Article

Derivation and Functional Analysis of Patient-Specific Induced Pluripotent Stem Cells as an In Vitro Model of Chronic Granulomatous Disease

Institute of Genetic Medicine, Newcastle University, Newcastle, UK.
Stem Cells (Impact Factor: 7.7). 04/2012; 30(4):599-611. DOI: 10.1002/stem.1053
Source: PubMed

ABSTRACT Chronic granulomatous disease (CGD) is an inherited disorder of phagocytes in which NADPH oxidase is defective in generating reactive oxygen species. In this study, we reprogrammed three normal unrelated patient's fibroblasts (p47phox and gp91phox) to pluripotency by lentiviral transduction with defined pluripotency factors. These induced pluripotent stem cells (iPSC) share the morphological features of human embryonic stem cells, express the key pluripotency factors, and possess high telomerase activity. Furthermore, all the iPSC lines formed embryoid bodies in vitro containing cells originating from all three germ layers and were capable of teratoma formation in vivo. They were isogenic with the original patient fibroblasts, exhibited normal karyotype, and retained the p47phox or gp91phox mutations found in the patient fibroblasts. We further demonstrated that these iPSC could be differentiated into monocytes and macrophages with a similar cytokine profile to blood-derived macrophages under resting conditions. Most importantly, CGD-patient-specific iPSC-derived macrophages showed normal phagocytic properties but lacked reactive oxygen species production, which correlates with clinical diagnosis of CGD in the patients. Together these results suggest that CGD-patient-specific iPSC lines represent an important tool for modeling CGD disease phenotypes, screening candidate drugs, and the development of gene therapy. Stem Cells
2012; 30:599–611

Download full-text

Full-text

Available from: Gabriele Saretzki, Jul 04, 2015
0 Followers
 · 
145 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Copyright: © 2012 Young W, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract The breakthrough development of induced pluripotent stem cell (iPSC) technology is not only revolutionizing basic stem cell science but is also spurring efforts to reprogram one somatic cell type directly into another. Induced pluripotent stem cells provide scientists with a self-renewing and, thus, unlimited, source of pluripotent cells for targeted differentiation, in principle, into the entire range of cell types found in the body. Therefore, iPSC technology and the increasingly refined abilities to differentiate iPSCs into disease-relevant mature cells has far reaching implications for understanding disease etiology and promoting drug discovery and other advances in regenerative medicine. In this review, we summarize the latest progress in the application of patient-specific iPSCs for disease modeling, drug screening and cell replacement therapy, and discuss their impact on precision personalized medicine.
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The function of the proteasome is essential for maintenance of cellular homeostasis, and in pluripotent stem cells, this has been extended to the removal of nascent proteins in a manner that restricts differentiation. In this study, we show enhanced expression of genes encoding subunits of the 20S proteasome in human embryonic stem cells (hESCs) coupled to their downregulation as the cells progress into differentiation. The decrease in expression is particularly marked for the alternative catalytic subunits of the 20S proteasome variant known as the immunoproteasome indicating the possibility of a hitherto unknown function for this proteasome variant in pluripotent cells. The immunoproteasome is normally associated with antigen-presenting cells where it provides peptides of an appropriate length for antibody generation; however, our data suggest that it may be involved in maintaining the pluripotency in hESCs. Selective inhibition of two immunoproteasome subunits (PSMB9 and PSMB8) results in downregulation of cell surface and transcriptional markers that characterize the pluripotent state, subtle cell accumulation in G1 at the expense of S-phase, and upregulation of various markers characterizing the differentiated primitive and definitive lineages arising from hESC. Our data also support a different function for each of these two subunits in hESC that may be linked to their selectivity in driving proteasome-mediated degradation of cell cycle regulatory components and/or differentiation inducing factors.
    Stem Cells 07/2012; 30(7):1373-84. DOI:10.1002/stem.1113 · 7.70 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In the past few years, cellular programming, whereby virtually all human cell types, including those deep within the brain or internal organs, can potentially be produced and propagated indefinitely in culture, has opened the door to a new type of disease modeling. Importantly, many diseases or disease predispositions have genetic components that vary from person to person. Now cells from individuals can be readily reprogrammed to form pluripotent cells, and then directed to differentiate into the lineage and the cell type in which the disease manifests. Those cells will contain the genetic contribution of the donor, providing an excellent model to delve into human disease at the level of individuals and their genomic variants. To date, over fifty such disease models have been reported, and while the field is young and hurdles remain, these tools promise to inform scientists about the cause and cellular-molecular mechanisms involved in pathology, unravel the role of environmental versus hereditary factors driving disease, and provide an unprecedented tool for screening therapeutic agents that might slow or halt disease progression.
    Current opinion in genetics & development 08/2012; DOI:10.1016/j.gde.2012.07.004 · 8.57 Impact Factor