Figure 1. ES-2 clusters demonstrate increased in vivo capacity for LNP uptake compared to normal tissues.

(A) Visual representation of the mRNA LNP structure. (B) A representative cryo-transmission electron microscopy (TEM) image shows the spherical shape and size distribution of LNPs. (C) Illustration of the experimental design used to create the ES-2-Luc mouse model of metastatic ovarian cancer. (D) Representative BLS images of ES-2-Luc bearing mouse (left) and sham mouse (right) 15 min after D-luciferin injection. (E) Light micrograph of a thin smear of ascites containing ES-2 clusters on a glass slide. Ascitic fluid was collected from ES-2-Luc mice. (F) Post-mortem photographs of a sham mouse and ES-2-Luc-bearing mice at the primary humane endpoint (~ day 12) showing distended abdomen due to accumulation of malignant ascites (blue arrows), decreased muscle bulk (red arrows and inset), metastases formed on omentum and mesentery (white arrows), and ovaries and gonadal fat pads (black arrows). (G) Experimental design depicting a biodistribution study with ES-2-WT-bearing mice and sham age-matched controls (not shown) administered Luc mRNA LNPs. (H) BLS images of ES-2-WT and sham mice before (top) and 4 h after (bottom) treatment with Luc mRNA LNPs. (I) Representative BLS images of tissues collected from ES-2-WT-bearing (ES-2-WT + Luc mRNA LNP) and sham (sham + Luc mRNA LNP) mice at 4 h-post injection with Luc mRNA LNP in comparison to negative (sham + PBS) and positive controls (ES-2-Luc-bearing mice + PBS). Organs are (left to right): liver, kidneys, spleen, lungs, gonadal fat pads. (J) BLS images of ascitic and physiological peritoneal fluids from ES-2-WT-bearing mice (middle) and sham mice (right), producing an active luciferase enzyme after Luc mRNA LNP treatment in comparison to a positive control (ascites of ES-2-Luc-bearing mice, left, treated with PBS).