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Angiogenic inhibition reduces germinal matrix hemorrhage

Nature Medicine volume 13, pages 477–485 (2007)Cite this article

Abstract

The germinal matrix of premature infants is selectively vulnerable to hemorrhage within the first 48 h of life. To assess the role of vascular immaturity in germinal matrix hemorrhage (GMH), we evaluated germinal matrix angiogenesis in human fetuses and premature infants, as well as in premature rabbit pups, and noted active vessel remodeling in all three. Vascular endothelial growth factor (VEGF), angiopoietin-2 and endothelial cell proliferation were present at consistently higher levels in the germinal matrix relative to the white matter anlagen and cortical mantle. On that basis, we asked whether prenatal treatment with either of two angiogenic inhibitors, the COX-2 inhibitor celecoxib, or the VEGFR2 inhibitor ZD6474, could suppress the incidence of GMH in premature rabbit pups. Celecoxib treatment decreased angiopoietin-2 and VEGF levels as well as germinal matrix endothelial proliferation. Furthermore, treatment with celecoxib or ZD6474 substantially decreased the incidence of GMH. Thus, by suppressing germinal matrix angiogenesis, prenatal celecoxib or ZD6474 treatment may be able to reduce both the incidence and severity of GMH in susceptible premature infants.

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Figure 1: VEGF expression is greater in germinal matrix than in cortex or white matter in premature infants and rabbit pups.
Figure 2: ANGPT-2 levels are greater in the germinal matrix than in the cortex or white matter in both humans and rabbits.
Figure 3: Endothelial proliferation was higher in GM than in cortex or WM in humans and rabbits.
Figure 4: PTGS2 levels, but not PTGS1 levels, are higher in the germinal matrix than in the cortex or white matter.
Figure 5: Germinal matrix hemorrhage in rabbit pups.

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Acknowledgements

We thank J. Etlinger for critical evaluation of the manuscript and J. Abrahams for technical and editorial assistance. We thank T. Takano and X. Han for assistance with confocal imaging. Supported by the US National Institutes of Health/National Institute of Neurological Disorders and Stroke grants NS050586 to P.B., R01NS29813 to S.G., and Ro130007 and the Philip Morris Organization to M.N.

Author information

Author notes

  1. Praveen Ballabh

    Present address: Present address: Regional Neonatal Center, Maria Fareri Children's Hospital, Westchester Medical Center, Valhalla, New York 10595, USA.,

Authors and Affiliations

  1. Department of Pediatrics, New York Medical College-Westchester Medical Center, Valhalla, 10595, New York, USA

    Praveen Ballabh, Hongmin Xu & Furong Hu

  2. Department of Anatomy & Cell Biology, New York Medical College-Westchester Medical Center, Valhalla, 10595, New York, USA

    Praveen Ballabh

  3. Department of Pathology, New York Medical College-Westchester Medical Center, Valhalla, 10595, New York, USA

    Alex Braun

  4. Department of Physiology, New York Medical College-Westchester Medical Center, Valhalla, 10595, New York, USA

    Kira Smith, Aracelie Rivera, Zoltan Ungvari & Anna Csiszar

  5. Department of Neurosurgery, University of Rochester, Rochester, 15642, New York, USA

    Nanhong Lou & Steven A Goldman

  6. Department of Neurology, University of Rochester, Rochester, 15642, New York, USA

    Steven A Goldman

  7. Center for Aging and Developmental Biology, University of Rochester, Rochester, 15642, New York, USA

    Maiken Nedergaard

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Contributions

P.B., in whose laboratory this study was performed, planned and supervised experiments, performed confocal microscopy, data analysis and co-wrote the manuscript; H.X. and F.H. performed the animal experiments, immunostaining and western blot analysis; A.B. chose and contributed samples and assisted in editing the manuscript; A.R., K.S., Z.U. and A.C. contributed to laser dissection of brain sections, and to PCR on human and rabbit brains, and also performed the related data analysis; N.L. performed intracranial pressure and cerebral blood flow monitoring of the rabbit pups; S.G. and M.N. assisted in experimental design and planning, and co-wrote the manuscript; M.N. provided direct support for the study.

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Correspondence to Praveen Ballabh.

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Supplementary information

Supplementary Fig. 1

VEGF expression in blood vessels and glial cells of human GM, cortex and WM. (PDF 969 kb)

Supplementary Fig. 2

VEGFR1 expression comparable among cortex, WM and GM in human and rabbit subjects. (PDF 113 kb)

Supplementary Fig. 3

VEGFR2 protein and mRNA expression similar in human and rabbit neonates. (PDF 143 kb)

Supplementary Fig. 4

Tie 2 expression comparable among cortex, WM and GM in humans and rabbits. (PDF 189 kb)

Supplementary Fig. 5

Greater expression of ANGPT-1 protein in fetuses than premature infants. (PDF 355 kb)

Supplementary Fig. 6

Intracranial cerebral pressure, cerebral blood flow, and mean arterial blood pressure on glycerol treatment in celecoxib treated as well as untreated pups. (PDF 55 kb)

Supplementary Fig. 7

Glycerol does not affect angiogenesis and angiogenic factors. (PDF 23 kb)

Supplementary Fig. 8

ZD6474 downregulates VEGFR2 and ERK1/2 phosphorylation. (PDF 170 kb)

Supplementary Methods (PDF 148 kb)

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Ballabh, P., Xu, H., Hu, F. et al. Angiogenic inhibition reduces germinal matrix hemorrhage. Nat Med 13, 477–485 (2007). https://doi.org/10.1038/nm1558

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