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Monoclonal antibodies that block Roundabout 1 and 2 signaling target pathological ocular neovascularization through myeloid cells

ABSTRACT: Monoclonal antibodies that block Roundabout 1 and 2 signaling target pathological ocular neovascularization through myeloid cells

PROVIDER: PRJNA1174746 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR31074320.fastq.gz	Fastqsanger.gz
	SRR31074321.fastq.gz	Fastqsanger.gz
	SRR31074322.fastq.gz	Fastqsanger.gz
	SRR31074323.fastq.gz	Fastqsanger.gz
	SRR31074324.fastq.gz	Fastqsanger.gz

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Monoclonal antibodies that block Roundabout 1 and 2 signaling target pathological ocular neovascularization through myeloid cells

Project description:Roundabout (ROBO) 1 and 2 are transmembrane receptors that bind secreted SLIT ligands through their extracellular domains (ECD), and signal through their cytoplasmic domains to modulate the cytoskeleton and regulate cell migration, adhesion, and proliferation. SLIT-ROBO signaling was discovered as a guidance cue for axons in the developing nervous system, but ROBO receptors are expressed by many additional cell types and more broadly regulate organ morphogenesis, as well as cancer and pathological ocular neovascularization. As pharmacological tools to prevent SLIT-ROBOR signaling are lacking, herein we developed human monoclonal antibodies (mAb ) against the ROBO1 and 2 ECD. One antibody that inhibited in vitro SLIT2 signaling through ROBO1/2 reduced ocular neovascularization in two pathological models in vivo . Single cell RNA sequencing revealed that antibody treatment affected several cell types relevant to angiogenesis, including endothelial cells, pericytes, and a heterogeneous population of myeloid cells. mAb treatment improved Blood-Retina Barrier integrity and prevented pathological pericyte activation . SLIT-ROBO signaling inhibition prevented pathological activation of myeloid cells, which was phenocopied by both myeloid cell specific ablation of Robo1 and 2 and by knockout of the downstream effector Pi3kγ . Anti-ROBO1/2 blocking antibodies may thus provide a new promising strategy to combat inflammation in blinding eye diseases.

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COCO/DAND5 inhibits developmental and pathological ocular angiogenesis

Project description:Neovascularization contributes to multiple visual disorders including age-related macular degeneration (AMD). Current therapies for treating ocular angiogenesis are centered on the inhibition of vascular endothelial growth factor (VEGF). While clinically effective, some AMD patients are refractory or develop resistance to anti-VEGF and concerns of increased risks of developing geographic atrophy following long-term treatment have been raised. Identification of alternative pathways to inhibit pathological angiogenesis is thus important. We have identified a novel inhibitor of angiogenesis, COCO/DAND5, a member of the Cerberus-related DAN family. We demonstrate that COCO inhibits sprouting, migration and cellular proliferation of cultured endothelial cells. Intravitreal injections of COCO inhibited retinal vascularization during development and in models of retinopathy of prematurity and AMD. COCO equally abrogated angiogenesis in choroid explants and in a model of choroidal neovascularization. Mechanistically, COCO inhibited the expression of TGFβ and BMP pathwaysand altered ATP production, glucose uptake and redox balance of endothelial cells. Together, these data show that COCO is an inhibitor of retinal and choroidal angiogenesis, possibly representing a therapeutic option for the treatment of neovascular ocular diseases.

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COCO/DAND5 inhibits developmental and pathological ocular angiogenesis

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Project description:PurposeThe administration of anti-vascular endothelial growth factor agents is the standard firs-line therapy for ocular vascular diseases, but some patients still have poor outcomes and drug resistance. This study investigated the role of DCZ19903, a small molecule multitarget kinase inhibitor, in ocular angiogenesis.MethodsThe toxicity of DCZ19903 was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assays, flow cytometry, Calcein-AM/PI staining, and terminal uridine nick-end labeling staining. Oxygen-induced retinopathy and laser-induced choroidal neovascularization models were adopted to assess the antiangiogenic effects of DCZ19903 by Isolectin B4 (GS-IB4) and hematoxylin-eosin staining. EdU assays, transwell migration assays, tube formation, and choroid sprouting assays were performed to determine the antiangiogenic effects of DCZ19903. The antiangiogenic mechanism of DCZ19903 was determined using network pharmacology approach and western blots.ResultsThere was no obvious cytotoxicity or tissue toxicity after DCZ19903 treatment. DCZ19903 exerted the antiangiogenic effects in OIR model and choroidal neovascularization model. DCZ19903 inhibited the proliferation, tube formation, migration ability of endothelial cells, and choroidal explant sprouting. DCZ19903 plus ranibizumab achieved greater antiangiogenetic effects than DCZ19903 or ranibizumab alone. DCZ19903 exerted its antiangiogenic effects via affecting the activation of ERK1/2 and p38 signaling.ConclusionsDCZ19903 is a promising drug for antiangiogenic treatment in ocular vascular diseases.Translational relevanceThese findings suggest that DCZ19903 possesses great antiangiogenic potential for treating ocular vascular diseases.

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