translational h is w - liverpool health partners bill... · torrent msrt-pcr identify mutations for...
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Primary)Tumour)
Pre?neoplas'c)lesion)
Metasta'c)Deposits)
Metasta'c)Deposits)
e.g.)K?Ras)p.G12D,)p53)p.R273H,)p16)(fs))
Targeted)therapy)
Targeted)therapy)
Circula'ng)Tumour))Cells)
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Prokyma(
Ion Torrent MSRT-PCR
Identify mutations for subsequent monitoring using <1K sequences with gene specific primers
Wash
Purify & Concentrate CTCs
5 CTCs outnumbered by
~ 50 billion other cells
Monitor CTC levels
And tailor patient specific therapy
CTC Capture Sample Processing
Deep Sequence Monitor
5 CTCs outnumbered by
< 1,000 other cells
Blood
Target mutation identified
Cancer cells monitored
• Can)iden'fy)single)SNP)from)20,000)wild)type)sequences.)))
• BUT)need)to)avoid)PCR)error)being)“read”)as)a)muta'on.)
Ion)Torrent)
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ATTAGCCTACTCTATATCTCTGGGCTCGATYCGATCCTAGCTCGATCGATATC)
ATTAGCCTACTCTATATCTCTGGGTTCGATYCGATCCTAGCTCGATCGATATC)
Limi'ng)Dilu'on)Next)Genera'on)Sequencing)
Limi'ng)Dilu'on)Next)Genera'on)Sequencing)
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Limi'ng)Dilu'on)Next)Genera'on)Sequencing)
3)sequences)with)10%)muta'on)
10)sequences)with)<)1%)PCR)error)
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" Prokyma((i4i)(
Acknowledgements(" Pancreas(Research(Group(
" University(of(Heidelberg(" Andrea(Bauer((" Jörg(D.(Hoheisel(" Markus(Büchler(
" Eithne(Costello(" John(Neoptolemos(" Paula(Ghaneh(" Robert(SuMon(" Chris(Halloran(" Dan(Palmer(" Victoria(Shaw(" Karen(Aughton(" Li(Yan(" Lucy(Oldfield(" Claire(Jenkinson(" Vicki(EllioM(
" Cold(Spring(Harbor(Laboratories(" David(Tuveson(
" Brian(Lane(" KaTe(Bullock(" Liz(Garner((" Sara(Harrison(" Nils(Elander(" James(Nicholson((" Fran(Atherton(" Thomas(Hanna(" Robert(Ferguson(" Wafa(AbuXAlainin(" Kulbir(Mann(" Sumit(Nandi(
The(paTents(and(volunteers(
" University(College(London(" John(F.(Timms(" Stephane(Camuzeaux(" Usha(Menon(" Stephen(Pereira((
" Damian(Bond(" John(Hunt(" Chris(Stanley(" Nick(Bryan(" Tom(Hanna(