Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis

Silberman, M.; Barac, Yaron D.; Yahav, Hava; Wolfovitz, Efrat; Einav, Shmuel; Resnick, Nitzan; Binah, Ofer

doi:10.1007/s10456-009-9143-7

Cited by 21 publications

(11 citation statements)

References 61 publications

(84 reference statements)

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“…The presence of promoter sequences termed shear stress response elements (SSREs) has been proposed to mediate the responsiveness of EC genes to shear stress. Although a single SSRE (the sequence GAGACC) was reported to encode shear stress responsiveness in endothelium (121,131), promoter regions of genes may encode for multiple SSREs (100,127). On the other hand, more recent studies have shown that transcriptional changes seen with flow also may be independent of SSREs (136).…”

Section: Onset Of Flow and Flow Adaptationmentioning

confidence: 99%

Mechanotransduction in the Endothelium: Role of Membrane Proteins and Reactive Oxygen Species in Sensing, Transduction, and Transmission of the Signal with Altered Blood Flow

Chatterjee

Fisher

2014

Antioxidants & Redox Signaling

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Although several elements of mechanosensing such as the sensing event, the transduction, transmission, and reception of the mechanosignal are now reasonably well understood, the links among these discrete steps in the pathway are not clear. Thus, identifying the mechanisms for the interaction of the K(ATP) channel, the kinases, and ROS to drive long-term adaptive responses in ECs is necessary. A critical re-examination of the signaling events associated with complex flow patterns (turbulent, oscillatory) under physiological conditions is also essential for the progress in the field. Since these complex shear patterns may be associated with an atherosclerosis susceptible phenotype, a specific challenge will be the pharmacological modulation of the responses to altered signaling events that occur at specific sites of disturbed or obstructed flow.

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Section: Onset Of Flow and Flow Adaptationmentioning

confidence: 99%

Mechanotransduction in the Endothelium: Role of Membrane Proteins and Reactive Oxygen Species in Sensing, Transduction, and Transmission of the Signal with Altered Blood Flow

Chatterjee

Fisher

2014

Antioxidants & Redox Signaling

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“…Promoters comprising AP1 response elements have been shown to respond to mechanical strain, as were elements comprising binding sites for SP1 or NFkB . A consensus shear stress responsive element SSRE (GAGACC) has been described to be activated especially in systems related to the vasculature (Inoue et al, 2004;Kletsas et al, 2002;Liedert et al, 2006b;Liedert et al, 2009;Miyagi et al, 2005;Ogasawara et al, 2001;Peverali et al, 2001;Silberman et al, 2009;Sumpio et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes

Seefried

Mueller-Deubert²,

Schwarz³

et al. 2010

eCM

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Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissuespecific offspring, as osteoblasts and chondrocytes, cells of cardiovascular tissues and lung cells are sensitive to mechanical loading but molecules and mechanisms involved have to be unraveled. It is well established that cellular mechanotransduction is mediated e.g. by activation of the transcription factor SP1 and by kinase signaling cascades resulting in the activation of the AP1 complex. To investigate cellular mechanisms involved in mechanotransduction and to analyze substances, which modulate cellular mechanosensitivity reporter gene constructs, which can be transfected into cells of interest might be helpful. Suitable small-scale bioreactor systems and mechanosensitive reporter gene constructs are lacking. To analyze the molecular mechanisms of mechanotransduction and its crosstalk with biochemically induced signal transduction, AP1 and SP1 luciferase reporter gene constructs were cloned and transfected into various cell lines and primary cells. A newly developed bioreactor and small-scale 24-well polyurethane dishes were used to apply cyclic stretching to the transfected cells. 1 Hz cyclic stretching for 30 min in this system resulted in a significant stimulation of AP1 and SP1 mediated luciferase activity compared to unstimulated cells. In summary we describe a small-scale cell culture/bioreactor system capable of analyzing subcellular crosstalk mechanisms in mechanotransduction, mechanosensitivity of primary cells and of screening the activity of putative mechanosensitizers as new targets, e.g. for the treatment of bone loss caused by both disuse and signal transduction related alterations of mechanotransduction.

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“…Moving forward, one possible approach to enhance the sensor sensitivity would be through optimizing inducible promoter region. It is important to note that shear stress pathways are complicated and the complex interplay of between shear stress response elements in shear sensitive gene promoters has not been fully elucidated 52 . However we anticipate that careful construction of the promoter using a concoction of shear inducible elements could develop the sensitivity for finer resolution sensing of shear induced pathway activation.…”

Section: Discussionmentioning

confidence: 99%

A cell-based sensor of fluid shear stress for microfluidics

Varma

Voldman

2015

Lab Chip

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Microsystems designed for cell-based studies or applications inherently require fluid handling. Flows within such systems inevitably generate fluid shear stress (FSS) that may adversely affect cell health. Simple assays of cell viability, morphology or growth are typically reported to indicate any gross disturbances to cell physiology. However, no straightforward metric exists to specifically evaluate physiological implications of FSS within microfluidic devices, or among competing microfluidic technologies. This paper presents the first genetically encoded cell sensors that fluoresce in a quantitative fashion upon FSS pathway activation. We picked a widely used cell line (NIH3T3s) and created a transcriptional cell-sensor where fluorescence turns on when transcription of a relevant FSS-induced protein is initiated. Specifically, we chose Early Growth Factor-1 (a mechanosensitive protein) upregulation as the node for FSS detection. We verified our sensor pathway specificity and functionality by noting induced fluorescence in response to chemical induction of the FSS pathway, seen both through microscopy and flow cytometry. Importantly, we found our cell sensors to be inducible by a range of FSS intensities and durations, with a limit of detection of 2 dynes/cm2 when applied for 30 minutes. Additionally, our cell-sensors proved their versatility by showing induction sensitivity when made to flow through an inertial microfluidic device environment with typical flow conditions. We anticipate these cell sensors to have wide application in the microsystems community, allowing the device designer to engineer systems with acceptable FSS, and enabling the end-user to evaluate the impact of FSS upon their assay of interest.

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Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis

Cited by 21 publications

References 61 publications

Mechanotransduction in the Endothelium: Role of Membrane Proteins and Reactive Oxygen Species in Sensing, Transduction, and Transmission of the Signal with Altered Blood Flow

Mechanotransduction in the Endothelium: Role of Membrane Proteins and Reactive Oxygen Species in Sensing, Transduction, and Transmission of the Signal with Altered Blood Flow

A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes

A cell-based sensor of fluid shear stress for microfluidics

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