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CXCR3 antagonism of SDF-1(5-67) restores trabecular function and prevents retinal neurodegeneration in a rat model of ocular hypertension.

Denoyer A, Godefroy D, Célérier I, Frugier J, Degardin J, Harrison JK, Brignole-Baudouin F, Picaud S, Baleux F, Sahel JA, Rostène W, Baudouin C - PLoS ONE (2012)

Bottom Line: Current antiglaucoma therapy does not target the causal trabecular pathology, which may explain why treatment failure is often observed.The protective effect of CXCR3 antagonism is related to restoration of the trabecular function.These data demonstrate that proteolytic cleavage of CXCL12 is involved in trabecular pathophysiology, and that local administration of a selective CXCR3 antagonist may be a beneficial therapeutic strategy for treating ocular hypertension and subsequent retinal degeneration.

View Article: PubMed Central - PubMed

Affiliation: UPMC University Paris 6, Institut de la Vision, UMRS968, Paris, France. alexandre.denoyer@gmail.com

ABSTRACT
Glaucoma, the most common cause of irreversible blindness, is a neuropathy commonly initiated by pathological ocular hypertension due to unknown mechanisms of trabecular meshwork degeneration. Current antiglaucoma therapy does not target the causal trabecular pathology, which may explain why treatment failure is often observed. Here we show that the chemokine CXCL12, its truncated form SDF-1(5-67), and the receptors CXCR4 and CXCR3 are expressed in human glaucomatous trabecular tissue and a human trabecular cell line. SDF-1(5-67) is produced under the control of matrix metallo-proteinases, TNF-α, and TGF-β2, factors known to be involved in glaucoma. CXCL12 protects in vitro trabecular cells from apoptotic death via CXCR4 whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase activation. Ocular administration of SDF-1(5-67) in the rat increases intraocular pressure. In contrast, administration of a selective CXCR3 antagonist in a rat model of ocular hypertension decreases intraocular pressure, prevents retinal neurodegeneration, and preserves visual function. The protective effect of CXCR3 antagonism is related to restoration of the trabecular function. These data demonstrate that proteolytic cleavage of CXCL12 is involved in trabecular pathophysiology, and that local administration of a selective CXCR3 antagonist may be a beneficial therapeutic strategy for treating ocular hypertension and subsequent retinal degeneration.

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CXCL12 protects trabecular cells from apoptosis via CXCR4, whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase-3 activation.(A) 24-h incubation with CXCL12 (10 ng/mL [1.3 nM]) protects HTM3 cells from apoptotic stress induced by 15-min exposure to 0.01% benzalkonium chloride (BAC), whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) increases apoptosis as assessed by microplate cytometry using Hoechst dye. ** P<0.05 vs. unstressed cells, § P<0.05 and §§ P<0.01 vs. BAC-exposed cells. (B) The protective effect of CXCL12 (10 ng/mL, 24 h) is reversed by CXCR4 antagonist (AMD-3100, 1 µM), whereas the apoptotic effect of SDF-1(5-67) (10 ng/mL, 24 h) is inhibited by CXCR3 antagonist (NBI-74330, 1 µM). CXCL10 (10 ng/mL [1.1 nM], 24 h), a conventional ligand for CXCR3, mimics the apoptotic effect of SDF-1(5-67). ** P<0.01. (C) Dose-dependent effect of 24-h incubation with SDF-1(5-67) or with CXCL12. ** P<0.01 vs. CXCL12. (D) SDF-1(5-67) (10 ng/mL) increases caspase-3 activation as assessed by immunoflowcytometry. CXCR3 antagonist (NBI-74330, 1 µM) inhibits SDF-1(5-67)-induced caspase 3 activation. ** P<0.01 vs. unstimulated, §§ P<0.01 vs. SDF-1(5-67)-stimulated. Data in graphs are presented as means ± SEM.
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pone-0037873-g003: CXCL12 protects trabecular cells from apoptosis via CXCR4, whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase-3 activation.(A) 24-h incubation with CXCL12 (10 ng/mL [1.3 nM]) protects HTM3 cells from apoptotic stress induced by 15-min exposure to 0.01% benzalkonium chloride (BAC), whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) increases apoptosis as assessed by microplate cytometry using Hoechst dye. ** P<0.05 vs. unstressed cells, § P<0.05 and §§ P<0.01 vs. BAC-exposed cells. (B) The protective effect of CXCL12 (10 ng/mL, 24 h) is reversed by CXCR4 antagonist (AMD-3100, 1 µM), whereas the apoptotic effect of SDF-1(5-67) (10 ng/mL, 24 h) is inhibited by CXCR3 antagonist (NBI-74330, 1 µM). CXCL10 (10 ng/mL [1.1 nM], 24 h), a conventional ligand for CXCR3, mimics the apoptotic effect of SDF-1(5-67). ** P<0.01. (C) Dose-dependent effect of 24-h incubation with SDF-1(5-67) or with CXCL12. ** P<0.01 vs. CXCL12. (D) SDF-1(5-67) (10 ng/mL) increases caspase-3 activation as assessed by immunoflowcytometry. CXCR3 antagonist (NBI-74330, 1 µM) inhibits SDF-1(5-67)-induced caspase 3 activation. ** P<0.01 vs. unstimulated, §§ P<0.01 vs. SDF-1(5-67)-stimulated. Data in graphs are presented as means ± SEM.

Mentions: In a TC model of toxic-induced apoptosis [34], addition of CXCL12 (10 ng/mL [1.3 nM]) decreased apoptosis, whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) potentiated apoptosis (Fig. 3A). Both the protective effect of CXCL12 and the deleterious effect of SDF-1(5-67) were concentration-dependent, with a maximal effect of either chemokine observed at 10 ng/mL (Fig. 3B).


CXCR3 antagonism of SDF-1(5-67) restores trabecular function and prevents retinal neurodegeneration in a rat model of ocular hypertension.

Denoyer A, Godefroy D, Célérier I, Frugier J, Degardin J, Harrison JK, Brignole-Baudouin F, Picaud S, Baleux F, Sahel JA, Rostène W, Baudouin C - PLoS ONE (2012)

CXCL12 protects trabecular cells from apoptosis via CXCR4, whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase-3 activation.(A) 24-h incubation with CXCL12 (10 ng/mL [1.3 nM]) protects HTM3 cells from apoptotic stress induced by 15-min exposure to 0.01% benzalkonium chloride (BAC), whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) increases apoptosis as assessed by microplate cytometry using Hoechst dye. ** P<0.05 vs. unstressed cells, § P<0.05 and §§ P<0.01 vs. BAC-exposed cells. (B) The protective effect of CXCL12 (10 ng/mL, 24 h) is reversed by CXCR4 antagonist (AMD-3100, 1 µM), whereas the apoptotic effect of SDF-1(5-67) (10 ng/mL, 24 h) is inhibited by CXCR3 antagonist (NBI-74330, 1 µM). CXCL10 (10 ng/mL [1.1 nM], 24 h), a conventional ligand for CXCR3, mimics the apoptotic effect of SDF-1(5-67). ** P<0.01. (C) Dose-dependent effect of 24-h incubation with SDF-1(5-67) or with CXCL12. ** P<0.01 vs. CXCL12. (D) SDF-1(5-67) (10 ng/mL) increases caspase-3 activation as assessed by immunoflowcytometry. CXCR3 antagonist (NBI-74330, 1 µM) inhibits SDF-1(5-67)-induced caspase 3 activation. ** P<0.01 vs. unstimulated, §§ P<0.01 vs. SDF-1(5-67)-stimulated. Data in graphs are presented as means ± SEM.
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pone-0037873-g003: CXCL12 protects trabecular cells from apoptosis via CXCR4, whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase-3 activation.(A) 24-h incubation with CXCL12 (10 ng/mL [1.3 nM]) protects HTM3 cells from apoptotic stress induced by 15-min exposure to 0.01% benzalkonium chloride (BAC), whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) increases apoptosis as assessed by microplate cytometry using Hoechst dye. ** P<0.05 vs. unstressed cells, § P<0.05 and §§ P<0.01 vs. BAC-exposed cells. (B) The protective effect of CXCL12 (10 ng/mL, 24 h) is reversed by CXCR4 antagonist (AMD-3100, 1 µM), whereas the apoptotic effect of SDF-1(5-67) (10 ng/mL, 24 h) is inhibited by CXCR3 antagonist (NBI-74330, 1 µM). CXCL10 (10 ng/mL [1.1 nM], 24 h), a conventional ligand for CXCR3, mimics the apoptotic effect of SDF-1(5-67). ** P<0.01. (C) Dose-dependent effect of 24-h incubation with SDF-1(5-67) or with CXCL12. ** P<0.01 vs. CXCL12. (D) SDF-1(5-67) (10 ng/mL) increases caspase-3 activation as assessed by immunoflowcytometry. CXCR3 antagonist (NBI-74330, 1 µM) inhibits SDF-1(5-67)-induced caspase 3 activation. ** P<0.01 vs. unstimulated, §§ P<0.01 vs. SDF-1(5-67)-stimulated. Data in graphs are presented as means ± SEM.
Mentions: In a TC model of toxic-induced apoptosis [34], addition of CXCL12 (10 ng/mL [1.3 nM]) decreased apoptosis, whereas SDF-1(5-67) (10 ng/mL [1.3 nM]) potentiated apoptosis (Fig. 3A). Both the protective effect of CXCL12 and the deleterious effect of SDF-1(5-67) were concentration-dependent, with a maximal effect of either chemokine observed at 10 ng/mL (Fig. 3B).

Bottom Line: Current antiglaucoma therapy does not target the causal trabecular pathology, which may explain why treatment failure is often observed.The protective effect of CXCR3 antagonism is related to restoration of the trabecular function.These data demonstrate that proteolytic cleavage of CXCL12 is involved in trabecular pathophysiology, and that local administration of a selective CXCR3 antagonist may be a beneficial therapeutic strategy for treating ocular hypertension and subsequent retinal degeneration.

View Article: PubMed Central - PubMed

Affiliation: UPMC University Paris 6, Institut de la Vision, UMRS968, Paris, France. alexandre.denoyer@gmail.com

ABSTRACT
Glaucoma, the most common cause of irreversible blindness, is a neuropathy commonly initiated by pathological ocular hypertension due to unknown mechanisms of trabecular meshwork degeneration. Current antiglaucoma therapy does not target the causal trabecular pathology, which may explain why treatment failure is often observed. Here we show that the chemokine CXCL12, its truncated form SDF-1(5-67), and the receptors CXCR4 and CXCR3 are expressed in human glaucomatous trabecular tissue and a human trabecular cell line. SDF-1(5-67) is produced under the control of matrix metallo-proteinases, TNF-α, and TGF-β2, factors known to be involved in glaucoma. CXCL12 protects in vitro trabecular cells from apoptotic death via CXCR4 whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase activation. Ocular administration of SDF-1(5-67) in the rat increases intraocular pressure. In contrast, administration of a selective CXCR3 antagonist in a rat model of ocular hypertension decreases intraocular pressure, prevents retinal neurodegeneration, and preserves visual function. The protective effect of CXCR3 antagonism is related to restoration of the trabecular function. These data demonstrate that proteolytic cleavage of CXCL12 is involved in trabecular pathophysiology, and that local administration of a selective CXCR3 antagonist may be a beneficial therapeutic strategy for treating ocular hypertension and subsequent retinal degeneration.

Show MeSH
Related in: MedlinePlus