About This Project
One of the major challenges in diagnosing diabetic retinopathy (DR) in its early stages is the absence of easily observable ophthalmic symptoms. Research suggests that retinal neurodegeneration is the earliest pathological feature of DR. Therefore, we propose to identify retinal neurodegeneration using potential blood-based biomarkers. We will obtain evidence through an in vitro model of diabetic retinopathy.
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What is the context of this research?
Diabetic retinopathy (DR) is a growing epidemic and the most common complication of diabetes, affecting vision. Currently, DR is only diagnosed after the onset of microvascular complications due to a lack of easily observable symptoms, and treatment is only available for the advanced stages of the disease. DR damages the neuronal and vascular retina, and it has been evident that neurodegeneration in the retina is the earliest alteration in the disease's progression. Therefore, it is necessary to identify biomarkers for early detection of retinopathy. The proposed project aims to create a proof of concept and translate the research into clinical practice for further identification of potential biomarkers.
What is the significance of this project?
The proposed in vitro study will be helpful for creating a proof of concept that may lead to the identification of novel biomarkers for screening diabetic retinopathy (DR) in the future. The detection of retinal neurodegeneration in diabetic subjects is crucial, and the data generated from this study could be useful for designing future studies using clinical samples. While clinical diagnostic tools such as optical coherence tomography and electroretinogram have been proven effective in detecting retinal neurodegeneration in its early stages, these techniques are often expensive and not easily accessible for screening purposes in many cases. Currently, there are no blood-based diagnostic tests for detecting diabetic retinopathy, highlighting the need for this study to address this gap.
What are the goals of the project?
The goal of my research is to identify potential biomarkers for detecting retinal neurodegeneration in diabetic subjects. I plan to create a proof-of-concept using an in vitro model of diabetic retinopathy, in which PC12 cell lines will be used to induce diabetes mellitus and diabetic retinopathy. I will isolate extracellular vesicles (EVs), specifically neuron-derived extracellular vesicles (NDEVs), from this in vitro model of retinopathy, and measure the potential proteins as neurodegenerative markers in these vesicles. I will use techniques such as enzyme-linked immunosorbent assay (ELISA), western blot, and transmission electron microscopy (TEM).
Cell Culture Work: In- vitro studies are very important to create proof of concept. The raised amount will be utilised for the creation of diabetic retinopathy model.
TEM Charges- These charges would be useful to pay fees for using transmission electron microscopy facility. This is necessary for the characterisation and validation of extracellular vesicles.
EVs validation and characterisation- This budget would be helpful to buy chemicals and consumables for the experiments.
Platform fee: Experiment.com charges an 8% platform fee plus payment processing fees.
June 1-20, 2023: In vitro model of diabetic retinopathy using PC12 cell lines will be created.
June 21- July 31, 2023: NDEVs will be isolated and characterized from the in vitro model of retinopathy.
August 1-31, 2023: Neurodegenerative markers present in isolated NDEVs will be quantified using ELISA and western blot.
Overall, the project team plans to complete the experimental work for this study within a period of 90 days, from June 1 to August 31, 2023.
May 11, 2023
Jun 20, 2023
Creation of disease model
Jul 31, 2023
Isolation, characterisation and validation of EVs
Aug 31, 2023
Quantification of biomarkers and data analysis
Meet the Team
Siddhita Amit Jadhav
"As an experienced biomedical scientist, Siddhita has spent over 8 years conducting research in the field of ophthalmology. During her time at the Eye Research Institute in Mumbai, India, she became particularly interested in the concept of neurodegeneration in diabetic retinopathy and began exploring this area in more detail as part of her PhD research at Maastricht University, Netherlands.
Through her work, Siddhita is striving to answer important questions about the role that retinal neurodegeneration plays in disease pathology. Using her expertise in molecular biology and culturing techniques, she is working to identify potential diagnostic biomarkers for retinopathy screening, with the goal of improving early detection and treatment of this condition.
Siddhita's dedication to advancing our understanding of neurodegeneration in diabetic retinopathy and her commitment to finding practical solutions to improve patient outcomes make her a valuable contributor to the scientific community."
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