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A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
Published on: September 11, 2020
Distinct MicroRNAs Identified in Rabbit Blood Arising from Induced Diabetes and a Surgically Simulated Diabetic
Girish J Kotwal1, Sabine Waigel2, Julia Chariker3
1Noveratech LLC, Louisville, KY, 40299, USA.
Background:
Diabetic complications have been studied extensively in recent years. There are very few biomarkers in body fluids that can pinpoint a distinct diabetic complication due to insufficient known specific biomarkers for ischemia.
Objective:
Identifying microRNA in animal models for each complication could enable early diagnosis of a given complication if verified in humans. MicroRNA (miRNA) profiling has been done in rodent models for a number of diabetic complications, like diabetic glomerular injury, atherosclerosis, cognitive impairment, diabetic wound healing, angiopathy and other complications. Due to multiple differences between rodents and humans, the changes in rabbit skin, considered closer to humans than even pigs, may better simulate human diabetic complications of ischemia.
Methods:
To study the miRNA profile of rabbits in which diabetes was induced or ischemia was surgically generated, we studied whether diabetes or ischemia-induced specific miRNA could be detected. MicroRNA from the blood of diabetic rabbits and rabbits with local ischemia was collected in PAXgene Blood RNA tubes specifically designed for miRNA isolation and extracted using the PAX gene miRNA extraction kit. The isolated RNA was quality controlled using an RNA analyzer, and further, using RNA seq technology, it was analyzed for distinct miRNAs that were detected in diabetic and non-diabetic rabbits induced with ischemia.
Results:
A miRNA that was found to be expressed in diabetic rabbits and ischemic rabbits but not in untreated rabbits was miRNA-183. Several miRNAs were differentially expressed across comparison groups, and several upregulated miRNAs were identified being unique to each comparison. In rabbits with a potential diabetic complication of a long-term ischemic model, there was one distinct microRNA, which was highly significantly upregulated in ischemia rabbit (miRNA-133-3p). One miRNA that was highly significantly upregulated in diabetic rabbit but not in ischemic rabbits was miRNA-3074-5p. Only statistically significant results have been considered and analyzed.
Conclusion:
These findings could lead to a precise and timely diagnosis of a potential single diabetic complication without invasive tissue biopsies and could be a novel tool in the management of diabetic patients developing complications due to the progression of diabetes.
Insights
Researchers identified specific microRNAs (miRNAs) in rabbits that could help diagnose diabetic complications like ischemia early. This discovery may lead to non-invasive diagnostic tools for diabetic patients.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Diabetic complications are a significant health concern.
- Biomarkers for specific diabetic complications, particularly ischemia, are lacking in body fluids.
- MicroRNA (miRNA) profiling in animal models shows promise for early diagnosis.
Purpose of the Study:
- To investigate miRNA profiles in rabbits with induced diabetes or ischemia.
- To identify specific miRNAs associated with diabetic complications, especially ischemia.
- To evaluate rabbits as a model for human diabetic complications due to physiological similarities.
Main Methods:
- Diabetes and local ischemia were surgically induced in rabbits.
- Blood samples were collected for microRNA isolation using specialized kits.
- RNA sequencing (RNA seq) technology was employed for miRNA analysis.
Main Results:
- MicroRNA-183 was detected in both diabetic and ischemic rabbits, but not in controls.
- Distinct miRNAs were upregulated in each comparison group.
- MicroRNA-133-3p was significantly upregulated in ischemic rabbits, while miRNA-3074-5p was upregulated in diabetic rabbits.
Conclusions:
- Specific miRNAs can serve as biomarkers for diabetic complications like ischemia.
- These findings may enable early, non-invasive diagnosis of diabetic complications.
- This research offers a potential new tool for managing diabetic patients with complications.
