In retinopathy of prematurity (ROP), compromised physiologic vascularity and delay in physiologic retinal vascular development (PRVD) in Phase I precede vasoproliferative intravitreal neovascularization (IVNV) in Phase II. During prior grant periods, we found that intravitreal antibodies that inhibit VEGF-signaling reorder angiogenesis, which reduces IVNV and allows some PRVD, also impair pup growth, reduce physiologic vascularity and disorganize retinal structure in rodent oxygen-induced retinopathy (OIR) models. Targeting over-expressed VEGF or activated proteins in the VEGF-signaling pathway with cell-specific shRNA inhibits IVNV without reducing pup growth or physiologic vascularity but does not completely extend PRVD. In human infants with severe ROP we found that recurrent IVNV occurs due to incomplete PRVD and compromised physiologic vascularity after intravitreal anti-VEGF antibody. Together, these findings suggest that physiologic reorientation of angiogenesis to treat pathologic IVNV is insufficient to extend PRVD, but complete PRVD - with preserved physiologic vascularity - is important to prevent recurrent IVNV. Our next goal is to identify mechanisms to extend PRVD and preserve physiologic vascularity to prevent severe ROP by exploiting protective mechanisms endogenously or through exogenous treatment. Using models of maternal uteroplacental insufficiency (UPI) and pup OIR, we identified upregulated systemic erythropoietin (EPO) as a potentially-protective factor, which extended PRVD. We also found the protein, mediator of ERBB2-driven motility (MEMO1), localized to IVNV but not to normal developing vessels and interfered with EPO-induced mechanisms to inhibit retinal endothelial cell (EC) apoptosis and quench reactive oxygen species (ROS). MEMO1 was necessary for VEGF-induced angiogenesis mediated through the transcription factor, STAT3, in ECs. Using a lentiviral gene therapy method to specifically knockdown STAT3 in retinal ECs in vivo, we found EC-STAT3 knockdown inhibited IVNV in rat OIR. These findings support our goal for the next grant period to test the hypothesis that PRVD and physiologic vascularity can be modulated by EPO signaling through EPO- receptor (EPOR) in angiogenesis.
Specific Aim 1 is to test if induced endogenous EPO signaling from UPI and pup-OIR is important for physiologic vascularity and regrowth by reducing ROS and apoptosis.
Specific Aim 2 is to test if exogenous EPO is sufficient to promote PRVD and improve retinal structure and function without increasing IVNV in the setting of knocked down EC EPOR.
Specific Aim 3 is to test if MEMO1 interferes with EPO-mediated mechanisms to promote PRVD, and if MEMO1 sustains VEGF-induced angiogenic signaling leading to IVNV. Methods include: novel lentiviral gene therapy in rat; oxygen-induced retinopathy models in rat and in novel composite murine dam UPI/pup OIR models; humanized knockin mice in which EPO signaling is hypoactive; Micron IV imaging; subretinal injections; optical coherence tomography; electroretinography; immunohistochemistry of flat mounts and sections; western blot, real-time PCR.

Public Health Relevance

Retinopathy of prematurity (ROP) is a leading cause of childhood blindness worldwide. Besides vascular endothelial growth factor (VEGF), erythropoietin appears can have either beneficial or detrimental effects on severe ROP and premature infants. Our lab is working to understand how to promote beneficial effects of erythropoietin and to inhibit the unwanted effects and ultimately to develop strategies to safely prevent severe ROP without adversely affecting infant development.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY015130-15
Application #
9656756
Study Section
Diseases and Pathophysiology of the Visual System Study Section (DPVS)
Program Officer
Shen, Grace L
Project Start
2004-12-02
Project End
2024-08-31
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
15
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Utah
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Simmons, Aaron B; Bretz, Colin A; Wang, Haibo et al. (2018) Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy. Angiogenesis 21:751-764
Bretz, Colin A; Divoky, Vladimir; Prchal, Josef et al. (2018) Erythropoietin Signaling Increases Choroidal Macrophages and Cytokine Expression, and Exacerbates Choroidal Neovascularization. Sci Rep 8:2161
Becker, Silke; Wang, Haibo; Simmons, Aaron B et al. (2018) Targeted Knockdown of Overexpressed VEGFA or VEGF164 in M├╝ller cells maintains retinal function by triggering different signaling mechanisms. Sci Rep 8:2003
Wang, Haibo; Hartnett, M Elizabeth (2017) Roles of Nicotinamide Adenine Dinucleotide Phosphate (NADPH) Oxidase in Angiogenesis: Isoform-Specific Effects. Antioxidants (Basel) 6:
Becker, Silke; Wang, Haibo; Yu, Baifeng et al. (2017) Protective effect of maternal uteroplacental insufficiency on oxygen-induced retinopathy in offspring: removing bias of premature birth. Sci Rep 7:42301
Zhu, Weiquan; Shi, Dallas S; Winter, Jacob M et al. (2017) Small GTPase ARF6 controls VEGFR2 trafficking and signaling in diabetic retinopathy. J Clin Invest 127:4569-4582
Todorich, Bozho; Thanos, Aristomenis; Yonekawa, Yoshihiro et al. (2017) Correspondence. Retina 37:e52-e54
Hartnett, Mary Elizabeth (2017) Advances in understanding and management of retinopathy of prematurity. Surv Ophthalmol 62:257-276
Shulman, Julia P; Weng, Cindy; Wilkes, Jacob et al. (2017) Association of Maternal Preeclampsia With Infant Risk of Premature Birth and Retinopathy of Prematurity. JAMA Ophthalmol 135:947-953
Becker, Silke; Wang, Haibo; Stoddard, Gregory J et al. (2017) Effect of subretinal injection on retinal structure and function in a rat oxygen-induced retinopathy model. Mol Vis 23:832-843

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