LZTR1 deficiency exerts high metastatic potential by enhancing sensitivity to EMT induction and controlling KLHL12-mediated collagen secretion

Abstract

Leucine zipper-like transcriptional regulator 1 (LZTR1), a substrate adaptor of Cullin 3 (CUL3)-based E3 ubiquitin ligase, regulates proteostasis of the RAS subfamily. Mutations in LZTR1 have been identified in patients with several types of cancer. However, the role of LZTR1 in tumor metastasis and the target molecules of LZTR1, excluding the RAS subfamily, are not clearly understood. Here, we show that LZTR1 deficiency increases tumor growth and metastasis. In lung adenocarcinoma cells, LZTR1 deficiency induced the accumulation of the RAS subfamily and enhanced cell proliferation, invasion, and xenograft tumor growth. Multi-omics analysis to clarify the pathways related to tumor progression showed that MAPK signaling, epithelial-mesenchymal transition (EMT), and extracellular matrix (ECM) remodeling-related gene ontology terms were enriched in LZTR1 knockout cells. Indeed, LZTR1 deficiency induced high expression of EMT markers under TGF-β1 treatment. Our search for novel substrates that interact with LZTR1 resulted in the discovery of a Kelch-like protein 12 (KLHL12), which is involved in collagen secretion. LZTR1 could inhibit KLHL12-mediated ubiquitination of SEC31A, a component of coat protein complex II (COPII), whereas LZTR1 deficiency promoted collagen secretion. LZTR1-RIT1 and LZTR1-KLHL12 worked independently regarding molecular interactions and did not directly interfere with each other. Further, we found that LZTR1 deficiency significantly increases lung metastasis and promotes ECM deposition around metastatic tumors. Since collagen-rich extracellular matrix act as pathways for migration and facilitate metastasis, increased expression of RAS and collagen deposition may exert synergistic or additive effects leading to tumor progression and metastasis. In conclusion, LZTR1 deficiency exerts high metastatic potential by enhancing sensitivity to EMT induction and promoting collagen secretion. The functional inhibition of KLHL12 by LZTR1 provides important evidence that LZTR1 may be a repressor of BTB-Kelch family members. These results provide clues to the mechanism of LZTR1-deficiency carcinogenesis.

Image_1.pdf

“…CREBBP encodes an acetyltransferase and is one of the most mutated genes in small cell lung cancer (48), however, comutations in this gene and NOTCH1 might negatively affect the benefit of adjuvant therapy in EGFR-mutated NSCLC tumors (49). LZTR1 encodes a protein that regulates polyubiquitination and degradation of RAS proteins and has been suggested to regulate the growth and invasion of lung adenocarcinoma cells through RAS/ MAPK signaling (50). IRF4 encodes a transcription factor important in regulating immune and has been shown to be overexpressed in NSCLC tumor tissue and may play a role in increasing cell proliferation rate and colony formation of NSCLC tumor cells through activation of the Notch-Akt signaling pathway (51).…”

Table_5.xlsx

“…CREBBP encodes an acetyltransferase and is one of the most mutated genes in small cell lung cancer (48), however, comutations in this gene and NOTCH1 might negatively affect the benefit of adjuvant therapy in EGFR-mutated NSCLC tumors (49). LZTR1 encodes a protein that regulates polyubiquitination and degradation of RAS proteins and has been suggested to regulate the growth and invasion of lung adenocarcinoma cells through RAS/ MAPK signaling (50). IRF4 encodes a transcription factor important in regulating immune and has been shown to be overexpressed in NSCLC tumor tissue and may play a role in increasing cell proliferation rate and colony formation of NSCLC tumor cells through activation of the Notch-Akt signaling pathway (51).…”

Table_7.xlsx

“…CREBBP encodes an acetyltransferase and is one of the most mutated genes in small cell lung cancer (48), however, comutations in this gene and NOTCH1 might negatively affect the benefit of adjuvant therapy in EGFR-mutated NSCLC tumors (49). LZTR1 encodes a protein that regulates polyubiquitination and degradation of RAS proteins and has been suggested to regulate the growth and invasion of lung adenocarcinoma cells through RAS/ MAPK signaling (50). IRF4 encodes a transcription factor important in regulating immune and has been shown to be overexpressed in NSCLC tumor tissue and may play a role in increasing cell proliferation rate and colony formation of NSCLC tumor cells through activation of the Notch-Akt signaling pathway (51).…”