCR、免疫共沉淀和其他标准技术来确定。将通过测定表面标记物以及各种激酶的活性及其表达来监测分化细胞。基础研究中获得的信息将用于指导新一代deltanoid和deltanoid与辅助诱导剂的组合的开发，而对白血病细胞的体外转化研究将用于鉴定最适合于启动临床试验的髓性白血病亚组。

DESCRIPTION (provided by applicant): Acute and chronic myeloid leukemias are responsible for approximately 15,000 deaths in the USA every year. These diseases frequently afflict people in the prime of their lives, and substantially less than half of all achieve long-term remission with current therapies, Most treatment options are both intense and toxic, requiring anti-neoplastic poly-chemotherapy, and, in some cases, therapy as radical as allogeneic hematopoietic cell transplantation, with its attendant risks. The overall goal of this proposal is to develop differentiation therapy to supplement the treatment regimens for this group of diseases. We will focus on the identification of the most effective analogs (deltanoids) of the physiological form of vitamin D, 1,25- dihydroxyvitamin D3 (1,25D3) administered at low concentrations to leukemia cells in culture to study alterations in gene expression and other changes in cell physiology. The deltanoids and 1,25D3 will also be administered in combination with nontoxic substances currently used as food preservatives or additives, or with potential for such use, to maximize the differentiation activity of the deltanoids. Established lines of leukemia as well as samples of leukemic cells freshly obtained from patients will be used for studies of cell differentiation. Insight into differentiation control will be obtained by examination of signaling pathways with particular attention to MAPK pathways and into cell cycle regulation by the expression of cyclin-dependent kinases as well as their activators and inhibitors. This will be accomplished by adding pharmacological agents, antisense oligonucleotides, transcription factor decoys, and transfected plasmid constructs to study the molecular consequences of these manipulations, which will be determined by immunoblotting, Northern analysis, RT-PCR, coimmunoprecipitation, and other standard techniques. Differentiating cells will be monitored by determination of surface makers as well as the activity of various kinases as and as their expression. The information obtained in basic studies will be utilized to guide development of a new generation of deltanoids and deltanoid combinations with co-inducers, while translational studies on leukemic cells ex vivo will serve to identify subgroups of myeloid leukemias most suitable for the initiation of clinical trials.