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Targeting HIF-activated collagen prolyl 4-hydroxylase expression disrupts collagen deposition and blocks primary and metastatic uveal melanoma growth

Abstract

Uveal melanoma (UM) is the most prevalent primary intraocular malignancy in adults, and patients that develop metastases (~50%) survive <1 year, highlighting the urgent need for new therapies. TCGA has recently revealed that a hypoxia gene signature is associated with poor UM patient prognosis. Here we show that expression of hypoxia-regulated collagen prolyl-4-hydroxylase genes P4HA1 and P4HA2 is significantly upregulated in UM patients with metastatic disease and correlates with poor prognosis, suggesting these enzymes might be key tumor drivers. We targeted hypoxia-induced expression of P4HA1/2 in UM with KCN1, a hypoxia inducible factor-1 (HIF-1) pathway inhibitor and found potent inhibition of primary and metastatic disease and extension of animal survival, without overt side effects. At the molecular level, KCN1 antagonized hypoxia-induced expression of P4HA1 and P4HA2, which regulate collagen maturation and deposition in the extracellular matrix. The treatment decreased prolyl hydroxylation, induced proteolytic cleavage and rendered a disordered structure to collagen VI, the main collagen produced by UM, and reduced UM cell invasion. Together, these data demonstrate that extracellular collagen matrix formation can be targeted in UM by inhibiting hypoxia-induced P4HA1 and P4HA2 expression, warranting further development of this strategy in patients with uveal melanoma.

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Fig. 1: Expression of hypoxia-inducible metastasis-related genes in UM patient eye tumors and cell lines.
Fig. 2: KCN1 inhibits primary ocular tumor growth and formation of hepatic metastases in vivo.
Fig. 3: KCN1 extends survival in orthotopic mouse models of uveal melanoma.
Fig. 4: Effect of KCN1 on hypoxia-induced collagen secretion and maturation by melanoma cells.

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Acknowledgements

We thank Dr Bruce Ksander (Schepens Eye Institute, Boston, MA) for providing Mel290 and OMM2-5 cells (OMM2-5 was established by Dr Timothy Murray, Bascom Palmer Eye Institute, Miami, FL); Dr Jerry Niederkorn (Department of Ophthalmology, UT Southwestern, Dallas, TX) for OMM1 and 92.1 cells (92.1 cells were established by Dr Gregorius P. Luyten, Department of Ophthalmology, Rotterdam University Hospital); Dr June Kan-Mitchel (Wayne State University, Detroit, MI) for OCM1 cells, and Dr Dario Rusciano (Friedrich Miescher Institute, Basel, Switzerland) for providing B16LS9 cells. We thank the Winship Research Pathology Core Lab (Emory University) for IHC and Masson’s staining. We also acknowledge Zhengjia Chen (Department of Biostatistics & Bioinformatics, Emory Rollins School of Public Health) for providing statistical consultation. We appreciate the helpful advice and assistance of all members of the Laboratory of Molecular Neuro-Oncology. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby-marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Funding

This work was supported in part by grants from the NIH (R01 CA116804, R01 CA176001, R01 CA180805, R24 EY017045, P30 EY06360, P30 CA138292 and P30 CA13148), the National Natural Science Foundation of China (NNS81201808), the Fight For Sight Postdoctoral Award, the Emory Melanoma Prevention and Research Discovery Fund, a Winship Cancer Institute pilot grant, a Central South University Lieying pilot grant, the V Foundation, the Max Cure Foundation, the Samuel Waxman Cancer Research Foundation, the Alan B. Slifka Foundation, and an unrestricted grant from Research to Prevent Blindness, Inc.

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Correspondence to Hans E. Grossniklaus or Erwin G. Van Meir.

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EGVM is an inventor on a patent jointly held by Emory University and Scripps Research Institute that includes the KCN1 compound used in this study. He is also a share-holder of OncoSpherix Inc.

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Kaluz, S., Zhang, Q., Kuranaga, Y. et al. Targeting HIF-activated collagen prolyl 4-hydroxylase expression disrupts collagen deposition and blocks primary and metastatic uveal melanoma growth. Oncogene 40, 5182–5191 (2021). https://doi.org/10.1038/s41388-021-01919-x

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