Post on 26-Mar-2015
Prof. Nicola Rosato
Dr. Massimo Bottini
1. Nanotechnology, nanodrugs and carbon nanotubes
2. “Bionano” research lines: the past
3. “Bionano” research lines: the present
4. “Bionano” research lines: nanodrugs for intratumor Treg targeting
5. “Bionano” research lines: the future
6. Conclusions
7. Acknowledgments
“Bionano” research lines
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3NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
The science of manipulating matter at the atomic and molecular level to obtain The science of manipulating matter at the atomic and molecular level to obtain materials with specifically enhanced chemical and physical properties.materials with specifically enhanced chemical and physical properties.
Information Technology
Energy
Medicine
Consumer Goods
• Smaller, faster, more energy efficient and powerful computing and other IT-based systems
• More efficient and cost effective technologies for energy production− Solar cells− Fuel cells− Batteries− Bio fuels
• Foods and beverages− Advanced packaging materials,
sensors, and lab-on-chips for food quality testing
• Appliances and textiles− Stain proof, water proof and wrinkle
free textiles• Household and cosmetics
− Self-cleaning and scratch free products, paints, and better cosmetics
• Cancer treatment• Bone treatment• Drug delivery• Appetite control• Drug development• Medical tools• Diagnostic tests• Imaging
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“Bionano” research lines
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7NAST Workshop, July 3, 2013, Rome
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Protein corona of PEG-modified carbon nanotubes
Biodegradation of PEG-modified carbon nanotubes
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Biocompatibility of PEG-modified carbon nanotubes
Intratumor Treg targeting by PEG-modified carbon nanotubes
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CD4+CD25highFoxP3+ regulatory T cells (Treg) are harnessed by tumors to protect themselves from host anti-tumor responses.
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Spleen (healthy) Spleen (tumor) Tumor
Immune cell relative abundance
NAST Workshop, July 3, 2013, Rome
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The manipulation of Treg function selectively into tumors might be the next frontier of cancer immunotherapy.
= carbon nanotube
= PEG
= monoclonal antibody
= cargo
= fluorochromeRegulatory T cellCD25
TCR/CD3
CD4
PEG-modified carbon nanotubes
RES
Tumor
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= SWCNT
= PEG
= targeting agent
= fluorochrome
= Treg-specific receptor
Spleen
Tumor
Length (nm)
0 100 200 300
Fre
qu
ency
(%
)
0
10
20
30
40 10141 nm
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Spleen(healthy)
Spleen(tumor)
Tumor
GITR FR4 CD39 CD103
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Effect of number of DTA-1 mAb/SWCNT
Tar
get
ing
eff
icie
ncy
(%
)
0102030405060
100pM300pM1nM
100pM300pM1nM
100pM300pM1nM
0 5
Unst.
Teff
Treg
DTA-1/SWCNT
*
* *
*
Effect of dose Effect of incubation time
Tar
get
ing
eff
icie
ncy
(%
)
0
10
20
30
40
50
60 01 5
Unst.T regT eff
10
# mAbs
*
*
*
Ta
rge
tin
g e
ffic
ien
cy
(%)
0102030405060
1h1+5h6h
1h1+5h6h
1h1+5h6h
0 5
Unst.
Teff
Treg
DTA-1/SWCNT
* *
** *
DTA-1/SWCNT
Tar
get
ing
sel
ecti
vity
0
4
8
12
16 100pM300pM1nM
100pM300pM1nM
0 5
Treg:Unst.
Treg:Teff
* *
*
Ta
rge
tin
g s
ele
ctiv
ity
0
4
8
12
16
T reg:Unst.
T reg:T eff
01 510
# mAbs* ** *
* *
DTA-1/SWCNT
Ta
rge
tin
g s
ele
cti
vit
y
0
4
8
12
16 1h1+5h6h
1h1+5h6h
0 5
Treg:Unst.
Treg:Teff
* *
*
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1 hour 1 hour + 5 hours 6 hours
NAST Workshop, July 3, 2013, Rome
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Anti-FR4 mAb
Tar
get
ing
eff
icie
ncy
(%
)
0
10
20
30
40
50
60 01 5
Unst.T regT eff
10
# mAbs
**
Tar
get
ing
eff
icie
ncy
(%
)
0
10
20
30
40
50
60 01 5
Unst.T regT eff
10
# mAbs*
Anti-CD39 mAb Anti-CD103 mAb
Tar
get
ing
eff
icie
ncy
(%
)
0
10
20
30
40
50
60 01 5
Unst.T regT eff
10
# mAbs
*
*
*
Ta
rge
tin
g s
ele
ctiv
ity
0
4
8
12
16
T reg:Unst.
T reg:T eff
01 510
# mAbs* *
Ta
rge
tin
g s
ele
cti
vit
y
0
4
8
12
16
T reg:Unst.
T reg:T eff
01 510
# mAbs
* *
*
Ta
rge
tin
g s
ele
cti
vit
y
0
4
8
12
16
T reg:Unst.
T reg:T eff
01 510
# mAbs
* *
*
**
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NAST Workshop, July 3, 2013, Rome
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Ta
rge
tin
g e
ffic
ien
cy
(%)
0
10
20
30
40
50
60 01 5
GITRCD39
FR4
10
# mAbs*
*
Treg targeting efficiency vs. receptor
Tar
get
ing
sel
ecti
vity
0
4
8
12
16 01 5
GITRCD39
FR4
10
# mAbs**
Tar
get
ing
sel
ecti
vity
0
4
8
12
16 01 5
GITRCD39
FR4
10
# mAbs**
Treg vs. Unstained Treg vs. Teff
Treg targeting selectivity vs. receptor
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Time (h)0 2 4 6 8
Blo
od
co
nc
. (
g m
L-1
)
0
5
10
15 HealthyB16-bearing
Pharmacokinetic profile
Ta
rge
tin
g e
ffic
ien
cy
(%)
02468
10121416 Unst.
Teff Treg
no mAb
DTA-1
anti-HA
**
Targeting efficiency (spleen of healthy mice)
Tar
get
ing
eff
icie
ncy
(%
)
02468
10121416 Unst.
Teff Treg
Tumor
Spleen
*
Targeting efficiency (B16-bearing mice)
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Aim 1. Assess tumor killing elicited by PNT-DTA1. We will assess attenuation of intra-tumor Treg function and efficient tumor-killing by PNT-DTA1 systemically administered to B16-bearing mice.
Aim 2. Assess increased intra-tumor delivery of PNT-DTA1 elicited by co-administered iRGD peptides. We will assess whether co-administration of PNT-DTA1 with iRGD peptides leads to a tumor-specific increase in nanoparticle accumulation and Treg targeting, and more efficient tumor killing in B16-bearing mice.
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2,5ug 5ug 10ug
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Treg-targeting efficiency and selectivity of PEG-SWCNT-DTA1 depend on number of ligands per nanotube, incubation time, dose, and targeted surface marker.
PEG-SWCNT-DTA were internalized by Treg through receptor-mediated endocytosis and transported into the cytoplasm and nucleus ex vivo and in vivo.
Injection of PEG-SWCNT-DTA1 in animals carrying tumors enabled very good targeting of T reg residing in the tumor microenvironment, while much less efficiency and almost no selectivity was evident in the spleen.
Preferential penetration of nanoparticles into the tumor microenvironment compared to other tissues ( i.e. spleen) was a consequence of 1.EPR effect2.naturally increased intra-tumor Treg vs. Teff ratio 3.use of markers that are enriched in intra-tumor vs. peripheral Treg (i.e. GITR)
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PEG-SWCNTs were degraded by activated neutrophils in approximately 3 hours
Prof. E. Ruoslahti
Prof. N. Bottini’s Lab Prof. A. Magrini
Prof. M. Mattei
Dr. A. Pietroiusti
Dr. L. Campagnolo
NovellaMassimo Cristiano
Arthritis National Research Foundation
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Prof. B. Fadeel
Prof. A. Star
The Prof.