Spatially resolved analysis of TGF/BMP signalling in pancreatic ductal adenocarcinoma by digital pathology identifies patient subgroups with adverse outcome.
Bräutigam K., Zens P., Reinhard S., Rohrbach JL., Leedham SJ., Wenning AS., Gloor B., Koelzer VH., Wartenberg M.
BACKGROUND: Transforming Growth Factor (TGF) and Bone Morphogenetic Protein (BMP) signalling critically influence pancreatic ductal adenocarcinoma (PDAC) progression, with TGF-B paradoxically exerting both tumour-promoting and -suppressive effects. Parallel to this observation, the specific context-dependent, spatial dynamics of these pathways and their interaction with the tumour microenvironment (TME) remain poorly understood. METHODS: We performed a spatially resolved analysis of PDAC on a multi-region tissue microarray cohort of 117 curatively resected PDAC specimens consisting of tumour centre (TC), tumour front (TF), and stromal(-predominant) tissue cores each. Protein (ID1, pSMAD2) and mRNA (TGF-A, TGF-B1/2, BMP4, GREM1) expression were assessed in each tissue compartment by immunohistochemistry and in situ hybridization, respectively, quantified by digital image analysis, and correlated with clinicopathologic features. RESULTS: ID1 was significantly overexpressed in PDAC cells compared to associated stroma (p < 0.01), while pSMAD2 was largely absent in PDAC cells, but preserved among associated stroma compartments, particularly in TF cores (p = 0.04). Higher stromal GREM1 signal correlated with reduced overall tumoural ID1 protein expression (p = 0.02), and TGF-B2high/TGF-Alow stroma was significantly associated with worse survival (p < 0.01). Intratumoural TGF-B2 was inversely correlated with stromal pSMAD2 expression (p = 0.03) and was associated with lymph node involvement (p = 0.02). FOXP3+ regulatory T-cells were significantly reduced in TGF-B2high tumours (p = 0.04), while higher tumoural TGF-B1 exhibited a trend towards increased FOXP3+ cells (p = 0.08). CONCLUSIONS: Our spatial analysis reveals intratumoural heterogeneity of TGF/BMP signalling and its significance for PDAC progression. Notably, stromal TGF-B2 emerges as a prognostic biomarker, while TGF-B1 and ID1 are implicated in adverse clinical and pathologic features. These findings highlight the importance of TGF/BMP signalling niches in the TME with implications for PDAC biology and can inform the development of future therapeutic strategies.