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Improvement of ACK1-targeted therapy efficacy in lung adenocarcinoma using chloroquine or bafilomycin A1
Jinhong Zhu1, Kui Cao2, Meng Zhao1
1Department of Clinical Laboratory, Biobank, Harbin Medical University Cancer Hospital, 150 Haping Road, Harbin, 150040, Heilongjiang, China.
Background:
Activated Cdc42-associated kinase 1 (ACK1) is a promising druggable target for cancer, but its inhibitors only showed moderate effects in clinical trials. The study aimed to investigate the underlying mechanisms and improve the antitumor efficacy of ACK1 inhibitors.
Methods:
RNA-seq was performed to determine the downstream pathways of ACK. Using Lasso Cox regression analysis, we built a risk signature with ACK1-related autophagy genes in the lung adenocarcinoma (LUAD) patients from The Cancer Genome Atlas (TCGA) project. The performance of the signature in predicting the tumor immune environment and response to immunotherapy and chemotherapy were assessed in LUAD. CCK8, mRFP-GFP-LC3 assay, western blot, colony formation, wound healing, and transwell migration assays were conducted to evaluate the effects of the ACK1 inhibitor on lung cancer cells. A subcutaneous NSCLC xenograft model was used for in vivo study.
Results:
RNA-seq revealed the regulatory role of ACK1 in autophagy. Furthermore, the risk signature separated LUAD patients into low- and high-risk groups with significantly different prognoses. The two groups displayed different tumor immune environments regarding 28 immune cell subsets. The low-risk groups showed high immune scores, high CTLA4 expression levels, high immunophenoscore, and low DNA mismatch repair capacity, suggesting a better response to immunotherapy. This signature also predicted sensitivity to commonly used chemotherapy and targeted drugs. In vitro, the ACK1 inhibitors (AIM-100 and Dasatinib) appeared to trigger adaptive autophagy-like response to protect lung cancer cells from apoptosis and activated the AMPK/mTOR signaling pathway, partially explaining its moderate antitumor efficacy. However, blocking lysosomal degradation with chloroquine/Bafilamycine A1 or inhibiting AMPK signaling with compound C/shPRKAA1 enhanced the ACK1 inhibitor's cytotoxic effects on lung cancer cells. The efficacy of the combined therapy was also verified using a mouse xenograft model.
Conclusions:
The resulting signature from ACK1-related autophagy genes robustly predicted survival and drug sensitivity in LUAD. The lysosomal degradation inhibition improved the therapeutic effects of the ACK1 inhibitor, suggesting a potential role for autophagy in therapy evasion.
Insights
Activated Cdc42-associated kinase 1 (ACK1) inhibitors show moderate effects in cancer. Blocking autophagy-related lysosomal degradation enhances their efficacy, suggesting a new therapeutic strategy for lung adenocarcinoma (LUAD).
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Activated Cdc42-associated kinase 1 (ACK1) is a potential cancer target, but its inhibitors have limited clinical efficacy.
- Understanding ACK1's mechanisms is crucial for improving cancer treatment outcomes.
Purpose of the Study:
- To investigate the role of ACK1 in autophagy and its impact on lung adenocarcinoma (LUAD) progression.
- To develop a prognostic and predictive signature based on ACK1-related autophagy genes.
- To explore strategies for enhancing the efficacy of ACK1 inhibitors.
Main Methods:
- RNA sequencing (RNA-seq) and Lasso Cox regression analysis to build a risk signature in LUAD patients (TCGA).
- In vitro assays (CCK8, LC3, western blot, colony formation, migration) to assess ACK1 inhibitor effects.
- In vivo xenograft model to evaluate combined therapy efficacy.
Main Results:
- A risk signature based on ACK1-related autophagy genes effectively predicted LUAD patient prognosis and immune microenvironment.
- ACK1 inhibitors induced an adaptive autophagy response, activating the AMPK/mTOR pathway and conferring resistance.
- Blocking lysosomal degradation or AMPK signaling significantly enhanced ACK1 inhibitor cytotoxicity in vitro and in vivo.
Conclusions:
- The ACK1-autophagy gene signature is a robust predictor of survival and drug sensitivity in LUAD.
- Inhibiting lysosomal degradation alongside ACK1 inhibition offers a promising strategy to overcome therapeutic resistance.
- Autophagy plays a key role in therapy evasion, highlighting its potential as a therapeutic target.
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