# Targeting Lymph Node Dependent Immune Tolerance in Cancer

> **NIH NIH R01** · STANFORD UNIVERSITY · 2022 · $523,629

## Abstract

Project Summary
Background: Malignant melanoma and pancreatic ductal adenocarcinoma (PDAC) typically spread to lymph
nodes (LNs) prior to outgrowth in distant tissues. While metastasis to LNs is frequently attributed to passive
drainage from tumor lymphatics, the mechanisms enabling LN metastasis and its functional role in tumor
progression remain poorly understood. LNs are education hubs of the adaptive immune response and harbor
the majority of potentially tumor-reactive lymphocytes. Recently, we discovered that in colonizing LNs, tumor
cells induce tumor-specific immune tolerance through their interactions with leukocytes that subsequently
circulate throughout the host, resulting in systemic tolerance that facilitates metastatic seeding of distant sites.
Hypothesis and objective: We hypothesize that by targeting the induction of immune tolerance in LNs,
we can both prevent distant metastasis and induce tumor regression. We will use multiple cutting-edge
methods to identify the cellular and molecular mechanisms of LN tolerance induction and develop approaches
for breaking it. These goals will be pursued in the following aims:
Specific Aims: Aim 1: Identify the mechanisms by which lymph node metastases induce tumor-specific
immune tolerance through their activation of immunosuppressive lymphocyte populations. Aim 2: Determine
the role of epigenetic regulation in LN metastasis and tolerance induction. Aim 3: Investigate
immunotherapeutic approaches targeting tumor immune tolerance in LNs.
Study design and methods: Using high-content multiplexed microscopy, mass cytometry, photoconversion-
based lineage tracing, TCR sequencing, and genetic mouse models of antigen presentation, we will dissect the
cellular interactions in LNs that we hypothesize are responsible for tolerance induction and dissemination, and
validate the findings in human tissues and datasets. LN metastatic tumor cells exhibit conserved and stable
transcriptional profiles indicative of epigenetic reprogramming. We will assess the role of epigenetic alterations
in conferring a pro-LN metastatic transcriptional signature using bisulfite sequencing and ATAC-seq, and
employ T-ATAC-seq to elucidate changes in the epigenetic landscape within tolerized T cells that recognize
tumor antigens. We will evaluate the ability of our novel immunostimulatory antibody conjugates, targeted
specifically to LNs, to reprogram APCs in metastatic LNs, employ HDAC inhibitors to reprogram both malignant
and lymphocyte populations away from tumor immune tolerance, and combine these strategies to elicit robust
anti-tumor combination therapies in mouse models of melanoma and PDAC.
Expected results and impact: Upon successful conclusion of this work, we will have identified the
mechanisms by which LN metastasis induces systemic tolerance, and evaluated novel immunotherapeutic
strategies to overcome these mechanisms.

## Key facts

- **NIH application ID:** 10366092
- **Project number:** 5R01CA251174-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** EDGAR G. ENGLEMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $523,629
- **Award type:** 5
- **Project period:** 2021-04-01 → 2026-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10366092

## Citation

> US National Institutes of Health, RePORTER application 10366092, Targeting Lymph Node Dependent Immune Tolerance in Cancer (5R01CA251174-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10366092. Licensed CC0.

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