SHAPE Society

1. Zahi A. Fayad, PhD
Associate Professor
Imaging Science Laboratories
Depts of Radiology and Medicine
The Zena and Michael A. Wiener Cardiovascular
Institute and The Marie-Josee and Henry R. Kravis
Cardiovascular Health Center
Mount Sinai School of Medicine, New York, NY
11-12-2005 AEHA - Shape Guidelines
Dallas, Tx
Noninvasive Molecular Imaging of Atherosclerosis
With MRI - The Future Lies in Molecular Imaging,
The question is which molecule

2. Choudhury RP; Fuster V; Fayad ZA Nature Drug Dev. 2004;3:913-925
Atherosclerosis: A Progressive Process
Imaging Science Laboratories

3. Noninvasive Detection of Plaque Instability
with Use of Radiolabeled (SPECT) Annexin A5 in Patients
with Carotid-Artery Atherosclerosis
Kietselaer BLJH et al. NEJM 350;14;April 1, 2004
•Left-Sided TIA 3 days before
imaging
•Annexin A5 hot spot
•Annexin A5 staining
•Right-sided TIA 3
months before
imaging
ªNo Annexin hot spot
•No Annexing A5
staining

4. Davies JR, Rudd JH, Stroke 2005 in press
Plaque Activity Assessment -18F-FDG- Macs
PET/CT or PET/MR
Rudd JHF et al. Circ 2002;105:2708-2711

5. • Paramagnetic (Iron)
• Superparamagnetic (Gd)
• Targeted Contrasts/Carriers
Current Contrast-Enhanced MR Limited
Plaque Activity Information
Gd-chelate enhanced
-non specific
Targeted enhanced contrast
agent - specific interaction with
plaque
Choudhury RP; Fuster V; Fayad ZA Nature Drug Dev. 2004
Aorta
IVC
Non specific Targeted

6. Molecular Imaging Contrast Agents
Choudhury RP; Fuster V; Fayad ZA Nature Drug Dev. 2004
Lipinski MJ; Fuster V; Fayad ZA Nature Cardiovascular Medicine 2004
• Improve MR detection sensitivity
• CA w high-relaxivity
- e.g., small peptides
• Increase payload of contrast agent
• Targeted/Functional
• Colloidal Carriers/Lipid based
- Liposomes
- microemulsions
- micelles
- lipoproteins
• Targets

7. Age Characterization Using Fibrin-Targeted
MR Contrast Agent in Chronic Thrombus
0
2
4
6
8
10
12
1
* P<0.01
Pre Acute 48 hours 1 week 2 weeks 3 weeks 4 weeks 6 weeks 8 weeks
ContrasttoNoiseRatio(CNR)
*
*
*
4
x
Acute Thrombus
4
x
1-week-old Thrombus
10
x
8-week-old Thrombus
4
x
6-week-old Thrombus
Sirol M; Fayad ZA et al. Atherosclerosis 2005:182:79-85
Sirol M; Fayad ZA et al. Circ 2005;112:1594-1600

8. MATRIX METALLOPROTEINASES TARGETING
- Gd-peptide Approach
IHC of atherosclerotic
plaque for MMP2
MMP Zymography of
aorta of ApoE mouse
40x
40x
P947 (Guerbet)
short peptide MMPs inhibitor
100 µmol/kg
Amirbekian V; Fayad ZA et al. AHA 2005;D220 @ NOV 15 @ 4:45PM
Imaging Science Laboratories
NER P947 vs. P1135
0
0.5
1
1.5
2
2.5
3
1 2 3 4 5
Time-Point (1= Pre-Contrast;2= 1hr post;3= 2
hrspost;4 = 3hrs post;5= 22 hrspost)
NER
P947 External
Standard
P947
control
P1135

9. Lipid-Based Particulates
Lipoproteins:
Chylo, VLDL, TG, LDL, HDL
• Gd, Iron, Fluorescents, etc.
• Ab, peptide, etc.

10. In Vivo Aortic Atherosclerotic Plaque in ApoE-KO Mice
Gd Immunomicelles Macrophages scavenger receptor types I&II
Murine (CD204)
9.4T
TR=500 ms; TE= 8.6 ms
101 µm in plane spatial resolution
500 µm slice thickness
Injected - 13.7µmol/kg
Mφ
A
C
Amirbekian V; Fayad ZA et al.; AHA 2005;Nov 15;D220 @2pm
Imaging Science Laboratories

11. Sirol M; Fayad ZA et al. Circulation. 2004;109:2890-2896
Sirol M; Fayad ZA et al. AHA 2005 Nov 15 D220 @ 2:45 pm
Atherosclerotic Plaques Detected by
Gadofluorine-Enhanced In Vivo MRI
Imaging Science Laboratories

12. Gadofluorine M Plaque MR
ApoE-KO mouse lesions
• Yellow - localization in ECM (tenascin)
• No cellular uptake
Mφ
Lymphocytes
Smooth muscle cells
mAb (green)
Gadofluorine-carbocyanin conjugate (red)
DAPI cell nuclei (blue)
Sirol M; Fayad ZA et al. AHA 2005 NOV 15 D220;4:30PM
9.4T
TR=500 ms; TE= 8.6 ms
101 µm in plane spatial resolution
500 µm slice thickness
100µmol/Kg (gadofluorine M)

13. Kelly KA, Weissleder R et al. Circ Res. 2005;96:327-336.
Multimodal CLIO-Cy5.5-VCAM-1 Peptide (Iron-peptide)
TARGETED SUPERPARAMAGNETIC NP
Amino-Cross-linked Iron Oxide
=Amino-CLIO

14. In vivo Macrophage Imaging Using Iron Oxide Particles in
Atherosclerotic Rabbits
GRASP (white spots)T2* GRE (dark spots)
24h POST CONTRAST

15.  Lipoprotein carrier
 Natural Affinity to plaques
 Small size (7-12 nm)
 Does not trigger immune reactions
 Easily reconstituted
 Efficient drug carrier
Frias JC et al. J. Am. Chem. Soc. 2004 126(50): p. 163-167
HDL Properties

16. ApoA-I
DMPE-NBD
(optical)
Unesterified
Cholesterol
Cholesterol
Ester
Gd-DTPA-
DMPE
(MRI)
Triglyceride
Phospholipid
r1p = 10.4 mM-1
s-1
(1.5 T, 25 °C)
Imaging Science LaboratoriesFrias JC et al. J. Am. Chem. Soc. 2004 126(50): p. 163-167

17. 119
98
52
37
30
22
apoA-I
H
D
L
rH
D
L
+
Gd-D
T
PA
-D
M
PE
kDa
GdGd-DTPA-DMPE-DTPA-DMPE
rr1p1p = 10.4 mM= 10.4 mM-1-1
ss-1-1
(1.5 T, 25(1.5 T, 25 °°C)C)
4-12% SDS-PAGE
DPPE-NBD
HN
O O
P
O
OH
O
H O
O
O
NO2
N
O
N
14
14
reconstitutiondelipidation
Control
N
N
H
O
O
O
P
O
OH
O
H O
O
O
N
O
N
O O
O
O O
OO
Gd
J. C. Frías et al., J. Am. Chem. Soc., 2004, 126, 16316
12
12

18. 0
10
20
30
40
50
60
0 10 20 30 40 50 60 70 80
Fraction number
Abs280nm
rHDL-sphe
rHDL-disc
apoE-KO plasma (x10)
incubated HDL
VLDL
LDL IDL HDL
Cholesterol estersTriglycerides
Gadolinium complex
Phospholipids
ApoA-I
Reconstituted hollow
spherical HDL
Reconstituted
discoidal HDL
Native incubated HDL

19. Pre-contrast 48 hours post-contrast1 hr post-contrast 24 hr post-contrast
ApoE-KO mice; 4.36 mmol/kg @ 9.4-T MRM rHDL-Gd-DTPA-DMPE
Frias JC et al. J. Am. Chem. Soc. 2004 126(50): p. 163-167
DAPI srHDL-NBD CD68:RPE
Mφ
rHDL for Plaque Detection
in ApoE KO Mice

20.  Silvia Aguiar, MD
 Gilbert Aguinaldo, MD
 Vardan Amirbekian
 Lucy Feliciano
 JC Frias, PhD
 Fabien Hyafil, MD
 Vitalii Itskovich, PhD
 Alex Leber, MD
 Enric Lopez, MD, PhD
 Michael Lipinski
 Frank Macaluso, RT
 NIH/NHLBI R01HL71021; R01HL78667; R43HL71470
 Intellectual Property Managed by Mount Sinai
 Venkatesh Mani, PhD
 Lena Mara
 Gabor Mizsei, MS
 Dan Samber, MS
 Karen Saebo, PhD
 Marc Sirol, MD
 Cheuk Tang, PhD
 Hiraoki Taniguchi, MD
 Esad Vucic, MD
 Karen Weinshelbaum
Imaging Science Laboratories (ISL)
Mount Sinai School of Medicine

21.  Valentin Fuster, MD, PhD
 Donald Smith, MD
 Jonathan Halperin, MD
 Pedro Moreno, MD
 K-R Purushothaman, MD
 Michael Farkouh, MD
 Mary Ann McLaughlin, MD
 Jeffrey Olin, MD
 Alison Schecter, MD
Radiology
 Burton Drayer, MD
 Zvi Lefkovitz, MD
 Bradley Delman, MD
 Emil Cohen, MD
CVI
 Jesse Weinberger, MD
 Stanley Tuhrim, MD
Neurology
 Sanjay Rajagopalan, MD
 Javier Sanz, MD
 Santo Dellegrottaglie, MD
Clinical CMR/CT
Mount Sinai School of Medicine
Collaborators
 Edward A. Fisher, MD, PhD
 John Postley, MD
 Carlton Boxhill, MD
 Lisa Hudgings, MD
 Christine Williams, MD
 Tom Starc, MD
 Sam Gidding, MD
 Kevin Williams, MD
 Yukihiko Momiyama, MD
 Jean-Francois Toussaint, MD, PhD
Pathology
 John T. Fallon, MD, PhD
Pediatrics
 Jeff Saland, MD

22. The following relationships exist related to this presentation:
SOME CONTRAST AGENTS DISCUSSED ARE NOT FDA APPROVED
Presenter Disclosure Information
 NIH/NHLBI R01HL71021; R01HL78667; R43HL71470
 Siemens Medical Solutions
 Pfizer/GE HealthCare
 Guerbet
 Schering
 Society of Atherosclerosis Imaging (www.SAI.ORG)
President
 Society of Cardiovascular Magnetic Resonance Board
Member
 Intellectual Property Managed by Mount Sinai