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Glucotoxic and diabetic conditions induce caspase 6-mediated degradation of nuclear lamin A in human islets, rodent islets and INS-1 832/13 cells

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Abstract

Nuclear lamins form the lamina on the interior surface of the nuclear envelope, and regulate nuclear metabolic events, including DNA replication and organization of chromatin. The current study is aimed at understanding the role of executioner caspase 6 on lamin A integrity in islet β-cells under duress of glucotoxic (20 mM glucose; 24 h) and diabetic conditions. Under glucotoxic conditions, glucose-stimulated insulin secretion and metabolic cell viability were significantly attenuated in INS-1 832/13 cells. Further, exposure of normal human islets, rat islets and INS-1 832/13 cells to glucotoxic conditions leads to caspase 6 activation and lamin A degradation, which is also observed in islets from the Zucker diabetic fatty rat, a model for type 2 diabetes (T2D), and in islets from a human donor with T2D. Z-Val-Glu-Ile-Asp-fluoromethylketone, a specific inhibitor of caspase 6, markedly attenuated high glucose-induced caspase 6 activation and lamin A degradation, confirming that caspase 6 mediates lamin A degradation under high glucose exposure conditions. Moreover, Z-Asp-Glu-Val-Asp-fluoromethylketone, a known caspase 3 inhibitor, significantly inhibited high glucose-induced caspase 6 activation and lamin A degradation, suggesting that activation of caspase 3 might be upstream to caspase 6 activation in the islet β-cell under glucotoxic conditions. Lastly, we report expression of ZMPSTE24, a zinc metallopeptidase involved in the processing of prelamin A to mature lamin A, in INS-1 832/13 cells and human islets; was unaffected by high glucose. We conclude that caspases 3 and 6 could contribute to alterations in the integrity of nuclear lamins leading to metabolic dysregulation and failure of the islet β-cell.

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Abbreviations

ELISA:

Enzyme-linked immunosorbent assay

ER:

Endoplasmic reticulum

FTI:

Farnesyl transferase inhibitor

GSIS:

Glucose stimulated insulin secretion

IFNγ:

Interferon gamma

IL-1β:

Interleukin-1beta

iNOS:

Inducible nitric oxide synthase

LMNA:

Lamin A gene

NO:

Nitric oxide

T2D:

Type 2 Diabetes

TNF α:

Tumor necrosis factor alpha

Z-DEVD-FMK:

Caspase 3 inhibitor (Z-Asp-Glu-Val-Asp-fluoromethylketone)

ZDF:

Zucker diabetic fatty

ZLC:

Zucker lean control

ZMPSTE24:

Zinc metallopeptidase (STE24 homolog)

Z-VEID-FMK:

Caspase 6 inhibitor (Z-Val-Glu-Ile-Asp-fluoromethylketone)

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Acknowledgments

This research was supported in part by a Merit Review award (to AK; 1BX000469) from the Department of Veterans Affairs, the National Institutes of Health (DK94201 and EY022230), the Juvenile Diabetes Research Foundation (5-2012-257), and Research Stimulation Funds from the Office of Vice President for Research-Wayne State University. AK is also the recipient of a Senior Research Career Scientist Award from the Department of VA (13S-RCS-006). KS is the recipient of Rumble Fellowship from Wayne State University. We thank Prof. Chris Newgard for providing INS-1 832/13 cells.

Conflict of interest

The authors declare no conflict of interests.

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Authors and Affiliations

  1. B-4237 Research Service, β-Cell Biochemistry Laboratory, John D. Dingell VA Medical Center, 4646 John R, Detroit, MI, 48201, USA

    Syeda Khadija, Rajakrishnan Veluthakal, Vaibhav Sidarala & Anjaneyulu Kowluru

  2. Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MI, 48201, USA

    Syeda Khadija, Rajakrishnan Veluthakal, Vaibhav Sidarala & Anjaneyulu Kowluru

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  1. Syeda Khadija
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  2. Rajakrishnan Veluthakal
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Correspondence to Anjaneyulu Kowluru.

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Khadija, S., Veluthakal, R., Sidarala, V. et al. Glucotoxic and diabetic conditions induce caspase 6-mediated degradation of nuclear lamin A in human islets, rodent islets and INS-1 832/13 cells. Apoptosis 19, 1691–1701 (2014). https://doi.org/10.1007/s10495-014-1038-4

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