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Vitamin D regulates glucose homeostasis pathways, but effects of vitamin D supplementation on insulin sensitivity and β-cell function remain unclear. To investigate the effects of vitamin D3 supplementation on insulin sensitivity and β-cell function. This is a pre-specified secondary analysis of the Vitamin D and type 2 diabetes (D2d) study. Overweight/obese adults at high risk for type 2 diabetes were randomly treated with vitamin D3 4000 IU or matching placebo daily for 24 months. Disposition index (DI), as an estimate of β-cell func