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WNK1-OSR1 Signaling Regulates Angiogenesis-Mediated Metastasis towards Developing a Combinatorial Anti-Cancer
Chia-Ying Hou1,2, Chung-Yung Ma1, Yu-Ju Lin1
1Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli County 35053, Taiwan.
Abstract:
Lysine-deficient protein kinase-1 (WNK1) is critical for both embryonic angiogenesis and tumor-induced angiogenesis. However, the downstream effectors of WNK1 during these processes remain ambiguous. In this study, we identified that oxidative stress responsive 1b (osr1b) is upregulated in endothelial cells in both embryonic and tumor-induced angiogenesis in zebrafish, accompanied by downregulation of protein phosphatase 2A (pp2a) subunit ppp2r1bb. In addition, wnk1a and osr1b are upregulated in two liver cancer transgenic fish models: [tert x p53-/-] and [HBx,src,p53-/-,RPIA], while ppp2r1bb is downregulated in [tert x p53-/-]. Furthermore, using HUVEC endothelial cells co-cultured with HepG2 hepatoma cells, we confirmed that WNK1 plays a critical role in the induction of hepatoma cell migration in both endothelial cells and hepatoma cells. Moreover, overexpression of OSR1 can rescue the reduced cell migration caused by shWNK1 knockdown in HUVEC cells, indicating OSR1 is downstream of WNK1 in endothelial cells promoting hepatoma cell migration. Overexpression of PPP2R1A can rescue the increased cell migration caused by WNK1 overexpression in HepG2, indicating that PPP2R1A is a downstream effector in hepatoma. The combinatorial treatment with WNK1 inhibitor (WNK463) and OSR1 inhibitor (Rafoxanide) plus oligo-fucoidan via oral gavage to feed [HBx,src,p53-/-,RPIA] transgenic fish exhibits much more significant anticancer efficacy than Regorafenib for advanced HCC. Importantly, oligo-fucoidan can reduce the cell senescence marker-IL-1β expression. Furthermore, oligo-fucoidan reduces the increased cell senescence-associated β-galactosidase activity in tert transgenic fish treated with WNK1-OSR1 inhibitors. Our results reveal the WNK1-OSR1-PPP2R1A axis plays a critical role in both endothelial and hepatoma cells during tumor-induced angiogenesis promoting cancer cell migration. By in vitro and in vivo experiments, we further uncover the molecular mechanisms of WNK1 and its downstream effectors during tumor-induced angiogenesis. Targeting WNK1-OSR1-mediated anti-angiogenesis and anti-cancer activity, the undesired inflammation response caused by inhibiting WNK1-OSR1 can be attenuated by the combination therapy with oligo-fucoidan and may improve the efficacy.
Insights
Lysine-deficient protein kinase-1 (WNK1) and oxidative stress responsive 1b (OSR1) drive cancer cell migration and angiogenesis. A combination therapy targeting WNK1 and OSR1, with oligo-fucoidan, shows superior efficacy against liver cancer in fish models.
Area of Science:
- Molecular Biology
- Cancer Research
- Angiogenesis
Background:
- Lysine-deficient protein kinase-1 (WNK1) is crucial for angiogenesis in embryonic development and tumors.
- The downstream targets of WNK1 in these processes, particularly in cancer, are not fully understood.
Purpose of the Study:
- To elucidate the downstream effectors of WNK1 in tumor-induced angiogenesis and cancer cell migration.
- To investigate the therapeutic potential of targeting the WNK1 pathway in hepatocellular carcinoma (HCC).
Main Methods:
- Utilized zebrafish models ([tert x p53] and [HBx,src,p53]) to study angiogenesis and cancer.
- Employed human umbilical vein endothelial cells (HUVECs) and HepG2 hepatoma cells for in vitro co-culture experiments.
- Performed gene knockdown and overexpression studies, inhibitor treatments (WNK1, OSR1), and assessed cell migration and senescence markers.
Main Results:
- Identified oxidative stress responsive 1b (OSR1) as upregulated and protein phosphatase 2A (PP2A) subunit PPP2R1B as downregulated in angiogenesis.
- Confirmed WNK1 promotes hepatoma cell migration, with OSR1 acting downstream in endothelial cells and PPP2R1A in hepatoma cells.
- A combination therapy of WNK1 inhibitor (WNK463), OSR1 inhibitor (Rafoxanide), and oligo-fucoidan demonstrated superior anticancer efficacy in HCC fish models compared to Regorafenib, also reducing inflammation and senescence.
Conclusions:
- The WNK1-OSR1-PPP2R1A axis is a critical regulator of tumor-induced angiogenesis and cancer cell migration.
- Combined inhibition of WNK1 and OSR1, supplemented with oligo-fucoidan, offers a promising therapeutic strategy for advanced HCC.
- Oligo-fucoidan may attenuate inflammation and senescence associated with WNK1-OSR1 inhibition, enhancing therapeutic outcomes.
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