二代測序?yàn)槟[瘤個體化治療提供選擇

二代測序?yàn)槟[瘤個體化治療提供更多選擇北京諾禾致源生物信息科技有限公司魏龍剛2011-2015

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Confidential基因列表亞洲肺腺癌患者突變頻率靶向藥物（FDA/CFDA批準(zhǔn)）臨床意義EGFR40%吉非替尼/厄洛替尼/?？颂婺?阿法替尼/AZD9291/CO1868同時檢測敏感位點(diǎn)與耐藥位點(diǎn)，綜合提示臨床用藥HER22%阿法替尼MET3%克唑替尼KRAS6%曲美替尼/司美替尼基因突變信息提示EGFRTKIs藥物不敏感;基因突變信息提示患者具有更差的預(yù)后PIK3CA3%替西莫司BRAF2%維羅非尼基因突變導(dǎo)致EGFRTKIs藥物不敏感ALK8%克唑替尼/色瑞替尼/艾樂替尼同時檢測融合突變與耐藥位點(diǎn)，綜合提示臨床用藥ROS13%克唑替尼基因融合突變提示臨床用藥RET2%卡博替尼/凡德他尼/舒尼替尼精準(zhǔn)醫(yī)療時代下的非小細(xì)胞肺癌治療：多靶點(diǎn)多藥物FDA/CFDA批準(zhǔn)用于肺癌；FDA/CFDA批準(zhǔn)用于其它癌種；肺癌臨床試驗(yàn)中現(xiàn)有腫瘤分子病理檢測方法面臨的挑戰(zhàn)一個患者，多種biomarker的結(jié)果需要同時給出多種檢測技術(shù)AMRS-PCR/IHC/FISH/RT-PCR….需要配備多種檢測流程、設(shè)備、空間、人員培訓(xùn)需要多種驗(yàn)證(validation)、試劑盒和QC樣本有限報告周期實(shí)驗(yàn)平臺的可擴(kuò)展性2011-2015

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Confidential下一代測序(NGS)能夠同時檢測多種biomarker一套檢測流程，檢測多種變異類型（SNP,indel,CNV,fusion)科研和臨床多次證明技術(shù)準(zhǔn)確性臨床應(yīng)用從外顯子到多基因多位點(diǎn)檢測NCCN指南推薦的技術(shù)2011-2015

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Confidential基因覆蓋17個癌種中至少70%以上的基因變異；Cosmic

數(shù)據(jù)庫基因所占比例2011-2015

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ConfidentialA

483

genes

cancer

panel

design細(xì)胞增殖通路23%腫瘤干細(xì)胞相關(guān)通路19%凋亡抑制相關(guān)通路11%炎癥相關(guān)通路11%細(xì)胞周期相關(guān)通路8%基因組不穩(wěn)定相關(guān)8%侵襲和轉(zhuǎn)移相關(guān)通路4%免疫相關(guān)通路4%血管生成相關(guān)通路4%能量代謝相關(guān)通路4%表觀遺傳相關(guān)通路4%483個基因全部歸類到11條信號通路A

483

genes

cancer

panel

design選擇483個基因全部7146個外顯子和19個基因常發(fā)生重排的部分2011-2015

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Confidential內(nèi)含子樣本收取樣本處理H&E染色文庫制備文庫庫檢上機(jī)測序數(shù)據(jù)質(zhì)控信息分析2011-2015

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ConfidentialWorkflow使用同一患者的組織/血液與其白細(xì)胞配對檢測使用白細(xì)胞可以增加體細(xì)胞突變檢測的靈敏度和特異性EGFR

exon19

c.2237_2254del2011-2015

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ConfidentialKRAS

exon2

c.G35A

p.G12DTP53

rs1042522tumornormalWorkflow-Key

point:Workflow-Key

point:otspot突變病理染色和診斷目的發(fā)現(xiàn)與腫瘤相關(guān)的更多的h提高h(yuǎn)otspot突變檢出率2011-2015

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ConfidentialLabelCount/Frequency%

of

genes

with

an

alteration

reported89.00%%

of

samples

with

at

least

one

clinically

relevantalteration31%Mean

number

of

alterations

per

sample3.09(range

0-28)Mean

number

of

clinically

relevant

alterations

persample0.7(range

0-10)2011-2015

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Confidential580例臨床樣本中，89%的樣本檢測到體細(xì)胞突變，每個樣本平均攜帶3.09個體細(xì)胞突變，通過數(shù)據(jù)庫篩選，只有31%的突變有臨床有效性證據(jù)。Summary2011-2015

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Confidential準(zhǔn)確性是二代測序進(jìn)行腫瘤相關(guān)檢測的第一步；如何更好的解讀基因變異和臨床意義，才是大范圍基因檢測的關(guān)鍵基因解讀(Variant

interpretation)Variant

callingRaw

dataClean

dataSNP+InDelCall:

mutect,StrelkaAnn:annovarFilter:dbSNP,cosmic,clivarCNVCall：ExomeCNVAnn：annovarFilter：DGV,dbVarSVCall:CRESTAnn:annovarNext:NovoFusionMappingDuplicationBWA，PICARDSomatic

mutation

analysis

(for

target

drug):Tumor

(solid

tumor

and

ctDNA)

+White

blood

cellCandidate

MutationsLow

qualityAdapterSNP+InDel+CNVCall:

samtoolsAnn:annovarFilter:pharmGKB,

BIC,

.etcGermline

mutation

analysis

(for

chemical

drug

and

genetic

risk)White

blood

cellACMG2011-2015

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ConfidentialVariant

annotationMutationDriver

mutationPassenger

mutationMini

driverLoss

of

functionGain

of

functionUnknownEtc.N-of-on.incOncomineTMNovoDatabaseLiterature

researchDrug

related

information:Sensitive/ResistanceClinical

informationPathway

analysisBiomarkersummaryBiology

relevanceClinical

relevanceClinical

evidenceRelevant

therapiesClinical

trailsReport2011-2015

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interpretationBiomarkersummaryBiology

relevanceClinical

relevanceClinical

evidenceRelevant

therapiesClinical

trails2011-2015

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ConfidentialSample

report-FGFR4

H673R(肺癌)Mutation：SNVDriver

mutationGain

of

functionDrug

related

information:Clinical

informationPathway

analysis突變對蛋白功能，信號通路的影響靶向藥物分析基因與腫瘤基因與NSCLC2011-2015

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Confidential基因功能分析Sample

report-FGFR4

H673R(肺癌)2011-2015

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solutions:

Key

differentiators變異上下游通路分析變異信號通路聚類分析多變異結(jié)果整合分析中國臨床實(shí)驗(yàn)單一通路：KRAS

p.Q61L(肺癌)Mutation：SNVDriver

mutationGain

of

functionDrug

related

information:Clinical

informationPathway

analysis2011-2015

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Confidential單一通路：KRAS

p.Q61L(肺癌)2011-2015

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Confidential單一通路：CTNNB1

p.T41A(纖維瘤)Proprietary

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Confidential?Proprietary

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Confidential用藥提示W(wǎng)nt

pathway

inhibitors,

such

as

PRI-724,

are

under

investigation

in

early

clinicaltrials

for

solid

tumors

(Tanwar

et

al.,

2011;

21695255,Kogan

et

al.,

2012;

22356261,Lachenmayer

et

al.,

2012;

22811581,Zhuet

al.,

2012;

22964660,Paripati

et

al.,2009;19812504).The

secreted

Frizzled-related

proteins

(sFRP)

are

another

class

of

Wnt

inhibitorsthat

have

been

proposed

to

block

tumor

growth

(Kogan

et

al.,

2012;

22356261).The

Wnt

antagonist

Dickkopf-related

protein

3

(Dkk3)

has

also

shown

tumorsuppressor

activity

(Veeck

and

Dahl,

2012;

21982838).Cox-2

inhibitors,

including

celecoxib

and

diclofenac,

have

been

reported

to

inhibitthe

Wnt/beta-catenin

pathway

and

may

also

be

relevant

in

a

tumor

with

beta-cateninactivation

(Zhang

et

al.,

2014;

24374173,

Sareddy

et

al.,

2013;

24013885).???單一通路：CTNNB1

p.T41A(纖維瘤)變異上下游通路分析變異信號通路聚類分析多變異結(jié)果整合分析中國臨床實(shí)驗(yàn)The

solutions:

Key

differentiators2011-2015

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ConfidentialTP53,KEAP1,MERTK,NCAM1,PDGFRB,WEE1

mutation細(xì)胞周期相關(guān)通路通路聚類:TP53,WEE12015

ASCO通路聚類:TP53,WEE12011-2015

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Confidential變異上下游通路分析變異信號通路聚類分析多變異結(jié)果整合分析中國臨床實(shí)驗(yàn)The

solutions:

Key

differentiators雙陽突變:EGFR

L858R,

T790M(肺癌)患者從確診(2013.6)到檢測(2015.8)，易瑞沙治療，沒有其他輔助治療下一步：Osimertinib樣本類型:cfDNA2011-2015

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Confidential變異上下游通路分析變異信號通路聚類分析多變異結(jié)果整合分析中國臨床實(shí)驗(yàn)The

solutions:

Key

differentiators中國臨床實(shí)驗(yàn)67歲男性臨床診斷：肺腺癌基因檢測：2011-2015

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Confidential解讀率提升，約75%的患者可能從基因變異提升的臨床信息中獲益基因解讀(Variant

interpretation)臨床應(yīng)用-常規(guī)檢測陰性，NGS檢測陽性74歲女性臨床診斷：肺腺癌基因檢測：HER2

indels鏡下描述：腫瘤細(xì)胞占比為50%-70%。2011-2015

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Confidential臨床應(yīng)用-常規(guī)檢測陰性，NGS檢測陽性HER2激活突變突變豐度19%2011-2015

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Confidential2011