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A novel method for visualizing and tracking endogenous mRNA in a specific cell population in pathological
Md Imam Uddin1,2, Tyler C Kilburn3, Sara Z Jamal3
1Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, AA1324 Medical Center North, Nashville, TN, 37232, USA. md.i.uddin@Vanderbilt.Edu.
Scientific Reports
|January 29, 2021
Summary
Researchers developed a novel imaging probe to detect endoglin mRNA, a marker for retinal neovascularization. This method allows real-time visualization of disease progression in conditions like oxygen-induced retinopathy (OIR).
Area of Science:
- Ophthalmology
- Molecular Imaging
- Biotechnology
Background:
- Potentially blinding conditions like diabetic retinopathy involve neovascularization (NV).
- Accurate assessment of retinal NV is crucial for patient management.
- Current imaging methods lack real-time molecular assessment of NV.
Purpose of the Study:
- To investigate endoglin mRNA as a marker for retinal NV.
- To develop and validate a novel molecular imaging probe for endoglin mRNA.
- To assess the probe's ability to visualize and quantify retinal NV in vivo.
Main Methods:
- Developed a fluorescently labeled anti-sense endoglin short hairpin RNA (AS-Eng shRNA) probe conjugated to a lipid for enhanced circulation.
- Administered the probe to a mouse model of oxygen-induced retinopathy (OIR).
- Performed ex vivo retinal imaging to detect endoglin mRNA-dependent fluorescence and co-localization with macrophage marker IBA1.
Main Results:
- The AS-Eng shRNA-lipid probe specifically detected endoglin mRNA in neovascular structures of OIR mouse retinas.
- Fluorescence signal intensity correlated with neovascularization burden.
- Endoglin mRNA-expressing cells co-localized with IBA1-positive cells, suggesting macrophage/microglia involvement.
Conclusions:
- Endoglin mRNA is a viable molecular target for imaging retinal neovascularization.
- The developed probe enables sensitive, real-time detection and quantification of retinal NV.
- This technology holds promise for assessing blinding retinal diseases in living systems.
