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着浓厚的兴趣。我们一直在积极研究种系遗传变异如何改变各种抗癌药物的药代动力学、反应和毒性，从而导致治疗窗口已经很窄的治疗中临床结果的个体间差异。我们已经在这些多态性及其与药物治疗相关的表型之间建立了分子联系。我们的大部分工作集中在药物代谢和转运候选基因中的遗传变异，例如ABCB1（P-糖蛋白，MDR1）、ABCG2（BCRP）、SLCO1B3（OATP1B3、OATP8）、CYP3A4、CYP3A5、CYP1B1、CYP2C19、CYP2D6、 UGT1A1、UGT1A9 和其他几个。 我们还对非候选基因方法感兴趣，其中探索大量多态性以建立与临床结果的关系，并进行实验以验证探索性扫描产生的潜在致病等位基因。我们与 Affymetrix 合作对 DMET 芯片进行 Beta 测试，该芯片包含 170 个药物处置基因的 1,256 个遗传变异，目前正在开展一项临床试验，将使用 DMET 芯片对在 NCI 接受治疗的患者进行基因分型，以探索这些基因之间的潜在联系以及多种癌症的各种治疗方法。目前，我们正在验证初始 DMET 芯片实验结果方面取得进展。 虽然许多这些研究是为了解释遗传对药代动力学变异性的影响而进行的，但我们也对阐明药效学的遗传标记和几种主要抗癌药物的治疗结果抱有浓厚的兴趣，因为该领域的研究相当少。 DMET：多西紫杉醇的药代动力学和毒性特征表现出显着的个体差异。沙利度胺是一种活性抗癌剂，并且还表现出广泛的药理学变化。过去的药物遗传学研究尚未解释这种变异。参加一项使用多西他赛和沙利度胺与单独使用多西他赛的随机 II 期试验的前列腺癌患者使用 Affymetrix DMET 1.0 平台进行基因分型，该平台测试 170 个药物处置基因的 1,256 个遗传变异。分析遗传多态性与临床反应和毒性的关联。三个基因中的 10 个 SNP 可能与治疗反应相关：PPARδ、SULT1C2 和 CHST3。 8 个基因中的 11 个 SNP 与治疗毒性相关：SPG7、CHST3、CYP2D6、NAT2、ABCC6、ATP7A、CYP4B1 和 SLC10A2。 DMET 和直接测序之间的基因分型结果显示出大于 96% 的一致性。这些发现强调了非 CYP450 代谢酶和转运蛋白在多西紫杉醇和沙利度胺的药理学中可能发挥的作用。 VEGFR2 多态性与贝伐单抗/索拉非尼：高血压 (HT) 和手足皮肤反应 (HFSR) 可能与贝伐单抗和索拉非尼的活性有关。我们假设这些毒性将对应于这些药物的有利结果，HT 和 HFSR 将一致，并且 VEGFR2 基因型变异将与毒性和临床结果相关。将毒性发生率以及 VEGFR2 H472Q 和 V297I 状态与临床结果进行比较。在使用贝伐单抗治疗前列腺癌患者的试验中，经历过 HT 的个体在贝伐单抗治疗后的 PFS 比没有这种毒性的患者更长（31.5 个月 vs 14.9 个月，n=60，P=0.0009），以及贝伐单抗和索拉非尼治疗实体瘤患者的试验（11.9 个月 vs 14.9 个月）。 3.7 个月，n=27，P=0.052）。索拉非尼和贝伐单抗联合治疗后，HT 还与 >5 倍的 OS 获益相关（5.7 个月与 29.0 个月，P=0.0068）。 HFSR 是索拉非尼治疗期间延长 PFS 的标志（6.1 个月 vs 3.7 个月，n=113，P=0.0003）。 HT 是接受贝伐珠单抗和/或索拉非尼治疗的患者发生 HFSR 的危险因素。 VEGFR2 H472Q 变异等位基因携带者患 HT 和 HFSR 的风险更大。这项研究表明，HT 和 HFSR 可能是有利临床结果的标志物，HT 的发展可能是 HFSR 的标志物，VEGFR2 等位基因可能与贝伐单抗和/或索拉非尼治疗期间毒性的发生有关。 XRCC1 多态性和放射：XRCC1 基因多态性与前列腺癌放射治疗相关：放射治疗是局部前列腺癌的一种潜在治愈性重要治疗选择。然而，在放射治疗后 8 年，即使在风险最高的患者亚组中，大约 10% 的患者也会出现临床疾病复发。在此，我们研究了 DNA 修复的五种分子标记。对513名去势抵抗性前列腺癌（CRPC）患者，包括284名接受放疗的患者、229名未接受放疗的患者以及152名健康个体，进行了DNA切除修复基因5个多态性的基因分型：ERCC1 N118N（500C>T）、XPD K751Q（ 2282A>C)、XRCC1 R194W (685C>T)、 XRCC1 R399Q (1301G>A) 和 PARP1 V762A (2446T>C)。评估的多态性没有显示出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。然而，在放射治疗亚组中，中位生存时间与 XRCC1 单倍型相关。 R399Q AA/R194W CC 单倍型患者的中位生存时间为 11.75 年，R399Q AG/R194W CC 单倍型患者的中位生存时间为 12.17 年，R399Q AG/R194W CT 单倍型患者的中位生存时间为 66.65 年，R399Q AG/R194W CT 单倍型患者的中位生存时间为 6.21 年。 R399Q GG/R194W CT 单倍型（p = 0.034）。当对所有患者进行调查时，并未发现这种关联。我们得出的结论是，XRCC1 的基因多态性可能会影响接受局限性前列腺癌放疗的患者的预后。 ERCC1 多态性和铂类化疗：ERCC1 的基因多态性被认为有助于改变对铂类化疗的敏感性。尽管ERCC1 N118N（500 C>T，rs11615）是研究最多的多态性，但这种多态性对铂类化疗的影响仍不清楚。这是第一项探讨 ERCC1 N118N 对基因表达和铂敏感性的功能影响的研究。本研究的目的是调查 AAT（包含沉默突变的变体等位基因）密码子使用频率的降低是否对控制良好的生物系统中的 ERCC1 产生功能影响。具体来说，将带有 C 或 T 等位基因的 ERCC1 cDNA 克隆引入 ERCC1 缺陷细胞系 UV20 中，并测定这两个等位基因对 ERCC1 转录、翻译和铂敏感性的影响。顺铂治疗后 ERCC1 mRNA 和蛋白表达水平均增加，在治疗后 4 小时达到峰值，但两个等位基因之间没有差异 (p>0.05)。尽管在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，但用ERCC1补充的细胞在铂暴露后表现出比亲本细胞系显着更高的存活率（p<0.0001）。这些数据表明，N118N 本身与 ERCC1 表达或功能的表型差异无关，但这种多态性可能与其他致病变异或单倍型有关。