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The impact of substrate stiffness on morphological, transcriptional and functional aspects in RPE

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Homo sapiens

PROVIDER: GSE225642 | GEO | 2024/03/11

REPOSITORIES: GEO

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The impact of substrate stiffness on morphological, transcriptional and functional aspects in RPE.

Project description:Alterations in the structure and composition of Bruch's membrane (BrM) and loss of retinal pigment epithelial (RPE) cells are associated with various ocular diseases, notably age-related macular degeneration (AMD) as well as several inherited retinal diseases (IRDs). We explored the influence of stiffness as a major BrM characteristic on the RPE transcriptome and morphology. ARPE-19 cells were plated on soft ( E=30kPa ) or stiff ( E=80kPa ) polyacrylamide gels (PA gels) or standard tissue culture plastic (TCP). Next-generation sequencing (NGS) data on differentially expressed small RNAs (sRNAs) and messenger RNAs (mRNAs) were validated by qPCR, immunofluorescence or western blotting. The microRNA (miRNA) fraction of sRNAs grew with substrate stiffness and distinct miRNAs such as miR-204 or miR-222 were differentially expressed. mRNA targets of differentially expressed miRNAs were stably expressed, suggesting a homeostatic effect of miRNAs. mRNA transcription patterns were substrate stiffness-dependent, including components of Wnt/beta-catenin signaling, Microphthalmia-Associated Transcription Factor (MITF) and Dicer. These findings highlight the relevance of mechanical properties of the extracellular matrix (ECM) in cell culture experiments, especially those focusing on ECM-related diseases, such as AMD.

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The impact of substrate stiffness on morphological, transcriptional and functional aspects in RPE [miRNA-Seq]

Project description:To investigate the influence of stiffness as a major Bruch's membrane characteristic on the RPE transcriptome and morphology. ARPE-19 cells were plated on soft or stiff polyacrylamide gels (PA gels) or standard tissue culture plastic (TCP). We then performed gene expression profiling analysis using data obtained from Next-generation sequencing of small RNAs of ARPE-19 cells cultured on 3 different biomechanical conditions.

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The impact of substrate stiffness on morphological, transcriptional and functional aspects in RPE

Project description:The impact of substrate stiffness on morphological, transcriptional and functional aspects in RPE

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Project description:This SuperSeries is composed of the SubSeries listed below.

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Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

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