Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional Regulation of HL60 Neutrophil Differentiation: Promyelocyte trajectory vs. Neutrophil trajectory

ABSTRACT: Using the HL60 multipotent promyelocytic leukemia cell line, we performed experiments that lead to two different cell fate attractors by treatments of varying dosage and duration of the differentiation agent all-trans-retinoic acid (ATRA). The dosage and duration combinations corresponding to the two treatments were chosen by means of of cytometric measurements of CD11b, a well-known early differentiation marker, such that the two populations are poised at the same stage of differentiation. Using the selected treatment combinations, one population proceeds toward the terminally differentiated neutrophil attractor while the other population reverts back toward the undifferentiated promyelocytic attractor. We monitored the gene expression changes in the two populations after their respective treatments over a period of five days and identified a set of genes that diverged in their expression; a subset of which promotes neutrophil differentiation while the other represses cell cycle progression. By employing promoter based transcription factor binding site analysis, we found enrichment of transcription factors functionally linked to tumor progression, cell cycle, and development. Keywords: Time course, dose response Untreated HL60 cells are hybrized on the channel 1 with Cy3, ATRA-Treated HL60 cells are hybridized on channel 2 with Cy5. HK60 cells were treated with high dosage/short duration and low dosage/long duration to achieve 55% of CD11b+ cells. These positive cells were then isolated and culture in fresh media and total RNA were isolated for comparison. Cells treated with high dosage/short duration treatment proceed toward the neutrophil attractor while cells treated with the low dosage/long duration treatment revert back toward the promyelocyte attractor.

ORGANISM(S): Homo sapiens

SUBMITTER: Albert Huang

PROVIDER: E-GEOD-13673 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:During commitment of a multipotent stem or progenitor cell to a particular lineage, a large number of genes alter their expression in a coordinated manner orchestrated by the gene regulatory network (GRN). The constraints imposed by the GRN govern how cells move in the high-dimensional gene expression state space and can be understood as a dynamical system in which phenotypic cell states (cell types) are attractors that stabilize the cell-type characteristic gene expression pattern against molecular noise. Despite insights from various theoretical models, it remains elusive how multipotent cells, when committing to a specific lineage, exit their attractor and enter a new distinct attractor. Here we show, using single-cell resolution monitoring of transcript patterns by qPCR that commitment of multipotent blood progenitor cells to either the erythroid or the myeloid lineage is preceded by a destabilization of the progenitorsâ?? attractor state and a slowing-down of relaxation of cells from outlier states, indicating a critical state transition (â??tipping pointâ??). The high-dimensionality of the system (many genes) and availability of individual trajectories of a large ensemble of systems (many cells) affords a novel signature for critical transition which can be predicted from theory: Decrease of correlation between cells and concomitant increase of correlation between genes as the cell population approaches the tipping point. Consistent with a destabilizing bifurcation that simultaneously opens access to the erythroid and myeloid attractors, differentiation signal for either lineage caused some cells to commit to the â??wrongâ?? fate; moreover providing conflicting signals resulted in a delayed decision at the bifurcation point that however was ultimately resolved by commitment to one fate. These results suggest that the theoretical framework of â??early-warning signsâ?? and critical transitions can be applied to ensembles of high-dimensional systems, offering a formal tool for analyzing single-cell omics data beyond current descriptive computational pattern recognition. Mouse blood progenitor cells (EML cell line) was exposed to EPO, IL-3/GM-CSF or a mixture of both cytokines and gene expression change was measured in sorted subpopulations wrt Sca1 progenitor surface marker expression. In total, there was 4 conditions (including control), three time points (including d0) and 20 samples (10 samples in duplicates) were analyzed. Two independent experiments were performed for each condition. The untreated progenitor cell population was used as control.

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Transcriptional Regulation of HL60 Neutrophil Differentiation: Promyelocyte trajectory vs. Neutrophil trajectory

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