An endothelial SOX18–mevalonate pathway axis enables repurposing of statins for infantile hemangioma
Annegret Holm, Matthew S. Graus, Jill Wylie-Sears, Jerry Wei Heng Tan, Maya Alvarez-Harmon, Luke Borgelt, Sana Nasim, Long Chung, Ashish Jain, Mingwei Sun, Liang Sun, Pascal Brouillard, Ramrada Lekwuttikarn, Yanfei Qi, Joyce Teng, Miikka Vikkula, Harry Kozakewich, John B. Mulliken, Mathias Francois, Joyce Bischoff
Annegret Holm, Matthew S. Graus, Jill Wylie-Sears, Jerry Wei Heng Tan, Maya Alvarez-Harmon, Luke Borgelt, Sana Nasim, Long Chung, Ashish Jain, Mingwei Sun, Liang Sun, Pascal Brouillard, Ramrada Lekwuttikarn, Yanfei Qi, Joyce Teng, Miikka Vikkula, Harry Kozakewich, John B. Mulliken, Mathias Francois, Joyce Bischoff
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Research Article Angiogenesis Vascular biology

An endothelial SOX18–mevalonate pathway axis enables repurposing of statins for infantile hemangioma

Abstract

Infantile hemangioma (IH) is the most common tumor in children and a paradigm for pathological vasculogenesis, angiogenesis, and regression. Propranolol, the mainstay of treatment, inhibits IH vessel formation via a β-adrenergic receptor-independent off-target effect of its R(+) enantiomer on endothelial SOX18 - a member of the SOX (SRY-related HMG-box) family of transcription factors. Transcriptomic profiling of patient-derived hemangioma stem cells uncovered the mevalonate pathway (MVP) as a target of R(+) propranolol. Loss and gain of function of SOX18 confirmed it is both necessary and sufficient for R(+) propranolol suppression of the MVP, including regulation of sterol regulatory element–binding protein 2 (SREBP2) and the rate-limiting enzyme HMG-CoA reductase (HMGCR). A biological relevance of the endothelial SOX18-MVP axis in IH patient tissue was demonstrated by nuclear colocalization of SOX18 and SREBP2. Functional validation in a preclinical IH xenograft model revealed that statins — competitive inhibitors of HMGCR — efficiently suppress IH vessel formation. We propose an endothelial SOX18-MVP axis as a central regulator of IH pathogenesis and suggest statin repurposing to treat IH. The pleiotropic effects of R(+) propranolol and statins along the SOX18-MVP axis to disable an endothelial cell–specific program may have therapeutic implications for other vascular disease entities involving pathological vasculogenesis and angiogenesis.

Authors

Annegret Holm, Matthew S. Graus, Jill Wylie-Sears, Jerry Wei Heng Tan, Maya Alvarez-Harmon, Luke Borgelt, Sana Nasim, Long Chung, Ashish Jain, Mingwei Sun, Liang Sun, Pascal Brouillard, Ramrada Lekwuttikarn, Yanfei Qi, Joyce Teng, Miikka Vikkula, Harry Kozakewich, John B. Mulliken, Mathias Francois, Joyce Bischoff

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Figure 3

Biosynthetic output of the MVP is mediated via a SOX18-/SREBP2-dependent mechanism.

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Biosynthetic output of the MVP is mediated via a SOX18-/SREBP2-dependent...
(A and B) HUVECs were incubated with MBCD for 16 hours, followed by treatment with or without R(+) propranolol or with or without Sm4 for 16 hours. Endogenous cholesterol levels were measured by mass spectrometry (n = 3 biological replicates). (C and D) Overexpression of ragged-opossum (RaOp) in HUVECs followed by immunofluorescence staining of HMGCS1 and HMGCR (n = 3 biological replicates). P values were calculated using a 2-tailed, unpaired t test (A and B) and by a 1-way ANOVA multiple-comparison test with Tukey’s correction (C and D). Data are shown as the mean ± SD. (E) Schematic of SREBP2 maturation by SCAP, site 1 protease (S1P), and S2P to produce the bHLH domain that translocates to the nucleus. (F and G) HemSCs undergoing endothelial differentiation for 6 days were treated for 2 hours with or without R(+) propranolol, lysed, and analyzed by WB with anti-SREBP2, anti-SOX18, and anti-GAPDH (n = 3 biological replicates; 1 of the biological replicates was analyzed in 2 independent experiments, yielding n = 4 data points). (H and J) Relative gene expression of SREBP2, SCAP, S1P, and S2P in HemSCCtr versus HemSCSOX18OE, with HemSCSOX18OE treated with or without R(+) propranolol for 24 hours. (I and K) Relative gene expression of SREBP2, SCAP, S1P, and S2P in HemECCtr versus HemECshSOX18. (L and M) Immunofluorescence staining of SOX18 and SREBP2 in HemECshSOX18 versus HemECCtr (n = 3 biological replicates). Scale bar: 25 μm. P values were calculated using a 2-tailed, unpaired t test (A–D and H–M) and a 1-way ANOVA multiple-comparison test with Šidák correction (F and G). Data represent sample sizes n = 4 (A and B) and n = 3 (C, D, I, K, and M) biological replicates; n = 3 biological replicates with 1 of the biological replicates analyzed in 2 independent experiments yielding n = 4 data points (F and G); and n = 3–5 independent experiments (H and J). Data are shown as the mean ± SD.