Implementing CRISPR/Cas9 Genomic Screens to Identify Genetic Risk Factors for Lung Radiation Injury
Abstract
Genetic factors that predispose to radiation-induced lung injury are poorly understood, and there are currently no genomic tests to identify persons at elevated risk for radiation toxicity. Symptomatic radiation pneumonitis, a type of lung injury characterized by an inflammatory response in normal lung tissue, occurs in approximately 30 percent of patients following definitive chemoradiation for lung cancer, and approximately 3 to 10 percent of these patients do not survive. There is also a long-term risk of radiation-induced pulmonary fibrosis, which can lead to late complications including cor pulmonale and respiratory failure. These risks also exist for occupational and military radiation exposures. Currently, it is not possible to proactively identify these high-risk patients. Modern strategies are urgently needed to identify persons at risk of radiation toxicity so that more effective precautionary measures or radiation treatment strategies may be proactively implemented. To address this goal, we recently validated a genome-wide CRISPR/Cas9 knockout screening platform in a lung tumor-derived cell line, which successfully identified approximately 250 genes that promote radioresistance and 200 genes that promote radiosensitization (including numerous expected hits such as ATM, LIG4, NHEJ1, and DNA-PKcs), which is a proof of concept for using CRISPR/Cas9 genome-scale screens as a novel radiogenomic approach. Over half of these genes have no defined role in the radiation or DNA damage response pathways, and our preliminary data suggests that most of these candidate genes have a causal role in the cellular response to ionizing radiation. Our CRISPR/Cas9 validation screening data indicate that we have the technical capability to employ genome-scale CRISPR/Cas9 screens to identify known and novel radiation response genes with high fidelity.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2021
- Accession Number
- AD1170599
Entities
People
- Neil T. Pfister
Organizations
- Emory University