Endothelin-1 traps potently reduce pathologic markers back to basal levels in an in vitro model of diabetes

Abstract

Background

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.

Methods

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).

Results

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.

Conclusion

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.

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Funding

This project was funded by private funding.

Author information

Affiliations

Authors

Contributions

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.

Corresponding author

Correspondence to Arjun Jain.

Ethics declarations

Conflict of interest

The author(s) declare that they have no competing interests.

Declaration statement

Any inquiries on data of this study can be directed to Dr. Arjun Jain.

Additional information

Key points

ET-traps have a very high binding affinity.

ET-traps significantly return to basal levels different pathologic markers both at the mRNA and at the protein level.

ET-traps are non-cytotoxic.

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Jain, A., Chen, S., Yong, H. et al. Endothelin-1 traps potently reduce pathologic markers back to basal levels in an in vitro model of diabetes. J Diabetes Metab Disord 17, 189–195 (2018). https://doi.org/10.1007/s40200-018-0360-8

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Keywords

  • DM
  • Diabetes mellitus; ECM
  • Extracellular matrix; ET-1
  • Endothelin-1; ETtr
  • Endothelin-1 traps; FFP
  • Fc-fusion protein; FN
  • Fibronectin