Fig. 5

Increased metastasis in ZTX models accurately predicts lymph node involvement in the patients. A Cartoon illustrating implantation of PDX models generated from non-small cell lung cancer patients with or without lymph node involvement (LN+, right part of the figure and LN-, left part of the figure respectively) in zebrafish larvae. B Representative fluorescent micrographs of tumor cells (shown in red) disseminated to the caudal hematopoietic plexus three days after implantation from patients having lymph node negative (LN-) or positive (LN+) disease upon diagnosis. C Quantification of the average number of tumor cells disseminated to the caudal hematopoietic plexus (metastasized cells) three days following implantation of PDX models generated from patients with lymph node negative (LN-) or positive (LN+) disease. Dashed line indicates the establish diagnostic cut-off of 5 disseminated cells. *: p < 0.05. n = 12–33 larvae per model. D Quantification of the diagnostic sensitivity (accuracy of predicting patients having lymph node positive disease) and selectivity (accuracy of predicting patients not having lymph node positive disease) using the cut-off value shown in C. n = 7 and 17 for sensitivity and specificity respectively. E Distribution of tumor stages associated with lymph node negative (LN-) or positive (LN+) disease. No stage was significantly associated with lymph node involvement. The number of patients associated with each group is given in Supplemental Table 1. F-I distribution of histological tumor sub-types (F), tumor differentiation stages (G), average age (H), and gender (I), among patients with lymph node negative (LN-) or positive (LN+) disease upon diagnosis. Female gender, younger age, large cell carcinoma and poorly differentiated cancer were significantly associated with higher risk of lymph node positive disease (p < 0.05). The number of patients in each group is given in Supplemental Table 1. LC: Large cell carcinoma. Sq: Squamous cell carcinoma. Ad: Adenocarcinoma. J-L Quantification of the average stromal content (J), vascularity (K) and EMT-score (L) of PDX models grown in mice derived from patients with either lymph node negative (LN-) or positive (LN+) disease upon diagnosis. n-values for each group are given in A. NS: non-significant. G: Good. M: Moderate. P: Poor. Hi: High. In: Intermediate. Lo: Low