Autophagy protects against human islet amyloid polypeptide-associated apoptosis.
Aims/Introduction:  Islets in type 2 diabetes are characterized by deposition of islet amyloid polypeptide (IAPP) as well as β-cell dysfunction. The unique amyloidogenic character of human (h)IAPP is associated with cytotoxicity. Autophagy is a ubiquitous system of cellular recycling that contributes to cell survival. Thus, we examined whether autophagy could ameliorate hIAPP-associated cytotoxicity.


METHODS
First, we used a COS-1 cell model, lacking endogenous IAPP that might affect cytotoxicity related to exogenous hIAPP. Next, we used the mouse β-cell line, MIN-6 cells. Both cells were transfected with hIAPP or rat (r)IAPP expression constructs, or transfected with bicistronic vectors expressing green fluorescent protein (GFP) and either hIAPP or rIAPP for flow cytometry analysis. Cell viability and apoptosis markers were studied in relation to chemical or genetic modulation of autophagy.


RESULTS
The viability of cells expressing hIAPP was significantly decreased as compared with those expressing rIAPP and the cleavage of pro-caspase-3 was elevated in hIAPP-transfected cells. The formation of autophagosomes and the conversion of microtubule-associated protein light chain 3B I to II were elevated in hIAPP-expressing cells. The viability of hIAPP-expressing cells was increased after treatment with rapamycin, an inducer of autophagy, and decreased after treatment with 3-methyladenine, an inhibitor of autophagy. In MIN-6 cells, annexin positive cells were increased by 3-methyladenine and decreased by rapamycin using flow cytometry. Knocking down of the autophagy protein 5 gene decreased hIAPP-transfected cell viability.


CONCLUSIONS
Autophagy is co-localized with hIAPP expression and it plays a protective role in hIAPP-associated apoptosis. (J Diabetes Invest, doi: 10.1111/j.2040-1124.2010.00065.x, 2010).
Departments of Clinical Laboratory Medicine.
Shuhei
S
Morita
The First Department of Medicine.
Setsuya
S
Sakagashira
Departments of Clinical Laboratory Medicine.
Yoshinori
Y
Shimajiri
Department of Medicine, Division of Endocrinology and Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, USA.
Norman L
NL
Eberhardt
Forensic Medicine.
Toshikazu
T
Kondo
Neurology, Wakayama Medical University, Wakayama, Japan.
Tomoyoshi
T
Kondo
Departments of Clinical Laboratory Medicine.
Tokio
T
Sanke
Autophagy; Islet amyloid polypeptide; Type 2 diabetes