3-D multiplex Histology Reveals Microenvironment of Early Duct Lesions in Human Donor Pancreas: An Untold Exocrine-endocrine Relationship

09:30 - 09:50, Room 104 (104講堂)

Abstract - 

3-D multiplex histology reveals microenvironment of early duct lesions in human donor pancreas: An untold exocrine-endocrine relationship

Back Background: Formation of low-grade pancreatic intraepithelial neoplasia (PanIN) lesions has been linked to the development of pancreatic ductal adenocarcinoma. However, at the early stage, i.e., when the pancreas appears to be normal, how the pancreatic exocrine, endocrine, neurovascular, and stromal tissues respond to the initial duct lesion formation remains unclear.

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Aims: To apply panoramic and 3-D multiplex histology to detect early pancreatic lesions (vs. normal parenchyma) and characterize the changes in the peri-lesional microenvironment, including the islet, stromal, and neurovascular remodeling, in human cadaveric donor pancreases. 

Methods: Tissue clearing was applied to the freshly perfused, paraformaldehyde-fixed donor pancreases for large-scale 3-D confocal microscopy. Serial vibratome sections (350 µm in thickness) of the specimens were immunostained with epithelium, vasculature (blood and lymphatic vessels), nerves, stellate cells, and islets to reveal the lesion microenvironment. 

Results: Large-scale tissue scanning led to detection of early-stage duct lesions (PanIN 1A/B) in the donor pancreases. High-definition 3-D image data of the lesion microenvironment reveal: (1) islet aggregation, (2) formation of islet-duct complex, (3) adipocyte association, (4) inflammation (CD45+) with lymphangiogenesis (D2-40+), (5) stromal activation, and (6) increase in nerve density. In particular, the formation of lesion-islet complex and direct islet-epithelium contact indicate that the endocrine islets are not bystanders in PanIN lesion progression. Also, because 3-D confocal imaging is noninvasive to the specimens, we integrated the 3-D approach with the standard 2-D pathological examination (H&E staining and microscopy) to confirm the PanIN lesions, which is crucial for the clinical assessment of duct lesion progression. 

Conclusions: We systematically developed a 3-D/2-D integrative imaging approach to detect and characterize the early duct lesion-islet association in human donor pancreases. The spatial information and the high-definition 3-D and 2-D images advance our understanding of the lesion microenvironment and lay the foundation for building an image database to depict the human pancreas in health, obesity, and cancer lesion progression.