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Persistent infiltration and pro-inflammatory differentiation of monocytes cause unresolved inflammation in brain arteriovenous malformation

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Abstract

An abnormally high number of macrophages are present in human brain arteriovenous malformations (bAVM) with or without evidence of prior hemorrhage, causing unresolved inflammation that may enhance abnormal vascular remodeling and exacerbate the bAVM phenotype. The reasons for macrophage accumulation at the bAVM sites are not known. We tested the hypothesis that persistent infiltration and pro-inflammatory differentiation of monocytes in angiogenic tissues increase the macrophage burden in bAVM using two mouse models and human monocytes. Mouse bAVM was induced through deletion of AVM causative genes, Endoglin (Eng) globally or Alk1 focally, plus brain focal angiogenic stimulation. An endothelial cell and vascular smooth muscle cell co-culture system was used to analyze monocyte differentiation in the angiogenic niche. After angiogenic stimulation, the Eng-deleted mice had fewer CD68+ cells at 2 weeks (P = 0.02), similar numbers at 4 weeks (P = 0.97), and more at 8 weeks (P = 0.01) in the brain angiogenic region compared with wild-type (WT) mice. Alk1-deficient mice also had a trend toward more macrophages/microglia 8 weeks (P = 0.064) after angiogenic stimulation and more RFP+ bone marrow-derived macrophages than WT mice (P = 0.01). More CD34+ cells isolated from peripheral blood of patients with ENG or ALK1 gene mutation differentiated into macrophages than those from healthy controls (P < 0.001). These data indicate that persistent infiltration and pro-inflammatory differentiation of monocytes might contribute to macrophage accumulation in bAVM. Blocking macrophage homing to bAVM lesions should be tested as a strategy to reduce the severity of bAVM.

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Acknowledgments

This study was supported by grants to H. Su from the National Institutes of Health (R01 NS027713, R01 HL122774 and R21 NS083788), and from the Michael Ryan Zodda Foundation and the UCSF Research Evaluation and Allocation Committee (REAC). We thank members of the UCSF BAVM Study Project (http://avm.ucsf.edu) for their support, and Voltaire Gungab for assistance with manuscript preparation.

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    Authors and Affiliations

    1. Department of Anesthesia and Perioperative Care, Center for Cerebrovascular Research, University of California, San Francisco, 1001 Potrero Avenue, Box 1363, San Francisco, CA, 94143, USA

      Rui Zhang, Zhenying Han, Vincent Degos, Fanxia Shen, Eun-Jung Choi, Shuai Kang, Michael Wong, Wan Zhu, Lei Zhan & Hua Su

    2. INSERM, U676, Hôpital Robert Debré, Paris, France

      Vincent Degos

    3. Department of Radiology, University of California, San Francisco, San Francisco, CA, USA

      Zhengda Sun

    4. Institute of Genetic Medicine, International Centre for Life, Newcastle University, Newcastle, UK

      Helen M. Arthur

    5. Department of Physiology and Functional Genomics, University of Florida, Gainesville, FL, USA

      S. Paul Oh

    6. Department of Medicine, University of Toronto, Toronto, ON, Canada

      Marie E. Faughnan

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    1. Rui Zhang
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    All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted.

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    Rui Zhang and Zhenying Han have contributed equally to this work.

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    Zhang, R., Han, Z., Degos, V. et al. Persistent infiltration and pro-inflammatory differentiation of monocytes cause unresolved inflammation in brain arteriovenous malformation. Angiogenesis 19, 451–461 (2016). https://doi.org/10.1007/s10456-016-9519-4

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