# Using CRISPR/Cas9 Pro-code technology to dissect the migratory dendritic cell signature

> **NIH NIH F30** · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · 2022 · $48,908

## Abstract

PROJECT SUMMARY
Immune checkpoint blockade (ICB) has revolutionized the treatment of cancer but only about 20% of patients
respond to treatment. The immune composition of the tumor microenvironment (TME) has been implicated in
determining response to ICB. In particular, myeloid cells (MC) have been associated with both tumor progression
and anti-tumor immunity. Among MC, classical type I dendritic cells (cDC1s) are uniquely capable of
incorporating microenvironmental cues in the context of antigen (Ag) uptake and migrating to the lymph node
(LN) where they present Ag and shape effector T cell responses. At the same time, cDC1s are also important in
central and peripheral tolerance and the induction of T regulatory cells (Tregs). Using bulk and single cell RNA
sequencing (scRNAseq) of mouse and human non-small cell lung cancer (NSCLC) lesions, we have identified
a common transcriptional program in cDC1s that is upregulated upon migration to the LN. This migratory
signature includes many immunoregulatory genes (e.g. PD-L1, CD200, ITGB8, SOCS2), therefore suggesting
that tumors may hijack cDC1 tolerogenic programs to evade the immune system. Though these data provide a
comprehensive transcriptional profile of migratory cDC1s, the function and relevance of many of the genes
remain unknown. Our central hypothesis is that the upregulated genes in this signature control cDC1
homeostasis and phenotype. In Aim1, we will be using CRISPR/Cas9 Protein Barcode (Pro-code) technology
to interrogate multiple KO to the migratory signature in steady state and in tumors, and performing high
dimensional phenotyping in vivo to identify and characterize the genes that regulate the unique phenotype and
molecular profile of cDC1s. In Aim 2, we will use in vitro OT-I/OT-II assays and tumor killing assays to determine
how the migratory signature influences cDC1-T cell priming and activation. And in Aim 3, we will perform single
KOs to the migratory cDC1 signature and determine changes to immune composition in the TME and tumor
progression. The outcome of these studies will be a mechanistic, functional, and contextual understanding of
the cDC1 transcriptional signature in the TME. This project may yield novel insights into DC biology and identify
new ways to modulate or target this compartment for anti-tumor immunity.

## Key facts

- **NIH application ID:** 10424518
- **Project number:** 5F30CA243210-04
- **Recipient organization:** ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
- **Principal Investigator:** Steven Tiwen Chen
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $48,908
- **Award type:** 5
- **Project period:** 2019-07-08 → 2023-05-26

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10424518

## Citation

> US National Institutes of Health, RePORTER application 10424518, Using CRISPR/Cas9 Pro-code technology to dissect the migratory dendritic cell signature (5F30CA243210-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10424518. Licensed CC0.

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