新一代分子診斷技術(shù)在腫瘤精準醫(yī)學中的應(yīng)用報告

1、精準醫(yī)學（腫瘤領(lǐng)域）檢測方案設(shè)計和質(zhì)控基于新一代測序（NGS）腫瘤精準醫(yī)學檢測產(chǎn)品設(shè)計全球多中心數(shù)據(jù)整合和數(shù)據(jù)質(zhì)控基于iFISH-CTC技術(shù)單細胞測序臨床應(yīng)用腫瘤早期檢測腫瘤個體化診療方案整合國際腫瘤數(shù)據(jù)庫基因Panel設(shè)計與應(yīng)用超高深度基因測序Panel設(shè)計腫瘤負荷、繼發(fā)耐藥監(jiān)控第1頁精準醫(yī)學（腫瘤領(lǐng)域）檢測方案設(shè)計和質(zhì)控基于新一代測序（NGS）腫瘤精準醫(yī)學檢測產(chǎn)品設(shè)計全球多中心數(shù)據(jù)整合和數(shù)據(jù)質(zhì)控第2頁新一代測序平臺互補融合性不一樣測序儀平臺能夠使用不一樣基因富集方法PCR捕捉PCR捕捉IonIonIlluminaIllumina第3頁基因變異類型檢測 I：突變基因全長（全外顯子）全部點突變

2、影響原因：基因富集覆蓋度（90%）影響原因：測序深度（200X）匯報中突變 %靈敏度：1%第4頁臨床研究發(fā)覺癌組織中CNV與Mutation共存腫瘤分型新概念（C型腫瘤，M型腫瘤）CSCO年會廈門 基因變異類型檢測 II：拷貝數(shù)變異（Copy Number Variation, CNV）第5頁基因變異類型檢測 III：融合基因NGS檢測融合基因RNA轉(zhuǎn)錄混正當NCI-MATCH Sample Test -Oncomine Comprehensive Panel第6頁腫瘤發(fā)病機制學說腫瘤器官發(fā)生學說腫瘤血液發(fā)生學說腫瘤泛感染發(fā)生學說及診療進展50例接收纈更昔洛韋（Valcyte）治療膠質(zhì)母細胞瘤

3、患者2年生存率為62%，對照組為18%（P0.001）。治療周期長亞組預(yù)后更加好，2年生存率可達70%和90%上海xx醫(yī)院使用xxx抗病毒藥品治療xxx瘤取得突破性進展N Engl J Med Sep 5;369(10):985-6第7頁基于iFISH-CTC技術(shù)單細胞測序臨床應(yīng)用腫瘤早期檢測腫瘤個體化診療方案8第8頁iFISH-CTC技術(shù)助力腫瘤精準醫(yī)學研究第9頁腫瘤早期iFISH-CTC檢測數(shù)據(jù)（.1-.8）5363例統(tǒng)計癌種靈敏度乳腺癌 95.93%結(jié)直腸癌93.18%肝癌81%胰腺癌 97.73%肺癌91.67%胃癌92.59%甲狀腺癌87%食管癌81.8%第10頁CSCO年會廈門 癌

4、癥早期篩查受檢者信息檢測路徑病理檢驗結(jié)果：低分化腺癌手術(shù)切除腺癌第11頁腫瘤早期綜合檢測及預(yù)防案例 -福建x州xx醫(yī)院循環(huán)腫瘤細胞檢測體內(nèi)存在腫瘤細胞提醒癌組織特征及用藥指導發(fā)覺 22 個腫瘤細胞影像學顯示右肺彌漫狀陰影（纖維化/癌變）激光顯微切割單個CTC腫瘤細胞單細胞全基因組測序分析12345定時隨訪和復查6癌標NSE ：72.6（ 99%每份樣本測序深度10000X第19頁Phase 1 results1,262 HCC related somatic mutations from original file (3,000,381 somatic mutations in total,

5、for all phenotypes).425 non-redundant HCC related somatic mutations, with unique genomic coordinates.These 425 mutations reside in 36 exons (covering 28 genes).Results would also be shown in figures later.第20頁Phase 1 resultsThe covered 28 genes are (by official symbol):AACS,ADAM22,AKAP13,ARVCF,ATR,C

6、DKN2A,CLCC1,CUL3,DMGDH,EGFR,ERBB2,GABBR1,GIF,GTPBP2,HNF1A,JAK2,LITAF,MET,MFSD8,MUC5B,MYCL,NOVA1,NTRK2,PCDH7,PLEKHG5,PSTK,RAF1,USP25.OR (by ensembl ID):ENSG00000008277.10,ENSG00000036257.8,ENSG00000081760.12,ENSG00000096968.8,ENSG00000099889.9,ENSG00000105976.10,ENSG00000116990.9,ENSG00000117983.13,E

7、NSG00000121940.11,ENSG00000132155.7,ENSG00000132837.10,ENSG00000134812.3,ENSG00000135100.13,ENSG00000139910.15,ENSG00000141736.9,ENSG00000146648.11,ENSG00000147889.12,ENSG00000148053.11,ENSG00000155313.11,ENSG00000164073.5,ENSG00000169851.11,ENSG00000170776.15,ENSG00000171680.16,ENSG00000172432.14,E

8、NSG00000175054.10,ENSG00000179988.9,ENSG00000189067.8,ENSG00000204681.6第21頁Phase 1 ResultsThese 36 exons covered 12,415bp in total, including:chr1 2061bpchr2 97bpchr3 201bpchr4 3218bpchr5 249bpchr6 322bpchr7 621bpchr8 0bpchr9 1986bpchr10 282bpchr11 350bpchr12 540bpchr13 0bpchr14 1400bpchr15 136bpchr

9、16 157bpchr17 342bpchr18 0bpchr19 0bpchr20 0bpchr21 325bpchr22 128bpchrX 0bpchrY 0bp第22頁6-phase selectorMethod description:Phase 2: max coverage “For each exon with SNVs covering 5 patients in LUAD and SCC, we selected the exon with highest RI that identified at least 1 new patient when compared to

10、the prior phase.” original paperData required: TCGA maf files (somatic mutation)第23頁6-phase selectorMethod description:Phase 3: RI = 30 & =3 patients “For each remaining exon with an RI 30 and with SNVs covering 3 patients in LUAD and SCC, we identified the exon that would result in the largest redu

11、ction in patients with only 1 SNV.” original paperData required: TCGA maf files (somatic mutation)第24頁6-phase selectorMethod description:Phase 4: RI = 20 & = 3 patients “Same procedure as phase 3, but using RI 20.” original paperData required: TCGA maf files (somatic mutation)第25頁6-phase selectorMet

12、hod description:Phase 5: Predicted drivers “We included all exons from additional genes previously predicted to harbor driver mutations in NSCLC.” original paperData required: predicted driver gene in HCC (from publication, 15 genes in reference paper)第26頁6-phase selectorMethod description:Phase 6:

13、Add fusions “For recurrent rearrangements in NSCLC involving the receptor tyrosine kinases ALK, ROS1, and RET, the introns most frequently implicated in the fusion event and the flanking exons were included.” original paperNot included (not under our consideration by far).第27頁ResultsThis panel inclu

14、des:604 genes (not necessarily have all exons of this gene included in panel)782 exons 120.1 kb in total, 154 bp per exon202 patients in total199 patients covered by this panel199 (99%), 181 (90%), 158 (78%) patients with at least 1, 2, 3 SNVsMean RI is 22.67 (Recurrence Index, the higher this number the more chance a SNV is found in patients)RI is