TRAIL Therapy for Rhabdomyosarcoma

横纹肌肉瘤的 TRAIL 疗法

• 批准号: 6331862
• 负责人: JANET A. HOUGHTON
• 金额: $ 24.73万
• 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 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6331862
• 关键词: SCID mouse; apoptosis; 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信号通路的关键调控因子，这些调控因子决定了4/7小儿横纹肌肉瘤（RMS）对TRAIL诱导的浓度< 1 ng/ml的细胞毒性和凋亡的敏感性。除Rh36外，所有培养的细胞系均表达高水平的DR5受体，但不表达DR4或诱饵受体DcR1或DcR2，并表达FADD和procaspase-8。c-FLIP在TRAIL敏感系和TRAIL抗性系中表达较高，因此与TRAIL敏感性无直接关系。procaspas8和-10在trail敏感的RD、Rh1和Rh30中表达最高。trail敏感的Rh18在缺乏procaspase-10和c-FLIP的情况下表达procaspase-8，而procaspase-10在trail抗性的Rh41中不表达。基于这些细胞特征，我们将验证以下假设：1)将TRAIL与DR5连接后，RMS细胞系中DR5、FADD、procaspase-8和c-FLIP之间形成DISC，导致I型（直接）或II型（依赖线粒体和procaspase-10）细胞死亡；2)Rh41中TRAIL的抗性是由于caspase-10的缺乏。第二个重点是阐明可以使TRAIL抗性RMS对TRAIL敏感的药物，或者可以引发与TRAIL的协同相互作用的药物。TRAIL与放线菌素- d或阿霉素联合在TRAIL敏感的RMS细胞系中显示出b>的加性效应，并且与重组人干扰素- γ (ifn - γ)联合在TRAIL耐药的HT29人中诱导了细胞毒性反应。结肠癌细胞。因此，我们将测试以下假设：1)TRAIL耐药的Rh28和Rh41可以通过环己烷酰亚胺、放线菌素- d或ifn - γ致敏TRAIL， 2)可以确定特异性化疗药物，在TRAIL敏感的RMS细胞系中引发与TRAIL的协同相互作用。第三，我们将根据组织培养的数据，验证TRAIL在单独给药或与化疗药物联合使用时诱导人类RMS异种移植物抗肿瘤反应的假设。该计划的长期目标是开发针对儿童RMS转移性疾病的高效治疗方法，包括：1)了解参与细胞死亡和凋亡调节的特定信号通路，2)确定新的分子靶点，以及3)基于特定分子特征开发新的治疗策略。