# Project 1 Mouse Models Analysis

> **NIH NIH U54** · STANFORD UNIVERSITY · 2024 · $503,335

## Abstract

SUMMARY/ABSTRACT: PROJECT 1
Lymph node (LN) metastasis precedes further dissemination for most solid malignancies and confers a stage III
diagnosis. Nonetheless, the mechanistic role of LN metastasis in further disease progression is poorly
understood. We discovered in mice that in colonizing LNs, tumor cells activate transcriptional programs that
enable their induction of tumor-specific immune tolerance through interactions with LN leukocytes. These
leukocytes subsequently recirculate throughout the host, resulting in systemic immune tolerance facilitating
metastatic seeding of distant sites. Importantly, we found the same conserved transcriptional signature in LN
metastases in humans, suggesting that the mechanisms driving LN metastasis and immune tolerance in our
mouse model are pertinent to human cancers. We hypothesize that local interactions between tumor cells,
leukocytes, and stroma within LNs before, during, and following LN colonization alter distant tissues to promote
disease progression. We expect these interactions to involve homotypic and heterotypic cell-cell interactions,
orchestration of architectural changes within LNs, tumors, and distant sites, and trafficking of various populations
throughout the host. We will use syngeneic mouse models to identify the mechanisms by which these interactions
facilitate disease progression through the following approaches. 1: Determine the kinetics by which tumors
generate systemic immune tolerance. Using a high-dimensional imaging platform (CODEX), we will uncover
longitudinal changes in local microenvironments across the host. 2: Interrogate the nature of heterotypic cell-cell
interactions within LNs and the mechanisms by which these interactions facilitate immune evasion and induction
of immune tolerance. We will use scRNA-seq to identify transcriptional changes in immune and stromal
populations interacting with malignant cells within LNs. Using spatial transcriptomics, we will determine how the
gene expression patterns of cells interacting with malignant cells differ from those at a distance. To confirm the
functional significance of these targets identified and those identified in Project 2, we will combine CRISPR-
mediated gene editing of LN metastatic tumors, along with cell depletion studies and knockout mice or inducible
mouse models. 3: Determine how trafficking of immune cells from tumor-involved LNs establish a pro-metastatic
microenvironment at distant sites. We will employ a novel cell labeling platform to evaluate trafficking of
leukocytes and stromal cells from tumor-involved LNs to distant sites and back to the primary tumor. These
studies will reveal the mechanisms whereby LN metastases promote tumor spread to distant sites and identify
novel targets for the prevention and treatment of metastatic disease.

## Key facts

- **NIH application ID:** 10931491
- **Project number:** 5U54CA274511-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** EDGAR G. ENGLEMAN
- **Activity code:** U54 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $503,335
- **Award type:** 5
- **Project period:** 2023-09-19 → 2028-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10931491

## Citation

> US National Institutes of Health, RePORTER application 10931491, Project 1 Mouse Models Analysis (5U54CA274511-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10931491. Licensed CC0.

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