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MITF is a driver oncogene and potential therapeutic target in kidney angiomyolipoma tumors through transcriptional regulation of CYR61

Abstract

Tuberous sclerosis complex (TSC) is an autosomal dominant tumor suppressor syndrome, characterized by tumor development in multiple organs, including renal angiomyolipoma. Biallelic loss of TSC1 or TSC2 is a known genetic driver of angiomyolipoma development, however, whether an altered transcriptional repertoire contributes to TSC-associated tumorigenesis is unknown. RNA-seq analyses showed that MITF A isoform (MITF-A) was consistently highly expressed in angiomyolipoma, immunohistochemistry showed microphthalmia-associated transcription factor nuclear localization, and Chromatin immuno-Precipitation Sequencing analysis showed that the MITF-A transcriptional start site was highly enriched with H3K27ac marks. Using the angiomyolipoma cell line 621-101, MITF knockout (MITF.KO) and MITF-A overexpressing (MITF.OE) cell lines were generated. MITF.KO cells showed markedly reduced growth and invasion in vitro, and were unable to form xenografted tumors. In contrast, MITF.OE cells grew faster in vitro and as xenografted tumors compared to control cells. RNA-Seq analysis showed that both ID2 and Cysteine-rich angiogenic inducer 61 (CYR61) expression levels were increased in the MITF.OE cells and reduced in the MITF.KO cells, and luciferase assays showed this was due to transcriptional effects. Importantly, CYR61 overexpression rescued MITF.KO cell growth in vitro and tumor growth in vivo. These findings suggest that MITF-A is a transcriptional oncogenic driver of angiomyolipoma tumor development, acting through regulation of CYR61.

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Fig. 1: MITF is highly expressed in kidney angiomyolipomas.
Fig. 2: MITF regulates cell growth, proliferation, migration, and invasion in 621-101 cells.
Fig. 3: Effects of MITF knockout/overexpression on tumor xenograft development by 621-101 angiomyolipoma cells.
Fig. 4: MITF inhibition induces metabolic alterations in 621-101 cells.
Fig. 5: RNA sequencing identifies ID2 and CYR61 as candidate targets of MITF.
Fig. 6: ID2 and CYR61 are expressed highly in kidney angiomyolipoma, and overexpression of CYR61 has growth effects.
Fig. 7: CYR61 rescues MITF-deficient AML tumor growth in vivo.

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Acknowledgements

This work was supported by the Engles Fund for TSC and LAM; the University of Pennsylvania Orphan Disease Center in partnership with the LAM Foundation; DOD-18-1-0592 (MZ), DOD-17-1-0205 (KG), and the NIH NCI 1P01CA120964 (DJK).

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Zarei, M., Giannikou, K., Du, H. et al. MITF is a driver oncogene and potential therapeutic target in kidney angiomyolipoma tumors through transcriptional regulation of CYR61. Oncogene 40, 112–126 (2021). https://doi.org/10.1038/s41388-020-01504-8

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