Stoichiometry of Transcription Factors Is Critical for Cardiac Reprogramming

Muraoka, Naoto; Ieda, Masaki

doi:10.1161/circresaha.114.305696

Cited by 18 publications

(14 citation statements)

References 19 publications

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“…Recently, a group used polycistronic vectors to express different ratios of the GMT factors and found that, if Mef2c expression is increased compared with Gata4 and Tbx5, then reprogramming efficiency of fibroblasts into cardiomyocytes increases three‐ to five‐fold (L. Wang et al, ). This study demonstrated that the stoichiometry of reprogramming factors is critical for reprogramming efficiency and may explain why other reports using the same factors demonstrate different reprogramming efficiencies (Muraoka and Ieda, ).…”

Section: Current and Preclinical Cellular Therapies For Cardiac Regensupporting

confidence: 60%

Navigating the labyrinth of cardiac regeneration

Lambers

Kume

2016

Developmental Dynamics

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Heart disease is the number one cause of morbidity and mortality in the world and is a major health and economic burden, costing the United States Health Care System more than $200 billion annually. A major cause of heart disease is the massive loss or dysfunction of cardiomyocytes caused by myocardial infarctions and hypertension. Due to the limited regenerative capacity of the heart, much research has focused on better understanding the process of differentiation toward cardiomyocytes. This review will highlight what is currently known about cardiac cell specification during mammalian development, areas of controversy, cellular sources of cardiomyocytes, and current and potential uses of stem cell derived cardiomyocytes for cardiac therapies. Developmental Dynamics 245:751-761,

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Section: Current and Preclinical Cellular Therapies For Cardiac Regensupporting

confidence: 60%

Navigating the labyrinth of cardiac regeneration

Lambers

Kume

2016

Developmental Dynamics

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“…We also found that retroviral pMX-GMT was more efficient for cardiac reprogramming than lentiviral pDox–GMT. Differences in the viral particles, the Dox-inducible gene expression system, and the stoichiometry of reprogramming factors might all contribute to the lower cardiac reprogramming efficiency seen with pDox-GMT [ 17 , 18 , 20 ]. However, it should be noted that the Dox-inducible fibroblast cell line might improve cardiac reprogramming [ 21 ].…”

Section: Discussionmentioning

confidence: 99%

Single-Construct Polycistronic Doxycycline-Inducible Vectors Improve Direct Cardiac Reprogramming and Can Be Used to Identify the Critical Timing of Transgene Expression

Umei

Yamakawa

Muraoka

et al. 2017

IJMS

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Direct reprogramming is a promising approach in regenerative medicine. Overexpression of the cardiac transcription factors Gata4, Mef2c, and Tbx5 (GMT) or GMT plus Hand2 (GHMT) directly reprogram fibroblasts into cardiomyocyte-like cells (iCMs). However, the critical timing of transgene expression and the molecular mechanisms for cardiac reprogramming remain unclear. The conventional doxycycline (Dox)-inducible temporal transgene expression systems require simultaneous transduction of two vectors (pLVX-rtTA/pLVX-cDNA) harboring the reverse tetracycline transactivator (rtTA) and the tetracycline response element (TRE)-controlled transgene, respectively, leading to inefficient cardiac reprogramming. Herein, we developed a single-construct-based polycistronic Dox-inducible vector (pDox-cDNA) expressing both the rtTA and TRE-controlled transgenes. Fluorescence activated cell sorting (FACS) analyses, quantitative RT-PCR, and immunostaining revealed that pDox-GMT increased cardiac reprogramming three-fold compared to the conventional pLVX-rtTA/pLVX-GMT. After four weeks, pDox-GMT-induced iCMs expressed multiple cardiac genes, produced sarcomeric structures, and beat spontaneously. Co-transduction of pDox-Hand2 with retroviral pMX-GMT increased cardiac reprogramming three-fold compared to pMX-GMT alone. Temporal Dox administration revealed that Hand2 transgene expression is critical during the first two weeks of cardiac reprogramming. Microarray analyses demonstrated that Hand2 represses cell cycle-promoting genes and enhances cardiac reprogramming. Thus, we have developed an efficient temporal transgene expression system, which could be invaluable in the study of cardiac reprogramming.

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“…The importance of TF stoichiometry is well established for efficient cellular reprogramming [58], and it is likely that efforts to improve axon growth by delivery of multiple TFs will similarly depend on optimal ratios of co-expression. For example, the observation that JUN-ATF3 heterodimers have been shown to drive strong transcriptional activation, while ATF3 homodimers act to repress transcription, raises the possibility that the phenotypic effects of forced co-expression will vary according to the relative levels.…”

Section: Physical Interactionmentioning

confidence: 99%

Selecting optimal combinations of transcription factors to promote axon regeneration: Why mechanisms matter

Venkatesh

Blackmore

2017

Neuroscience Letters

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Recovery from injuries to the central nervous system, including spinal cord injury, is constrained in part by the intrinsically low ability of many CNS neurons to mount an effective regenerative growth response. To improve outcomes, it is essential to understand and ultimately reverse these neuron-intrinsic constraints. Genetic manipulation of key transcription factors (TFs), which act to orchestrate production of multiple regeneration-associated genes, has emerged as a promising strategy. It is likely that no single TF will be sufficient to fully restore neuron-intrinsic growth potential, and that multiple, functionally interacting factors will be needed. An extensive literature, mostly from non-neural cell types, has identified potential mechanisms by which TFs can functionally synergize. Here we examine four potential mechanisms of TF/TF interaction; physical interaction, transcriptional cross-regulation, signaling-based cross regulation, and co-occupancy of regulatory DNA. For each mechanism, we consider how existing knowledge can be used to guide the discovery and effective use of TF combinations in the context of regenerative neuroscience. This mechanistic insight into TF interactions is needed to accelerate the design of effective TF-based interventions to relieve neuron-intrinsic constraints to regeneration and to foster recovery from CNS injury.

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Stoichiometry of Transcription Factors Is Critical for Cardiac Reprogramming

Cited by 18 publications

References 19 publications

Navigating the labyrinth of cardiac regeneration

Navigating the labyrinth of cardiac regeneration

Single-Construct Polycistronic Doxycycline-Inducible Vectors Improve Direct Cardiac Reprogramming and Can Be Used to Identify the Critical Timing of Transgene Expression

Selecting optimal combinations of transcription factors to promote axon regeneration: Why mechanisms matter

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