Development Therapy for Metastatic Colorectal Cancer

描述(申请人提供)：Fura/LV和重组人干扰素-γ在人结肠癌细胞中的相互作用是协同的，依赖于Fura/LV诱导的DNA损伤，Fas信号通路，而不依赖于P53。初步研究还表明，除了Fas外，还存在涉及caspase、IAP蛋白Survivin和p21Cip1的相互作用部位。Fura/LV联合干扰素-伽马的I期试验在经过大量治疗和未经治疗的患者中显示了反应。第二阶段试验现在将在以前未治疗或以前治疗的转移性结直肠癌患者中进行，总体目标是阐明影响Fura/LV、Fas和干扰素-γ信号转导的基因表达的预后意义，以及这些途径之间在诱导细胞凋亡、细胞毒性和反应方面的协同作用。 我们将通过组织芯片(蛋白质)或实时定量聚合酶链式反应(MRNA)来阐明已知的决定结肠癌细胞对Fura/LV敏感性的基因(TS、DPD、TP、COX-2)或影响Fas或干扰素-γ信号通路(Bcl2、Bax、P53、p21Cip1、Fas、caspase-1、-3、-8、Survivin、STAT1)的预后意义。在Fura/LV/干扰素-伽马治疗前(样本2)和治疗后4天(样本3)以及同一患者的原发肿瘤(样本1)中，将确定转移瘤中的基因表达。转移瘤中基因表达的变化在预测疾病进展时间和反应方面的意义，以及Fas表达变化与CD15+和CD56+细胞表达升高的相关性，以及毒性和干扰素-γ的药代动力学，将被确定。 使用寡核苷酸微阵列分析(Affymetrix)，假设我们将阐明额外的基因，这些基因是预测疾病进展时间、8周应答、总体应答、固有耐药或Fura/LV/干扰素-伽马治疗后获得性耐药的独立分子决定因素。 预计拟议的研究将确定以下关键靶点：1)死亡受体、Fura/LV和干扰素-γ信号通路之间的趋同；2)确定Fura/LV和干扰素-γ在转移性结直肠癌中的协同作用；3)预测Fura/LV/干扰素-γ治疗的应答、内在耐药或获得性耐药；4)在合理的临床前设计的基础上，进一步了解Fura/LV/干扰素-γ治疗的新靶点，并开发出治疗干预的新靶点，开发高效的治疗方法。

DESCRIPTION (provided by applicant): The interaction between FUra/LV and recombinant human IFN-gamma is synergistic in human colon carcinoma cells, dependent on Fura/LV-induced DNA damage, the Fas signaling pathway, and independent of p53. Preliminary studies also demonstrate sites of interaction in addition to Fas that involve caspases, the IAP protein survivin, and p21Cip1. A Phase I trial of FUra/LV combined with IFN-gamma has demonstrated responses in heavily pretreated and untreated patients. A Phase II trial will now be conducted in patients with previously untreated or previously treated metastatic colorectal cancer, with the overall goal to elucidate the prognostic significance of expression of genes that influence FUra/LV, Fas and IFN-gamma signaling and the synergistic interaction between these pathways in the induction of apoptosis, cellular cytotoxicity and response. We will elucidate, by tissue microarray (protein) or Real Time PCR (mRNA), the prognostic significance of genes known to determine sensitivity to FUra/LV (TS,DPD, TP, Cox-2) or influence the Fas or IFN-gamma signaling pathways (Bcl-2, Bax, p53, p21Cip1, Fas, caspases -1, -3, -8, survivin, STAT1) in colon carcinoma cells. Gene expression will be determined in metastases both before (sample 2) and at 4 days after (sample 3) FUra/LV/IFN-gamma treatment, and in primary tumors (sample 1) from the same patients. The significance of changes in gene expression in metastases in predicting time to disease progression, response, and how changes in Fas expression correlate with elevated expression in CD15+ and CD56+ cells, as well as toxicity and IFN-gamma pharmacokinetics, will be determined. Using oligonucleotide microarray analyses (Affymetrix), the hypothesis is that we will elucidate additional genes that are independent molecular determinants for predicting time to disease progression, response at 8 weeks, overall response, intrinsic drug resistance or acquired drug resistance following FUra/LV/IFN-gamma therapy. It is anticipated that the proposed studies will identify critical targets that 1) are convergent between death receptor, FUra/LV and IFN-gamma signaling pathways, 2) determine synergistic interactions between FUra/LV and IFN-gamma in metastatic colorectal carcinoma, 3) predict for response, intrinsic drug resistance or acquired drug resistance in FUra/LV/IFN-gamma therapy, and 4) will further our knowledge in the identification of new targets for therapeutic intervention, and in the development of highly effective therapeutic approaches in the treatment of this disease, based upon rational preclinical design.