Large polyacrylamide hydrogels for large-batch cell culture and mechanobiological studies

Shi, Xuechen; Janmey, Paul A.

doi:10.1101/2023.01.27.525967

Cited by 2 publications

(2 citation statements)

References 34 publications

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“…Subsequently, the round coverslip was gently removed, leaving the resulting gel disk firmly attached to the square coverslip, with the embedded beads positioned within 2 lm below the gel surface. To allow for the increased cell numbers needed for RNA testing, PAA gels were prepared onto 75 9 51 mm microscope slides (Electron Microscopy Sciences, CAS# 71862-01) according to previously described protocol [63].…”

Section: Polyacrylamide Substrate Preparationmentioning

confidence: 99%

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

Vessella,

Xiang,

Xiao

et al. 2024

FEBS Open Bio

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The extracellular matrix (ECM) regulates carcinogenesis by interacting with cancer cells via cell surface receptors. Discoidin Domain Receptor 2 (DDR2) is a collagen‐activated receptor implicated in cell survival, growth, and differentiation. Dysregulated DDR2 expression has been identified in various cancer types, making it as a promising therapeutic target. Additionally, cancer cells exhibit mechanosensing abilities, detecting changes in ECM stiffness, which is particularly important for carcinogenesis given the observed ECM stiffening in numerous cancer types. Despite these, whether collagen‐activated DDR2 signaling and ECM stiffness‐induced mechanosensing exert similar effects on cancer cell behavior and whether they operate through analogous mechanisms remain elusive. To address these questions, we performed bulk RNA sequencing (RNA‐seq) on human SH‐SY5Y neuroblastoma cells cultured on collagen‐coated substrates. Our results show that DDR2 downregulation induces significant changes in the cell transcriptome, with changes in expression of 15% of the genome, specifically affecting the genes associated with cell division and differentiation. We validated the RNA‐seq results by showing that DDR2 knockdown redirects the cell fate from proliferation to senescence. Like DDR2 knockdown, increasing substrate stiffness diminishes cell proliferation. Surprisingly, RNA‐seq indicates that substrate stiffness has no detectable effect on the transcriptome. Furthermore, DDR2 knockdown influences cellular responses to substrate stiffness changes, highlighting a crosstalk between these two ECM‐induced signaling pathways. Based on our results, we propose that the ECM could activate DDR2 signaling and mechanosensing in cancer cells to orchestrate their cell fate through distinct mechanisms, with or without involving gene expression, thus providing novel mechanistic insights into cancer progression.

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Section: Polyacrylamide Substrate Preparationmentioning

confidence: 99%

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

Vessella,

Xiang,

Xiao

et al. 2024

FEBS Open Bio

View full text Add to dashboard Cite

show abstract

“…Subsequently, the round coverslip was gently removed, leaving the resulting gel disk firmly attached to the square coverslip, with the embedded beads positioned within 2 μm below the gel surface. To allow for the increased cell numbers needed for RNA testing, PAA gels were prepared onto 71 x51 mm microscope slides (electron microscopy sciences) according to previously described protocol (48).…”

Section: Polyacrylamide Substrate Preparationmentioning

confidence: 99%

The roles of DDR2 and substrate stiffness on cancer cell transcriptome and proliferation

Vessella,

Xiang,

Xiao

et al. 2023

Preprint

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The interactions between cancer cells and the ECM regulate carcinogenesis. The collagen receptor kinase DDR2 is dysregulated in certain cancer cells, but its precise role in these malignancies remains unclear. In this study, we perform RNA-seq to determine how DDR2 and the biomechanical environment regulate cancer cell behaviors. We show that DDR2 knockdown in SH-SY5Y neuroblastoma cells inhibits proliferation and promotes senescence by regulating relevant genes. Increasing substrate stiffness reduces proliferation and promotes cell spreading but does not change senescence or transcriptome. Furthermore, DDR2 knockdown modulates cellular responses to substrate stiffness changes, unraveling a crosstalk between DDR2 and mechanosensing. These findings indicate DDR2 and ECM biomechanics regulate cancer cell behavior through distinct mechanisms, providing new mechanistic insights of cancer progression.

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Large polyacrylamide hydrogels for large-batch cell culture and mechanobiological studies

Cited by 2 publications

References 34 publications

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

The roles of DDR2 and substrate stiffness on cancer cell transcriptome and proliferation

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