An MRI-based classification scheme to predict passive access of 5 to 50 nm large nanoparticles to tumors. Anastassia Karageorgis1, 2§, Sandrine Dufort1, 2, 3§, Lucie Sancey1,2 §#, Maxime Henry1, 2,

Samuli Hirsjärvi4, Catherine Passirani4, Jean-Pierre Benoit4, Julien Gravier5, 6Δ, Isabelle

Texier5, 6, Olivier Montigon2, 7, Mériem Benmerad1, 2, Valérie Siroux1, 2, Emmanuel L.

Barbier2, 7, Jean-Luc Coll1, 2 *

Supplementary Figure 1: Examples of images obtained with the different MRI sequences

for each tumor model. Anatomical view (T2-weighted); Apparent Diffusion Coefficient

(ADC) map; Vessel Size Index (VSI); Blood Volume fraction (BVf).

Supplementary Figure 2: Blood vessel immuno-staining performed on tumor sections using an

anti-CD31 antibody (Scale bar 200 µm).

Subcutaneous Orthotopic

  HUH-7 HEK293(β3) IGROV1 U87MG HT29 TS/A-pc TS/A-pc U87MGMRI vessel diameter 13.24 ± 1.14 13.12 ± 2.23 15.07 ± 1.85 12.84 ± 1.19 14.25 ± 1.54 12.72 ± 1.17 16.34 ± 1.63 13.19 ± 1.63

IHC vessel diameter 14.53 ± 0.27 11.71 ± 1.60 15.08 ± 1.98 10.40 ± 1.18 11.52 ± 0.86 8.93 ± 0.75 16.43 ± 3.87 9.96 ± 0.7

number of vessels 17.1 ± 6.6 17.9 ± 6.4 18.4 ± 6.4 31.4 ± 6.4 34.5 ± 8.6 40.3 ± 17.2 14.41 ± 6.92 46.56 ± 12.7

A

B

HUH-7

TS/a-pcU87MG HT29

HEK293(β3) IGROV1

Subcutaneous Orthotopic

TS/a-pc

U87MG

Supplementary Figure 3:

Characterization of the different nanoparticles’ properties. In addition to the average size and

hydrodynamic diameter, the electric charge is presented. Using fluorescence imaging after

systemic tail vein administration, the blood half-life and tissue accumulation were measured

in the mice’s organs 24 hours after LNCs, LNEs or USRPs intravenous injections (10 nmol of

dye).

Supplementary Figure 4: Semi-quantifications of the quantity of fluorescent LNC 24 hours after

intravenous injection in the different tumor models. The fluorescent intensities in organs (A) were

similar among the different groups but varied largely depending on the tumor type (B). n: number of

animals per group

Supplementary Figure 5: Correlation between each tumor parameter evaluated by MRI and

fluorescence imaging. Fluorescence intensities were compared to the permeability (A), water diffusion

coefficient (B), vessel diameter (C) and blood volume fraction (D) obtained by MRI. The correlation

factor R² was evaluated for each parameter.

Supplementary Figure 6: Determination of a predictive equation between MRI parameters and

fluorescence signals. An equation was determined using the SAS® software (A). Two of the four

studied MRI parameters have a significant impact on the equation (B). The predictability of the

equation is assessed by a cross validation test (C).