Fig. 7
The expression of Mac-TIMP-1 and Epi-CD63 predicts poor prognosis in PDAC. (A) The simple workflow of DSP. Antibodies covalently linked to DNA indexing oligonucleotide are used to stain tissue section. Several repeats of UV light liberates indexing oligonucleotide from the region of interest (ROI), and oligonucleotides are hybridized to NanoString fluorescent barcodes and quantified by NGS (NovaSeq 10B 150PE) (upper). Fluorescent imaging establishes the overall architecture of the tissue. Tissue-specific morphological features were highlighted by PanCK (green), CD45 (red) and SYTO13 (blue). PanCK, CD45, and SYTP13 identify epithelium, immune cells, and nucleic acid respectively. Representative ROI for PanCK and CD45 are shown (lower). (B) Cell-type enrichment analysis (xCell) was performed for the prediction of cell types within the CD45+ population. (C) TIMP1 expression in M1 and M2 macrophage. M1 and M2 macrophages was predicted by webtool xCell. (D) Expression correlation between TIMP1 or TIMP2 in CD45+ immune population and CD63 in PanCK+ epithelial population. (E) Kaplan-Meier analysis of overall survival based on the expression of TIMP1 and CD63 levels in NCKUH cohort. (F) Waterfall plot (oncoplot) for the proportion of patient cases with disease status (stage, differentiation, lymph node, liver metastasis, chemotherapy response) in different levels of TIMP1/CD63. Others: TIMP1highCD63low or TIMP1lowCD63high. The mean value was used as the optimal cut-off values of TIMP1 and CD63. chi-square test was used for the correlation analysis. (G) A schematic illustration showing the interaction between macrophage and pancreatic cancer cells by the TIMP-1/CD63/ERKactive axis, and the leading consequence of the interaction. The working model was created in https://BioRender.com