NGS用于指导晚期恶性肿瘤治疗2016 (3)

NGS用于指导晚期恶性肿瘤治疗2016 ASCO新进展-精准检测，精准治疗Insight on Novel Mechanisms and Precision Care,方勇浙江大学医学院附属邵逸夫医院肿瘤内科,基因检测一览,Presented By Claudine Isaacs at 2016 ASCO Annual Meeting,遗传性肿瘤重叠表型,同病异治异病同治,Sequencing is ,Presented By David Hayes at 2016 ASCO Annual Meeting,测序的优势价格更低廉品质更高适用范围更广 机器 试剂盒 软件组织灵活性更高 石蜡标本 无需细胞 甚至RNA！,近8年，ASCO年会上与NGS相关的摘要数目投稿,内容：NGS指导下的肿瘤免疫治疗cfDNA检测与肿瘤治疗NGS在多种实体肿瘤治疗中的应用,Cancer is a disease of ,Presented By David Hayes at 2016 ASCO Annual Meeting,癌症是一种的疾病?,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,以杂交捕获法为基础的二代基因测序法可作为预测抗PD-1/PD-L1治疗黑色素瘤疗效的标记物,Introduction,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,简介在进展期黑色素瘤患者中，抗PD-1治疗有30-45%的有效率；反应可能持续存在；较难辨别有效人群；因此需要有效预测标记物。,Candidate biomarkers for anti-PD-1/PD-L1,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,抗PD-1/PD-L1的候选标记物,克隆性增殖，突变或新的抗原,Aims,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,目标：假设：小片段基因能精确地代替基因突变负荷以杂交捕获法为基础的二代基因测序法检测236-315个基因编码区序列 突变负荷和特殊突变与临床结果的相关性 与全外显子测序所得突变负荷相比较T细胞受体测序与结果的相关性,Methods,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,方法：,Mutation load and response,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,突变负荷和反应,Mutation load and response,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,突变负荷和反应,Mutation load and response,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,突变负荷和反应,Mutation load and survival,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,突变负荷和生存率,Specific mutations,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,特定突变,Specific mutations,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,特定突变,Mutation load comparisons,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,突变负荷比较,Mutation load and survival in unselected patients,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,在未选择患者中的突变负荷及生存率,在未选择患者中突变负荷与生存率无关,TCR clonality,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,TCR克隆,Potential implications,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,潜在临床价值及意义,Conclusions,Presented By Douglas Johnson at 2016 ASCO Annual Meeting,结论二代杂交捕获法为基础的NGS检测突变负荷与抗PD-1/PD-L1疗效相关236-315个基因突变负荷的检测结果与全外显子测序结果一致通过此法检测突变负荷可能有助于临床的治疗决策和预测疗效。,内容：NGS指导下的肿瘤免疫治疗cfDNA检测与肿瘤治疗NGS在多种实体肿瘤治疗中的应用,ctDNA PPV vs. tissue biopsy (NSCLC, CRC, Breast, Others),Presented By David Hayes at 2016 ASCO Annual Meeting,背景：,Serial Next Generation Sequencing (NGS) of cell free DNA (cfDNA) reveals clonal evolution of AKT1 E17K mutant tumors: analyses from patients enrolled on a phase I basket study of an AKT inhibitor (AZD5363),Presented By Lillian Smyth at 2016 ASCO Annual Meeting,血浆游离DNA（cfDNA)的NGS揭示AKT1 E17K突变肿瘤的克隆演变：一项AKT抑制剂（AZD5363）I期篮子研究的患者分析,Background Targetable Driver Oncogenes,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,背景 靶向驱动致癌基因大量的肿瘤基因型分析已显著提高了肿瘤驱动基因的认识，其中一部分已成为靶向基因其中一个驱动基因，AKT1 E17K突变 反复的（2%），人类癌症的致癌事件 一项研究治疗意义的I期篮子研究正在进行中，其中一种pan-AKT抑制剂（AZD5363）的靶向治疗具有较好的前景 当然，目前还不清楚是否同时存在其他突变，导致肿瘤细胞对AKT1 E17K依赖的改变。,Background- Liquid biopsies,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,背景 液体活检对肿瘤发展及耐药产生的理解很大程度上依赖于取得新鲜肿瘤组织而单个病灶的活检存在很多挑战（异质性，局限的可行性，侵袭性以及价格昂贵）都可能被液体活检克服肿瘤来源的血浆游离DNA,Background- Liquid biopsies,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,检测肿瘤基因组的改变发现疾病基因组的复杂性（异质性）监测治疗中的肿瘤动力学追踪现实中耐药机制的演变（监测分子性疾病）,背景 液体活检,Objectives,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,目标在基因富集（含有AKT1 E17K突变）的人群中，正在接受靶向治疗，尝试运用cfDNA以：1.发现及追踪AKT1 E17K突变2.描述具有AKT1 E17K突变的肿瘤中同时发生的突变3.监测分子耐药机制的演变，通过 平行追踪所有肿瘤突变 辨别新近获得的突变的出现,Study design,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,研究设计前瞻性，单中心，相关性研究合适的患者 AKT1 E17K突变阳性的实体肿瘤 入组参加AZD5363的1期多中心篮子研究，一种口服的催化性pan-AKT抑制剂，作用于AKT1突变的进展期实体肿瘤（NCT01226316） -已知具有RAS/RAF突变的肿瘤患者被排除 -根据RECISTv1.1标准定义，具有可测量病灶,Study Schema,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,研究纲要,Sample Collection,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,血样本收集,Patient demographics,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,患者统计学资料,AKT1 E17K detected and monitored in cfDNA by ddPCR,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,在22/25个（88%）患者中， ddPCR检测到AKT1在D21，AKT1的减少预示PFS 的升高,以ddPCR检测及监控cfDNA中的AKT1 E17K,AKT1追踪肿瘤反应 持续的（21天）AKT1清除 与持续的RECIST反应相关（P=0.025）,AKT1 mutant allele fraction (MAF) in cfDNA highly concordant between ddPCR and NGS,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,cfDNA的AKT1突变等位基因分数（MAF）在ddPCR和NGS中高度一致,NGS of Pre-treatment cfDNA (n=25) captured tumor heterogeneity,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,治疗前cfDNA（n=25）的NGS发现肿瘤异质性在8个（38%）病人中检测到17种额外的改变（共21个病人具有可比较的治疗前肿瘤）发现3个（12%）病人中同时出现了相关的突变（ARAF，RAF1，KRAS），但是所有都对药物治疗没有反应,Slide 13,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,治疗前最常见的突变基因,Pre-treatment cfDNA captured tumor heterogeneity,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,治疗前的cfDNA发现肿瘤异质性,Conclusions,Presented By Lillian Smyth at 2016 ASCO Annual Meeting,结论在这项探索性研究中，对通过基因panel捕获的cfDNA进行二代测序是可行的，并且与ddPCR得到的结果高度一致发现疾病异质性能够监测肿瘤克隆动力学/发现克隆演变/突出反应的不同模式辨别在治疗过程中获得的或富集的潜在相关的基因组改变为前期分子靶向药物的研发提供了一种有效方法。,内容：NGS指导下的肿瘤免疫治疗cfDNA检测与肿瘤治疗NGS在多种实体肿瘤治疗中的应用,二代基因测序在未治疗、转移性EGFR突变肺癌中发现并发的基因改变,TITLE,二代基因测序胆管肿瘤揭示肿瘤体系变异和化疗耐药性的联系,二代基因测序胆管肿瘤揭示肿瘤体系变异和化疗耐药性的联系,TP53, CDK N2A, ARID1A基因突变,TITLE,对主要的中枢神经系统肿瘤进行二代基因测序，作为个体化治疗肿瘤研究项目的一部分，发现与靶向抑制剂治疗敏感性相关的变异,对主要的中枢神经系统肿瘤进行二代基因测序，作为个体化治疗肿瘤研究项目的一部分，发现与靶向抑制剂治疗敏感性相关的变异,Slide 1,BRCA1/2阴性的乳腺及卵