Reprogramming towards pluripotency requires AID-dependent DNA demethylation

By N. Bhutani, Jennifer J. Brady, M. Damian, A. Sacco, S. Corbel, Helen M. Blau. Nature, 2010

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Reprogramming of somatic cell nuclei to yield induced pluripotent stem (iPS) cells makes possible derivation of patient-specific stem cells for regenerative medicine. However, iPS cell generation is asynchronous and slow (2–3 weeks), the frequency is low (<0.1%), and DNA demethylation constitutes a bottleneck. To determine regulatory mechanisms involved in reprogramming, we generated interspecies heterokaryons (fused mouse embryonic stem (ES) cells and human fibroblasts) that induce reprogramming synchronously, frequently and fast. Here we show that reprogramming towards pluripotency in single heterokaryons is initiated without cell division or DNA replication, rapidly (1 day) and efficiently (70%). Short interfering RNA (siRNA)-mediated knockdown showed that activation-induced cytidine deaminase (AID, also known as AICDA) is required for promoter demethylation and induction of OCT4 (also known as POU5F1) and NANOG gene expression. AID protein bound silent methylated OCT4 and NANOG promoters in fibroblasts, but not active demethylated promoters in ES cells. These data provide new evidence that mammalian AID is required for active DNA demethylation and initiation of nuclear reprogramming towards pluripotency in human somatic cells.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/ff2360ee384d263dc84c620658210751423de8f0

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Citations in the corpus

Reversing DNA Methylation: Mechanisms, Genomics, and Biological Functions (2014, neutral): “, 2010), mouse ESC/human fibroblast fused heterokaryons (Bhutani et al., 2010), and iPSCs (Kumar et al.”
DNA Methylation and Its Basic Function (2013, background): “Overexpression of AID/APOBEC promotes DNA demethylation in zebrafish (Rai et al, 2008), whereas knockdown or knockout inhibits the DNA demethylation of various genes necessary for cellular reprogramming and development (Bhutani et al, 2010; Muramatsu et al, 2000; Popp et al, 2010).”
Highly efficient miRNA-mediated reprogramming of mouse and human somatic cells to pluripotency (2011, positive): “Several recent studies have demonstrated the importance of other chromatin remodeling processes in iPSC reprogramming (Bhutani et al., 2010; Lagarkova et al., 2010; Mali et al., 2010).”
Thymine DNA Glycosylase Is Essential for Active DNA Demethylation by Linked Deamination-Base Excision Repair (2011, neutral): “The identification of a ternary complex containing TDG, AID and GADD45a is consistent with the recent recognition of AID as a factor required for DNA demethylation during reprogramming of somatic cells (Bhutani et al., 2010) and erasure of DNA methylation at imprinted and other loci in PGCs (Popp et”
Senescence impairs successful reprogramming to pluripotent stem cells. (2009, background): “(Bhutani et al., 2010).”
Induced pluripotency: history, mechanisms, and applications. (2010, background): “The deaminase Aid has been proposed recently to play a role in the demethylation of the NANOG and OCT4 promoters after cell fusion of mouse ESCs and human fibroblasts (Bhutani et al. 2010; Popp et al. 2010).”
DNA Demethylation Dynamics (2011, background): “We postulate here, based on analyses of recent evidence from our and other laboratories (Bhutani et al., 2010; Cortellino et al., 2011; Guo et al., 2011; He et al., 2011), that DNA methylation and demethylation can be a two-way street, characterized by multiple pathways (Figure 1).”
Nuclear reprogramming to a pluripotent state by three approaches (2010, result): “In contrast to expectations, results from two laboratories using mixed-species heterokaryons have shown that pluripotency genes such as Oct4 and Nanog are activated and their promoters demethylated within 1 day of fusion when mouse ES cells are fused with human B cells or with human fibroblast”
Single-cell expression analyses during cellular reprogramming reveal an early stochastic and a late hierarchic phase. (2012, background): “, 2002) or cell fusion (Bhutani et al., 2010) induce reprogramming rapidly and possibly as a single event with little heterogeneity observed in somatic cells, possibly consistent with a deterministic process (Hanna et al.”
Targets and dynamics of promoter DNA methylation during early mouse development (2010, positive): “This is supported by the observations that demethylation of pluripotency genes is required to reprogram somatic cells to a pluripotent state and that interfering with DNA methylation improves the rate of reprogrammin”