Diagnostic Capabilities Beyond the Tumor
Cell Genome
Gregory J. Tsongalis, PhD, HCLD
Professor of Pathology
Director, Molecular Pathology
Geisel School of Medicine at Dartmouth
Dartmouth Hitchcock Medical Center
Norris Cotton Cancer Center
Lebanon, NH
gregory.j.tsongalis@hitchcock.org
Outline
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Technology and Personalized Medicine
miRNAs
The liquid biopsy
Beyond that…….
Technology and Therapeutics as Drivers of Medical Care
Translational Research
Basic
Science
Technology
Therapeutics
Clinical Laboratory Medicine
Clinical
Medicine
The Genome Or The Human Karyotype
Massively Parallel or Next Generation Sequencing at DHMC
MiSeq
Sequence by Synthesis
PGM
Personalized Medicine
(A) Before crizotinib; (B) after 7 weeks of crizotinib.
Shaw A T and Engelman J A JCO 2013;31:1105-1111
Specificity
(miRNAs as Biomarkers)
Exportin 5
Dicer
Pri-miRNA
DroshaDGCR8
Pre-miRNA
miRNA:miR
NA* duplex
Unwinding
Nucleus
RISC
Cytoplasm
Ribosome
Translational Repression
mRNA Cleavage
C. L. Bartels and G.J. Tsongalis, Clin Chem, 2009;55: 623-631
miRNA Profiling from Pancreatic FNAs
mir148a
8
Expression Level
(log22-ΔCT)
7
6
5
NP
CP
PanIN
AC
5
*
4
4
3
2
1
**
5
**
16
0
24 FFPE matched tumor and normal HNSCC
(6 laryngeal, 6 oral tongue)
miRs: 21, 205, 12746 upregulated
miRs:451, 125b downregulated
Szafranska, Doleshal, Edmunds , Gordon , Luttges, Mundingv Barth Jr , Gutmann, Siriawinata , Pipas ,
Tannapfel , Korc, Hahn, Labourier , Tsongalis . Clin Chem 54(10):1716-1724, 2008.
Hanoun, Delpu, Suriawinata, Bournet, Bureau, Selves, Tsongalis ,
Dufresne, Buscail, Cordelier, Torrisani. Clin Chem 56(7):1107-1118, 2010.
Issues With The Tissues
– Tumor tissue has inherent limitations
– Tumor tissue is a single snapshot in time
– Selection bias due to tumor heterogeneity
– Invasive procedure
A Huge Problem
Resected Specimen
Small Needle Core
Biopsy
Exhausted Block
The Liquid Biopsy
• Cell Free DNA (cfDNA)
– Cells undergoing apoptosis or necrosis
– Amount of cfDNA correlates with tumor staging
and prognosis
Xenograft Model
Kerrington Smith, MD
Promega PlexorHY
Breath-based Diagnostics
J. E. Hill, PhD
Thayer School of Engineering at Dartmouth
Breath-based Diagnostics
Conclusions
• Circulating nucleic acids become future biomarkers
of choice
• More advanced analysis of simpler specimens
Molecular Pathology Laboratory
and Translational Research Program
Samantha Allen
Paul Burchard
Francine de Abreu, Ph.D.
Leanne Cook
Betty Dokus
Amber Erskine
Torrey Gallagher
Arnold Hawk
Joel Lefferts, Ph.D.
Emmeline Liu
Rebecca O’Meara
Jason Peterson
Elizabeth Reader
Barbara Soares
Heather Steinmetz
Laura Tafe, M.D.
Scott Turner, PhD
Wendy Wells, M.D.
Terri Wilson
Eric York