Clinical Pharmacogenetics

临床药物遗传学

• 批准号: 8349079
• 负责人: William Douglas Figg
• 金额: $ 59.52万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349079
• 关键词: ABCB1 gene; ABCG2 gene; Affect; Aftercare; Alleles; Anti-Retroviral Agents; Antiepileptic Agents; Antineoplastic Agents; BAY 54-9085; Biological Assay; CYP1B1 gene; CYP2C19 gene; CYP2D6 gene; CYP3A4 gene; CYP3A5 gene; CYP4B1 gene; Candidate Disease Gene; Cell Line; Cells; Chemotherapy-Oncologic Procedure; Cisplatin; Clinical; Clinical Trials; Codon Nucleotides; Complement; Complementary DNA; DNA; DNA Repair; Data; Development; Disease; Drug Delivery Systems; Drug Kinetics; Drug Transport; ERCC1 gene; Enrollment; Enzymes; Excision Repair; Exhibits; Favorable Clinical Outcome; Frequencies; Gene Expression; Genes; Genetic; Genetic Markers; Genetic Polymorphism; Genetic Transcription; Genetic Variation; Genotype; Goals; Hand; Haplotypes; Hypertension; Immunosuppressive Agents; Incidence; Individual; Inherited; Knowledge; Laboratories; Link; Malignant Neoplasms; Malignant neoplasm of prostate; Molecular; NAT2 gene; Outcome; P-Glycoprotein; PPAR delta; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pharmacogenetics; Pharmacology; Phase II Clinical Trials; Phenotype; Platinum; Play; Process; Radiation; Radiation Genetics; Radiation therapy; Randomized; Reaction; Recurrence; Research; Resistance; Risk; Risk Factors; Role; Scanning; Silent Mutation; Skin; Solid Neoplasm; Subgroup; Testing; Thalidomide; Therapeutic; Therapeutic Index; Time; Toxic effect; Translations; UGT1A1 gene; Variant; Vascular Endothelial Growth Factor Receptor-2; Work; XRCC1 gene; base; bevacizumab; biological systems; cancer radiation therapy; cancer therapy; chemotherapy; docetaxel; drug development; drug metabolism; experience; foot; genetic association; genetic variant; interest; mRNA Expression; molecular marker; novel; protein expression; research study; response; therapeutic protein; trend

Our laboratory has a strong interest in pharmacogenetics. We have been active in studying how germline genetic variants can alter pharmacokinetics, response, and toxicity of various anticancer agents, thereby contributing to inter-individual variation in clinical outcomes in therapies with an already narrow therapeutic window. We have established a molecular link between these polymorphisms and their phenotype as it relates to drug treatment. Most of our work has been focused on genetic variations in drug metabolism and transporting candidate genes such as ABCB1 (P-glycoprotein, MDR1), ABCG2 (BCRP), SLCO1B3 (OATP1B3, OATP8), CYP3A4, CYP3A5, CYP1B1, CYP2C19, CYP2D6, UGT1A1, UGT1A9 and several others. We are also interested in non-candidate gene approaches where large numbers of polymorphisms are explored to establish a relationship with clinical outcome, and experiments are conducted to validate potential causative alleles resulting from exploratory scanning. We have worked with Affymetrix to beta-test the DMET chip that contains 1,256 genetic variations in 170 drug disposition genes, and are currently establishing a clinical trial where patients treated at the NCI will be genotyped with the DMET chip to explore potential links between these genes and various treatments of several cancers. We are currently making progress in validating the results from the initial DMET chip experiments. While many of these studies have been conducted in order to explain some of the genetic influence on pharmacokinetic variability, we also have a strong interest in clarifying genetic markers of pharmacodynamics and therapeutic outcome of several major anticancer agents since this field has been rather poorly studied. DMET: Docetaxel exhibits significant inter-individual variation in its pharmacokinetic and toxicity profile. Thalidomide is an active anticancer agent and also exhibits wide pharmacologic variation. Past pharmacogenetic research has not explained this variation. Patients with prostate cancer enrolled in a randomized phase II trial using docetaxel and thalidomide versus docetaxel alone were genotyped using the Affymetrix DMET 1.0 platform, which tests for 1,256 genetic variations in 170 drug disposition genes. Genetic polymorphisms were analyzed for associations with clinical response and toxicity. Ten SNPs in three genes were potentially associated with response to therapy: PPARdelta, SULT1C2, and CHST3. Eleven SNPs in eight genes were associated with toxicities to treatment: SPG7, CHST3, CYP2D6, NAT2, ABCC6, ATP7A, CYP4B1, and SLC10A2. Genotyping results between DMET and direct sequencing showed greater than 96% concordance. These findings highlight the role that non-CYP450 metabolizing enzymes and transporters may play in the pharmacology of docetaxel and thalidomide. VEGFR2 polymorphism & bevacizumab/sorafenib: Hypertension (HT) and hand-foot skin reactions (HFSR) may be related to the activity of bevacizumab and sorafenib. We hypothesized that these toxicities would correspond to favorable outcome in these drugs, that HT and HFSR would coincide, and that VEGFR2 genotypic variation would be related to toxicity and clinical outcomes. Toxicity incidence and VEGFR2 H472Q and V297I status were compared to clinical outcomes. Individuals experiencing HT had longer PFS following bevacizumab therapy than those without this toxicity in trials utilizing bevacizumab in patients with prostate cancer (31.5 vs 14.9mo, n=60, P=0.0009), and bevacizumab and sorafenib in patients with solid tumors (11.9 vs 3.7 mo, n=27, P=0.052). HT was also linked to a >5-fold OS benefit after sorafenib and bevacizumab cotherapy (5.7 versus 29.0 mo, P=0.0068). HFSR was a marker for prolonged PFS during sorafenib therapy (6.1 vs 3.7 mo, n=113, P=0.0003). HT was a risk factor for HFSR in patients treated with bevacizumab and/or sorafenib. Carriers of variant alleles at VEGFR2 H472Q experienced greater risk of developing HT and HFSR. This study suggests that HT and HFSR may be markers for favorable clinical outcome, HT development may be a marker for HFSR, and VEGFR2 alleles may be related to the development of toxicities during therapy with bevacizumab and/or sorafenib. XRCC1 polymorphism & radiation: Genetic polymorphisms in XRCC1 associated with radiation therapy in prostate cancer: Radiation therapy is a potentially curative, important treatment option in localized prostate cancer. However, at 8 years after radiation therapy, even in the best risk subset of patients, approximately 10% of patients will experience clinical disease recurrence. Herein, we investigated five molecular markers of DNA repair. 513 patients with castrate-resistant prostate cancer (CRPC), including 284 patients who received radiotherapy, 229 patients without radiotherapy, and 152 healthy individuals were genotyped for 5 polymorphisms in DNA excision repair genes: ERCC1 N118N (500C>T), XPD K751Q (2282A>C), XRCC1 R194W (685C>T), XRCC1 R399Q (1301G>A) and PARP1 V762A (2446T>C). The polymorphisms evaluated did not show differences between the patient group and the healthy controls, nor did they show a trend toward an association with survival. However, in the radiation treated subgroup, the median survival time was associated with the XRCC1 haplotype. The median survival time was 11.75 years for patients with the R399Q AA/R194W CC haplotype, 12.17 years for patients with the R399Q AG/R194W CC haplotype, 66.65 years for patients with the R399Q AG/R194W CT haplotype, and 6.21 years for patients with the R399Q GG/R194W CT haplotype (p = 0.034). This association was not found when all patients were investigated. We conclude that the genetic polymorphisms in XRCC1 may affect the outcome in patients who received radiotherapy for localized prostate cancer. ERCC1 polymorphism & platinum-base chemotherapy: Genetic polymorphisms in ERCC1 are thought to contribute to altered sensitivity to platinum-based chemotherapy. Although ERCC1 N118N (500 C>T, rs11615) is the most studied polymorphism, the impact of this polymorphism on platinum-based chemotherapy remains unclear. This is the first study in which the functional impact of ERCC1 N118N on gene expression and platinum sensitivity was explored. The aim of this study is to investigate if the reduced codon usage frequency of AAT, which contains the variant allele of the silent mutation, has functional impact on ERCC1 in a well-controlled biological system. Specifically, the ERCC1 cDNA clone with either the C or T allele was introduced into an ERCC1 deficient cell line, UV20, and assayed for the effect of the two alleles on ERCC1 transcription, translation and platinum sensitivity. Both ERCC1 mRNA and protein expression levels increased upon cisplatin treatment, peaking at 4h post-treatment, however there were no differences between the two alleles (p>0.05). Cells complemented with ERCC1 showed significantly higher survival proportion than the parental cell line following platinum exposure (p<0.0001), although no differences were observed between the cells transfected with the wild type or the polymorphic allele. These data suggest that N118N itself is not related to the phenotypic differences in ERCC1 expression or function, but rather this polymorphism may be linked to other causative variants or haplotypes.

我们的实验室对药物遗传学有浓厚的兴趣。我们一直积极研究种系遗传变异，可以改变各种抗癌剂的药代动力学，反应和毒性，从而有助于具有已经狭窄的治疗窗口的治疗中临床结果的个体差异。我们已经建立了与药物治疗有关的这些多态性与它们的表型之间的分子联系。 Most of our work has been focused on genetic variations in drug metabolism and transporting candidate genes such as ABCB1 (P-glycoprotein, MDR1), ABCG2 (BCRP), SLCO1B3 (OATP1B3, OATP8), CYP3A4, CYP3A5, CYP1B1, CYP2C19, CYP2D6, UGT1A1, UGT1A9还有其他几个。 我们还对探索大量多态性的非候选基因方法感兴趣，以建立与临床结果的关系，并进行实验以验证探索性扫描引起的潜在致病等位基因。我们已经与Affymetrix合作，为170种药物处置基因中包含1,256种遗传变异的DMET芯片进行了β测试，目前正在建立一项临床试验，在该试验中，在NCI接受治疗的患者将用DMET芯片进行基因分型，以探索这些基因和多个Cancers的各种治疗方法之间的潜在联系。我们目前正在验证初始DMET芯片实验的结果方面取得了进展。 尽管已经进行了许多研究以解释对药代动力学变异性的某些遗传影响，但由于该领域的研究很差，因此我们也有兴趣阐明药物学和治疗结果的遗传标志和治疗结果。 DMET：多西他赛在其药代动力学和毒性特征上表现出显着的个体间变化。沙利度胺是一种活跃的抗癌剂，也表现出广泛的药理变异。过去的药物遗传学研究尚未解释这种变化。使用多西他胺和单独的多西他赛的前列腺毒素进行了前列腺癌的患者，使用Affymetrix DMET 1.0平台进行了基因分型，该平台在170个药物处置基因中测试了1,256个遗传变异。分析了与临床反应和毒性的关联分析遗传多态性。三个基因中的十个SNP可能与对治疗的反应有关：ppardelta，sult1c2和chst3。八个基因中的11个SNP与治疗的毒性有关：SPG7，CHST3，CYP2D6，NAT2，ABCC6，ATP7A，CYP4B1和SLC10A2。 DMET和直接测序之间的基因分型结果表现出大于96％的一致性。这些发现突出了非CYP450代谢酶和转运蛋白在多西他赛和沙利度胺的药理学中起作用的作用。 VEGFR2多态性和贝伐单抗/索拉非尼：高血压（HT）和手脚皮肤反应（HFSR）可能与贝伐单抗和索拉非尼的活性有关。我们假设这些毒性与这些药物中的有利结果相对应，HT和HFSR将重合，并且VEGFR2基因型变异将与毒性和临床结果有关。将毒性发生率和VEGFR2 H472Q和V297I状态与临床结果进行了比较。在贝伐单抗治疗后，患有HT的个体比没有这种毒性的个体在使用贝伐单抗患者的试验中没有这种毒性（31.5 vs 14.9mo，n = 60，p = 0.0009），以及贝伐单抗和贝伐单抗和Sorafenib的实体肿瘤患者（11.9 vs 3.7 mo 3.7 Mo，n = 0.052） HT还与索拉非尼和贝伐单抗结构疗法后的OS益处相关（5.7对29.0 MO，P = 0.0068）。 HFSR是索拉非尼治疗期间长时间PFS的标记（6.1 vs 3.7 mo，n = 113，p = 0.0003）。 HT是HFSR的危险因素，在接受贝伐单抗和/或索拉非尼治疗的患者中。 VEGFR2 H472Q的变异等位基因的载体发生了更大的HT和HFSR风险。这项研究表明，HT和HFSR可能是有利临床结果的标志物，HT发育可能是HFSR的标志物，而VEGFR2等位基因可能与贝伐单抗和/或索拉非尼治疗期间的毒性发展有关。 XRCC1多态性与辐射：XRCC1中与前列腺癌的放射治疗相关的遗传多态性：放射治疗是局部前列腺癌的潜在治愈性，重要的治疗选择。但是，在放射治疗后的8年中，即使是患者的最佳风险，大约10％的患者也会出现临床疾病复发。在此，我们研究了五个DNA修复的分子标记。 513例耐Castrate前列腺癌患者（CRPC），包括284例接受放射疗法的患者，229例没有放疗的患者，152名健康个体在DNA切除修复基因中进行了5种多态性的基因分型：ERCC1 N118N（500C> T），500C> T），XPD K751Q（500C> T） （685c> t），XRCC1 R399Q（1301G> a）和PARP1 V762A（2446T> c）。评估的多态性并未显示患者组和健康对照之间的差异，也没有显示出与生存相关的趋势。但是，在辐射处理的亚组中，中位生存时间与XRCC1单倍型有关。患有R399Q AA/R194W CC单倍型患者的中位生存时间为11。75年，患有R399Q AG/R194W CC单倍型的患者为12。17年，患有R39999999Q AG/R194W CT/6.64的患者的患者为66.65岁，并具有6.294W CT/6.21的患者。单倍型（p = 0.034）。研究所有患者时，找不到这种关联。我们得出的结论是，XRCC1中的遗传多态性可能会影响接受局部前列腺癌放疗的患者的结果。 ERCC1多态性和铂基化学疗法：ERCC1中的遗传多态性被认为有助于改变对基于铂的化学疗法的敏感性。尽管ERCC1 N118N（500 C> T，RS11615）是研究最多的多态性，但这种多态性对铂基化学疗法的影响尚不清楚。这是探索ERCC1 N118N对基因表达和铂敏感性的功能影响的第一个研究。这项研究的目的是研究AAT的密码子使用频率降低（包含静音突变的变异等位基因）是否对良好控制的生物系统中的ERCC1具有功能影响。具体而言，将带有C或T等位基因的ERCC1 cDNA克隆引入了ERCC1缺陷细胞系UV20中，并测定了两个等位基因对ERCC1转录，翻译和铂敏感性的影响。顺铂治疗后ERCC1 mRNA和蛋白质表达水平都升高，在处理后4H峰值，但是两个等位基因之间没有差异（p> 0.05）。与铂暴露后的亲本细胞系相比，与ERCC1相辅相成（P <0.0001）的生存率明显高，尽管用野生型或多态性等位基因转染的细胞之间没有观察到差异。这些数据表明N118N本身与ERCC1表达或功能的表型差异无关，而是该多态性可能与其他病因变体或单倍型有关。