# Identification of Novel Regulators of Natural Killer Cell Activity

> **NIH NIH R21** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2022 · $180,970

## Abstract

Project Summary/Abstract:
Natural killer (NK) cells are a key part of the innate immune system with the ability to kill both hematological
malignancies and solid tumors. NK cell-based therapies are rapidly gaining clinical interest. However,
mechanisms that NK cells use to mediate anti-tumor activity remain unclear. Here, we specifically investigate a
novel CRISPR/Cas9-mediated screening method to identify new immunologic (NK cell) targets in head and neck
squamous cells carcinoma (HNSCC) cells. We chose HNSCC for these studies as NK cells are known to highly
infiltrate HNSCC and a high degree of NK cells infiltration positively correlates with HNSCC patient survival.
Additionally, the anti-EGFR antibody cetuximab and checkpoint inhibitor antibodies are immune therapies
approved for HNSCC. Despite these therapeutic advances, this malignancy often becomes refractory to these
immune-based therapies as selective pressures push tumor cells to develop resistance mechanisms to escape
NK cell-mediated killing. For this project we will determine how somatic mutations in tumor cells can change their
response to NK cell-based immunotherapy. We hypothesize that this CRISPR/Cas9-based genetic screening
system will identify mechanisms of resistance that lead to better targeting by NK cells. We will use a genome
wide CRISPR/Cas9 KO library to mimic loss of function mutations in HNSCC. After a round of selection with NK
cells, those genes that increase resistance or sensitivity of tumor cells to NK cells will be profiled. Afterwards the
top targets will be validated in vitro and RNA sequencing will be performed to define the molecular pathways
involved. These studies will also use top hits obtained from studies of CRISPR/Cas9 screening on glioblastoma
stem cells that identified novel regulators of NK cell-mediated killing. This combined analysis of HNSCC and
glioblastoma increases the power to identify key genes that regulate NK cell activity against diverse tumors. We
also aim to identify and specifically prioritize novel targets where pharmacological agents impact molecular
pathways that enhance NK cell-mediated killing. The most promising genetic hits will be further studied in vivo
using both a xenograft mouse model of engineered human tumors treated with human NK cells and a syngeneic
system that allows us to validate the ability of proposed genes to regulate NK cell-mediated activity against
HNSCC in an immunocompetent mouse model. These complementary in vitro, in silico and in vivo approaches
will allow us to test pharmacological and genetic strategies to up or down-regulate tumor antigens and NK cell
receptors that can be translated into clinical trials to improve anti-HNSCC and likely other anti-tumor activity.

## Key facts

- **NIH application ID:** 10312800
- **Project number:** 5R21CA241006-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Dan S. Kaufman
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $180,970
- **Award type:** 5
- **Project period:** 2020-12-07 → 2022-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10312800

## Citation

> US National Institutes of Health, RePORTER application 10312800, Identification of Novel Regulators of Natural Killer Cell Activity (5R21CA241006-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10312800. Licensed CC0.

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