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Micropillar-based phenotypic screening platform uncovers involvement of HDAC2 in nuclear deformability

ABSTRACT: Micropillar-based phenotypic screening platform uncovers involvement of HDAC2 in nuclear deformability

PROVIDER: PRJNA785487 | ENA |

REPOSITORIES: ENA

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Micropillar-based phenotypic screening platform uncovers involvement of HDAC2 in nuclear deformability

Project description:Nuclear deformability is an essential feature of migratory cells. Intercellular movement and migration in the extracellular space requires cells to squeeze through constrictions up to an order of magnitude smaller than the average diameter of their biggest and stiffest organelle. One field of study in which aberrant nuclear shapes and deformability are of particular interest is cancer, with abnormal nuclear structures described as a hallmark of the disease and an indicator of pathological progression, while increased nuclear deformability correlates with increased metastatic potential. While most previous studies focused on the description of nuclear morphology in adaptation to different 3D geometries, and mechanistic investigations of nuclear deformation have been limited to contributions of the cytoskeleton, we tap for the first time into the potential of micropillared substrates to serve as drug screening platforms for compounds that influence nuclear deformability. To this end, we combine a micropillared culture substrate with image analyses to test the effect of various drugs on nuclear deformability and successfully identify histone deacetylases inhibitors (iHDACs) as active compounds in nuclear shape regulation on micropillars. Our experiments show that HDAC-activity as gene transcription regulators appears to activate a nuclear shape-rescue program in osteosarcoma cells, providing an excellent platform for identification of novel nuclear shape-determinants and possibly druggable targets for anti-cancer therapies.

2022-11-21 | GSE189982 | GEO

A phenotypic screening platform for identifying chemical modulators of astrocyte reactivity

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HDAC2 in primary sensory neurons constitutively restrains chronic pain by repressing α2δ-1 expression and associated NMDA receptor activity

Project description:Purpose: To investigate genome-wide gene regulation by HDAC2 in dorsal root ganglion neurons in mice. Methods: The mRNA was extracted from the mouse dosal root ganglion of 8-week-old HDAC2 condition knockout (Advillin-Cre+/HDAC2loxP++, hdac2-cko) and littermates controls (WT, without Cre). After quality control, library was constructed and sequenced on the Illumina Platform with pared end 150 bp (Novogene Corporation Inc. Sacramento CA). Three replicates for each sample were done. Results: Hdac2-cKO DRGs shows a significant increase in 210 genes and a decrease in 455 genes (P<0.05).

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Effect of HDAC2 siRNA on HCC

Project description:Determine the effect and specificity of HDAC2 siRNA compared to SAHA inhibition of HDAC2 in hepatocellular carcinoma (HCC) Profile of treated cells compared to control Transcriptomic profiling

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Quantitative proteomic SILAC analysis of enteroids deleted for Hdac1 or Hdac2

Project description:We have exploited organoid SILAC approaches that we have previously developed (A SILAC-Based Method for Quantitative Proteomic Analysis of Intestinal Organoids.- Gonneaud A, Jones C, Turgeon N, Lévesque D, Asselin C, Boudreau F, Boisvert FM. -Sci Rep. 2016 Nov 30;6:38195. doi: 10.1038/srep38195) to investigate proteomic changes after deletion of epigenetic eraser genes Hdac1 and Hdac2 in enteroids. Both HDAC1 and HDAC2 are epigenetic erasers that drive specific and redundant gene expression patterns, in part by removing acetyl groups on histones. Deletion of these Hdac in intestinal epithelial cell (IEC) in vivo alters intestinal homeostasis, dependent on the Hdac deleted and the level of expression of both. To determine the intrinsic specific IEC function of HDAC1 and HDAC2, we have performed transcriptomic and quantitative proteomic approaches on enteroids deficient in Hdac1 or Hdac2. We have defined changes in both mRNA and protein expression patterns affecting IEC differentiation. We have identified IEC Hdac1- and Hdac2-dependent common as well as specific pathways and biological processes. These findings uncover unrecognized similarities and differences between Hdac1 and Hdac2 in IEC.

2019-11-14 | PXD011869 | Pride

Effect of HDAC2 siRNA on HCC

Project description:Determine the effect and specificity of HDAC2 siRNA compared to SAHA inhibition of HDAC2 in hepatocellular carcinoma (HCC)

2014-10-02 | GSE52232 | GEO

Role of Hdac2 in pancreatic ductal adenocarcinoma

Project description:Microarrays were used to determine transcriptional differences between Hdac2 knockout versus Hdac2 wildtype pancreatic cancer cells.

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A phenotypic screening platform for chronic pain therapeutics using all-optical electrophysiology

Project description:A phenotypic screening platform for chronic pain therapeutics using all-optical electrophysiology

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A phenotypic screening platform for identifying chemical modulators of astrocyte reactivity [BulkRNAseq.ONC]

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