Derivation of Human Induced Pluripotent Stem Cells for Cardiovascular Disease Modeling

Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Circulation Research (Impact Factor: 11.02). 04/2011; 108(9):1146-56. DOI: 10.1161/CIRCRESAHA.111.240374
Source: PubMed


The successful derivation of human induced pluripotent stem cells (hiPSCs) by dedifferentiation of somatic cells offers significant potential to overcome obstacles in the field of cardiovascular disease. hiPSC derivatives offer incredible potential for new disease models and regenerative medicine therapies. However, many questions remain regarding the optimal starting materials and methods to enable safe, efficient derivation of hiPSCs suitable for clinical applications. Initial reprogramming experiments were carried out using lentiviral or retroviral gene delivery methods. More recently, various nonviral methods that avoid permanent and random transgene insertion have emerged as alternatives. These include transient DNA transfection using plasmids or minicircles, protein transduction, or RNA transfection. In addition, several small molecules have been found to significantly augment hiPSC derivation efficiency, allowing the use of a fewer number of genes during pluripotency induction. We review these various methods for the derivation of hiPSCs, focusing on their ultimate clinical applicability, with an emphasis on their potential for use as cardiovascular therapies and disease-modeling platforms.

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Available from: Joseph C Wu, Mar 23, 2015
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    • "Suicide genes have also been employed as cytotoxic strategy, in vitro and in vivo models (Huber et al., 1991; Clark et al., 1997; Nor et al., 2002; Nakayama et al., 2005; Hodish et al., 2009; Evans and Dey, 2011; Duarte et al., 2012; Mazor et al., 2012), including combination with replication competent oncolytic viruses (Ahn et al., 2009; Kaur et al., 2009), with some evidence of clinical benefit in solid tumors (Pandha et al., 1999; Freytag et al., 2003, 2007; Nemunaitis et al., 2003; Voges et al., 2003; Li et al., 2007; Xu et al., 2009). Finally, non-integrating vectors (Banasik and McCray, 2010), strategies for the replacement or correction of defective genes (Narsinh et al., 2011; Mukherjee and Thrasher, 2013; Li et al., 2014), together with effective suicide gene strategies, may lead to a more broad application of stem cell or inducible pluripotent stem cell based applications in cancer and regenerative medicine. "
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    ABSTRACT: Adoptive T-cell therapy can involve donor lymphocyte infusion (DLI) after allogeneic hematopoietic stem cell transplantation, the administration of tumor infiltrating lymphocyte (TILs) expanded ex-vivo, or more recently the use of T cell receptor (TCR) or chimeric antigen receptor (CAR) redirected T cells. However cellular therapies can pose significant risks, including graft-versus-host-disease and other on and off-target effects, and therefore strategies need to be implemented to permanently reverse any sign of toxicity. A suicide gene is a genetically encoded molecule that allows selective destruction of adoptively transferred cells. Suicide gene addition to cellular therapeutic products can lead to selective ablation of gene-modified cells, preventing collateral damage to contiguous cells and/or tissues. The ‘ideal’ suicide gene would ensure the safety of gene modified cellular applications by granting irreversible elimination of ‘all’ and ‘only’ the cells responsible for the unwanted toxicity. This review presents the suicide gene safety systems reported to date, with a focus on the state-of-the-art and potential applications regarding two of the most extensively validated suicide genes, including the clinical setting: herpes-simplex-thymidine-kinase (HSV-TK) and inducible-caspase-9 (iCasp9).
    Frontiers in Pharmacology 10/2014; 5(524). DOI:10.3389/fphar.2014.00254 · 3.80 Impact Factor
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    • "iPS cells, which closely resemble embryonic stem (ES) cells, can be derived from human somatic tissues from a variety of diseases to recapitulate complex physiological phenotypes. Patient-specific iPS cells may provide an additional tool for studying human heart disease [6]. Here, to identify the key elements responsible for hypoplasia of left heart development, we generated disease-specific iPS cells in patients with congenital heart malformation, identified unique genes differentially expressed in HLHS, and explored the responsible regulatory network involved in myocardial patterning and morphogenesis during cardiac development with respect to LV hypoplasia in humans. "
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    ABSTRACT: The genetic basis of hypoplastic left heart syndrome (HLHS) remains unknown, and the lack of animal models to reconstitute the cardiac maldevelopment has hampered the study of this disease. This study investigated the altered control of transcriptional and epigenetic programs that may affect the development of HLHS by using disease-specific induced pluripotent stem (iPS) cells. Cardiac progenitor cells (CPCs) were isolated from patients with congenital heart diseases to generate patient-specific iPS cells. Comparative gene expression analysis of HLHS- and biventricle (BV) heart-derived iPS cells was performed to dissect the complex genetic circuits that may promote the disease phenotype. Both HLHS- and BV heart-derived CPCs were reprogrammed to generate disease-specific iPS cells, which showed characteristic human embryonic stem cell signatures, expressed pluripotency markers, and could give rise to cardiomyocytes. However, HLHS-iPS cells exhibited lower cardiomyogenic differentiation potential than BV-iPS cells. Quantitative gene expression analysis demonstrated that HLHS-derived iPS cells showed transcriptional repression of NKX2-5, reduced levels of TBX2 and NOTCH/HEY signaling, and inhibited HAND1/2 transcripts compared with control cells. Although both HLHS-derived CPCs and iPS cells showed reduced SRE and TNNT2 transcriptional activation compared with BV-derived cells, co-transfection of NKX2-5, HAND1, and NOTCH1 into HLHS-derived cells resulted in synergistic restoration of these promoters activation. Notably, gain- and loss-of-function studies revealed that NKX2-5 had a predominant impact on NPPA transcriptional activation. Moreover, differentiated HLHS-derived iPS cells showed reduced H3K4 dimethylation as well as histone H3 acetylation but increased H3K27 trimethylation to inhibit transcriptional activation on the NKX2-5 promoter. These findings suggest that patient-specific iPS cells may provide molecular insights into complex transcriptional and epigenetic mechanisms, at least in part, through combinatorial expression of NKX2-5, HAND1, and NOTCH1 that coordinately contribute to cardiac malformations in HLHS.
    PLoS ONE 07/2014; 9(7):e102796. DOI:10.1371/journal.pone.0102796 · 3.23 Impact Factor
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    • "Induced pluripotent stem cells (hiPS) offers a great opportunity for the generation of hCMC and therefore, the MCA based system are highly suitable for the functional investigation of hiPS-derived hCMC. Moreover, patient derived hiPS allows the establishment of specific hCMC pathology models that could be used for sophisticated screening systems [32]. "
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    ABSTRACT: Unexpected adverse effects on the cardiovascular system remain a major challenge in the development of novel active pharmaceutical ingredients (API). To overcome the current limitations of animal-based in vitro and in vivo test systems, stem cell derived human cardiomyocyte clusters (hCMC) offer the opportunity for highly predictable pre-clinical testing. The three-dimensional structure of hCMC appears more representative of tissue milieu than traditional monolayer cell culture. However, there is a lack of long-term, real time monitoring systems for tissue-like cardiac material. To address this issue, we have developed a microcavity array (MCA)-based label-free monitoring system that eliminates the need for critical hCMC adhesion and outgrowth steps. In contrast, feasible field potential derived action potential recording is possible immediately after positioning within the microcavity. Moreover, this approach allows extended observation of adverse effects on hCMC. For the first time, we describe herein the monitoring of hCMC over 35 days while preserving the hCMC structure and electrophysiological characteristics. Furthermore, we demonstrated the sensitive detection and quantification of adverse API effects using E4031, doxorubicin, and noradrenaline directly on unaltered 3D cultures. The MCA system provides multi-parameter analysis capabilities incorporating field potential recording, impedance spectroscopy, and optical read-outs on individual clusters giving a comprehensive insight into induced cellular alterations within a complex cardiac culture over days or even weeks.
    PLoS ONE 07/2013; 8(7):e68971. DOI:10.1371/journal.pone.0068971 · 3.23 Impact Factor
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