Expanding Cancer and Therapeutic
Biomarker Research Opportunities
at MUSC
Richard R. Drake, Ph.D.
Professor and Director, MUSC Proteomics Center
SmartState Endowed Chair in Proteomics
Benign
PCa
stroma
100
0
Biomarker Research – The Past 7 Years
Translational Human Biomarker Discovery
Using Mass Spectrometry-based Proteomic
and Glycomic Methods
Cancers Studied:
Prostate*
Kidney*
Breast**
Colon*
Head and Neck
Specimens
Tissue
Esophageal**
Serum
Liver**
Urine
Bladder
EPS
*Current Funding and **Previous EDRN funding
Prostate Cancer Challenges
Normal
HGPIN
Epithelium
Localized
PCa
Indolent
VS
Aggressive
Over
Under
Risk
stratification
Metastatic
PCa
PSA
Biochemical
recurrence
???
Prognosis
Hormone
Refractory
PCa
Inefficient monitoring
of therapy control
Limited therapeutics
Prediction
New therapeutic
targets
BIOMARKERS!!!
FDA-Approved Cancer Biomarkers
Biomarker
α-Fetoprotein
α-Fetoprotein-L3
DCP
Human chorionic
gonadotropin-β
CA19-9
CA125
Pap smear
CEA
EGF receptor
Type
Glycoprotein
Glycoprotein
Protein
Source
Serum
Serum
Serum
Cancer Type
Liver
Liver
Liver
Clinical Use
Monitoring
Risk
Risk
Glycoprotein
Carbohydrate
Glycoprotein
Cervical smear
Glycoprotein
Glycoprotein
Serum
Serum
Serum
Cervix
Serum
Colon
KIT
Thyroglobulin
PSA
CA15-3
CA27-29
Cytokeratins
Estrogen & progesterone receptors
Protein (IHC)
Glycoprotein
Glycoprotein
Glycoprotein
Glycoprotein
Protein (IHC)
GI tumor
Serum
Serum
Serum
Serum
Breast tumor
Testicular
Pancreatic
Ovarian
Cervical
Colon
Colon
GI stromal
tumors
Thyroid
Prostate
Breast
Breast
Breast
Staging
Monitoring
Monitoring
Screening
Monitoring
Selection of therapy
Diagnosis &
selection of therapy
Monitoring
Monitoring
Monitoring
Monitoring
Prognosis
Protein (IHC)
Breast tumor
Breast
HER2/NEU
HER2/NEU
Glycoprotein (IHC)Breast tumor
Glycoprotein
Serum
Breast
Breast
HER2/NEU
Chromosomes 3,
7, 9, and 17
DNA (FISH)
Breast tumor
Breast
DNA (FISH)
Urine
Bladder
NMP22
Fibrin/FDP
BTA
CEA and mucin
Protein
Protein
Protein
Glycoprotein
Urine
Urine
Urine
Urine
Bladder
Bladder
Bladder
Bladder
Selection of therapy
Prognosis &
selection of therapy
Monitoring
Prognosis &
selection of therapy
Screening &
monitoring
Screening &
monitoring
Monitoring
Monitoring
Monitoring
HCC Supported
Shared Resources
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Lipidomics
Flow Cytometry & Cell Sorting
Cell & Molecular Imaging
Biostatistics
Clinical Trials
Tissue Biorepository
Small Animal Imaging
Cellular Therapy
Gene Knockout
Drug Discovery/Screening
Translational Research Lab
Drug Metabolism/Clinical Pharmacology
HCC’s Research Programs
(Membership = 150)
BIOMARKERS
BIOMARKERS
Proteomics
and
Glycomics
BIOMARKERS
Integrated Biomarker Discovery Approach
Tissue Proteomics:
MALDI MS Imaging
Prostatic Secretions:
EPS Urine/Direct EPS
Metabolic Labeling:
Cell Lines & Click
Chemistry Capture
Bruker Solarix 7T Dual Source FT-MS
Bruker Instrument Demo
In Bremen - August
Delivery and
Installation at MUSC
First week of
December 2011
* 1000
500
400
300
200
100
843. 50
843. 55
843. 60
843. 65
843. 70
843. 75
8
CASI
Continuous Accumulation of Selected Ions
CASI Example – Olanzapine Imaging in Kidney
The Cancer Tissue Proteome
Anderson and Quaranta, Nature Reviews Cancer 8, 227-234 (2008)
Tissue Preparation and Analysis Workflow for MALDI-MSI
Slice frozen embedded
tissue on cryostat
at 10 μm
Adjacent section H & E stained
Serial sections acquired
conductive
slide for
MALDI-MSI
ImagePrep
Pathologist defines
areas of interest
Spray
with matrix
Compare molecular image to stained
serial section
Acquire
mass spectra
Intens. [a.u.]
UltraFlex III TOF/TOF
w/ Smartbeam
x104
TestWCXreflectronQC\0_C4\1\1SRef
2.0
1.5
1.0
0.5
0.0
Molecular profile
500
1000
1500
2000
2500
3000
3500
m/z
Molecular image
MALDI-IMS Utilizing m/z 4355 can Identify
PCa Specific Regions of Prostate
Histology
MALDI imaging of m/z 4355
2mm
Benign
Benign
PCa
PCa
stroma
100
stroma
0
Relative Intensity
m/z 4355
Tumor
Benign
4000
6000
8000
10000
12000
Mass to charge (m/z)
14000
16000
18000
Cazares, L.H., Troyer, D.A., et al. (2009)Clin. Cancer Res., 15, 5541-5551.
Biomarker Performance Across Validation Set
A.
80
n = 23
B.
1.0
0.9
0.8
0.7
Sensitivity
Averaged Normalized Intensity m/z 4355
60
40
n = 31**
n = 14*
0.6
0.5
0.4
0.3
0.2
0.1
20
0.0
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1-Specificity
0
PCa
benign benign
adjacent distal
Currently being assessed in TMA’s
Cell Line Based Metabolic Labeling with
Azide-Modified Sugar Tags
AC4ManNAz – Sialic Acid
Labeling
Biotin-alkyne
N3
N3
SAv beads
Cu2+
N3
1
2
3
4
5
6
10
9
8
7
Expression of CDCP1 in prostate patient tissues
B
CDCP1+PI
CDCP1
CDCP1+PI
b
c
d
e
f
Tumor
Normal
a
Currently being assessed in TMA’s
CA19-9
Boronolectins to Sialyl Lewis Carbohydrate Antigens
+ Magnetic Bead
Carbohydrate Antigen
Affinity Capture System
Glycoprotein Biomarker
Identification
Non Cancer
-
+
Blnk
NonAggressive
Aggressive
PCa
+
+
SLeX-Boronolectin-biotin
976249-
?
PAP?
?
3828-
PSA
Streptavidin
Cell Lines,
Clinical Tissues
and Fluids
Glycoprotein
Biomarkers
Human
Subjects and
Animal Models
Imaging
Biomarkers
HCC Supported
Shared Resources
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







Lipidomics
Flow Cytometry & Cell Sorting
Cell & Molecular Imaging
Biostatistics
Clinical Trials
Tissue Biorepository
Small Animal Imaging
Cellular Therapy
Gene Knockout
Drug Discovery/Screening
Translational Research Lab
Drug Metabolism/Clinical Pharmacology
*Proteomics and Glycomics
Conclusions and Challenges



The core resources are present at the
HCC; coordination of these resources
to develop therapeutic biomarkers is
needed
Challenge: establishing an IT
infrastructure to coordinate the data
analysis and linkage of sample
information to individual patients
Opportunity: coordinating with the
cancer genetics emphasis and new
hires