A Small Molecule Primes Embryonic Stem Cells for Differentiation

The Skaggs Institute of Chemical Biology and the Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Cell stem cell (Impact Factor: 22.27). 06/2009; 4(5):416-26. DOI: 10.1016/j.stem.2009.04.001
Source: PubMed


Embryonic stem cells (ESCs) are an attractive source of cells for disease modeling in vitro and may eventually provide access to cells/tissues for the treatment of many degenerative diseases. However, applications of ESC-derived cell types are largely hindered by the lack of highly efficient methods for lineage-specific differentiation. Using a high-content screen, we have identified a small molecule, named stauprimide, that increases the efficiency of the directed differentiation of mouse and human ESCs in synergy with defined extracellular signaling cues. Affinity-based methods revealed that stauprimide interacts with NME2 and inhibits its nuclear localization. This, in turn, leads to downregulation of c-Myc, a key regulator of the pluripotent state. Thus, our findings identify a chemical tool that primes ESCs for efficient differentiation through a mechanism that affects c-Myc expression, and this study points to an important role for NME2 in ESC self-renewal.

Download full-text


Available from: Costas A Lyssiotis,
  • Source
    • "Advances in chemistry have returned unparalleled sensitivity and scope for detecting, isolating and identifying not only the major metabolites (as was the norm only two decades ago), but the full array of structurally related minor cometabolites – Nature " s equivalent of combinatorial chemistry (Chen et al., 2009). Empowered by ready access to such pre-programmed bioactive chemical diversity, structure activity relationship (SAR) investigations by co-metabolite have become a powerful paradigm for accelerating the evaluation and characterization of new drug classes (pharmacophores) (Zhan et al., 2008).More recently the differentiation of embryonic stem cells for disease modeling in-vitro by using "Stauprimide" a family of indolecarbazoles from Streptomyces staurosporeus (Zhu et al., 2009). A new group of obligate marine actinomycete produced "Salinosporamide-A" shows cytotoxic towards HCT-116 human colon carcinoma cell lines (Feeling et al., 2003). "
    [Show abstract] [Hide abstract]
    ABSTRACT: o maintain and improve the quality of life offered by modern healthcare requires an ongoing commitment to the development of new drugs, to improve and replace those that have become less effective, and to bring to the community safer treatments for an ever-wider array of important diseases. Irrespective of the specific medical need, the drug discovery pipeline is critically dependent on access to diverse, high-quality molecular libraries capable of inspiring drug-led discovery, and ultimately new drugs. A poor choice of chemistry leads to wasted resources and no drugs. Historically the pharmaceutical industry has relied heavily on microbial natural products, which represent an extraordinarily diverse, preassembled pool of biologically active molecules, programmed to be potent and selective modulators of key biopolymers, cells, tissues, organs and animals. Knowledge of Nature’s intellectual property, gleaned from the evolutionary equivalent of a billion-year global drug discovery program, with an unlimited budget and a workforce of trillions, can disclose privileged bioactive structures that inform, guide and inspire modern drug discovery, re-purposing ecological advantage to pharmaceutical benefit. Although we cannot see them, microbes are essential for every part of human life; indeed all life on Earth. The emerging field of metagenomics provides a new way of viewing the microbial world that will not only transform modern microbiology, but also may revolutionize understanding of the entire living world.
  • Source
    • "Author's personal copy resulting in downregulation of c-Myc, a key factor in the maintenance of ESC self-renewal (Zhu et al., 2009). Thus, Spd is dependent on the subsequent treatment with Activin A for the differentiation of Spd-primed ESC to definitive endoderm. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Diabetes is a group of metabolic diseases, rising globally at an alarming rate. Type 1 (juvenile diabetes) is the autoimmune version of diabetes where the pancreas is unable to produce insulin, whereas Type 2 (adult onset diabetes) is caused due to insulin resistance of the cells. In either of the cases, elevated blood glucose levels are observed which leads to progressive comorbidity like renal failure, cardiovascular disease, retinopathy, etc. Metformin, sulphonyl urea group of drugs, as well as insulin injections are the available therapies. In advanced cases of diabetes, the drug alone or drug in combination with insulin injections are not able to maintain a steady level of blood glucose. Moreover, frequent insulin injections are rather cumbersome for the patient. So, regenerative medicine could be a permanent solution for fighting diabetes. Islet transplantation has been tried with a limited amount of success on a large population of diabetics because of the shortage of cadaveric pancreas. Therefore, the best proposed alternative is regenerative medicine involving human pluripotent stem cell (hPSC)-derived beta islet transplantation which can be obtained in large quantities. Efficient protocols for in vitro differentiation of hPSC into a large number of sustained insulin-producing beta cells for transplantation will be considered to be a giant leap to address global rise in diabetic cases. Although most of the protocols mimic in vivo pancreatic development in humans, considerable amount of lacuna persists for near-perfect differentiation strategies. Moreover, beta islets differentiated from hPSC have not yet been successfully translated under clinical scenario.
    Vitamins & Hormones 02/2014; 95:223-48. DOI:10.1016/B978-0-12-800174-5.00009-0 · 2.04 Impact Factor
  • Source
    • "As reported before, Spd has been shown to inhibit nuclear localization of nucleoside diphosphate kinase B (DNPK B), which led to the down regulation of c-MYC a known main factor in hESC self-renewal. Thus it could prime hESCs for differentiation (26). As we previously reported (27), pre-treatment of hESCs with SM suppressors of pluripotency could be a strategy to improve the efficiency of DE induction. "
    [Show abstract] [Hide abstract]
    ABSTRACT: In vitro production of a definitive endoderm (DE) is an important issue in stem cell-related differentiation studies and it can assist with the production of more efficient endoderm derivatives for therapeutic applications. Despite tremendous progress in DE differentiation of human embryonic stem cells (hESCs), researchers have yet to discover universal, efficient and cost-effective protocols. In this experimental study, we have treated hESCs with 200 nM of Stauprimide (Spd) for one day followed by activin A (50 ng/ml; A50) for the next three days (Spd-A50). In the positive control group, hESCs were treated with Wnt3a (25 ng/ml) and activin A (100 ng/ml) for the first day followed by activin A for the next three days (100 ng/ml; W/A100-A100). Gene expression analysis showed up regulation of DE-specific marker genes (SOX17, FOXA2 and CXCR4) comparable to that observed in the positive control group. Expression of the other lineage specific markers did not significantly change (p<0.05). We also obtained the same gene expression results using another hESC line. The use of higher concentrations of Spd (400 and 800 nM) in the Spd-A50 protocol caused an increase in the expression SOX17 as well as a dramatic increase in mortality rate of the hESCs. A lower concentration of activin A (25 ng/ml) was not able to up regulate the DE-specific marker genes. Then, A50 was replaced by inducers of definitive endoderm; IDE1/2 (IDE1 and IDE2), two previously reported small molecule (SM) inducers of DE, in our protocol (Spd-IDE1/2). This replacement resulted in the up regulation of visceral endoderm (VE) marker (SOX7) but not DE-specific markers. Therefore, while the Spd-A50 protocol led to DE production, we have shown that IDE1/2 could not fully replace activin A in DE induction of hESCs. These findings can assist with the design of more efficient chemically-defined protocols for DE induction of hESCs and lead to a better understanding of the different signaling networks that are involved in DE differentiation of hESCs.
    Cell Journal 02/2014; 16(1):63-72. · 1.11 Impact Factor
Show more