HIFU, Drug Delivery, and Immunotherapyamos3.aapm.org/abstracts/pdf/99-30864-365478-117357.pdfCedars...
Transcript of HIFU, Drug Delivery, and Immunotherapyamos3.aapm.org/abstracts/pdf/99-30864-365478-117357.pdfCedars...
7142015
1
HIFU Drug Delivery and
Immunotherapy
Katherine W Ferrara
UC Davis
Minimally toxic activatable particle
Treat accessible lesions Big hairy audacious goal (BHAG)
Immune therapy is effective for
melanoma emerging as effective for
many cancers
bull FDA-approved
ndash Interleukin-2 (RCC melanoma)
ndash CTLA-4 antibody (melanoma)
ndash PD-1 Ab (melanoma)
bull Investigational
ndash CpG as adjuvant to radiotherapy (melanoma
companion animals)
ndash
Durable response in
significant (15) of patients
7142015
2
Complexation of Cu(II) and Dox within
liposomes
Ammonium sulfate
method
Copper gluconate
TEA method
6
3
7
5 4
pH
Problem Even liposomal doxorubicin has substantial
cardiac toxicity and dose cannot exceed 500 mgm2 in
lifetime
Solution Create a doxorubicin salt that is very stable in
circulation
Kheirolomoom et al Molecular Pharmaceutics
0
50
100
3 5 7 8
Do
x r
ela
tive
Fl
()
pH
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Temperature sensitive particles
paired with mild heating
Achieve hyperthermia Inject and release drug 0-40 minutes later Alter microenvironment Cell death M1 macrophages- 24 hours and later
7142015
3
CuDox-LTSLs
40 min US
US+
US+
CuDox-LTSLs
20 min US
Heart
Heart
Tumor
Thermally-activated vehicles
Below Tc Above Tc
DSPE-PEG-2000 DPPC Lyso-palmitoyl PC Drug
bull Direct and local release can
increase concentration and
decrease systemic toxicity
bull Small molecule can penetrate
tissue
Kheirolomoom et al JCR 2013
US-
US-
Tissue
Autofl
Dox
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles
Tissue
Autofl
Increasing
Dox
Particles US
C u D o x -L T S L s AS D o x -L T S L s
0
1
2
3
4
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
M u s c le + U S T u m o r
C u D o x -L T S L s AS D o x -L T S L s
0
2
4
6
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
T u m o r + U S M u s c le
Cu-dox temperature-
sensitive particles Conventional temperature-
sensitive particles
plt005 compared to control
Plt0001 compared to control
CuDox-TSL (8 treatments)
0
20
40
60
80
100
120
0 50 100 150 200 250
Su
rviv
al
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
-500
500
1500
2500
3500
4500
5500
0 10 20 30 40 50
Tu
mo
r g
row
th
Day post treatment
Control
Control+US
CuDox-LTSLs
CuDox-LTSLs+US
200 250
~
~
US
Treat 2xweek 4 weeks
6 mgkg
Kheirolomoom et al JCR 2013
7142015
4
1 mm
100 microm
Days
Kheirolomoom et al JCR 2013
2 Weeks
100 microm
1 mm
(Fe)
100 microm
(F480)
4 Weeks
1 mm
100 microm
001
01
1
10
100
03 07 10 20 40
ID
Time insonation (hrs)
05 cm
1 cm
2 cm
4 cm
Assumes 5 blood volume in tumor
10 sec tumor blood refresh
5 L blood volume
Diameter
ID delivered
depends on volume insonified
time of insonation
Why do we favor thermally-sensitive
nanoparticles
-10
-5
0
5
10
15
20
25
30
0 5 10 15 20 25 30We
igh
t c
ha
ng
e
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
0
5
10
15
20
25
WBC KmicroL RBCMmicroL
Blo
od
ce
ll c
ou
nt
CuDox-LTSLs
Control
0
2
4
6
8
10
Albumin Total protein
Pro
tein
g
dL
CuDox-LTSLs
Control
0
400
800
1200
1600
Org
an
we
igh
t m
g
CuDox-LTSLs
Control
Toxicity is low
7142015
5
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Use of TLR agonists to enhance Teffector
cell function
adapted from Immunotherapy 2009 November 1 1(6) 949ndash964
DC Dendritic cell iDC Immature dendritic cell MDSC Myeloid-derived suppressor cell Macro Monocytemacrophage TAM Tumor-associated macrophage Teff Effector T cell Treg Regulatory T cell US ultrasound
TLR
activation
iDC
MDSC TAM Treg
Tumor
Teff
Teff
Macro
Th1
Mature
DC
IL-12
IFN-a TNF-a
Tumor Tumor
antigen
IL-12
IFN-g
IL-2
Copper-doxorubicin temperature-sensitive
particles + CpG (3 treatments)
0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 4 5 5 0
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
1 0 0 0 0
1 2 0 0 0
1 4 0 0 0
1 6 0 0 0
1 8 0 0 0
2 0 0 0 0
D a y p o s t tre a tm e n t
ND
L T
um
or
gro
wth
(
)
C o n tro l
C p G
C p G C o n tra la te ra l
C u D o x + U S + C p G
C u D o x + U S + C p G C o n tra la te ra l
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
2
Complexation of Cu(II) and Dox within
liposomes
Ammonium sulfate
method
Copper gluconate
TEA method
6
3
7
5 4
pH
Problem Even liposomal doxorubicin has substantial
cardiac toxicity and dose cannot exceed 500 mgm2 in
lifetime
Solution Create a doxorubicin salt that is very stable in
circulation
Kheirolomoom et al Molecular Pharmaceutics
0
50
100
3 5 7 8
Do
x r
ela
tive
Fl
()
pH
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Temperature sensitive particles
paired with mild heating
Achieve hyperthermia Inject and release drug 0-40 minutes later Alter microenvironment Cell death M1 macrophages- 24 hours and later
7142015
3
CuDox-LTSLs
40 min US
US+
US+
CuDox-LTSLs
20 min US
Heart
Heart
Tumor
Thermally-activated vehicles
Below Tc Above Tc
DSPE-PEG-2000 DPPC Lyso-palmitoyl PC Drug
bull Direct and local release can
increase concentration and
decrease systemic toxicity
bull Small molecule can penetrate
tissue
Kheirolomoom et al JCR 2013
US-
US-
Tissue
Autofl
Dox
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles
Tissue
Autofl
Increasing
Dox
Particles US
C u D o x -L T S L s AS D o x -L T S L s
0
1
2
3
4
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
M u s c le + U S T u m o r
C u D o x -L T S L s AS D o x -L T S L s
0
2
4
6
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
T u m o r + U S M u s c le
Cu-dox temperature-
sensitive particles Conventional temperature-
sensitive particles
plt005 compared to control
Plt0001 compared to control
CuDox-TSL (8 treatments)
0
20
40
60
80
100
120
0 50 100 150 200 250
Su
rviv
al
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
-500
500
1500
2500
3500
4500
5500
0 10 20 30 40 50
Tu
mo
r g
row
th
Day post treatment
Control
Control+US
CuDox-LTSLs
CuDox-LTSLs+US
200 250
~
~
US
Treat 2xweek 4 weeks
6 mgkg
Kheirolomoom et al JCR 2013
7142015
4
1 mm
100 microm
Days
Kheirolomoom et al JCR 2013
2 Weeks
100 microm
1 mm
(Fe)
100 microm
(F480)
4 Weeks
1 mm
100 microm
001
01
1
10
100
03 07 10 20 40
ID
Time insonation (hrs)
05 cm
1 cm
2 cm
4 cm
Assumes 5 blood volume in tumor
10 sec tumor blood refresh
5 L blood volume
Diameter
ID delivered
depends on volume insonified
time of insonation
Why do we favor thermally-sensitive
nanoparticles
-10
-5
0
5
10
15
20
25
30
0 5 10 15 20 25 30We
igh
t c
ha
ng
e
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
0
5
10
15
20
25
WBC KmicroL RBCMmicroL
Blo
od
ce
ll c
ou
nt
CuDox-LTSLs
Control
0
2
4
6
8
10
Albumin Total protein
Pro
tein
g
dL
CuDox-LTSLs
Control
0
400
800
1200
1600
Org
an
we
igh
t m
g
CuDox-LTSLs
Control
Toxicity is low
7142015
5
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Use of TLR agonists to enhance Teffector
cell function
adapted from Immunotherapy 2009 November 1 1(6) 949ndash964
DC Dendritic cell iDC Immature dendritic cell MDSC Myeloid-derived suppressor cell Macro Monocytemacrophage TAM Tumor-associated macrophage Teff Effector T cell Treg Regulatory T cell US ultrasound
TLR
activation
iDC
MDSC TAM Treg
Tumor
Teff
Teff
Macro
Th1
Mature
DC
IL-12
IFN-a TNF-a
Tumor Tumor
antigen
IL-12
IFN-g
IL-2
Copper-doxorubicin temperature-sensitive
particles + CpG (3 treatments)
0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 4 5 5 0
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
1 0 0 0 0
1 2 0 0 0
1 4 0 0 0
1 6 0 0 0
1 8 0 0 0
2 0 0 0 0
D a y p o s t tre a tm e n t
ND
L T
um
or
gro
wth
(
)
C o n tro l
C p G
C p G C o n tra la te ra l
C u D o x + U S + C p G
C u D o x + U S + C p G C o n tra la te ra l
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
3
CuDox-LTSLs
40 min US
US+
US+
CuDox-LTSLs
20 min US
Heart
Heart
Tumor
Thermally-activated vehicles
Below Tc Above Tc
DSPE-PEG-2000 DPPC Lyso-palmitoyl PC Drug
bull Direct and local release can
increase concentration and
decrease systemic toxicity
bull Small molecule can penetrate
tissue
Kheirolomoom et al JCR 2013
US-
US-
Tissue
Autofl
Dox
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles
Tissue
Autofl
Increasing
Dox
Particles US
C u D o x -L T S L s AS D o x -L T S L s
0
1
2
3
4
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
M u s c le + U S T u m o r
C u D o x -L T S L s AS D o x -L T S L s
0
2
4
6
Do
xo
ru
bic
in f
luo
re
sc
en
ce
ra
tio
T u m o r + U S M u s c le
Cu-dox temperature-
sensitive particles Conventional temperature-
sensitive particles
plt005 compared to control
Plt0001 compared to control
CuDox-TSL (8 treatments)
0
20
40
60
80
100
120
0 50 100 150 200 250
Su
rviv
al
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
-500
500
1500
2500
3500
4500
5500
0 10 20 30 40 50
Tu
mo
r g
row
th
Day post treatment
Control
Control+US
CuDox-LTSLs
CuDox-LTSLs+US
200 250
~
~
US
Treat 2xweek 4 weeks
6 mgkg
Kheirolomoom et al JCR 2013
7142015
4
1 mm
100 microm
Days
Kheirolomoom et al JCR 2013
2 Weeks
100 microm
1 mm
(Fe)
100 microm
(F480)
4 Weeks
1 mm
100 microm
001
01
1
10
100
03 07 10 20 40
ID
Time insonation (hrs)
05 cm
1 cm
2 cm
4 cm
Assumes 5 blood volume in tumor
10 sec tumor blood refresh
5 L blood volume
Diameter
ID delivered
depends on volume insonified
time of insonation
Why do we favor thermally-sensitive
nanoparticles
-10
-5
0
5
10
15
20
25
30
0 5 10 15 20 25 30We
igh
t c
ha
ng
e
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
0
5
10
15
20
25
WBC KmicroL RBCMmicroL
Blo
od
ce
ll c
ou
nt
CuDox-LTSLs
Control
0
2
4
6
8
10
Albumin Total protein
Pro
tein
g
dL
CuDox-LTSLs
Control
0
400
800
1200
1600
Org
an
we
igh
t m
g
CuDox-LTSLs
Control
Toxicity is low
7142015
5
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Use of TLR agonists to enhance Teffector
cell function
adapted from Immunotherapy 2009 November 1 1(6) 949ndash964
DC Dendritic cell iDC Immature dendritic cell MDSC Myeloid-derived suppressor cell Macro Monocytemacrophage TAM Tumor-associated macrophage Teff Effector T cell Treg Regulatory T cell US ultrasound
TLR
activation
iDC
MDSC TAM Treg
Tumor
Teff
Teff
Macro
Th1
Mature
DC
IL-12
IFN-a TNF-a
Tumor Tumor
antigen
IL-12
IFN-g
IL-2
Copper-doxorubicin temperature-sensitive
particles + CpG (3 treatments)
0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 4 5 5 0
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
1 0 0 0 0
1 2 0 0 0
1 4 0 0 0
1 6 0 0 0
1 8 0 0 0
2 0 0 0 0
D a y p o s t tre a tm e n t
ND
L T
um
or
gro
wth
(
)
C o n tro l
C p G
C p G C o n tra la te ra l
C u D o x + U S + C p G
C u D o x + U S + C p G C o n tra la te ra l
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
4
1 mm
100 microm
Days
Kheirolomoom et al JCR 2013
2 Weeks
100 microm
1 mm
(Fe)
100 microm
(F480)
4 Weeks
1 mm
100 microm
001
01
1
10
100
03 07 10 20 40
ID
Time insonation (hrs)
05 cm
1 cm
2 cm
4 cm
Assumes 5 blood volume in tumor
10 sec tumor blood refresh
5 L blood volume
Diameter
ID delivered
depends on volume insonified
time of insonation
Why do we favor thermally-sensitive
nanoparticles
-10
-5
0
5
10
15
20
25
30
0 5 10 15 20 25 30We
igh
t c
ha
ng
e
Day post treatment
Control
CuDox-LTSLs
CuDox-LTSLs+US
0
5
10
15
20
25
WBC KmicroL RBCMmicroL
Blo
od
ce
ll c
ou
nt
CuDox-LTSLs
Control
0
2
4
6
8
10
Albumin Total protein
Pro
tein
g
dL
CuDox-LTSLs
Control
0
400
800
1200
1600
Org
an
we
igh
t m
g
CuDox-LTSLs
Control
Toxicity is low
7142015
5
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Use of TLR agonists to enhance Teffector
cell function
adapted from Immunotherapy 2009 November 1 1(6) 949ndash964
DC Dendritic cell iDC Immature dendritic cell MDSC Myeloid-derived suppressor cell Macro Monocytemacrophage TAM Tumor-associated macrophage Teff Effector T cell Treg Regulatory T cell US ultrasound
TLR
activation
iDC
MDSC TAM Treg
Tumor
Teff
Teff
Macro
Th1
Mature
DC
IL-12
IFN-a TNF-a
Tumor Tumor
antigen
IL-12
IFN-g
IL-2
Copper-doxorubicin temperature-sensitive
particles + CpG (3 treatments)
0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 4 5 5 0
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
1 0 0 0 0
1 2 0 0 0
1 4 0 0 0
1 6 0 0 0
1 8 0 0 0
2 0 0 0 0
D a y p o s t tre a tm e n t
ND
L T
um
or
gro
wth
(
)
C o n tro l
C p G
C p G C o n tra la te ra l
C u D o x + U S + C p G
C u D o x + U S + C p G C o n tra la te ra l
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
5
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
Use of TLR agonists to enhance Teffector
cell function
adapted from Immunotherapy 2009 November 1 1(6) 949ndash964
DC Dendritic cell iDC Immature dendritic cell MDSC Myeloid-derived suppressor cell Macro Monocytemacrophage TAM Tumor-associated macrophage Teff Effector T cell Treg Regulatory T cell US ultrasound
TLR
activation
iDC
MDSC TAM Treg
Tumor
Teff
Teff
Macro
Th1
Mature
DC
IL-12
IFN-a TNF-a
Tumor Tumor
antigen
IL-12
IFN-g
IL-2
Copper-doxorubicin temperature-sensitive
particles + CpG (3 treatments)
0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 4 5 5 0
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
1 0 0 0 0
1 2 0 0 0
1 4 0 0 0
1 6 0 0 0
1 8 0 0 0
2 0 0 0 0
D a y p o s t tre a tm e n t
ND
L T
um
or
gro
wth
(
)
C o n tro l
C p G
C p G C o n tra la te ra l
C u D o x + U S + C p G
C u D o x + U S + C p G C o n tra la te ra l
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
6
CpG CpG
Contralateral
CuDox+US+CpG
Contralateral
CuDox+US+CpG
Control
2 mm
200 microm
CuDox
Abscopal response
Immune cell
populations
0
04
08
12
16
2
I
FN
-g+
C
D8
+ T
ce
lls
of
Le
uk
oc
yte
s
0
1
2
3
MD
SC
s
Outline
Activatable particles
+ hyperthermia
Particles + ablation
+ adjuvants
Activatable particles +
hyperthermia +
adjuvants
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
7
Long circulating
nanoparticles
Ablation + 64Cu-LCL
20
0 IDcc
25 IDcc
1cm
6 hours 20 hours 48 hours
0
1
0 2 4 6 8Op
tical
Den
sit
y
(Arb
itra
ry U
nit
s)
Location (mm)
Circle 1 Circle 2
Circle 3 No Tx
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
8
Ablate fraction of tumor + drug
0
02
04
06
08
1
12
14
0 50 100 150
Tu
mo
r V
olu
me (
cc)
Days after First Treatment
No Tx LCL
Single Single + LCL
Circle Circle + LCL
2 mm
4 mm
4 Protocols 100 Curative for
local disease
bull Hyperthermia +
TSL in 8 treatments
bull Hyperthermia+TSL
+ CpG in lt3
treatments
bull Hyperthermia +
Ablation + TSL in 3
treatments
bull Ablation + LCL
Wong et al in review
0
25
50
75
100
0 100 200
Su
rviv
al
Days After First Treatment
Hyp+TSL+Circle
TSL+Circle
TSL+Single
No Tx
0
25
50
75
100
0 40 80 120 160
Surv
ival
Time (days)
NT
LCL
Single
Single + LCL
Circle
Circle + LCL
None
TSL
LCL
Drug
US None Comb Hypertherm Ablation
lt100 cure
lt100 cure wCpG
100 cure wo CpG
100 cure w CpG
NA
NA
Curative Protocols
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
9
1 Protocol- extending survival in
systemic disease needs
optimization
bull Hyperthermia+TSL
+ CpG in lt3
treatments
Kheirolomoom et al in review
0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8 5 2 5 6 6 0 6 4
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
D a y p o s t tre a tm e n t
Su
rv
iva
l (
)
C o n tro l
C p G
C u D o x + U S + C p G
p=002
Conclusion
bull Imaging essential in developing new
nanotherapies identifying cell populations
evaluating efficacy
bull Activatable nanotherapies can cure local
disease enhance systemic therapy
bull Addition of immune adjuvants may allow
the accessible lesions to be treated
resulting in systemic benefits
Charles Caskey James Chomas Paul Dayton Lisa Even Brett Fite Karen Gagnon Xiaowen Hu Elizabeth Ingham Sarah Johnson Azadeh Kheirolomoom Dustin Kruse Jiro Kusunose Chun-Yen Lai Yu Liu Aaron Lum Karen Morgan Eric Paoli Shengping Qin Jai Woong Seo Susanne Stieger Kate Watson Andrew Wong Guodong Zhang Hua Zhang
U Bergen Frits Thorsen Cecilie Brekke Rolf Reed
UCD John Boone Sandy Borowsky Bob Cardiff
Nipavan Chiamvimonvat Roy Curry Greg Farwell
Bruce Hammock C Lebrilla Claude Meares Jinyi Qi
Tingrui Pan Scott Simon C Tepper So Yamada
LLNL Paul Hoeprich
G TechEmory Hanjoong Jo
Northwestern Tom Meade
BurnhamUCSB Juliana Hamzah Erkki Ruoslahti
UCB He Dong Ting Xu
Cedars Sinai Gadi Palled Dan Gazit Galina Shapiro
CMGI (Dave Kukis)
Pfizer Oncology
Targeson (Jack Rychak)
Image-Guided Therapy (Erik Dumont)
Siemens Medical Solutions
NCI and NIBIB
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
10
Manzoor AA et al Cancer Res 2012 Nov 172(21)5566-75
Dewhirst laboratory Duke University
Free dox
Dox-TSL
0 2 5 min
1 5 15 min of heat
Before
10 min after
heat ends
Intensity map (0 0 55) mm in x-y plane
Intensity map (0 5 55) mm in x-y plane
Thermal effects are very significant
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
11
Fite BZ et al PLoS ONE 7(4) e35509
Thermal and mechanical
oversight are feasible
MRgFUS insert from
Image Guided Therapy
7T 3 MHz ultrasound in
Bruker magnet
Heating is ON
from 0 to 20 s
With probe motion
05 1 15 2 25 3 35 4 45
05
1
15
2
25
3
35
4
45 1
15
2
25
3
35
4degC
Foiret et al submitted
US+ temperature sensitive
liposomes (TSL) or long circulating
liposomes (LCL)
bull US hyperthermia+TSL= local cure (100 in 8
treatments)
bull US hyperthermia+CpG+TSL= local cure (100
in 3 treatments) + slow systemic disease
ndash Needs optimization many alternatives
bull US ablation + LCL or TSL slow disease or cure
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
12
200 microm 200 microm
4mm 200 microm
After ablation
Canine melanoma - Combined radiotherapy
and immunotherapy using CPG
oligodeoxynucleotides and indolamine 23
dioxygenase (IDO) blockade
Monjazeb et al J Immunother Cancer 2013 1(Suppl 1) P256
Why nanoparticles Why ultrasound
Target accumulation
0
2
4
6
8
10
12
14
16
18
20
IDg
Paoli et al JCR 2010
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
13
Ultrasound used to guide hyperthermia
-eg Multi-frequency arrays
Therapy
Imaging
Therapy 128 elements at 5 - 7 MHz 027 mm pitch 384mm 4mm element
64 elements at 15 MHz 08 mm pitch 384 mm 5mm element
43 mm
24 mm
Stephens
et al
PMB
2008
IEEE
UFFC
2011
MRfUS
Image-Guided Therapy (IGT) system
bull 16 element annular array
bull Diameter 45mm
bull Radius of curvature 35mm
bull Aperture 85 degrees
bull Operating frequency 3MHz
bull Acoustic efficiency gt 65
bull Power rating 50Wac
bull Bandwidth 300KHz
bull Focal spot size (approx) 15mm x 15mm x 2mm
Temperature-sensitive particles
+ ultrasound= local cure
Temperature-sensitive particles
+ ultrasound + adjuvant = systemic cure
CuDox-lipo
100 nm
Copper-doxorubicin
temperature-sensitive
particles + CpG
Particles
US
Adjuvant
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor
7142015
14
Day 7 Foxp3+ (Treg) cells
Contralateral tumors- in rim
Directly treated-
None in rim or tumor
Lymph node
Tumor