Targeting myristoylated alanine-rich C kinase substrate phosphorylation site domain in lung cancer. Mechanisms and therapeutic implications

• Published in: American journal of respiratory and critical care medicine Vol. 190; no. 10; pp. 1127 - 1138
• Main Authors: Chen, Ching-Hsien, Statt, Sarah, Chiu, Chun-Lung, Thai, Philip, Arif, Muhammad, Adler, Kenneth B, Wu, Reen
• Format: Journal Article
• Language: English
• Published: United States American Thoracic Society 15-11-2014
• Subjects: Animals; Cell Culture Techniques; Cell Line, Tumor; Disease Models, Animal; Erlotinib Hydrochloride; Humans; Intracellular Signaling Peptides and Proteins - metabolism; Lung cancer; Lung Neoplasms - drug therapy; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Membrane Proteins - metabolism; Mice; Myristoylated Alanine-Rich C Kinase Substrate; Original; Phosphatidylinositol 3-Kinase - metabolism; Phosphatidylinositol Phosphates - metabolism; Phosphorylation - physiology; Protein Kinase Inhibitors - therapeutic use; Quinazolines - therapeutic use; Signal Transduction - physiology; lung cancer; MARCKS phosphorylation; PIP3; erlotinib; PI3K/AKT
• ISSN: 1073-449X; 1535-4970
• DOI: 10.1164/rccm.201408-1505oc

Phosphorylation of myristoylated alanine-rich C kinase substrate (phospho-MARCKS) at the phosphorylation site domain (PSD) is crucial for mucus granule secretion and cell motility, but little is known concerning its function in lung cancer.
We aimed to determine if MARCKS PSD activity can serve as a therapeutic target and to elucidate the molecular basis of this potential.
The clinical relevance of phospho-MARCKS was first confirmed. Next, we used genetic approaches to verify the functionality and molecular mechanism of phospho-MARCKS. Finally, cancer cells were pharmacologically inhibited for MARCKS activity and subjected to functional bioassays.
We demonstrated that higher phospho-MARCKS levels were correlated with shorter overall survival of lung cancer patients. Using shRNA silencing and ectopic expression of wild-type and PSD-mutated (S159/163A) MARCKS, we showed that elevated phospho-MARCKS promoted cancer growth and erlotinib resistance. Further studies demonstrated an interaction of phosphoinositide 3-kinase with MARCKS, but not with phospho-MARCKS. Interestingly, phospho-MARCKS acted in parallel with increased phosphatidylinositol (3,4,5)-triphosphate pools and AKT activation in cells. Through treatment with a 25-mer peptide targeting the MARCKS PSD motif (MPS peptide), we were able to suppress tumor growth and metastasis in vivo, and reduced levels of phospho-MARCKS, phosphatidylinositol (3,4,5)-triphosphate, and AKT activity. This peptide also enhanced the sensitivity of lung cancer cells to erlotinib treatment, especially those with sustained activation of phosphoinositide 3-kinase/AKT signaling.
These results suggest a key role for MARCKS PSD in cancer disease and provide a unique strategy for inhibiting the activity of MARCKS PSD as a treatment for lung cancer.