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Genome-Wide CRISPR Screening Identifies DCK and CCNL1 as Genes That Contribute to Gemcitabine Resistance in
Hai Yang1, Bin Liu2, Dongxue Liu1
1Department of Surgery, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, Germany.
Abstract:
Pancreatic cancer is one of the most lethal cancers. Due to the difficulty of early diagnosis, most patients are diagnosed with metastasis or advanced-stage cancer, limiting the possibility of surgical treatment. Therefore, chemotherapy is applied to improve patient outcomes, and gemcitabine has been the primary chemotherapy drug for pancreatic cancer for over a decade. However, drug resistance poses a significant challenge to the efficacy of chemotherapy. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) gene-editing system is a powerful tool, and researchers have developed CRISPR/Cas9 library screening as a means to identify the genes associated with specific phenotype changes. We performed genome-wide CRISPR/Cas9 knockout screening in the mouse pancreatic cancer cell line TB32047 with gemcitabine treatment and identified deoxycytidine kinase (DCK) and cyclin L1 (CCNL1) as the top hits. We knocked out DCK and CCNL1 in the TB32047 and PANC1 cell lines and confirmed that the loss of DCK or CCNL1 enhanced gemcitabine resistance in pancreatic cells. Many researchers have addressed the mechanism of DCK-related gemcitabine resistance; however, no study has focused on CCNL1 and gemcitabine resistance. Therefore, we explored the mechanism of CCNL1-related gemcitabine resistance and found that the loss of CCNL1 activates the ERK/AKT/STAT3 survival pathway, causing cell resistance to gemcitabine treatment.
Insights
Researchers identified cyclin L1 (CCNL1) as a key factor in pancreatic cancer gemcitabine resistance. Loss of CCNL1 activates survival pathways, enhancing resistance and offering new therapeutic targets for this lethal cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Pancreatic cancer is highly lethal, often diagnosed at advanced stages, necessitating chemotherapy.
- Gemcitabine is a primary chemotherapy drug, but resistance significantly limits its effectiveness.
- CRISPR/Cas9 gene editing facilitates identifying genes involved in drug resistance phenotypes.
Purpose of the Study:
- To identify genes conferring gemcitabine resistance in pancreatic cancer using genome-wide CRISPR/Cas9 screening.
- To elucidate the mechanism underlying cyclin L1 (CCNL1)-mediated gemcitabine resistance.
Main Methods:
- Genome-wide CRISPR/Cas9 knockout screening was performed on mouse pancreatic cancer cells (TB32047) with gemcitabine treatment.
- Deoxycytidine kinase (DCK) and cyclin L1 (CCNL1) were identified as top hits and subsequently knocked out in TB32047 and PANC1 cell lines.
- Mechanistic studies investigated CCNL1's role in gemcitabine resistance, focusing on signaling pathways.
Main Results:
- Loss of DCK or CCNL1 significantly enhanced gemcitabine resistance in pancreatic cancer cell lines.
- CCNL1 was identified as a novel contributor to gemcitabine resistance, distinct from previously studied DCK.
- Loss of CCNL1 was found to activate the ERK/AKT/STAT3 survival pathway, promoting gemcitabine resistance.
Conclusions:
- CCNL1 is a critical mediator of gemcitabine resistance in pancreatic cancer.
- The ERK/AKT/STAT3 pathway activation upon CCNL1 loss provides a mechanistic explanation for gemcitabine resistance.
- Targeting CCNL1 or its downstream pathways may offer novel therapeutic strategies for overcoming gemcitabine resistance in pancreatic cancer.
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