University of Angers MiNt: Micro et Nanomedicines Biomimetiques France Instituto Oncologico Veneto Italy Dr. Giovanna Lollo, MiNT, France Dr. Ilaria Marigo, IOV, Italy Nano Immuno Chemotherapy to treat cancers
University of Angers MiNt: Micro et Nanomedicines Biomimetiques France
Instituto Oncologico Veneto Italy
Dr. Giovanna Lollo, MiNT, France
Dr. Ilaria Marigo, IOV, Italy
Nano Immuno Chemotherapy to treat cancers
Lymphonanocarriers for the treatment of Metastatic Cancer
LYMPHOTARG
The LYMPHOTARG project proposes to develop specifically targeted anticancer treatments, by associating anticancer drug to specific nanostructures composed of lipids and polymers, which have a specific affinity for the lymph nodes. In this way, we expect to prevent the process of metastatic spreading through lymphatic vessels.
50 nm
PEG-HS (Solutol)
Surfactant (Span 80)
Triglyceride (labrafac)
Cooling
Heating
60-85°C
Dilution
(Water 20°C)
Heating
W/O
PIZ
Temperature
O/W
Water
Oil
Design and development of Gemcitabine loaded LNC
Gem-C12
Gem-C12: Modified Gemcitabine
Biodistribution Studies
n=5
Biodistribution studies on healthy mice following i.v. and s.c. administration of fluorescent labelled LNC
LNC reach the lymph nodes and remain there for a while (4 days for i.v. and 2 weeks for s.c.). Accumulation in liver and spleen following i.v. and s.c. administration was also evidenced.
Biodistribution Studies
0 10 20 30 40
Day post tumor-graft
0.0
0.2
0.4
0.6
0.8
1.0
Cu
mu
lativ
e P
rop
ortio
n
Su
rviv
ing
salin iv
gemzar iv
gemC12
LNC blank iv
LNC gemC12 iv
LNC blank sc
LNC gemC12 sc
Antitumor efficacy C
um
ula
tive p
rop
ort
ion
su
rviv
ing
mic
e
SCID-CB17 mice grafted at day 0 with 2.104 Ma44-3 cells and treated at day 5, 7 and 9 for IV treatment and at day 5 and 9 for SC treatment (N = 10). A total maximal tolerated dose of 40mg/kg (equivalent molar of gemcitabine) was used to treat groups with Gemzar, GemC12 micelles (GemC12) or LNC GemC12.
P < 0.05
Evaluation of systemic side effects
• Hematologic toxicity
Complete blood count (CBC) complete granulocyte count (WBC)
platelet count (PLT)
WBC (G/L) RBC (T/L) HGB (g/dL) HCT (%) PLT (G/L)
Saline IV 0,2 ± 0,1 9,8 ± 0,3 15,1 ± 0,3 47 ± 1 469 ± 50
Gemzar IV 0,2 ± 0,1 9,2 ± 0,3 14,0 ± 0,4 45 ± 1 245 ± 63
GemC12 IV 0,3 ± 0,2 9,1 ± 0,6 14 ± 1 45 ± 2 275 ± 63
Lnc Blank IV 0,6 ± 0,3 9,8 ± 0,2 15,1 ± 0,3 47,7 ± 0,9 465 ± 44
Lnc GemC12 IV 0,14 ± 0,03 8,9 ± 0,2 13,7 ± 0,3 43,1 ± 0,9 284 ± 24
Lnc Blank SC 0,4 ± 0,1 9,4 ± 0,7 14 ± 1 46 ± 3 453 ± 20
Lnc GemC12 SC 0,26 ± 0,06 9,4 ± 0,5 14,5 ± 0,8 45 ± 2 281 ± 83
*
* P < 0,05 (Kruskall-
Wallis test)
Severe combined immunodefiency mice very low WBC level Deficient lymphocytes B and T Normal natural killer cells, macrophages, granulocytes Abnormal thymus and lymph nodes
Gemzar induce significant myelosuppression on SCID-CB17 mice as revealed by platelet count in comparaison to saline group. Neither blank LNC or LNC loaded by GemC12 delivered by i.v. or s.c. induced significant myelosuppresion
Next step....
The global objective of the project is to develop a novel, synergistic strategy between nanomedicine and immunotherapy to treat cancer. A double approach combining the relief of tumor-induced immunosuppression and the stimulation of a specific immune response against the tumor is proposed.
NICHE: Nano Immuno CHEmotherapy
Good nanocarrier (LNC) prototype from an immunological point of view was developed. Gem-loaded LNC suppress BM-MDSC differentiation in vitro and in vivo
Nano Immuno Carriers
Cross talk between neoplastic and remote sites, such as the bone marrow and spleen, determine the release of soluble factors that drive the accumulation of myeloid cells (MDSC); these myeloid cells subsequently promote neovascularization and metastasis.
Bronte and Gabrilovich, Nat. Rev. Immunol., 2010
LNC accumulate into spleen and could probably interact with the MDSC
Immunesuppressive cells impair cancer immunotherapy
Adoptive Cell Transfer therapy (ACT)
T T
Adoptively
transferred T cells
Endogenous suppressive
cells
Adapted from Restifo, Dudley and Rosenberg. Nature reviews in Immunology 2012
Primary tumor
Mechanism of action
Stimulate immune response
Spleen
Lymph node
Nanomedicine
Target
Revert immunosuppression
MDSC
Monocyte
Tumor microenvironment
TAM
Antigens
Cross Presentation Activation of CD8+ and CD4+
CRT HMGB-1 ATP
DC
CD8+ CD4+
Tumor cell
ICD
NICHE Project