Ago2 regulation for K+ channel-mediated human adipose tissue-derived stromal cell self-renewal and survival in the nucleus

Abstract

Argonaute2 (Ago2) is a well-known factor that has intrinsic endonuclease activity and is a part of the fundamental gene regulatory machinery. Recently, we showed that nuclear Ago2 regulates voltage-gated potassium (Kv) channels and that Ago2/Kv1.3 has crucial functions in the self-renewal and cell de-aging processes in adipose tissue-derived stromal cells (ATSCs). In the nucleus, Ago2 bound to the promoter regions of calcium-activated potassium channel 3, potassium channel subfamily K member 1 (KCNK1), and voltage-gated potassium channel 2, and the expression of these genes was significantly upregulated at the level of transcription. We detected an active K+ channel that plays a critical role in Ago2-mediated ATSC self-renewal through the control of membrane potential during cell self-renewal and differentiation. Among the several regulatory subunits of voltage-dependent K+ (Kv) channels, Kv1.3 and Kv1.5 have been shown to impact tissue differentiation and cell growth in cultured ATSCs following their direct binding to the regulatory region of the Kv channel gene. In ATSCs, interference with Ago2 or K+ channel gene expression or treatment with tetraethylammonium significantly downregulated stemness gene expression, induced cell cycle arrest, and inhibited the ability of cells to transdifferentiate into neurons or beta-cells via Oct4 knockdown. Blockage of the K+ channel significantly induced protein kinase C (PKC) alpha, beta, and delta phosphorylation and negatively affected Ago2 and Oct4 expression. This K+ channel blockage also resulted in the upregulation of p53 and p21 expression and the inactivation of mitogen-activated protein kinase (MEK), extracellular signal-regulated kinase 1/2 (ERK 1/2), AKT, beta-catenin, and STAT3. Our results suggest that the nuclear Ago2 regulation of the K+ channel or stemness-related gene expression plays a critical role in adult stem cell self-renewal and differentiation.