Vitamin D Analogs as Adjuvants in Chemotherapy

描述（由申请人提供）： 该提案的总体目标是开发分化治疗，以补充人类髓系白血病的治疗方案。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 D）以低浓度给予培养中的白血病细胞，并研究基因表达和其他细胞变化的改变。类三角洲和1，25 D诱导单核细胞/巨噬细胞样分化，并且还将与目前用作食品防腐剂或添加剂或具有这种用途的潜力的无毒物质组合施用，以使类三角洲的分化活性最大化。这些分化剂的活性的进一步增强将通过加入细胞内信号传导途径的抗炎抑制剂来探索。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的研究。通过先前的观察提供了基本原理，即抗氧化剂提供了还原环境并诱导基因的表达，所述基因补充了类Deltanoid响应基因的分化诱导作用。深入了解分化控制将通过检查信号通路，特别注意MAPK通路和转录因子。这将通过添加药理学试剂、反义寡核苷酸、siRNA、转录因子诱饵和转染的质粒构建体来完成，以研究这些操作的分子结果，这将通过免疫印迹、定量RT-PCR、免疫沉淀和其他标准技术来确定。将通过测定表面标志物以及各种酶的活性和表达来监测分化细胞。在基础研究中获得的信息将用于指导新一代deltanoid和deltanoid与共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。公共卫生相关性：分化疗法依赖于激活现有的细胞程序，而不是有毒的药物来对抗恶性肿瘤，已经有效地治疗了一些癌症。我们建议将其开发为被称为骨髓性白血病的血液恶性肿瘤的治疗方法，虽然每年在美国造成约15，000人死亡，但不太可能获得商业支持以找到其治愈方法。因此，髓性白血病可以被认为是一种孤儿病，值得公共资源支持开发新的治疗方法。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop differentiation therapy to supplement the treatment regimens for human myeloid leukemia. We will focus on the identification of the most effective analogs (deltanoids) of the physiological form of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D) administered at low concentrations to leukemia cells in culture and study alterations in gene expression and other cellular changes. The deltanoids and 1,25D induce monocytic/macrophage-like differentiation and 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 of the deltanoids. Further enhancement of the activity of these differentiation agents will be explored by the addition of an anti-inflammatory inhibitor of an intracellular signaling pathway. Established lines of leukemia, as well as samples of leukemic cells freshly obtained from patients, will be used for studies of of cell differentiation. The rationale is provided by previous observation that antioxidants provide a reducing environment and induce expression of genes which complement the differentiation-inducing actions of deltanoid-responsive genes. Insight into differentiation control will be obtained by examination of signaling pathways with particular attention to MAPK pathways and to transcription factors. This will be accomplished by adding pharmacological agents, antisense oligonucleotides, siRNAs, transcription factor decoys, and transfected plasmid constructs to study the molecular consequences of these manipulations , which will be determined by immunoblotting, quantitative RT-PCR, immunoprecipitation, and other standard techniques. Differentiating cells will be monitored by determination of surface markers as well as the activity and the expression of various enzymes. 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. PUBLIC HEALTH RELEVANCE: Differentiation therapy, which depends on the activation of existing cellular programs rather than on toxic drugs to combat malignant tumors, is already effective as the treatment of some cancers. We propose to develop it as therapy for blood malignancy known as myeloid leukemia, which although kills approximately 15,000 people in USA every year, is unlikely to receive attention from commercial support for finding its cure. Thus, myeloid leukemia can be considered an orphan disease, and merits support from public sources for the development of novel therapy.