8/2/2012
Image-Guided Oncologic Surgery:
Mass Spectrometry Imaging
Nathalie Y.R. Agar, Ph.D.
Assistant Professor of Surgery
Assistant Professor of Radiology
Harvard Medical School
Director, Surgical Molecular Imaging Laboratory
Department of Neurosurgery
Brigham and Women’s Hospital
Assistant Professor, Department of Cancer Biology
Dana-Farber Cancer Institute
AAPM August 2, 2012
Boston Lying-In Hospital
1923 Operating Room
Brigham and Women’s Hospital
2011 Operating Room
Advanced Multimodality Image Guided Operating Suite (AMIGO)
1
8/2/2012
Mass Spectrometry
Ion Formation
Ion Separation
Ion Source
Mass Analyzer
Electron Impact (EI)
Chemical Ionization (CI)
Fast Atom Bombardment (FAB)
Electrospray Ionization (ESI)
Matrix Assisted Laser
Desorption/Ionization (MALDI)
Quadrupole
Magnetic Sector
Electric Sector
Ion Trap
Time-of-Flight
Ion Cyclotron
Ion Detection
Detector
Faraday Cup
Secondary Electron Multiplier (SEM)
Scintillator
Micro Channel Plate (MCP)
Mass Spectrometry Imaging
Applications
Surgical Margins
Tumor versus non-tumor tissue
Molecular Classification of Brain Tumors
Tumor type
Grade
Behavior (recurrence/progression)
Intraoperative
… prediction of response to targeted therapy
Drug Distribution and Metabolism
Pre-clinical and upcoming clinical studies
To access diseased tissue while preserving healthy surrounding brain.
SURGICAL MARGINS
2
8/2/2012
Surgical Handheld Probe for
Molecular Imaging
Navigation
optical sensors
2
Mass spectrometry
interface
1
International Application PCT/US09/53967 “Integrated Surgical Sampling Probe”. Jolesz, F.A., Golby, A.J., and Agar, N. Y. R.
International Application PCT/US2010/029586 “Method and System for Surface Sampling” Agar, N.Y.R., and Wiseman, J.M.
Desorption Electrospray Ionization
(DESI) Mass Spectrometry
No sample
preparation
Directly from
tissue
Limited to small
molecules
Takats, Z., Wiseman, J.M., Gologan, B., Cooks, G.R. (2004) Science. 306:471-3
Stereotactic Mass Spectrometry Imaging
a
b
c
100
219.1
90
239.1
80
70
263.0
60
319.1
50
888.7
40
362.9
862.5
30
788.5
916.8
564.8
20
384.8 441.3
520.8
588.7
848.5
656.6
932.7
950.4
766.5
10
0
200
300
400
500
600
m/z
700
800
900
1000
Agar, NYR., Golby, AJ, Ligon, KL, Norton, I, Mohan, V, Wiseman, JM, Tannenbaum, A, and Jolesz, FA (2011) Neurosurgery.
Development of Stereotactic Mass Spectrometry for Brain Tumor Surgery. 68(2):280-89; discussion 290
3
8/2/2012
Tumor Cell Concentration
by DESI Mass Spectrometry
263.0
100
1
794.4
100
838.6
90
90
80
90%
Relative Abundance
Relative Abundance
80
70
60
50
279.0
70
768.4
60
766.4
796.4
50
822.3
40
846.3
750.5
731.1
30
20
362.9
40
1778.9
10
30
0
700
313.0
20
462.8
10
559.0
676.0
794.4 838.6
768.4
750
800
850
1512.2 1609.8
1653.1
1374.8
1485.6
m/z
1131.6
968.9
1259.7
1872.0
1938.1
0
200
400
600
800
1000
1200
1400
1600
1800
2000
m/z
255.4
100
90
2
794.6
100
90
60
50
263.1
40
70
834.4
768.5
60
818.5
766.5
50
750.5
40
845.4
740.5
30
20
269.3
30
10
363.0
90%
838.6
80
70
Relative Abundance
Relative Abundance
80
20
0
700
301.1
10
750
800
850
m/z
462.9 511.1
750.5 794.6 838.6
584.9
1303.3 1394.2 1462.0 1579.4
981.0 1089.2 1148.8
1694.3 1789.8
1889.3 1969.3
0
200
400
600
800
1000
1200
1400
1600
1800
2000
m/z
891.0
100
789.5
905.2
90
80
Relative Abundance
Relative Abundance
80
70
919.1
60
255.4
50
839.0
40
70
60
50
839.0
40
795.3
815.4
30
933.1
849.2
769.2
20
363.0
751.5
703.0
10
30
283.4
769.2
30%
Recurrent left frontal
oligodendroglioma,
W.H.O. grade II with
chromosome 1p/19q
co-deletion
789.5
100
90
7
20
462.9
540.5 601.1
10
0
200
400
950.0
965.1
703.0
600
0
700
800
1110.5
750
800
850
m/z
1166.1 1293.4 1371.9
1000
1200
1514.9 1573.7 1660.0
1768.1
1400
1600
1891.3 1947.2
1800
2000
m/z
889.2
100
9
90
789.0
80
Relative Abundance
Relative Abundance
50%
789.0
100
263.7
90
80
70
363.5
60
50
905.2
279.6
284.0
40
70
60
50
40
30
863.2
816.9
727.3
765.9
10
0
463.5
700
877.2
20
565.5
10
0
200
786.8
20
917.2
30
400
933.1
1024.8
669.8 727.3
600
800
750
800
850
m/z
1444.3 1503.8
1281.4
1110.2
1000
1200
1400
1643.4 1699.8
1600
1820.3 1912.3
1800
2000
m/z
Correlation of Molecular Changes with
Histopathology
T: ITMS - p NSI Full ms[50.00-2000.00]
100
90
80
Intensity
70
60
50
50 %
infiltrated
889.17
789.00
891.17
789.92
40
905.17
892.08
30
10
786.83
698.83 704.00
713.67
721.92
727.25 731.17 737.17 741.92
751.00 757.67 760.83 765.92
775.00
794.92
785.25
802.92
807.00 810.83 816.92
821.17
829.67
834.92 839.00
863.17
841.83 849.08
858.83
877.17
867.00
908.17
894.08
901.00
886.17
917.17
903.17
893.17
20
918.17
921.08
909.08
933.08
923.17
935.92
0
700
710
720
730
740
750
760
770
780
790
800
810
820
m/z
830
840
850
860
870
880
890
900
910
920
930
940
T: ITMS - p NSI Full ms[50.00-2000.00]
100
90
80
Intensity
70
60
50
50 %
infiltrated and heterogenous
40
20
768.42
766.42
745.58
30
750.58
700.50
709.08 712.67
722.42
757.92 760.58
728.67 734.58 738.50
788.50
888.67
834.42
794.33
796.42
772.67
778.50
786.42
782.58
838.42
797.42 806.50
810.42 814.50
885.58
862.58
840.42
844.50 848.50
822.33
864.67
860.67
826.58
890.58
904.75
884.17
916.75
918.75
902.75
892.75
876.75
873.83
858.58
907.83
914.00
894.75
932.92
928.75
936.67
10
0
700
710
720
730
740
750
760
770
780
790
800
810
820
m/z
830
840
850
860
870
880
890
900
910
920
930
940
T: ITMS - p NSI Full ms[150.00-2000.00]
794.42
100
90
80
60
50
885.58
768.42
20
750.50
700.58
709.50
795.42
766.42
40
30
716.42
722.75
731.08
740.50
796.42
769.42
770.42
758.75
788.42
792.42
778.50
839.42
834.33
797.42
798.33
786.58
752.08
748.58
886.50
840.42
822.33
820.50
810.50
846.25
818.42
804.67
852.50 857.50 862.42
844.58
833.42
883.33
865.50 872.42
887.50
898.17
892.17
879.33
901.33
909.25
914.67 919.42
922.25
931.50
936.67 939.83
10
0
730
740
750
760
770
780
790
800
810
820
m/z
830
840
850
860
870
880
890
900
910
920
930
940
Molecular Assignment
T: ITMS - p NSI Full ms [150.00-2000.00]
794.42
838.58
100
90
885.58
80
70
Intensity
720
768.42
60
834.33
50
788.42
40
676.00
30
669.42
20
778.50
750.50
731.08 740.50
682.83 692.75 709.50
822.33
846.25
810.50
857.50
889.75
919.42
939.83
10
0
660
680
700
720
740
760
780
800
m/z
820
840
860
880
900
920
940
305.4
100
90
768.3
80
Relative Abundance
710
70
718.3
m/z 768
283.3
60
333.3
50
40
718.6
699.7
30
20
255.3
347.2
453.6
479.9
482.1
436.4
10
531.2
483.9
347.3
0
250
300
350
400
450
615.9 631.7
534.4 571.3 604.8
534.6
572.1
500
550
600
768.0
691.7
650
700
750
Phosphatidylcholine
16:0/16:0
m/z
751.4
100
90
80
80
70
60
50
70
m/z 838
60
50
40
419.0 419.5
30
751.7
283.3
20
40
437.2
283.4
331.1
10
284.2 297.2
30
0
280
0
250
418.8
327.1 331.5
309.5
300
320
340
420.3
375.3 379.6
360
m/z
380
400
420
445.5
440
419.3
20
10
-Serine
419.3
100
90
Relative Abundance
700
Relative Abundance
Intensity
70
838.58
90 %
solid
838.3
283.3
300
331.1
350
437.2 467.2
375.3 418.8
400
450
524.3 558.6 582.2 597.6 643.9
500
550
m/z
600
650
750.5
712.2 724.0
700
770.3 802.4
750
800
Phosphatidylserine
18:0/18:1
4
8/2/2012
3D Visualization in Slicer
Histopathology
Tumor cell concentration
0
DESI Mass Spectrometry
m/z 768.3 +/- 0.5
Phosphatidylcholine 16:0/16:0
95 %
0
100
Agar, NYR., Golby, AJ, Ligon, KL, Norton, I, Mohan, V, Wiseman, JM, Tannenbaum, A, and Jolesz, FA (2011) Neurosurgery.
Development of Stereotactic Mass Spectrometry for Brain Tumor Surgery. 68(2):280-89; discussion 290
MOLECULAR CLASSIFICATION OF BRAIN
TUMORS
Lipid Analysis for Glioma
Classification
Eberlin, LS, Dill, AL, Golby, AJ, Ligon, KL, Wiseman, JM, Cooks, RG, and Agar, NYR (2010) Angewandte Chemie. Discrimination
of Human Astrocytoma Subtypes by Lipid Analysis using Desorption Electrospray Ionization Imaging Mass Spectrometry. 49,
873.
5
8/2/2012
Grades of Astrocytomas
PS (36:1)
m/z 788.8
PS (40:6)
m/z 834.2
PI (38:4)
m/z 885.4
ST (24:1)
m/z 888.4
ST (26:1)
m/z 916.5
H&E
stained
Grade II
3 mm
0
100%
0
100%
0
100%
Grade III
3 mm
Grade IV
3 mm
Able to distinguish all 3 grades on basis of lipid profiles – “spray and tell”
Gemistocytic Gliomas
Glioma Subtypes
Grade III
LS Eberlin, I Norton, AL Dill, AJ Golby, KL Ligon, S Santagata, RG Cooks and NYR Agar (2012) Cancer Research. 72(3):64554. Classifying Human Brain Tumors using Lipid Imaging by Mass Spectrometry.
6
8/2/2012
Classification Models
Subtype
Grade
Concentration
Overall Recognition Capability
Cross Validation*
Subtype
99.3 %
97.9 %
Grade
99.7 %
99.2 %
Concentration
99.0 %
97.5 %
*leave-out 20%/ 10 iterations
Classification Results
Subtype
VALIDATION SAMPLES
Name
G4
G6
G8
*G26
G28
WHO Diagnosis
A3; med
A4; high
A4; recurrent A4
A4; recurrent OA3
A4
G32
II
Grade
%
III
IV
Concentration
0
0
0
8
99
92
16
33
21
2
36
91
A
OA
O
low
3
1
100
98
100
0
0
0
0
2
0
med
96
84
high
1
15
A4; recurrent A4
94
0
6
*G33
G48
G35
G30
G46
A4; high
A4; focal gliosarcoma
A4; high
A4-O; recurrent; high
A4-O recurrent A3
45
98
5
1
49
1
51
48
9
43
43
7
1
19
4
1
20
14
98
61
82
100
30
0
60
0
10
2
12
86
G29
A4-O; recurrent OA3
69
0
31
32
36
32
G5
G47_O
G47_1
G42_0
G49
G41
G20
G13
A4-O; high (80%)
A4-O; recurrent O2
A4-O; recurrent O2
O2 focal O3; High
O2; gemistocytes
O2/3; recurrent O2
OA2; residual A2; med (30%)
OA2-3; recurrent OA2-3
42
43
55
32
17
0
26
40
45
65
100
0
0
35
0
77
17
6
12
84
4
37
3
0
14
2
0
49
96
100
90
33
90
2
10
67
5
98
0
0
5
0
1
10
89
0
89
0
2
100
9
79% in Agreement Over 3 Classes
Correlation to Histopathology
Oligodendrioglioma II, Infiltrated
Subtype
Grade
Concentration
7
8/2/2012
Right Frontal Craniotomy
Tumor Cell Concentration
med
low
SampleID
Class
low
Class
med
high
Class
high
A
23
33
0
B
41
55
0
C
5
59
0
D
3
37
2
med
low
SampleID
Class GL
Class MG
Class A
Class OA
Class gr2
Class O
Class gr3
Class gr4
high
Class low
Class med
Class high
case2_S10
(B)
96
0
0
0
96
14
57
25
41
55
0
case2_S11
(A)
56
0
0
0
56
15
29
12
23
33
0
case2_S12
(C)
59
5
29
0
35
64
0
0
5
59
0
case2_S13
(D)
40
2
1
0
41
42
0
0
3
37
2
case2_S14
84
257
20
20
11
320
320
327
00
14
14
00
6
335
6
case2_S15
11
4
6
0
9
8
4
3
0
9
case2_S7
223
11
8
0
226
227
7
0
3
231
0
case2_S8
239
61
38
4
258
173
95
32
145
133
22
808
340
102
5
1041
870
192
86
220
563
365
Manuscript in preparation.
Pathology Review
→ Dura region surrounded by oligodendrioglioma
8
8/2/2012
Astrocytoma Grade IV (Glioblastoma)
Grade
III
III
II
IV
II
IV
Astrocytoma Grade IV (Glioblastoma)
Tumor Cell Concentration
Case 8: Tumor Cell Concentration
med
low
high
med
low
high
CHEMICAL BRAIN MAPPING
9
8/2/2012
17 Brain Regions
4 Subjects
1,768 tissue sections!!
Inferior Temporal
Calcarine
Hippocampus
Medial Orbital Frontal
Substantia Nigra
Mid Frontal
Caudate Head
Superior Frontal
Cingulate
Motor
Posterior Putamen
Ventromedial Prefrontal
Amygdala
Posterior Parietal
Anterior Insula
Angular Gyrus
Inferior Rostral
v
Hippocampus
Caudate nucleus
Medial frontal
Cingulate gyrus
Substantia nigra
Inferior temporal
INTRAOPERATIVE MASS
SPECTROMETRY
10
8/2/2012
Continued development…
MENINGIOMAS
BRAIN METASTASES
DRUG DISTRIBUTION AND METABOLISM
BREAST CANCER
Agar Lab
Xiaohui Liu
Behnood Gholami
Isaiah Norton
Revaz Machaidze
Jennifer Ide
Daniel Feldman
Neurosurgery
Alexandra Golby
Ian Dunn
Daniel Orringer
OR Nurses
Radiology
Ferenc Jolesz
Tina Kapur
Ron Kikinis
Pathology
Sandro Santagata
Keith Ligon
Shakti Ramkissoon
Kristi Gill
Margot Burns
Neurology
Philip DeJager
Psychiatry
David Silbersweig
DFCI
Charles Stiles
John Alberta
Claire Sauvageot
Mark Marchionni
Erin Davis
Thank you!
Purdue University
Graham Cooks
Livia Eberlin
Allison Dill
Boston University
Allen Tannenbaum
Rush University
David Bennett
Brain Science Foundation
Daniel E. Ponton Fund for the Neurosciences
Pilocytic Low-Grade Astrocytoma Fund, DFCI
James S. McDonnell Foundation
NIH Director’s New Innovator Award
National Center for Image Guided Therapy
(NCIGT)
Prosolia Inc.
Justin Wiseman
Bruker Daltonics
11