Inhibition of Lnk in Mouse Induced Pluripotent Stem Cells Promotes Hematopoietic Cell Generation

Tashiro, Kentaro; Ōmori, Makoto; Kono, Kenji; Hirata, Nobue; Yamaguchi, Tomoko; Sakurai, Fuminori; Takaki, Satoshi; Mizuguchi, Hiroyuki

doi:10.1089/scd.2012.0100

Cited by 2 publications

(3 citation statements)

References 50 publications

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“…Various prohypertrophic stimuli function via the activation of ERKs, c-Jun N-terminal kinases, and p38, which trigger numerous intracellular targets and, in turn, launch the hypertrophy process. In terms of ERK1/2, studies performed in different systems have yielded contradictory conclusions: SH2B3 is a negative modulator of the ERK1/2 signaling pathway downstream of c-kit, 5,18 EpoR, 19 c-Mpl, 28,29 and FLT3 17 on engagement with their specific ligands. Whereas in macrophages, SH2B3 deficiency diminishes ERK1/2 activation when stimulated with M-CSF.…”

Section: Discussionmentioning

confidence: 99%

Exacerbating Pressure Overload–Induced Cardiac Hypertrophy

et al. 2015

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P athological cardiac remodeling is the process by which a compensatory response is transformed to a maladaptive response on reacting to mechanical load (eg, valvular regurgitation or stenosis) or humoral factors (eg, catecholamine) and is the strongest predictor of heart failure, arrhythmia, and sudden death. In the early stage of this pathological process, to enhance cardiac pump function, the heart undergoes hypertrophic growth via the enlargement of individual myocytes without effects on cardiac function. However, continued insult would ultimately leads to cardiac chamber enlargement and fibrosis with perturbed contractile performance. In addition to histological and functional deterioration, cardiac remodeling is a composite of cellular and molecular changes, including protein synthesis, sarcomeric organization, fibrosis, cell death, and energy metabolism. Although the pathogenesis and development of pathological hypertrophy have been studied extensively, the complete molecular network has not been sufficiently dissected. Moreover, we have recently demonstrated that a number of protein molecules traditionally regarded as immunologic molecules exert key regulatory effects on this process. [1][2][3] Given the prominent and omnipresent role of the immune network in cell biology, the identification of new immune regulators in cardiac hypertrophy will provide additional insights into the mechanism and treatment of this life-threatening disease.Since its discovery, adaptor protein Src homology 2-B3 (SH2B3) has been highlighted as a nonredundant negative modulator in hematopoietic and B-cell development. [4][5][6] Adaptor proteins, which function as molecular scaffolds to orchestrate cellular signals, possess multiple functions, including organization and control of protein translocation, mediation of protein-protein interactions, and recruitment of required enzyme substrates. Based on structural similarities, SH2B3 (also referred to as Lnk) belongs to the SH2B family Abstract-The adaptor protein Src homology 2-B3 (SH2B3), which belongs to a subfamily of Src homology 2 proteins, is a broad inhibitor of growth factors and cytokine signaling in hematopoietic cells. However, the role of SH2B3 in nonhematopoietic systems, particularly cardiomyocytes, has not been defined. In this study, we observed noticeable increase in SH2B3 protein expression during pathological cardiac remodeling in both humans and rodents. Follow-up in vitro gain-and loss-of-function studies suggested that SH2B3 promotes the cardiomyocyte hypertrophy response. Consistent with the cell phenotype, SH2B3 knockout (SH2B3 −/− ) mice exhibited attenuated cardiac remodeling with preserved cardiac function after chronic pressure overload. Conversely, cardiac-specific SH2B3 overexpression aggravated pressure overload-triggered cardiac hypertrophy, fibrosis, and dysfunction. Mechanistically, SH2B3 accelerates and exacerbates cardiac remodeling through the activation of focal adhesion kinase, which, in turn, activates the prohypertrophic downstream phosph...

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Section: Discussionmentioning

confidence: 99%

Exacerbating Pressure Overload–Induced Cardiac Hypertrophy

et al. 2015

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“…EB differentiation of mouse ESCs and iPSCs was performed as reported previously [18, 26]. In brief, to initiate differentiation, mouse ESCs and iPSCs were trypsinized and plated on a culture dish for 30–60 minutes to remove the MEFs.…”

Section: Methodsmentioning

confidence: 99%

“…To form EBs, the cells were then suspended in differentiation medium (Dulbecco's modified Eagle's medium; Wako Chemical, Osaka, Japan, http://www.wako-chem.co.jp/english) supplemented with 15% FBS, NEAA, penicillin/streptomycin, 2 mM l ‐glutamine, and 100 μM 2‐mercaptoethanol (2‐ME; Nacalai Tesque Inc., Kyoto, Japan, http://www.nacalai.co.jp/en) and plated on a round‐bottom Lipidure‐coated 96‐well plate (Thermo Fisher Scientific, Yokohama, Japan, http://www.thermofisher.co.jp/) at 3 × 10 3 cells (ESCs) or 1 × 10 3 cells (iPSCs) per well. We mainly used 7‐day‐cultured EBs, except as indicated, because the proportion of Flk1‐expressing cells in EBs increased to a peak on day 7 and decreased over the next 2 days under our culture conditions, as reported previously [26]. To evaluate the differentiation potential of day 7 EB‐derived cells, the EBs were harvested and subjected to fluorescence‐activated cell sorting (FACS).…”

Section: Methodsmentioning

confidence: 99%

Expression of Coxsackievirus and Adenovirus Receptor Separates Hematopoietic and Cardiac Progenitor Cells in Fetal Liver Kinase 1-Expressing Mesoderm

Tashiro

Hirata

Okada

et al. 2015

Stem Cells Translational Medicine

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In developing embryos or in vitro differentiation cultures using pluripotent stem cells (PSCs), such as embryonic stem cells and induced pluripotent stem cells, fetal liver kinase 1 (Flk1)‐expressing mesodermal cells are thought to be a heterogeneous population that includes hematopoietic progenitors, endothelial progenitors, and cardiac progenitors. However, information on cell surface markers for separating these progenitors in Flk1+ cells is currently limited. In the present study, we show that distinct types of progenitor cells in Flk1+ cells could be separated according to the expression of coxsackievirus and adenovirus receptor (CAR, also known as CXADR), a tight junction component molecule. We found that mouse and human PSC‐ and mouse embryo‐derived Flk1+ cells could be subdivided into Flk1+CAR+ cells and Flk1+CAR− cells. The progenitor cells with cardiac potential were almost entirely restricted to Flk1+CAR+ cells, and Flk1+CAR− cells efficiently differentiated into hematopoietic cells. Endothelial differentiation potential was observed in both populations. Furthermore, from the expression of CAR, Flk1, and platelet‐derived growth factor receptor‐α (PDGFRα), Flk1+ cells could be separated into three populations (Flk1+PDGFRα−CAR− cells, Flk1+PDGFRα−CAR+ cells, and Flk1+PDGFRα+CAR+ cells). Flk1+PDGFRα+ cells and Flk1+PDGFRα− cells have been reported as cardiac and hematopoietic progenitor cells, respectively. We identified a novel population (Flk1+PDGFRα−CAR+ cells) with the potential to differentiate into not only hematopoietic cells and endothelial cells but also cardiomyocytes. Our findings indicate that CAR would be a novel and prominent marker for separating PSC‐ and embryo‐derived Flk1+ mesodermal cells with distinct differentiation potentials.

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Inhibition of Lnk in Mouse Induced Pluripotent Stem Cells Promotes Hematopoietic Cell Generation

Cited by 2 publications

References 50 publications

Exacerbating Pressure Overload–Induced Cardiac Hypertrophy

Exacerbating Pressure Overload–Induced Cardiac Hypertrophy

Expression of Coxsackievirus and Adenovirus Receptor Separates Hematopoietic and Cardiac Progenitor Cells in Fetal Liver Kinase 1-Expressing Mesoderm

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