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Targeting neovascular pericytes in neurofibromatosis type 1

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Abstract

Apart from tumor-driven neovascularization, a less-appreciated consequence of neurofibromatosis type 1 (NF1) is the hyperproliferation of vascular mural cells (pericytes). This study aims at establishing a role for pericytes in NF1, and determining whether interference with the function of a key pericyte component (NG2 proteoglycan) inhibits NF1 tumor neovascularization. Neovascularization in NF1 was studied in Nf+/+(control), Nf1+/−, and Nf1−/−embryos at E-10, ischemia-induced retinal angiogenesis model in 24 eyes of Nf1+/−, Nf1+/+mice, and in malignant peripheral nerve sheath tumors (MPNSTs) derived from NF1 patients (ST88-14, NMS-2PC) orthotopically grown in nude mice (Crl: nu/nu). The anti-angiogenic effect of intracorneal polymer pellets containing anti-NG2 neutralizing antibody was quantified in the nude-mouse corneal angiogenesis model in which angiogenesis was induced by xenografting NMS-2PC tumor into the corneal stroma of 22 eyes. By using confocal microscopy, immunohistochemistry, and BrdU proliferation assay, the pericyte/endothelium ratios and proliferation rates were measured. Activated pericytes were present at the leading tip of the angiogenic sprouts. Pericytes showed continuous investment of endothelium in both NMS-2PC and ST88-14 MPNST tumor xenografts. Mean corneal angiogenesis induced by NMS-2PC tumor grafts in NG2-antibody treated eyes was 1.491 and 3.186 mm 2 in isotype-matched non-immunoglobulin treated eyes (control) (P=0.0002). A total of 193.8 vascular nuclei (a measure of ischemia-induced retinal angiogenesis) was present in angiogenic retinal tufts in Nf1+/− mice compared to 89.23 in Nf1+/+ mice (control) (P < 0.0001). Mean pericyte/endothelium investment ratios were 1.015, 1.380, and 2.084 in control, Nf1+/−, and Nf1−/−embryos, respectively. Pericytes were 23% (control), 49% (Nf1+/−), and 69% (Nf1−/−) BrdU-positive. Endothelial cells from the same embryos were 29% (control), 47% (Nf1+/−), and 62% (Nf1−/−) BrdU-positive. Angiogenesis is accelerated in NF1 due to hyperproliferation of pericytes and endothelial cells. Mitotically activated NG2-positive pericytes, and endothelial cells may serve as potential therapeutic targets in NF1.

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Abbreviations

BrdU:

bromodeoxyuridine

CD31:

PECAM-1

CD105:

endoglin

flk 1:

VEGF receptor-2

MPNST:

malignant peripheral nerve sheath tumor

NG2:

nerve/glial antigen 2

NF1:

neurofibromatosis type 1

Nf1 :

neurofibromatosis type1 gene

PAS:

periodic acid-Schiff method

PBS:

phosphate-buffered saline

PDGF β-receptor:

platelet-derived growth factor beta receptor

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  1. Vascular Biology Division, La Jolla Institute for Molecular Medicine, 4570 Executive Drive, Suite 100, San Diego, California, 92121, USA

    Ugur Ozerdem

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Ozerdem, U. Targeting neovascular pericytes in neurofibromatosis type 1. Angiogenesis 7, 307–311 (2004). https://doi.org/10.1007/s10456-004-6643-3

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