Genetic determinants associated with pemetrexed response and toxicity

摘要 培美曲塞是一种多靶点抗叶酸药，最近被批准用于非小细胞肺癌的二线治疗 癌症（NSCLC）。在临床可达到的浓度下，最显著的相互作用是与胸苷酸 叶酸合成酶（TYMS），尽管已知该药物靶向至少两种其他叶酸依赖性酶。目标 该提案的目的是确定影响培美塞相关毒性的其他遗传变异， 通过采用全基因组的综合方法来测试遗传学的假设， 多态性显著影响对培美曲塞相关毒性和反应的易感性。到 为此，淋巴母细胞样细胞系，来源于大型参考文献人类多态性研究中心（Centre d'EtudyduPolymorphismeHumain 将对谱系和HapMap（高加索人、约鲁巴人、亚洲人）样本进行培美曲塞细胞毒性表型分析 和凋亡。基因型信息是在公共领域，使我们能够应用家族遗传学 策略，包括连锁分析，以检查基因组中携带遗传变异的区域 负责对培美塞诱导的细胞毒性和细胞凋亡的易感性。此外，个人和 将使用国际HapMap比较培美塞诱导细胞毒性的种族间差异 来自非洲人、欧洲人和亚洲人后裔的样本。外显子表达阵列产生在我们的 实验室将被纳入，以确定遗传变异通过其对基线基因的影响而起作用。 与培美曲塞敏感性相关的表达。通过整个基因组识别的遗传变异 该方法将在单独的LCL样本集中通过定量PCR和siRNA进行验证。我们将 评估TYMS、GARFT、DHFR中的新SNP和已知候选变体，以确定基因型 与组间（NCCTG、CALGB、ECOG、SWOG、NCI-C）研究中的反应或毒性相关， 培美曲塞与厄洛替尼作为NSCLC二线治疗的比较。我们的长期目标是确定一个 “药物遗传学特征”，将预测患者的缓解机会降低或缓解机会增加 培美曲塞不良反应的风险。具体目标是：1。为了确定基因的贡献， 对培美曲塞的细胞毒性和凋亡作用的敏感性。2.检查基线和时间依赖性 培美曲塞治疗后的表达模式。3.通过可获得的数据识别SNP， 与培美曲塞细胞毒性和细胞系基因表达相关的国际HapMap项目 来自欧洲人（CEU），非洲人（YRI）和亚洲人（CHB，JPT）。4.为了确定一个共同的 TYMS、GARFT和DHFR中的多态性，以及在特定目标1- 3例与培美曲塞治疗的晚期NSCLC患者的疗效和/或毒性相关的多中心研究 审判

ABSTRACT Pemetrexed is a multitargeted antifolate recently approved for second line therapy of non-small cell lung cancer (NSCLC). The most significant interaction at clinically achievable concentrations is with thymidylate synthase (TYMS), although the drug is known to target at least two other folate dependent enzymes. The goal of this proposal is to identify additional genetic variants that influence pemetrexed-associated toxicity and response by employing a genome-wide, comprehensive approach to test the hypothesis that genetic polymorphisms significantly influence susceptibility to toxicities and response associated with pemetrexed. To this end, lymphoblastoid cell lines, derived from large reference Centre d' Etude du Polymorphisme Humain pedigrees and HapMap (Caucasian, Yoruban, Asian) samples, will be phenotyped for pemetrexed cytotoxicity and apoptosis. Genotypic information is available in the public domain enabling us to apply familial genetic strategies, including linkage analysis, to examine regions in the genome harboring the genetic variants responsible for susceptibility to pemetrexed-induced cytotoxicity and apoptosis. In addition, inter-individual and inter-ethnic variation in pemetrexed-induced cytotoxicity will be compared using the International HapMap samples from populations of African, European and Asian descent. Exon expression array generated in our laboratory will be incorporated to identify genetic variants acting through their effect on baseline gene expression that associate with sensitivity to pemetrexed. Genetic variants identified through this whole genome approach will be validated in a separate sample set of LCLs and by quantitative PCR and siRNA. We will evaluate both novel SNPs and known candidate variants in TYMS, GARFT, DHFR to determine genotypes associated with response or toxicity in an intergroup (NCCTG, CALGB, ECOG, SWOG, NCI-C) study of pemetrexed compared to erlotinib as second line therapy for NSCLC. Our long-term goal is to identify a "pharmacogenetic signature" that will predict patients with a decreased chance for response or an increased risk for adverse reactions to pemetrexed. Specific aims are: 1. To determine the genetic contribution of susceptibility to the cytotoxic and apoptotic effects of pemetrexed. 2. To examine baseline and time dependent patterns of expression following pemetrexed treatment. 3. Identify SNPs, through data available from the International HapMap project, associated with both pemetrexed cytotoxicity and gene expression in cell lines derived from Europeans (CEU), Africans (YRI) and Asians (CHB, JPT). 4. To determine if a common polymorphism in TYMS, GARFT, and DHFR, as well as candidate polymorphisms identified in specific aims 1- 3 correlate with response and/or toxicity in advanced NSCLC patients treated with pemetrexed in a multicenter trial.