1. “Bionano” research lines
Prof. Nicola Rosato
Dr. Massimo Bottini

2. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Summary
Nanotechnology, nanodrugs and carbon nanotubes
“Bionano” research lines: the past
“Bionano” research lines: the present
“Bionano” research lines: nanodrugs for intratumor Treg targeting
“Bionano” research lines: the future
Conclusions
Acknowledgments
NAST Workshop, July 3, 2013, Rome

3. Nanotechnology, nanodrugs and carbon nanotubes
“Bionano” research lines 1.
Nanotechnology, nanodrugs and carbon nanotubes
NAST Workshop, July 3, 2013, Rome

4. Information Technology
“Bionano” research lines
Nanotechnology
The science of manipulating matter at the atomic and molecular level to obtain materials with specifically enhanced chemical and physical properties.
Energy
Information Technology
More efficient and cost effective technologies for energy production
Solar cells
Fuel cells
Batteries
Bio fuels
Smaller, faster, more energy efficient and powerful computing and other IT-based systems
Consumer Goods
Medicine
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
NAST Workshop, July 3, 2013, Rome

5. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Nanodrugs
NAST Workshop, July 3, 2013, Rome

6. Single-walled carbon nanotubes (SWCNTs)
“Bionano” research lines
Single-walled carbon nanotubes (SWCNTs)
NAST Workshop, July 3, 2013, Rome

7. “Bionano” research lines: the past2.
“Bionano” research lines: the past
NAST Workshop, July 3, 2013, Rome

8. Chemically-modified nanoparticles
“Bionano” research lines
Chemically-modified nanoparticles
NAST Workshop, July 3, 2013, Rome

9. Chemically-modified nanoparticles
“Bionano” research lines
Chemically-modified nanoparticles
NAST Workshop, July 3, 2013, Rome

10. PEG-modified carbon nanotubes in bio-medicine
“Bionano” research lines
PEG-modified carbon nanotubes in bio-medicine
NAST Workshop, July 3, 2013, Rome

11. “Bionano” research lines: the present3.
“Bionano” research lines: the present
NAST Workshop, July 3, 2013, Rome

12. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Present projects
Protein corona of PEG-modified carbon nanotubes
Biodegradation of PEG-modified carbon nanotubes
NAST Workshop, July 3, 2013, Rome

13. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Present projects
Biocompatibility of PEG-modified carbon nanotubes
Intratumor Treg targeting by PEG-modified carbon nanotubes
NAST Workshop, July 3, 2013, Rome

14. Nanodrugs for intratumor Treg targeting
“Bionano” research lines 4.
Nanodrugs for intratumor Treg targeting
NAST Workshop, July 3, 2013, Rome

15. Background: Treg in tumor development
“Bionano” research lines
Background: Treg in tumor development
CD4+CD25highFoxP3+ regulatory T cells (Treg) are harnessed by tumors to protect themselves from host anti-tumor responses.
Immune cell relative abundance
Spleen (healthy)
Spleen (tumor)
Tumor
NAST Workshop, July 3, 2013, Rome

16. Background: Treg in tumor development
“Bionano” research lines
Background: Treg in tumor development
The manipulation of Treg function selectively into tumors might be the next frontier of cancer immunotherapy.
RES
PEG-modified carbon nanotubes
Tumor
CD4
= carbon nanotube
TCR/CD3
= monoclonal antibody
CD25
= PEG
= fluorochrome
Regulatory T cell
= cargo
NAST Workshop, July 3, 2013, Rome

17. PEG-SWCNTs for intratumor Treg targeting
“Bionano” research lines
PEG-SWCNTs for intratumor Treg targeting
10141 nm
Spleen
= SWCNT
= PEG
= targeting agent
= fluorochrome
= Treg-specific receptor
Tumor
NAST Workshop, July 3, 2013, Rome

18. Choosing the target marker
“Bionano” research lines
Choosing the target marker
GITR
FR4
CD39
CD103
Spleen
(healthy)
Spleen
(tumor)
Tumor
NAST Workshop, July 3, 2013, Rome

19. Ex vivo PEG-SWCNT-DTA1 Treg targeting efficiency and selectivity
“Bionano” research lines
Ex vivo PEG-SWCNT-DTA1 Treg targeting efficiency and selectivity
Effect of number of DTA-1 mAb/SWCNT
Effect of incubation time
Effect of dose * * * * * * *
NAST Workshop, July 3, 2013, Rome

20. Ex vivo PEG-SWCNT-DTA1 trafficking
“Bionano” research lines
Ex vivo PEG-SWCNT-DTA1 trafficking
1 hour
1 hour + 5 hours
6 hours
NAST Workshop, July 3, 2013, Rome

21. Ex vivo PEG-SWCNT Treg targeting efficiency and selectivity
“Bionano” research lines
Ex vivo PEG-SWCNT Treg targeting efficiency and selectivity
Anti-FR4 mAb
Anti-CD39 mAb
Anti-CD103 mAb * * * * * * * *
NAST Workshop, July 3, 2013, Rome

22. Ex vivo PEG-SWCNT Treg targeting efficiency and selectivity
“Bionano” research lines
Ex vivo PEG-SWCNT Treg targeting efficiency and selectivity
Treg targeting efficiency vs. receptor
Treg targeting selectivity vs. receptor
Treg vs. Unstained
Treg vs. Teff * * *
NAST Workshop, July 3, 2013, Rome

23. In vivo studies “Bionano” research lines
Targeting efficiency (spleen of healthy mice)
Targeting efficiency (B16-bearing mice)
Pharmacokinetic profile * *
NAST Workshop, July 3, 2013, Rome

24. “Bionano” research lines: the future5.
“Bionano” research lines: the future
NAST Workshop, July 3, 2013, Rome

25. Nanodrugs for Treg-specific cancer immunotherapies
“Bionano” research lines
Nanodrugs for Treg-specific cancer immunotherapies
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.
NAST Workshop, July 3, 2013, Rome

26. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Nanodrugs for RA
2,5ug ug ug
NAST Workshop, July 3, 2013, Rome

27. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Conclusions 
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 Treg 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
EPR effect
naturally increased intra-tumor Treg vs. Teff ratio
use of markers that are enriched in intra-tumor vs. peripheral Treg (i.e. GITR)  
PEG-SWCNTs were degraded by activated neutrophils in approximately 3 hours
NAST Workshop, July 3, 2013, Rome

28. NAST Workshop, July 3, 2013, Rome
“Bionano” research lines
Acknowledgements
The Prof.
Massimo
Cristiano
Novella
Prof. N. Bottini’s Lab
Prof. A. Magrini
Prof. M. Mattei
Dr. A. Pietroiusti
Dr. L. Campagnolo
Prof. E. Ruoslahti
Prof. B. Fadeel
Arthritis National Research Foundation
Prof. A. Star
NAST Workshop, July 3, 2013, Rome