June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
A non-invasive imaging method to detect retinal hypoxia in a rat model of 50/10 oxygen-induced retinopathy using HYPOX4
Author Affiliations & Notes
  • Sara Jamal
    ophthalmology, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • John Penn
    ophthalmology, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • MD Imam Uddin
    ophthalmology, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Footnotes
    Commercial Relationships   Sara Jamal, None; John Penn, None; MD Uddin, None
  • Footnotes
    Support  R01EY029693, R01EY023397, BrightFocus Foundation GRANT ID: M2019023, & Reeves foundation
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 370. doi:
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      Sara Jamal, John Penn, MD Imam Uddin; A non-invasive imaging method to detect retinal hypoxia in a rat model of 50/10 oxygen-induced retinopathy using HYPOX4. Invest. Ophthalmol. Vis. Sci. 2021;62(8):370.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Neovascularization a condition that might arise due to retinal hypoxia is a leading cause of vision loss in a variety of retinopathies. The development of hypoxia detecting probes will provide a platform for early disease diagnosis and therapy monitoring. This study demonstrates the utility of HYPOX4 which is a hypoxia sensitive fluorescein-based imaging probe for early detection of retinal hypoxia in a 50/10 rat model of oxygen-induced retinopathy (OIR) in real-time.

Methods : OIR rat model was induced by exposing newborn Brown Norway rats (BN) to a 24 hrs alternate cycle of 50% and 10% oxygen for 14 days. The pups were returned to normoxia for 2 hrs and HYPOX4 was injected intraperitoneally for in vivo imaging of retinal hypoxia in 50/10 OIR model compared to age-matched control (exposed to normoxia throughout the duration of the experiment). A Separate group of OIR pups were injected with pimonidazole and sacrificed 90 min post-injection. Retinas were harvested and immune-stained for pimonidazole-adducts as an established method for ex vivo detection of retinal hypoxia. Post in vivo imaging, HYPOX4 injected pups were euthanized after 24 hrs for ex vivo imaging of retinal hypoxia. Vascular structures were counterstained using Isolectin B4 (IB4).

Results : Retinal hypoxia was observed within the avascular area in the peripheral retina that was detected with HYPOX4. The observation was validated using pimonidazole immune-staining. Confocal imaging allowed us to detect this retinal hypoxia in the inner retina hypoxic region.

Conclusions : Retinal hypoxia plays a major role in the onset and progression of neovascularization. HYPOX-4 was successfully used as a highly sensitive molecular imaging probe to detect retinal hypoxia in BN 50/10 OIR. Thus, HYPOX-4 could be utilized as a promising and noninvasive imaging tool for early-stage diagnosis and tracing of hypoxia in OIR and potentially in other vascular diseases.

This is a 2021 ARVO Annual Meeting abstract.

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