Decreased proliferation of aged rat beta cells corresponds with enhanced expression of the cell cycle inhibitor p27KIP1

Abstract

Background

Over 400 million people are diabetic. Type 1 and type 2 diabetes are characterized by decreased functional β-cell mass and, consequently, decreased glucose-stimulated insulin secretion. A potential intervention is transplantation of β-cell containing islets from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant-ready β-cells. Therefore, inducing β-cell proliferation ex vivo could be used to expand functional β-cell mass prior to transplantation. Various molecular pathways are sufficient to induce proliferation of young β-cells; however, aged β-cells are refractory to these proliferative signals. Given that the majority of cadaveric donors fit an aged demographic, defining the mechanisms that impede aged β-cell proliferation is imperative.

Results

We demonstrate that aged rat (5-month-old) β-cells are refractory to mitogenic stimuli that otherwise induce young rat (5-week-old) β-cell proliferation. We hypothesized that this change in proliferative capacity could be due to differences in cyclin-dependent kinase inhibitor expression. We measured levels of p16^INK4a, p15^INK4b, p18^INK4c, p19^INK4d, p21^CIP1, p27^KIP1 and p57^KIP2 by immunofluorescence analysis. Our data demonstrates an age-dependent increase of p27^KIP1 in rat β-cells by immunofluorescence and was validated by increased p27^KIP1 protein levels by western blot analysis. Interestingly, HDAC1, which modulates the p27^KIP1 promoter acetylation state, is downregulated in aged rat islets. These data demonstrate increased p27^KIP1 protein levels at 5 months of age, which may be due to decreased HDAC1 mediated repression of p27^KIP1 expression.

Significance

As the majority of transplant-ready β-cells come from aged donors, it is imperative that we understand why aged β-cells are refractory to mitogenic stimuli. Our findings demonstrate that increased p27^KIP1 expression occurs early in β-cell aging, which corresponds with impaired β-cell proliferation. Furthermore, the correlation between HDAC1 and p27 levels suggests that pathways that activate HDAC1 in aged β-cells could be leveraged to decrease p27^KIP1 levels and enhance β-cell proliferation.