Journal: Molecular cancer therapeutics
Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer
doi: 10.1158/1535-7163.MCT-25-1184
Figure Lengend Snippet: In vivo efficacy of L1CAM ADC against triple negative breast cancer and lung adenocarcinoma (LUAD) metastasis. A,B , Single-cell suspensions of MDA-MB-231 LM2 cells transduced with luciferase were injected into NSG mice via the tail vein. After 1 week of metastatic engraftment and outgrowth in the lungs, the mice received control ADC (1 mg/kg) or 13F04 ADC (1 mg/kg) by intraperitoneal injection once weekly for four weeks at the indicated times ( red arrows ). Tumor burden was monitored by BLI weekly. A , Metastatic burden of mice inoculated with MDA-MB-231 cells and treated with control ADC or 13F04 anti-L1CAM ADC. n = 5 mice per group. *** P = 0.0006. B , Overall survival plots of mice in the experiment of panel ( A ) ** P = 0.0035. C , NSG mice were inoculated via the tail vein with GFP-expressing Ru631 LUAD PDX cells. One week later, mice were treated with a single dose of 13F04 ADC or IgG control ADC, and lungs were harvested and analyzed by IF staining for GFP (cancer cells) and L1CAM in one week after ADC treatment. White arrows, cytoplasmic L1CAM staining upon anti-L1CAM ADC treatment. Scale bar, 10 μm. D , Quantification of L1CAM − or L1CAM + fraction of Ru631 PDX treated with ADC in the experiment of panel ( C ). n = 3,195 cells (Control); 2,110 cells (13F04). ** P = 0.0079. E , H&E staining of lung sections before or one week after a single dose of ADC treatment, administered four weeks after inoculation of Ru631 PDX cells into NSG mice via the tail vein. Magnified regions are shown ( red boxes ). Scale bar, 200 μm. F , Bar graph showing the percent area of metastatic lesions per lung in the experiments of panel ( E ). n = 3 per experimental condition. * P = 0.0239. G , IF staining for human pan-cytokeratin (cancer cells) and cleaved caspase 3 in lung metastatic colonies one week after a single dose of ADC treatment, administered four weeks post-inoculation of Ru631 PDX cancer cells into NSG mice via the tail vein. Scale bar, 50 μm. H , Fraction of cleaved caspase 3 + (apoptotic) cells detected by IF in the experiment of panel ( G ). n = 3,257 cells (Control); 788 cells (13F04). **** P < 0.0001. I , IF staining for GFP (cancer cells) and SOX2 one week after a single dose of ADC treatment, administered one week after inoculation of Ru631 PDX cancer cells into NSG mice via the tail vein. Scale bar, 10 μm. J , Percentage of SOX2 + cancer cells detected by IF in the experiment of panel ( I ). n = 5 per experimental condition. * P = 0.0317. K-N , Therapeutic assessment of LUAD metastasis treated with L1CAM or IgG ADCs. Single-cell suspensions from LUAD PDX tumoroids transduced with luciferase were injected into NSG mice via the tail vein. After 4 to 5.5 weeks of metastatic engraftment and outgrowth in the lungs, the mice received control ADC (1 mg/kg) or 13F04 ADC (1 mg/kg) by intraperitoneal injection once weekly for four weeks at the indicated times ( red arrows ). Tumor burden was monitored by BLI weekly. K , Metastatic burden of mice inoculated with Ru323 PDX tumoroid cells and treated with control ADC or 13F04 anti-L1CAM ADC. n = 5 mice per group. **** P < 0.0001. L , Overall survival plots of mice in the experiment of panel ( K ) ** P = 0.0018. M , Metastatic burden of mice inoculated with Ru631 PDX tumoroid cells and treated with control ADC or 13F04 anti-L1CAM ADC. Control, n = 14 mice; 13F04, n = 15 mice. **** P < 0.0001. N , Overall survival plots of mice in the experiment of panel ( M ). **** P < 0.0001. The bar graph indicates mean ± S.E.M. ( D,F,J ). Data are shown as a box (median ± 25-75%) and whisker (maximum to minimum values) plot ( H ). Statistical significance was assessed using the two-tailed Mann-Whitney test ( D,H,J ), two-tailed t test after passing the Shapiro-Wilk normality test ( F ), Kolmogorov-Smirnov test ( A,K,M ) or log-rank (Mantel-Cox) test ( B,L,N ).
Article Snippet: The extracellular domain of human L1CAM (isoform 1, residues 1-1120) fused to a poly-histidine tag via its C-terminus was obtained from Creative Biomart (L1CAM-His).
Techniques: In Vivo, Single Cell, Transduction, Luciferase, Injection, Control, Expressing, Staining, Whisker Assay, Two Tailed Test, MANN-WHITNEY
