# Multiplexed nanoparticle delivery to increase CRISPR/Cas gene editing for enhanced cancer therapy

> **NIH NIH R01** · UT SOUTHWESTERN MEDICAL CENTER · 2022 · $391,721

## Abstract

Project Summary
The programmable CRISPR/Cas gene editing system has great potential for cancer treatment due to the ability
to precisely inactivate or repair cancer-related genes. However, delivery of CRISPR to solid tumors for efficient
cancer therapy remains limited by the uniquely stiff and fibrotic tumor microenvironment that acts as a barrier to
nanoparticle uptake. Here, we propose to directly target tumor tissue mechanics via a multiplexed lipid
nanoparticle (LNP) approach involving co-delivery of focal adhesion kinase (FAK) siRNA, Cas9 mRNA, and
sgRNA (siFAK + CRISPR LNPs) to enable tumor delivery and enhance gene editing efficacy. We will leverage
our recently developed non-viral Selective ORgan Targeting (SORT) LNP platform that enables tissue-specific
nucleic acid delivery, protein delivery, and genome editing following intravenous (IV) administration. The
proposed approach involves a unique combination of siRNA-mediated gene silencing of FAK, a key modulator
of tumor ECM, and CRISPR-mediated gene editing (deletion) of tumor-related genes via a single all-in-one
nanoparticle approach. We will further leverage Liver and Lung SORT LNPs for to evaluate gene editing as a
strategy for permanent inactivation of programmed death-ligand 1 (PD-L1), supported by the clinical limitations
of anti-PD-L1 antibody atezolizumab therapy in hepatocellular carcinoma (HCC) and non-small cell lung cancer
(NSCLC). In this grant proposal, we Aim to (1) establish how FAK knockdown enhances SORT LNP-mediated
mRNA delivery and CRISPR gene editing, (2) optimize Liver and Lung SORT LNP formulations for multiplexed
siRNA + Cas9 mRNA + sgRNA delivery, and (3) evaluate the therapeutic efficacy of Liver and Lung SORT siFAK
+ Cas9 mRNA + sgPD-L1 delivery in orthotopic and genetically engineered mouse models of cancer. Regulating
the mechanical properties of tumor cells/ECM for enhancing the genetic suppression in tumor tissues provides
an innovative strategy for treating cancer using CRISPR. We anticipate that this general approach could further
synergize with additional types of therapeutics in the future.

## Key facts

- **NIH application ID:** 10419618
- **Project number:** 1R01CA269787-01
- **Recipient organization:** UT SOUTHWESTERN MEDICAL CENTER
- **Principal Investigator:** Daniel John Siegwart
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $391,721
- **Award type:** 1
- **Project period:** 2022-03-01 → 2027-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10419618

## Citation

> US National Institutes of Health, RePORTER application 10419618, Multiplexed nanoparticle delivery to increase CRISPR/Cas gene editing for enhanced cancer therapy (1R01CA269787-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10419618. Licensed CC0.

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