Endothelin-1 traps potently reduce pathologic markers back to basal levels in an in vitro model of diabetes
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Diabetes mellitus is a group of metabolic disorders in which there are high blood glucose levels over a prolonged period. Diabetes is one of many diseases associated with pathologically elevated levels of endothelin (ET)-1. We have recently proposed the development of ET-traps, which are an antibody – based fusion protein that potently bind and sequester pathologically elevated levels of endothelin-1.
We constructed ET-traps that were found to be very potent binders to ET-1, with a KD of 32.5ρM. We then treated human retinal microvascular endothelial cells (HRMECs), which are an in vitro model of glucose induced cellular damage, with 10 nM ET-1 or high glucose levels (25 mM).
In this study, we investigated the effects of our ET-trap constructs on the expression levels of both collagen 4α1 and fibronectin, which are both important pathologic markers in diabetes. Treating HRMECs with 10 nM ET-1 or 25 mM glucose significantly induces the expression of the ECM proteins fibronectin and collagen 4α1, as is found in chronic diabetic complications; Incubation of the cells with the ET-traps significantly prevented the increased expression of fibronectin and collagen 4α1 back to basal levels. This was found with both mRNA and protein expression levels of the two ECM proteins.
Our results provide the first evidence of the efficacy of ET-traps in reducing pathologic markers in an in vitro model (of diabetes). Further research is warranted to determine the efficacy of ET-traps as a therapeutic tool for diabetes, which is a major public health burden around the world.
KeywordsDM Diabetes mellitus; ECM Extracellular matrix; ET-1 Endothelin-1; ETtr Endothelin-1 traps; FFP Fc-fusion protein; FN Fibronectin
AJ and SC participated in the research design. HY and SC conducted the experiments. AJ and SC contributed new reagents or analytic tools. SC performed the data analysis. AJ, SC and SC wrote or contributed to the writing of the manuscript.
This project was funded by private funding.
Compliance with ethical standards
Conflict of interest
The author(s) declare that they have no competing interests.
Any inquiries on data of this study can be directed to Dr. Arjun Jain.
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