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Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

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Spatial Transcriptomics Reveals Cancer and Stromal Cell Heterogeneity Between Center and Invasive Front of Pancreatic

Tine Norman Alver¹, Helga Bergholtz¹, Maia Blomhoff Holm²

¹Department of Cancer Genetics, Institute for Cancer Research, Oslo University Hospital, Norway.

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc

|January 31, 2025

Summary

Pancreatic ductal adenocarcinoma (PDAC) shows spatial heterogeneity. Cancer cells and cancer-associated fibroblasts at the invasive front exhibit increased stress, proliferation, and epithelial-mesenchymal transition (EMT) pathway activity compared to the tumor center.

Keywords:

cancer associated fibroblasts pancreatic cancer spatial transcriptomics tumor biology

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Area of Science:

Oncology
Molecular Biology
Cancer Research

Background:

Intratumor heterogeneity is a key factor in pancreatic ductal adenocarcinoma (PDAC) treatment failure.
While genomic and transcriptional heterogeneity are known, the spatial distribution of these cell populations remains understudied.
The invasive front of tumors is hypothesized to harbor cells with enhanced migratory and invasive properties, including cancer-associated fibroblasts (CAFs), but this requires further characterization.

Purpose of the Study:

To compare the transcriptional profiles of cancer cells and CAFs between the tumor center and invasive front of human PDAC.
To investigate the spatial heterogeneity of cellular pathways and molecular markers in PDAC.

Main Methods:

Digital spatial profiling using the GeoMx system (Nanostring) was employed.
Transcriptional profiles of cancer cells and CAFs were analyzed separately in the tumor center and invasive front regions of four PDAC samples.
Immunohistochemical validation was performed on 17 PDAC samples.

Main Results:

Significant inter-regional heterogeneity was identified in PDAC.
Pathways related to cellular stress (TNFα-signaling, hypoxia, P53), proliferation (MYC targets), glycolysis, and epithelial-mesenchymal transition (EMT) showed increased activity at the invasive front in both cancer cells and CAFs.
Immunohistochemistry confirmed spatial heterogeneity in EMT and glycolysis markers.

Conclusions:

Transcriptional profiles of both cancer cells and CAFs differ significantly between the tumor center and invasive front in PDAC.
This spatial heterogeneity contributes to understanding PDAC progression and treatment resistance.
Targeting invasive front characteristics may offer new therapeutic strategies for PDAC.