Scot Wolfe Awarded NIH Grant for CRISPR Cancer Research

Scot Wolfe, PhD, has received an R01 grant from the National Cancer Institute to develop an innovative strategy for selectively killing cancer cells that harbor focal gene amplifications.

The approach leverages a CRISPR-Cas9 nickase to introduce site-specific single-strand DNA nicks. During DNA replication, these nicks can be converted into single-ended double-strand breaks when encountered by replication forks. Although cells can typically repair an isolated break, a large number of replication-associated breaks can overwhelm repair pathways and trigger cell death.

Wolfe aims to harness this mechanism as a cancer cell-selective genotoxic strategy. By directing multiple nicks to genomic regions that are highly amplified in cancer cells, the approach is designed to generate a toxic burden of DNA damage in tumor cells while sparing normal cells that carry only one or two copies of the same sequence.

As proof of concept, the project will focus on neuroblastoma, a childhood cancer in which MYCN amplification occurs in about 20% of cases and defines a high-risk, aggressive form with poor prognosis. The study builds on the Wolfe lab’s recent published work showing Cas9 nickases targeting MYCN can selectively kill neuroblastoma cells harboring amplified loci (Hanlon et al. Nature Communications 2025). In the NCI-funded work, Wolfe’s group will further optimize Cas9 nickases to enhance their toxicity against MYCN-amplified neuroblastoma cells. They will also evaluate the therapeutic potential of this approach in vivo using an orthotopic xenograft mouse model, and investigate whether inhibiting DNA repair pathways can further increase the effectiveness of Cas9 nickase.

This work seeks to establish a new framework for targeting cancers driven by gene amplification. Because CRISPR-Cas9 targeting is programmable, the platform could be adapted to treat a broad range of cancer types that harbor focal gene amplifications.