# Assessing the Transcriptional and Signaling Basis of Heterogeneity in the Epithelial-Mesenchymal Transition in Pancreatic Ductal Adenocarcinoma

> **NIH NIH F31** · UNIVERSITY OF VIRGINIA · 2024 · $40,415

## Abstract

PROJECT SUMMARY
The epithelial-mesenchymal transition (EMT) is a developmental process that is aberrantly reactivated in
pancreatic ductal adenocarcinoma (PDAC) to promote disease progression and chemoresistance. PDAC tumors
and cell lines typically contain a heterogeneous mixture of transformed cells displaying epithelial or mesenchymal
characteristics, complicating efforts to understand the regulatory mechanisms that govern this important
phenotypic switching. The observation that EMT can be initiated by a variety of different growth factors, low
oxygen tension, and matrix-mediated signaling strongly suggests that multiple signaling pathways cooperate to
drive robust EMT and raises the possibility that EMT heterogeneity is explained by the ability of only some cells
to activate robustly the pathways that cooperate to drive EMT. Another potential, but not necessarily mutually
exclusive, explanation for phenotypic heterogeneity is that some PDAC cells are primed to undergo EMT due to
transcriptional differences that enable utilization of specific transcription factors or signaling pathways. Based on
our preliminary data, we hypothesize that certain PDAC cells are transcriptionally primed to undergo EMT
and that EMT heterogeneity further depends upon cell-to-cell variations in kinase-regulated signaling
processes within cell populations. The objective of the work proposed here is to test these hypotheses through
the development of quantitative systems biology methods to study the basis of EMT heterogeneity regulation via
transcriptional and kinase-mediated signaling processes. In Aim 1, an iterative immunofluorescence imaging
pipeline will be developed to gather multiplexed signaling data on populations of PDAC cells treated with different
EMT agonists. Based on preliminary studies, we propose to measure markers for seven distinct signaling
pathway nodes and two EMT markers to create a dataset with nine features measured for thousands of cells for
each experimental condition. We will then apply a mutual information data science approach for the quantitative
identification of the signaling pathways that cooperate to drive robust EMT. Model predictions will be tested using
small molecule inhibitors and siRNA-mediated knockdowns. In Aim 2, we will use genetic barcoding for the
transcriptomic profiling of EMT-resistant or -compliant lineages within cell populations. Single-cell RNA
sequencing data from cells before and after EMT induction will be analyzed to identify transcriptional states that
preferentially enable PDAC cells to undergo the mesenchymal transition. The relevance of candidate transcripts
for explaining EMT priming will be tested through knockdown experiments. The methods developed in this work
will be broadly applicable to the study of EMT in other cancer settings and to the study of alternative types of
phenotypic switching. Moreover, the specific results will have implications for the design of novel combination
therapies for PDAC ba...

## Key facts

- **NIH application ID:** 10906025
- **Project number:** 5F31CA275364-02
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Michelle C Barbeau
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $40,415
- **Award type:** 5
- **Project period:** 2023-07-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10906025

## Citation

> US National Institutes of Health, RePORTER application 10906025, Assessing the Transcriptional and Signaling Basis of Heterogeneity in the Epithelial-Mesenchymal Transition in Pancreatic Ductal Adenocarcinoma (5F31CA275364-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10906025. Licensed CC0.

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