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Huck, W.

Publications and source records attributed to Huck, W..

2 recordsLinked to original sources

Small molecule inhibitor combination treatment effectively represses global B-cell signaling in diffuse large B-cell lymphoma

Cells sense their environment via signaling networks and malignant cells often hijack signaling pathways for their growth, which complicates defining the effects of drugs. Here, we map the state of the signaling network of B-cell lymphoma cells via simultaneous quantification of 111 (phospho-)proteins. We demonstrate that the B-cell signaling network can be disrupted with specific clinical small molecule inhibitors (iBTK, iSYK, iNF{kappa}B), thereby inducing a repressed state of the network. Principal component analysis identifies how the three inhibitors work along subtly different repression axes through the signaling state landscape. Finally, we observe that 1 {micro}M combination treatment with all three inhibitors is more effective in inducing the repressed state than 10 {micro}M of the single inhibitors. These results emphasize that cellular signaling occurs in complex networks, and underscore how quantification of the signaling state landscape provides insights into how combination drug treatments can bring cells in a desired network state.

molecular biology↗

ARTseq-FISH reveals position-dependent fate decisions driven by cell cycle changes

Differences in gene-expression profiles between individual cells can give rise to distinct cell fate decisions. Yet how localisation on a micropattern impacts initial changes in mRNA, protein, and phosphoprotein abundance remains unclear. To identify the effect of cellular position on gene expression, we developed a scalable antibody and mRNA targeting sequential fluorescence in situ hybridization (ARTseq-FISH) method capable of simultaneously profiling mRNAs, proteins, and phosphoproteins in single cells. We studied 67 (phospho-)protein and mRNA targets in individual mouse embryonic stem cells (mESCs) cultured on circular micropatterns. ARTseq-FISH reveals relative changes in both abundance and localization of mRNAs and (phospho-)proteins during the first 48 hours of exit from pluripotency. We confirm these changes by conventional immunofluorescence and time-lapse microscopy. Chemical labelling, immunofluorescence, and single-cell time-lapse microscopy further show that cells closer to the edge of the micropattern exhibit increased proliferation compared to cells at the centre. Together these data suggest that while gene expression is still highly heterogeneous position-dependent differences in mRNA and protein levels emerge as early as 12 hours after LIF withdrawal.

systems biology↗