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Sponsored by:
Mirosław Janowski,
MD, PhD
Associate Professor,
Diagnostic Radiology and
Nuclear Medicine
University of Maryland School
of Medicine
Piotr Walczak,
MD, PhD
Professor, Diagnostic
Radiology and
Nuclear Medicine
University of Maryland
School of Medicine
Wojciech
Lesniak, PhD
Research Associate,
Department of
Radiology
John Hopkins School
of Medicine
A New Frontier of Precision
Medicine: Using PET for Image-
Guided Neurointerventions
Sponsored by:
A New Frontier of Precision
Medicine: Using PET for Image-
Guided Neurointerventions
Experts discuss how PET/CT imaging
can be used to enable image-guided
neurointerventions and to study
targeted delivery and clearance of
therapeutic agents.
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Miroslaw Janowski, MD, PhD
Piotr Walczak, MD, PhD
Wojciech Lesniak, PhD
A new frontier of precision medicine: using PET for
image-guided neurointerventions
•MJ and PW: co-founders and co-owners of IntraART, LLC
•MJ and PW: co-founders and co-owners of Ti-Com, LLC
Disclosures
Routes for delivery of therapeutic agent to the brain
Systemic delivery
Pros: Easy drug administration
Cons: 1) Whole-body toxicity
2) Need for high dose of drug
3) Limited penetration
of blood-brain barrier (BBB)
Intra-arterial delivery
Pros: 1) Focal BBB opening
2) Wide spatial distribution
3) Minimal invasiveness
4) Facile repeatability
Cons: 1) Need for advanced
imaging
Intracerebral delivery
Pros: Beyond BBB
Cons: 1) Skull and brain puncture
2) Limited spatial distribution
3) Need for long-lasting
infusions (days) and for
special devices
Intrathecal/CSF delivery
Pros: 1) Beyond BBB
2) Minimal invasiveness
3) Wide spatial distribution
4) Facile repeatibility
Cons: 1) Limited drug penetration
2) Limited possibility for focal
targeting
Historical perspective
Surgery, 1970
10 patients, 2 hemiplegia, 2 hemiparesis
Arch Neurol. 1964;11(6):618-625
French, J.D., et al., Effects of intracarotid administration of nitrogen mustard
on normal brain and brain tumors. J Neurosurg, 1952. 9(4): p. 378-89.
Zylber-Katz E. et al. Clin Pharmacol Ther
67:631–641, 2000
Variability!!!
Historical perspective
Cancer, 2000
• Between March 1994 and November 1997, 5 universities treated 221 adult
patients with intra-arterial chemotherapy with or without osmotic opening of
the BBB (2464 procedures).
• Of evaluable patients with PCNSL, 40 of 53 (75%) achieved complete
response (CR)
• All evaluable patients with PNET (n = 17), metastatic disease (n = 12), or germ
cell tumor (n = 4) achieved stable disease (SD) or better.
• Of 57 evaluable patients with glioblastoma multiforme, 45 (79%) achieved SD
or better.
Historical perspective
Super-selective intra-arterial drug delivery to the
brain – toward re-invention
FROM BED TO BENCH AND BACK AGAIN
our journey towards effective delivery of therapeutics to the brain
Intra-arterial route with transient opening of the blood brain barrier and image
guidance holds great promise
Gupta et al. Science Trans. Med. 4;155; 2012
Global dysmyelination
Children with PMD transplanted
with stem cells
T2w MRI @ 1 year
Uchida et al. Science Trans. Med. 4;155; 2012
Neonatal shiverer mice transplanted
with stem cells
WHY INTRA-ARTERIAL ROUTE?
WHY INTRA-ARTERIAL ROUTE?
WHY INTRA-ARTERIAL ROUTE?
Zylber-Katz E. et al. Clin Pharmacol Ther 67:631–641
THE CHALLENGE OF IA DELIVERY OF THERAPEUTIC
AGENTS TO THE BRAIN
Walczak et al. Stroke. 2008 May;39(5):1569-74.
• Cranial window implantation
• Catheterization of ICA
• Visualization of trans-catheter
perfusion and osmotic opening
cortical BBB guided by MRI
• Intravital multiphoton microscopy
of IA IgG extravasation
• PET imaging
MULTIMODALITY IMAGING PLATFORM FOR GUIDING
IA DELIVERY IN MICE
GE-EPI
fast (2s/volume), highly sensitive to
MRI contrast
FastSlow
Walczak P, et al. J Cereb Blood Flow Metab 2017.
VISUALIZATION TRANS-CATHETER PERFUSION WITH
INTERVENTIONAL MRI
EXCESSIVE INFUSED SPEED INTO ICA (0.2ml/min)
CAUSED BRAIN DAMAGE
Pterygopalatine Artery NOT Ligated
VISUALIZATON OF IA CELL INFUSION UNDER REAL
TIME MRI
Pterygopalatine Artery Ligated
VISUALIZATON OF IA CELL INFUSION UNDER REAL
TIME MRI
Small cells (mGRPs) Large cells (rMSCs)
VISUALIZATON OF IA CELL INFUSION UNDER REAL TIME
MRI
3-dayoldouabainstroke
Beforetransplantation
Aftertransplantation
Red– transplantedcells
Green– vessels(vWF)
DAPI
Intraarterialinjection ofGRPs
Jablonska et al. J Cereb Blood Flow
Metab 2018 May;38(5):835-846
CAN CELLS EXTRAVASATE?
30 min
after TX
120 min
after TX
50 min
after TX
90 min
after TX
VISUALIZATION OF DIAPEDESIS BY TWO PHOTON
MICROSCOPY
Jablonska et al. J Cereb Blood Flow
Metab 2018 May;38(5):835-846
• Impermeable to water soluble drugs >180 Da
• Most therapeutic agents are larger
Most capillaries Brain capillaries
Hyperosmotic
agent (Mannitol)
BLOOD BRAIN BARRIER AND ITS CONTROLLED OPENING
Disruption of BBB
with Intraarterial
Mannitol (25%)
Transcatheter DSC MRI
GE-EPI
T1 MRI +Gd for
assessment of BBB
integrity
Catheter placed in ICA
Animal placed in 11.7T
MRI scanner
MRI-GUIDED BBBD IN MICE
Mouse
Mri-guided intraarterial catheter-based method for
predicting territory of local blood brain barrier opening
US Patent: WO2015179258A3
MRI-GUIDED OBBBO IN MICE
Chu et al. Front Neurol. 2018; 9: 921
SAFETY OF OBBBO IN MICE
Chu et al. Front Neurol. 2018; 9: 921
VARIABILITY OF CORTICAL PERFUSION DURING
CONTRAST AGENT INFUSION VIA ICA (O.5ml/min)
Chu et al. J Control Release
2020 Jan 10;317:312-321
Dynamic EPI
TEMPORARY CLOSURE OF THE CONTRALATERAL CCA
OPENS CORTICAL PERFUSION
Chu et al. J Control Release
2020 Jan 10;317:312-321
cCCA
closure
Mannitol
Gd
Dynamic imaging of IgG extravasation under 2 photon microscopy
Rhodamine
(0.53 kDa)
FITC-antibody
(1.53 kDa)
Chu et al. J Control Release
2020 Jan 10;317:312-321
Chu et al. J Control Release
2020 Jan 10;317:312-321
DYNAMIC IMAGING OF IgG EXTRAVASATION UNDER
TWO PHOTON MICROSCOPY
Chu et al. J Control Release
2020 Jan 10;317:312-321
HISTOLOGICAL ASSESSMENT OF BV EXTRAVASATION
MRI is excellent for visualization trans-catheter perfusion, monitoring BBB status but
sensitivity is rather low and insufficient for detection of injected drugs
Intravital 2PM offers subcellular resolution and high sensitivity, but very small
imaging area and is hardly quantitative
PET is highly sensitive enables quantitative, whole body imaging of injected drugs
BOTTOM LINE
Technique Resolution Sensitivity Quantification
MRI 10 - 100 µm µ - m Mol Non-
quantitative
PET 1 - 2 mm p - n Mol Quantitative
Comparison of MRI and PET
89Zr or 64Cu
p - nMol
PET contrast agent
SPIO
mMol
MRI contrast agent
Positron Emission Tomography (PET)
https://en.wikipedia.org/wiki/Positron_emission_tomography
“Organic-atom-like”: 18F (t1/2=109.7 min), 11C
(t1/2=20.33 min), 13N (t1/2=9.97 min), 15O (t1/2=2.03
min), 124I (t1/2=4.18 d)
Radiometals: 68Ga (t1/2=68 min), 64Cu (t1/2=12.7
h), 89Zr (t1/2=78.4 h)
1 2 .5 2 5 .0
-1 0
0
1 0
2 0
3 0
4 0
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
tim e [m in ]
mV
Abs.at280nm
0 5 1 0 1 5 2 0 2 5
0
5 .0 1 0 5
1 .0 1 0 6
1 .5 1 0 6
0
1 0
2 0
3 0
4 0
5 0
NH2 +
O
N
HN
O
O N
NH
OH HO
O
O
N
HOH
N
S
H
N
SCN
NH
O
N
H
N
O
O N
NH
OH HO
O
O
N
HOH
N
S
H
N
N
H
S
NH
O
N
H
N
O
O N
NH
OH
HO
O
O
N
HOH
N
O
H
N
N
H
S
+ 89Zr4+
NH
O
N
HN
O
O N
NH
O
O
O
O
N
OH
N
S
H
N
N
H
S
89
Zr4+
m /z
1 4 0 0 0 0 1 5 0 0 0 0 1 6 0 0 0 0
B V
B V -D F O
150215
152410
Radiolabeling of Bevacizumab with Zirconium-89
Bevacizumab (BV) DFO-Bz-NCS
HEPES, pH=7.2
1 h, RT
[89Zr]BV
BV-DFO
pH=9, 1 h, 37 oC
1:5 molar ratio
Abs.at450nm
2 5  g /m L 5 0  g /m L 1 0 0  g /m L
0 .2
0 .4
0 .6
0 .8
1 .0
B V
B V -D F O
BV
[89Zr]BV
Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622
MALDI-TOF Binding to VEGF SEC
Experimental Setup
I - intra-arterial infusion with intact blood brain barrier (IA/IBBB)
II - osmotic blood brain barrier opening and intra-arterial infusion (OBBBO/IA)
III - osmotic blood brain barrier opening and intravenous infusion (OBBBO/IV)
Evaluated macromolecules: Antibodies, Nanobodies and
Poly(amidoamine) dendrimers
T im e [m in ]
%ID/cc
0 5 1 0 1 5 2 0 2 5 3 0
0
5
1 0
1 5
2 0
2 5
3 0
3 5
Time [min]%ID/cc
0 5 10 15 20 25 30
0
10
20
30
40
ipsilateral hemisphere heart
OBBBO and IA infusionIA infusion with IBBB OBBBO and IV infusion
0
75
*
*
Kinetics of [89Zr]BV Uptake in Ipsilateral Hemisphere
Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622
0
75
Distribution of [89Zr]BV in Brain
R
S
T
R
L
S
T
R
C
T
X
R
H
IP
L
H
IP
T
H
A
C
B
B
F
S
H
Y
P
R
A
M
Y
L
A
M
Y
B
S
C
G
S
C
O
L
F
R
M
ID
L
M
ID
L
IC
R
IC
0
5
1 0
1 5
2 0
%ID/cc
G ro u p I
G ro u p II
G ro u p III
STR striatum
CTX cerebral cortex
HIP hippocampus
THA thalamus
CB cerebellum
BFS basal forebrain septum
HYP hippocampus
AMY amygdala
BS brain stem
CG central gray
SC superior colliculi
OLF olfactory bulb
MID midbrain
IC inferior colliculi
*
*
*
*
*
*
*
**
*
*
OBBBO and IA infusionIA infusion with intact BBB IV infusion and OBBBO
Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622
40
0
40
0
40
0
1 h pi 1 h pi 24 h pi
1 h pi 24 h pi
%ID/cc
1
h
2
4
h
1
h
2
4
h
1
h
2
4
h
0
5
1 0
1 5
2 0
2 5
IA /B B B I
B B B O /IA
IV /B B B O
OBBBO/IAIA/BBBI
IV/OBBBO
Whole Body PET/CT Imaging of [89Zr]BV
24 h pi
ipsilateral hemisphere
Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622. doi: 10.2967/jnumed.118.218792
Ex Vivo Biodistribution of [89Zr]BV
*
*
OBBBO/IA
IA/IBBB
OBBBO/IV
Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622. doi: 10.2967/jnumed.118.218792
Perspectives
Radiolabeling of Anti-gelsolin Nanobody with 89Zr
NB
DFO-Bz-NCS
HEPES, pH=7.2
1 h, RT
89ZrNB
pH=9, 1 h, 37 oC
1:5 molar ratio
NB-DFO
NB-DFO
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
T im e [m in ]
%ID/cc
0 5 1 0 1 5 2 0 2 5 3 0
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
O B B B O /IA
IA /B B B I
IV /O B B B O
OBBBO/IA IA/BBBI OBBBO/IV
0
100
%ID/cc
Kinetics of [89Zr]NB Uptake in Ipsilateral Hemisphere
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
OBBBO/IA 1 h pi
OBBBO/IA 24 h pi
IA/BBBI 1 h pi
IA/BBBI 24 h pi
OBBBO/IV 1 h pi
OBBBO/IV 24 h
pi
Whole Body PET/CT Imaging of [89Zr]NB11
100
0
100
0
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
Synthesis of Poly(amidoamine) Dendrimers
NH
Generation 2
Generation 1
N N
N
N
N
N
NH2
NH2
H2N
NH2
NH2
H2N
H2
N
2
N N
N
N
N
N
N
N
N
N
NH2
NH2
NH2
H2N
N
N
N
N
H2N
H2N
H2N
H2N
H2N NH2
NH2
H2N
NH2
H2N
NH2
NH2
(2)
NH2
H2N
(1)
O
OMe
(1)
O
OMe
(2)
NH2
H2N
Generation 3, etc.
Up to generation 12
Core
NN
O
N
O
N
N
H2N NH2
Generation 0
N N
H2N
H2N
NH2
NH2
N N
NH
N
O
H
H
HN
H2
O
H
H2 H
(1)
O
OMe
(2) NH2H2N
2
2
PAMAM Dendrimer Based Nanoparticles
therapeutics
contrast agents (X-ray, MRI)
targeting
agents
NHCCH3
O
CH2CHCH2OH
OH
CH2CHCH2OH
OH
N
NHCCH2CH2C
O
O
O-
NHC(OCH2CH2)nOH
O
Applications:
• Selective and controlled drug deliver
• Gene transfection
• Contrast agents
• Immunoassay and molecular diagnostic
platforms
• Nanocomposites
surface modification
HN
(NH2)64
1) DFO-Bz-NCS, 2) Glycidol
O
N
NH
O
ON
HN
HOOH
O
O
N
OH H
N
S
H
N
N
H
S
OH
OH
NH
12
89
Zr4+
O
N
NH
O
ON
HN
O
O
O
O
N
O H
N
S
H
N
N
H
S
89
Zr4+
NH OH
OH
NH
12
3 3
OH
OH
N
49
OH
OH
OH
OH
N
49
OH
OH
Modifications of Dendrimer and Radiolabeling
with Zirconium-89
89ZrG4(DFO)3(Bdiol)110G4(DFO)3(Bdiol)110G4(NH2)64
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
OBBBO/IA
IA/BBBI
OBBBO/IV
PET Imaging and Kinetics of 89ZrG4(DFO)3(Bdiol)110
Uptake in Ipsilateral Hemisphere
0
30
0
40
0
6
1 h piFrames 5 to 10 min
%ID/cc
%ID/cc
%ID/cc
T im e [m in ]
%ID/cc
0 5 1 0 1 5 2 0 2 5 3 0
0
1
2
3
4
5
6
O B B B O /IA
IA /B B B I
IV /O B B B O
N
S
*
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019,
doi: 10.1007/s00259-019-04347-y.
OBBBO / IA 24 h pi
IA/BBBI 1 h pi
IA / BBBI 24 h pi
IV/BBBI 1 h pi
IV / BBBI 24 h pi
100
0
100
0
OBBBO/IA 1 h pi
Whole Body PET-CT Imaging of 89ZrG4(DFO)3(Bdiol)110
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
Ex Vivo Biodistribution of 89ZrG4(DFO)3(Bdiol)110
at 24 h After Infusion
B
lood
Ipsilateralhem
isphere
C
ontralateralhem
isphere
H
eart
Lung
Liver
Thym
us
S
tom
ach
P
ancreas
S
pleen
K
idney
Fat
M
uscle
B
ladder
0
5
10
15
20
25
30
35
40
%ID/g
O B B B O /IA
IV /O B B B O
IA /B B B I
Ipsilateral hem
isphere
C
ontralateral hem
isphere
0 .0 0
0 .0 5
0 .1 0
0 .1 5
%ID/g
Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
Conclusions
 IA delivery of antibodies or nanobodies might be an attractive therapeutic
platform for treatment of CNS disorders
 Appropriate surface modification of PAMAM dendrimers with targeting agents
may facilitate specific uptake for selective drug delivery
Evaluation of PSMA-Targeted PAMAM Dendrimer
Lesniak WG et al., Mol Pharm. 2019, doi: 10.1021/acs.molpharmaceut.9b00181
Rhodamine
64Cu
PAMAM
G4
Lys-Glu-urea
PSMA+
PSMA-
Optical imaging
25
0
PET-CT
PSMA+ PSMA-
Optical and Photoacoustic Imaging of PSMA-targeted Dendrimer
5 hrs ai
PIP flu
dorsal ventraldorsal dorsal ventraldorsal
24 hrs ai
ventraldorsal
48 hrs ai
ventraldorsal
72 hrs ai
flu
1 2
3
4 5 6 7
8 9 10
11 12
13
14
15
PIP
PIPflu
1 - heart, 2 - lung, 3 - liver, 4 - pancreas, 5 - spleen, 6 - fat, 7 - kidneys,
8 -muscle,9 - small intestines, 10 - salivary glands, 11 - bladder, 12 - bone,
13 - lacrimal gland,14 - PSMA+ PC3 PIP, 15 - PSMA- PC3 flu
First-in-man real-time MRI-guided endovascular neurointervention
Zawadzki et al., JNISfaster infusion slower infusion
Publication: JNIS/BMJ Case REPORTS
• Intra-arterial route of delivery of therapeutic agents to the brain is becoming increasingly attractive.
• Advanced imaging is essential to precisely guide the procedure
• Advances
• Progress in imaging of intra-arterial interventions (MRI + PET)
• Drug design especially related to safety aspects (antibodies + macromolecules)
• Catheter design – reaching distal vessels allows for precise interventions
• Advantages
• Decreasing exposure of therapeutic agents to the peripheral organs
• No skull opening
• Easily repeatable
• Future directions:
• Design of MRI-visible microcatheters for endovascular neurointerventions
• Engineering of cells and molecules to better benefit from intra-arterial delivery
• Application of PET-MRI in interventional suites to benefit from advantages of high spatial resolution of MRI and high
sensitivity of PET during the same procedure to allow for unprecedented precision
Summary
Acknowledgments
GROUP:
Anna Jablonska
Yajie Liang
Chengyan Chu
Xiaoyan Lan
Yue Gao
Jipeng Zhang
Dariush Aligholizadeh
COLLABORATORS:
Graeme Woodworth
Rao Gullapalli
Linda Chang
Thomas Ernst
Ze Wang
Collaborators:
Monica Pearl
Martin Pomper
Guanshu Liu
Collaborators:
Izabela Malysz-Cymborska
Dominika Golubczyk
Lukasz Kalkowski
Wojciech Maksymowicz
Collaborators:
Barbara Lukomska
Luiza Stanaszek
Anna Andrzejewska
Katarzyna Drela
Sylwia Dabrowska
Piotr Rogujski
Collaborators:
Michal Zawadzki
Malgorzata Dorobek
Blazej Nowak
Collaborators:
Miguel Oliveira
Eduarda Oliveira
Johannes Boltze
Tim Magnus
Matthias Gelderblom
Peter Ludewig
Jan Gettemans
Olivier Zwaenepoel
Pedro Ramos Cabrer
Jesus Ruiz-Cabello
Supported by MSCRFII-0193, MSCRFII-0052, MSCRFE-0178, RO1 NS076573, 2RO1 NS045062, EUREKA
RO1DA026299, R21NS106436, R01NS091100, R01NS091110, P41 EB024495 EXPLORE ME
SIGN 2019
SIGN 2021
The University of Nottingham, UK
David Walker, Chair
Ruman Rahman, Deputy Chair
Richard Grundy
Emma Campbell, Project Manager
University of Strathclyde, UK
Marie Boyd
Alexander Mullen
Newcastle University, UK
Gareth Veal
Bristol Royal Hospital for Children, UK
Stephen Lowis
UCL Institute of Child Health
Darren Hargrave
Johns Hopkins University, US
Henry Brem
Monica Pearl
Jordan Green
Miroslaw Janowski
Kenneth Cohen
Piotr Walczak
National Cancer Institute
Katherine Warren
VU University Medical Centre, Amsterdam
Dannis van Vuurden
Fast Track Pharma Ltd
Steven Powell
Consortium – invitation to register
Sponsored by:
Mirosław Janowski,
MD, PhD
Associate Professor,
Diagnostic Radiology and
Nuclear Medicine
University of Maryland School
of Medicine
Piotr Walczak,
MD, PhD
Professor, Diagnostic
Radiology and
Nuclear Medicine
University of Maryland
School of Medicine
Wojciech
Lesniak, PhD
Research Associate,
Department of
Radiology
John Hopkins School
of Medicine
To learn more about Scintica’s solutions for
preclinical PET-CT imaging, please visit:
https://www.scintica.com/products/superargus-
sedecal-pet-ct-preclinical-systems/
Thank you!

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A New Frontier of Precision Medicine: Using PET for Image-Guided Neurointerventions

  • 1. Sponsored by: Mirosław Janowski, MD, PhD Associate Professor, Diagnostic Radiology and Nuclear Medicine University of Maryland School of Medicine Piotr Walczak, MD, PhD Professor, Diagnostic Radiology and Nuclear Medicine University of Maryland School of Medicine Wojciech Lesniak, PhD Research Associate, Department of Radiology John Hopkins School of Medicine A New Frontier of Precision Medicine: Using PET for Image- Guided Neurointerventions
  • 2. Sponsored by: A New Frontier of Precision Medicine: Using PET for Image- Guided Neurointerventions Experts discuss how PET/CT imaging can be used to enable image-guided neurointerventions and to study targeted delivery and clearance of therapeutic agents.
  • 3. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services
  • 4. To access webinar content, Q&A reports, FAQ documents, and information on lab workshops, subscribe to our mail list
  • 5. Miroslaw Janowski, MD, PhD Piotr Walczak, MD, PhD Wojciech Lesniak, PhD A new frontier of precision medicine: using PET for image-guided neurointerventions
  • 6. •MJ and PW: co-founders and co-owners of IntraART, LLC •MJ and PW: co-founders and co-owners of Ti-Com, LLC Disclosures
  • 7. Routes for delivery of therapeutic agent to the brain Systemic delivery Pros: Easy drug administration Cons: 1) Whole-body toxicity 2) Need for high dose of drug 3) Limited penetration of blood-brain barrier (BBB) Intra-arterial delivery Pros: 1) Focal BBB opening 2) Wide spatial distribution 3) Minimal invasiveness 4) Facile repeatability Cons: 1) Need for advanced imaging Intracerebral delivery Pros: Beyond BBB Cons: 1) Skull and brain puncture 2) Limited spatial distribution 3) Need for long-lasting infusions (days) and for special devices Intrathecal/CSF delivery Pros: 1) Beyond BBB 2) Minimal invasiveness 3) Wide spatial distribution 4) Facile repeatibility Cons: 1) Limited drug penetration 2) Limited possibility for focal targeting
  • 8. Historical perspective Surgery, 1970 10 patients, 2 hemiplegia, 2 hemiparesis Arch Neurol. 1964;11(6):618-625 French, J.D., et al., Effects of intracarotid administration of nitrogen mustard on normal brain and brain tumors. J Neurosurg, 1952. 9(4): p. 378-89.
  • 9. Zylber-Katz E. et al. Clin Pharmacol Ther 67:631–641, 2000 Variability!!! Historical perspective Cancer, 2000 • Between March 1994 and November 1997, 5 universities treated 221 adult patients with intra-arterial chemotherapy with or without osmotic opening of the BBB (2464 procedures). • Of evaluable patients with PCNSL, 40 of 53 (75%) achieved complete response (CR) • All evaluable patients with PNET (n = 17), metastatic disease (n = 12), or germ cell tumor (n = 4) achieved stable disease (SD) or better. • Of 57 evaluable patients with glioblastoma multiforme, 45 (79%) achieved SD or better.
  • 11. Super-selective intra-arterial drug delivery to the brain – toward re-invention
  • 12. FROM BED TO BENCH AND BACK AGAIN our journey towards effective delivery of therapeutics to the brain Intra-arterial route with transient opening of the blood brain barrier and image guidance holds great promise
  • 13. Gupta et al. Science Trans. Med. 4;155; 2012 Global dysmyelination Children with PMD transplanted with stem cells T2w MRI @ 1 year Uchida et al. Science Trans. Med. 4;155; 2012 Neonatal shiverer mice transplanted with stem cells WHY INTRA-ARTERIAL ROUTE?
  • 16. Zylber-Katz E. et al. Clin Pharmacol Ther 67:631–641 THE CHALLENGE OF IA DELIVERY OF THERAPEUTIC AGENTS TO THE BRAIN Walczak et al. Stroke. 2008 May;39(5):1569-74.
  • 17. • Cranial window implantation • Catheterization of ICA • Visualization of trans-catheter perfusion and osmotic opening cortical BBB guided by MRI • Intravital multiphoton microscopy of IA IgG extravasation • PET imaging MULTIMODALITY IMAGING PLATFORM FOR GUIDING IA DELIVERY IN MICE
  • 18. GE-EPI fast (2s/volume), highly sensitive to MRI contrast FastSlow Walczak P, et al. J Cereb Blood Flow Metab 2017. VISUALIZATION TRANS-CATHETER PERFUSION WITH INTERVENTIONAL MRI
  • 19. EXCESSIVE INFUSED SPEED INTO ICA (0.2ml/min) CAUSED BRAIN DAMAGE
  • 20. Pterygopalatine Artery NOT Ligated VISUALIZATON OF IA CELL INFUSION UNDER REAL TIME MRI
  • 21. Pterygopalatine Artery Ligated VISUALIZATON OF IA CELL INFUSION UNDER REAL TIME MRI
  • 22. Small cells (mGRPs) Large cells (rMSCs) VISUALIZATON OF IA CELL INFUSION UNDER REAL TIME MRI
  • 23. 3-dayoldouabainstroke Beforetransplantation Aftertransplantation Red– transplantedcells Green– vessels(vWF) DAPI Intraarterialinjection ofGRPs Jablonska et al. J Cereb Blood Flow Metab 2018 May;38(5):835-846 CAN CELLS EXTRAVASATE?
  • 24. 30 min after TX 120 min after TX 50 min after TX 90 min after TX VISUALIZATION OF DIAPEDESIS BY TWO PHOTON MICROSCOPY Jablonska et al. J Cereb Blood Flow Metab 2018 May;38(5):835-846
  • 25. • Impermeable to water soluble drugs >180 Da • Most therapeutic agents are larger Most capillaries Brain capillaries Hyperosmotic agent (Mannitol) BLOOD BRAIN BARRIER AND ITS CONTROLLED OPENING
  • 26. Disruption of BBB with Intraarterial Mannitol (25%) Transcatheter DSC MRI GE-EPI T1 MRI +Gd for assessment of BBB integrity Catheter placed in ICA Animal placed in 11.7T MRI scanner MRI-GUIDED BBBD IN MICE Mouse
  • 27. Mri-guided intraarterial catheter-based method for predicting territory of local blood brain barrier opening US Patent: WO2015179258A3 MRI-GUIDED OBBBO IN MICE Chu et al. Front Neurol. 2018; 9: 921
  • 28. SAFETY OF OBBBO IN MICE Chu et al. Front Neurol. 2018; 9: 921
  • 29. VARIABILITY OF CORTICAL PERFUSION DURING CONTRAST AGENT INFUSION VIA ICA (O.5ml/min) Chu et al. J Control Release 2020 Jan 10;317:312-321
  • 30. Dynamic EPI TEMPORARY CLOSURE OF THE CONTRALATERAL CCA OPENS CORTICAL PERFUSION Chu et al. J Control Release 2020 Jan 10;317:312-321
  • 31. cCCA closure Mannitol Gd Dynamic imaging of IgG extravasation under 2 photon microscopy Rhodamine (0.53 kDa) FITC-antibody (1.53 kDa) Chu et al. J Control Release 2020 Jan 10;317:312-321
  • 32. Chu et al. J Control Release 2020 Jan 10;317:312-321 DYNAMIC IMAGING OF IgG EXTRAVASATION UNDER TWO PHOTON MICROSCOPY
  • 33. Chu et al. J Control Release 2020 Jan 10;317:312-321 HISTOLOGICAL ASSESSMENT OF BV EXTRAVASATION
  • 34. MRI is excellent for visualization trans-catheter perfusion, monitoring BBB status but sensitivity is rather low and insufficient for detection of injected drugs Intravital 2PM offers subcellular resolution and high sensitivity, but very small imaging area and is hardly quantitative PET is highly sensitive enables quantitative, whole body imaging of injected drugs BOTTOM LINE
  • 35. Technique Resolution Sensitivity Quantification MRI 10 - 100 µm µ - m Mol Non- quantitative PET 1 - 2 mm p - n Mol Quantitative Comparison of MRI and PET 89Zr or 64Cu p - nMol PET contrast agent SPIO mMol MRI contrast agent
  • 36. Positron Emission Tomography (PET) https://en.wikipedia.org/wiki/Positron_emission_tomography “Organic-atom-like”: 18F (t1/2=109.7 min), 11C (t1/2=20.33 min), 13N (t1/2=9.97 min), 15O (t1/2=2.03 min), 124I (t1/2=4.18 d) Radiometals: 68Ga (t1/2=68 min), 64Cu (t1/2=12.7 h), 89Zr (t1/2=78.4 h)
  • 37. 1 2 .5 2 5 .0 -1 0 0 1 0 2 0 3 0 4 0 0 .0 0 .2 0 .4 0 .6 0 .8 1 .0 tim e [m in ] mV Abs.at280nm 0 5 1 0 1 5 2 0 2 5 0 5 .0 1 0 5 1 .0 1 0 6 1 .5 1 0 6 0 1 0 2 0 3 0 4 0 5 0 NH2 + O N HN O O N NH OH HO O O N HOH N S H N SCN NH O N H N O O N NH OH HO O O N HOH N S H N N H S NH O N H N O O N NH OH HO O O N HOH N O H N N H S + 89Zr4+ NH O N HN O O N NH O O O O N OH N S H N N H S 89 Zr4+ m /z 1 4 0 0 0 0 1 5 0 0 0 0 1 6 0 0 0 0 B V B V -D F O 150215 152410 Radiolabeling of Bevacizumab with Zirconium-89 Bevacizumab (BV) DFO-Bz-NCS HEPES, pH=7.2 1 h, RT [89Zr]BV BV-DFO pH=9, 1 h, 37 oC 1:5 molar ratio Abs.at450nm 2 5  g /m L 5 0  g /m L 1 0 0  g /m L 0 .2 0 .4 0 .6 0 .8 1 .0 B V B V -D F O BV [89Zr]BV Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622 MALDI-TOF Binding to VEGF SEC
  • 38. Experimental Setup I - intra-arterial infusion with intact blood brain barrier (IA/IBBB) II - osmotic blood brain barrier opening and intra-arterial infusion (OBBBO/IA) III - osmotic blood brain barrier opening and intravenous infusion (OBBBO/IV) Evaluated macromolecules: Antibodies, Nanobodies and Poly(amidoamine) dendrimers
  • 39. T im e [m in ] %ID/cc 0 5 1 0 1 5 2 0 2 5 3 0 0 5 1 0 1 5 2 0 2 5 3 0 3 5 Time [min]%ID/cc 0 5 10 15 20 25 30 0 10 20 30 40 ipsilateral hemisphere heart OBBBO and IA infusionIA infusion with IBBB OBBBO and IV infusion 0 75 * * Kinetics of [89Zr]BV Uptake in Ipsilateral Hemisphere Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622
  • 40. 0 75 Distribution of [89Zr]BV in Brain R S T R L S T R C T X R H IP L H IP T H A C B B F S H Y P R A M Y L A M Y B S C G S C O L F R M ID L M ID L IC R IC 0 5 1 0 1 5 2 0 %ID/cc G ro u p I G ro u p II G ro u p III STR striatum CTX cerebral cortex HIP hippocampus THA thalamus CB cerebellum BFS basal forebrain septum HYP hippocampus AMY amygdala BS brain stem CG central gray SC superior colliculi OLF olfactory bulb MID midbrain IC inferior colliculi * * * * * * * ** * * OBBBO and IA infusionIA infusion with intact BBB IV infusion and OBBBO Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622
  • 41. 40 0 40 0 40 0 1 h pi 1 h pi 24 h pi 1 h pi 24 h pi %ID/cc 1 h 2 4 h 1 h 2 4 h 1 h 2 4 h 0 5 1 0 1 5 2 0 2 5 IA /B B B I B B B O /IA IV /B B B O OBBBO/IAIA/BBBI IV/OBBBO Whole Body PET/CT Imaging of [89Zr]BV 24 h pi ipsilateral hemisphere Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622. doi: 10.2967/jnumed.118.218792
  • 42. Ex Vivo Biodistribution of [89Zr]BV * * OBBBO/IA IA/IBBB OBBBO/IV Lesniak WG, Chu C, et al. J. Nucl. Med. 2019, 60(5):617-622. doi: 10.2967/jnumed.118.218792
  • 44. Radiolabeling of Anti-gelsolin Nanobody with 89Zr NB DFO-Bz-NCS HEPES, pH=7.2 1 h, RT 89ZrNB pH=9, 1 h, 37 oC 1:5 molar ratio NB-DFO NB-DFO Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 45. T im e [m in ] %ID/cc 0 5 1 0 1 5 2 0 2 5 3 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 O B B B O /IA IA /B B B I IV /O B B B O OBBBO/IA IA/BBBI OBBBO/IV 0 100 %ID/cc Kinetics of [89Zr]NB Uptake in Ipsilateral Hemisphere Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 46. OBBBO/IA 1 h pi OBBBO/IA 24 h pi IA/BBBI 1 h pi IA/BBBI 24 h pi OBBBO/IV 1 h pi OBBBO/IV 24 h pi Whole Body PET/CT Imaging of [89Zr]NB11 100 0 100 0 Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 47. Synthesis of Poly(amidoamine) Dendrimers NH Generation 2 Generation 1 N N N N N N NH2 NH2 H2N NH2 NH2 H2N H2 N 2 N N N N N N N N N N NH2 NH2 NH2 H2N N N N N H2N H2N H2N H2N H2N NH2 NH2 H2N NH2 H2N NH2 NH2 (2) NH2 H2N (1) O OMe (1) O OMe (2) NH2 H2N Generation 3, etc. Up to generation 12 Core NN O N O N N H2N NH2 Generation 0 N N H2N H2N NH2 NH2 N N NH N O H H HN H2 O H H2 H (1) O OMe (2) NH2H2N 2 2
  • 48. PAMAM Dendrimer Based Nanoparticles therapeutics contrast agents (X-ray, MRI) targeting agents NHCCH3 O CH2CHCH2OH OH CH2CHCH2OH OH N NHCCH2CH2C O O O- NHC(OCH2CH2)nOH O Applications: • Selective and controlled drug deliver • Gene transfection • Contrast agents • Immunoassay and molecular diagnostic platforms • Nanocomposites surface modification
  • 49. HN (NH2)64 1) DFO-Bz-NCS, 2) Glycidol O N NH O ON HN HOOH O O N OH H N S H N N H S OH OH NH 12 89 Zr4+ O N NH O ON HN O O O O N O H N S H N N H S 89 Zr4+ NH OH OH NH 12 3 3 OH OH N 49 OH OH OH OH N 49 OH OH Modifications of Dendrimer and Radiolabeling with Zirconium-89 89ZrG4(DFO)3(Bdiol)110G4(DFO)3(Bdiol)110G4(NH2)64 Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 50. OBBBO/IA IA/BBBI OBBBO/IV PET Imaging and Kinetics of 89ZrG4(DFO)3(Bdiol)110 Uptake in Ipsilateral Hemisphere 0 30 0 40 0 6 1 h piFrames 5 to 10 min %ID/cc %ID/cc %ID/cc T im e [m in ] %ID/cc 0 5 1 0 1 5 2 0 2 5 3 0 0 1 2 3 4 5 6 O B B B O /IA IA /B B B I IV /O B B B O N S * Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 51. OBBBO / IA 24 h pi IA/BBBI 1 h pi IA / BBBI 24 h pi IV/BBBI 1 h pi IV / BBBI 24 h pi 100 0 100 0 OBBBO/IA 1 h pi Whole Body PET-CT Imaging of 89ZrG4(DFO)3(Bdiol)110 Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 52. Ex Vivo Biodistribution of 89ZrG4(DFO)3(Bdiol)110 at 24 h After Infusion B lood Ipsilateralhem isphere C ontralateralhem isphere H eart Lung Liver Thym us S tom ach P ancreas S pleen K idney Fat M uscle B ladder 0 5 10 15 20 25 30 35 40 %ID/g O B B B O /IA IV /O B B B O IA /B B B I Ipsilateral hem isphere C ontralateral hem isphere 0 .0 0 0 .0 5 0 .1 0 0 .1 5 %ID/g Lesniak WG, Chu C, et al. Eur J Nucl Med Mol Imaging. 2019, doi: 10.1007/s00259-019-04347-y.
  • 53. Conclusions  IA delivery of antibodies or nanobodies might be an attractive therapeutic platform for treatment of CNS disorders  Appropriate surface modification of PAMAM dendrimers with targeting agents may facilitate specific uptake for selective drug delivery
  • 54. Evaluation of PSMA-Targeted PAMAM Dendrimer Lesniak WG et al., Mol Pharm. 2019, doi: 10.1021/acs.molpharmaceut.9b00181 Rhodamine 64Cu PAMAM G4 Lys-Glu-urea PSMA+ PSMA- Optical imaging 25 0 PET-CT PSMA+ PSMA-
  • 55. Optical and Photoacoustic Imaging of PSMA-targeted Dendrimer 5 hrs ai PIP flu dorsal ventraldorsal dorsal ventraldorsal 24 hrs ai ventraldorsal 48 hrs ai ventraldorsal 72 hrs ai flu 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 PIP PIPflu 1 - heart, 2 - lung, 3 - liver, 4 - pancreas, 5 - spleen, 6 - fat, 7 - kidneys, 8 -muscle,9 - small intestines, 10 - salivary glands, 11 - bladder, 12 - bone, 13 - lacrimal gland,14 - PSMA+ PC3 PIP, 15 - PSMA- PC3 flu
  • 56. First-in-man real-time MRI-guided endovascular neurointervention Zawadzki et al., JNISfaster infusion slower infusion
  • 58. • Intra-arterial route of delivery of therapeutic agents to the brain is becoming increasingly attractive. • Advanced imaging is essential to precisely guide the procedure • Advances • Progress in imaging of intra-arterial interventions (MRI + PET) • Drug design especially related to safety aspects (antibodies + macromolecules) • Catheter design – reaching distal vessels allows for precise interventions • Advantages • Decreasing exposure of therapeutic agents to the peripheral organs • No skull opening • Easily repeatable • Future directions: • Design of MRI-visible microcatheters for endovascular neurointerventions • Engineering of cells and molecules to better benefit from intra-arterial delivery • Application of PET-MRI in interventional suites to benefit from advantages of high spatial resolution of MRI and high sensitivity of PET during the same procedure to allow for unprecedented precision Summary
  • 59. Acknowledgments GROUP: Anna Jablonska Yajie Liang Chengyan Chu Xiaoyan Lan Yue Gao Jipeng Zhang Dariush Aligholizadeh COLLABORATORS: Graeme Woodworth Rao Gullapalli Linda Chang Thomas Ernst Ze Wang Collaborators: Monica Pearl Martin Pomper Guanshu Liu Collaborators: Izabela Malysz-Cymborska Dominika Golubczyk Lukasz Kalkowski Wojciech Maksymowicz Collaborators: Barbara Lukomska Luiza Stanaszek Anna Andrzejewska Katarzyna Drela Sylwia Dabrowska Piotr Rogujski Collaborators: Michal Zawadzki Malgorzata Dorobek Blazej Nowak Collaborators: Miguel Oliveira Eduarda Oliveira Johannes Boltze Tim Magnus Matthias Gelderblom Peter Ludewig Jan Gettemans Olivier Zwaenepoel Pedro Ramos Cabrer Jesus Ruiz-Cabello Supported by MSCRFII-0193, MSCRFII-0052, MSCRFE-0178, RO1 NS076573, 2RO1 NS045062, EUREKA RO1DA026299, R21NS106436, R01NS091100, R01NS091110, P41 EB024495 EXPLORE ME
  • 62. The University of Nottingham, UK David Walker, Chair Ruman Rahman, Deputy Chair Richard Grundy Emma Campbell, Project Manager University of Strathclyde, UK Marie Boyd Alexander Mullen Newcastle University, UK Gareth Veal Bristol Royal Hospital for Children, UK Stephen Lowis UCL Institute of Child Health Darren Hargrave Johns Hopkins University, US Henry Brem Monica Pearl Jordan Green Miroslaw Janowski Kenneth Cohen Piotr Walczak National Cancer Institute Katherine Warren VU University Medical Centre, Amsterdam Dannis van Vuurden Fast Track Pharma Ltd Steven Powell Consortium – invitation to register
  • 63. Sponsored by: Mirosław Janowski, MD, PhD Associate Professor, Diagnostic Radiology and Nuclear Medicine University of Maryland School of Medicine Piotr Walczak, MD, PhD Professor, Diagnostic Radiology and Nuclear Medicine University of Maryland School of Medicine Wojciech Lesniak, PhD Research Associate, Department of Radiology John Hopkins School of Medicine To learn more about Scintica’s solutions for preclinical PET-CT imaging, please visit: https://www.scintica.com/products/superargus- sedecal-pet-ct-preclinical-systems/ Thank you!