Abstract
Purpose :
To investigate the effect of collagen type X encoding gene COL10A1 in the development of CNV and its mechanism. Then to verify whether the inhibition of collagen type X by antibody could be a new target for the treatment for CNV in a mouse model.
Methods :
C57BL/6J mice were treated with 532 nm argon laser for the formation of CNV model. The total RNA of retina-choroid tissue in either CNV mouse model group and normal control mouse group was collected for RNA sequencing and bioinformatic analysis to compare the difference of COL10A1 transcriptome expression. Then immunofluorescence of retina-choroid cryosections was carried out comparing the expression of collagen type X between CNV mouse model and normal controls. Next, human retinal microvascular endothelial cells (HRMEC) and human umbilical vein endothelial cells (HUVEC) in a hypoxic environment of 150 µM CoCl2 was used for an in vitro cell model of angiogenesis. Further more, COL10A1 small interfering RNA (siRNA) was applied to HRMEC and HUVEC under hypoxic conditions and the proliferation and tube formation ability were analyzed. Finally, anti-collagen type X mAb was intravitreally injected to the CNV mouse model on day 7 after laser, the leakage area and the CNV area in choroidal flat mounts were analyzed on day 14.
Results :
Collagen type X was increasingly expressed in retina-choroid tissue of mice with CNV by immunohistofluorescence (P < 0.01). COL10A1 mRNA and protein were also upregulated in HRMEC and HUVEC under hypoxic conditions (P < 0.02). siRNA knockdown of COL10A1 suppressed the proliferation (P < 0.001) and tube formation (P < 0.02) ability of HRMEC and HUVEC under hypoxic conditions. Snail family transcriptional repressor 1 (SNAIL1) and angiopoietin-2 (ANGPT2) were downregulated (P < 0.03) in COL10A1 knockdown HRMEC and HUVEC under hypoxic conditions. Thereafter, CNV lesion area by immunofluorescence of choroidal flat mounts was significantly decreased (P < 0.02) in a CNV mouse model intravitreally injected with anti-collagen type X mAb compared to controls.
Conclusions :
1. COL10A1 was increasingly expressed in the retina-choroid CNV tissue in mice; 2. In a hypoxic cell model, COL10A1 was also increasingly expressed and promotes angiogenesis of endothelial cells by upregulation of SNAIL1 and ANGPT2; 3. Intravitreal injection of collagen type X mAb could suppress CNV in mouse CNV model. COL10A1 maybe a new target for treatment of nAMD.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.