TRAIL Therapy for Rhabdomyosarcoma

横纹肌肉瘤的 TRAIL 疗法

• 批准号: 6862647
• 负责人: JANET A. HOUGHTON
• 金额: $ 24.9万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-09 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6862647
• 关键词: SCID mouse; actinomycin; apoptosis; clinical research; combination chemotherapy; complementary DNA; cycloheximide; cysteine endopeptidases; cytokine receptors; enzyme activity; human tissue; interferon gamma; neoplasm /cancer chemotherapy; nonhuman therapy evaluation; pediatric neoplasm /cancer; rhabdomyosarcoma; tumor necrosis factor alpha

The proposal will elucidate critical regulators of the TRAIL signaling pathway that determine the exquisite sensitivity of 4/7 pediatric rhabdomyosarcomas (RMS) to TRAIL-induced cytotoxicity and apoptosis at concentrations < 1 ng/ml. All cultured cell lines express high levels of receptor DR5, but not DR4 or the decoy receptors DcR1 or DcR2, and express FADD and procaspase-8, with the exception of Rh36. Expression of c-FLIP is high in TRAIL-sensitive and -resistant lines, thereby not correlating directly with TRAIL sensitivity. Expression of procaspases-8 and -10 is highest in TRAIL-sensitive RD, Rh1 and Rh30. TRAIL-sensitive Rh18 expresses procaspase-8 in the absence of procaspase-10 and c-FLIP, and procaspase-10 is not expressed in TRAIL-resistant Rh41. Based upon these cellular characteristics the following hypotheses will be tested: 1) Following ligation of TRAIL to DR5, a DISC is formed among DR5, FADD, procaspase-8 and c-FLIP in RMS cell lines resulting in type I (direct) or type II (mitochondria- and procaspase-10- dependent) cell death, and 2) TRAIL resistance in Rh41 is due to deficiency in caspase-10. The second focus is to elucidate agents that can sensitize TRAIL- resistant RMS to TRAIL, or that can elicit synergistic interactions with TRAIL. Combination of TRAIL with actinomycin-D or doxorubicin has demonstrated > additive effects in TRAIL-sensitive RMS lines, and with recombinant human interferon-gamma (IFN-gamma) has induced cytotoxic response in TRAIL-resistant HT29 human .colon carcinoma cells. We will therefore test the following hypotheses: 1) TRAIL- resistant Rh28 and Rh41 can be sensitized to TRAIL by cycloheximide, actinomycin-D or IFN-gamma, and 2) specific chemotherapeutic agents can be identified that elicit synergistic interactions with TRAIL in TRAIL-sensitive RMS cell lines. Thirdly, we will test the hypothesis that TRAIL induces antitumor responses in human RMS xenografts when administered alone, or in combination with chemotherapeutic agents based upon data derived in tissue culture. The long-term objectives of the proposal are to develop highly effective therapy for metastatic disease in pediatric RMS from: 1) understanding specific signaling pathways involved in the regulation of cell death and apoptosis, 2) the identification of new molecular targets, and 3) developing new therapeutic strategies based upon specific molecular characteristics.

该提案将阐明TRAIL信号通路的关键调节因子，这些调节因子决定了4/7儿童横纹肌肉瘤(RMS)对TRAIL诱导的细胞毒性和凋亡的敏感度。除Rh36外，所有培养的细胞株都表达高水平的DR5受体，但不表达DR4或诱骗受体DcR1或DcR2，并表达FADD和proaspase-8。C-flip在TRAIL敏感和抗性品系中高表达，因此与TRAIL敏感性不直接相关。在TRAIL敏感的RD、Rh1和Rh30中，原天冬氨酸酶-8和-10的表达最高。TRAIL敏感的Rh18在缺乏proaspase-10和c-flip的情况下表达proaspase-8，而在TRAIL敏感的Rh41中不表达proaspase-10。基于这些细胞特性，将检验以下假设：1)TRAIL与DR5连接后，在RMS细胞系中，DR5、FADD、proaspase-8和c-flip之间形成圆盘，导致I型(直接)或II型(依赖线粒体和Proaspase-10)细胞死亡；2)Rh41中的TRAIL抗性是由于caspase-10缺乏所致。第二个重点是阐明哪些药物可以使TRAIL耐药的RMS对TRAIL敏感，或者哪些药物可以与TRAIL产生协同作用。TRAIL与放线菌素-D或阿霉素联合应用在TRAIL敏感的RMS株中表现出相加效应，与重组人干扰素-γ(IFN-γ)在TRAIL耐药的HT29人结肠癌细胞中诱导细胞毒反应。因此，我们将检验以下假设：1)TRAIL耐药的Rh28和Rh41可以被放线菌亚胺、放线菌素-D或干扰素-γ增敏；2)在TRAIL敏感的RMS细胞系中，可以确定特定的化疗药物与TRAIL协同作用。第三，基于组织培养的数据，我们将测试TRAIL单独给药或与化疗药物联合使用时在人RMS异种移植中诱导抗肿瘤反应的假设。该提案的长期目标是从以下方面开发治疗儿童RMS转移疾病的高效方法：1)了解参与调节细胞死亡和凋亡的特定信号通路；2)识别新的分子靶点；3)基于特定的分子特征开发新的治疗策略。