Non–beta blocker enantiomers of propranolol and atenolol inhibit vasculogenesis in infantile hemangioma
Caroline T. Seebauer, … , Joyce Bischoff, Mathias Francois
Caroline T. Seebauer, … , Joyce Bischoff, Mathias Francois
Published December 7, 2021
Citation Information: J Clin Invest. 2022;132(3):e151109. https://doi.org/10.1172/JCI151109.
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Research Article Angiogenesis Vascular biology

Non–beta blocker enantiomers of propranolol and atenolol inhibit vasculogenesis in infantile hemangioma

Abstract

Propranolol and atenolol, current therapies for problematic infantile hemangioma (IH), are composed of R(+) and S(–) enantiomers: the R(+) enantiomer is largely devoid of beta blocker activity. We investigated the effect of R(+) enantiomers of propranolol and atenolol on the formation of IH-like blood vessels from hemangioma stem cells (HemSCs) in a murine xenograft model. Both R(+) enantiomers inhibited HemSC vessel formation in vivo. In vitro, similar to R(+) propranolol, both atenolol and its R(+) enantiomer inhibited HemSC to endothelial cell differentiation. As our previous work implicated the transcription factor sex-determining region Y (SRY) box transcription factor 18 (SOX18) in propranolol-mediated inhibition of HemSC to endothelial differentiation, we tested in parallel a known SOX18 small-molecule inhibitor (Sm4) and show that this compound inhibited HemSC vessel formation in vivo with efficacy similar to that seen with the R(+) enantiomers. We next examined how R(+) propranolol alters SOX18 transcriptional activity. Using a suite of biochemical, biophysical, and quantitative molecular imaging assays, we show that R(+) propranolol directly interfered with SOX18 target gene trans-activation, disrupted SOX18-chromatin binding dynamics, and reduced SOX18 dimer formation. We propose that the R(+) enantiomers of widely used beta blockers could be repurposed to increase the efficiency of current IH treatment and lower adverse associated side effects.

Authors

Caroline T. Seebauer, Matthew S. Graus, Lan Huang, Alex McCann, Jill Wylie-Sears, Frank Fontaine, Tara Karnezis, David Zurakowski, Steven J. Staffa, Frédéric Meunier, John B. Mulliken, Joyce Bischoff, Mathias Francois

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Figure 1

R(+) propranolol inhibits vessel formation in a murine model for IH.

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R(+) propranolol inhibits vessel formation in a murine model for IH. (A)...
(A) HemSCs were pretreated with PBS or 10 μM R(+) propranolol for 24 hours, suspended in Matrigel with PBS or 5 μM R(+) propranolol, and then injected into nude mice, with 2 implants/mouse (n = 8 mice). The mice were treated with 5 mg/kg R(+) propranolol or an equivalent volume of PBS twice a day as depicted in the schematic. Matrigel implants harvested after 7 days are shown in the top panel of the images. Scale bars: 10 mm. H&E staining indicated fewer blood vessels in the implants of R(+) propranolol–treated mice compared with implants in the control mice (middle panels). Scale bars: 100 μm. Anti–human CD31 staining (red) confirmed the reduced vessel density in R(+) propranolol–treated mice compared with vessel density in control mice (bottom panels). Nuclei were counterstained with DAPI (blue). Scale bars: 100 μm. Graphs show quantification of vessels/mm2 in the H&E-stained sections (left) and human CD31+ vessels/mm2 (right). (B) HemSCs were treated as described in A. Mice were treated with 12.5 mg/kg R(+) propranolol or the equivalent volume of PBS twice a day. Matrigel implants harvested after 7 days are displayed in the top panel of the images, with 2 implants/mouse (n = 8 mice). Scale bars: 10 mm. H&E staining (middle panels) and anti–human CD31 staining (red; bottom panels) showed a significant reduction in vessel density in the implants of R(+) propranolol–treated mice compared with control mice. Scale bars: 100 μm. P values were calculated using a 2-tailed, unpaired Student’s t test. Data show the mean ± SD. Data were collected for 2 implants in each of 4 mice, leading to an observation sample size of 8 per group.