scholarly journals Improving leukemic CD34+/CD38−blasts characterization with single-cell transcriptome sequencing

2017 ◽  
Author(s):  
Ambra Sartori ◽  
Phil Cheng ◽  
Emilie Falconnet ◽  
Pascale Ribaux ◽  
Jean-Pierre Aubry-Lachainaye ◽  
...  
Keyword(s):  
Single Cell ◽  
Gene Networks ◽  
Transcriptional Level ◽  
Self Renewal ◽  
Core Set ◽  
Cellular Signaling Pathway ◽  
Blast Cells ◽  
Cell Transcriptome ◽  
Single Cell Transcriptome ◽  
First Time

SUMMARY / ABSTRACTAcute myeloid leukemia (AML) is a particularly aggressive blood cancer that is difficult to treat because of the incomplete eradication of rare blast cells that possess self-renewal and leukemia-initiating properties. To characterize resistant blasts, we analyzed for the first time the transcriptomes of individual CD34+/CD38−blasts by single-cell mRNA sequencing of 359 CD33+/CD34+/CD38−/+sorted cells from two patients with AML and four unaffected individuals. We demonstrated that the captured blasts possess the transcriptomic hallmarks of self-renewal and leukemia-initiating ability. The effects of somatic mutations on the cancer cells are visible at the transcriptional level, and the cellular signaling pathway activity of the blasts is altered, revealing disease-associated gene networks. We also identified a core set of transcription factors that were co-activated in blasts, which suggests a joint transcription program among blasts. Finally, we revealed that leukemogenesis and putative prognostic gene-expression signatures are present at diagnosis in leukemic CD33+/CD34+/CD38−cells and can be detected using a single-cell RNA sequencing approach.

scholarly journals High fidelity hypothermic preservation of primary tissues in organ transplant preservative for single cell transcriptome analysis

2017 ◽  
Author(s):  
Wanxin Wang ◽  
Lolita Penland ◽  
Ozgun Gokce ◽  
Derek Croote ◽  
Stephen Quake
Keyword(s):  
Single Cell ◽  
Organ Transplant ◽  
Population Heterogeneity ◽  
High Fidelity ◽  
Intact Mouse ◽  
New Strategy ◽  
Cell Transcriptome ◽  
Definition Of ◽  
Single Cell Transcriptome ◽  
First Time

AbstractBackgroundHigh-fidelity preservation strategies for primary tissues are in great demand in the single cell RNAseq community. A reliable method will greatly expand the scope of feasible collaborations and maximize the utilization of technical expertise. When choosing a method, standardizability is as important a factor to consider as fidelity due to the susceptibility of single-cell RNAseq analysis to technical noises. Existing approaches such as cryopreservation and chemical fixation are less than ideal for failing to satisfy either or both of these standards.ResultsHere we propose a new strategy that leverages preservation schemes developed for organ transplantation. We evaluated the strategy by storing intact mouse kidneys in organ transplant preservative solution at hypothermic temperature for up to 4 days (6 hrs, 1, 2, 3, and 4 days), and comparing the quality of preserved and fresh samples using FACS and single cell RNAseq. We demonstrate that the strategy effectively maintained cell viability, transcriptome integrity, cell population heterogeneity, and transcriptome landscape stability for samples after up to 3 days of preservation. The strategy also facilitated the definition of the diverse spectrum of kidney resident immune cells, to our knowledge the first time at single cell resolution.ConclusionsHypothermic storage of intact primary tissues in organ transplant preservative maintains the quality and stability of the transcriptome of cells for single cell RNAseq analysis. The strategy is readily generalizable to primary specimens from other tissue types for single cell RNAseq analysis.

scholarly journals Enriched expression of genes associated with autism spectrum disorders in human inhibitory neurons

2017 ◽  
Author(s):  
Ping Wang ◽  
Dejian Zhao ◽  
Herbert M. Lachman ◽  
Deyou Zheng
Keyword(s):  
Single Cell ◽  
Gene Networks ◽  
Developmental Stages ◽  
Cell Types ◽  
Autism Spectrum ◽  
Inhibitory Neurons ◽  
Expression Of Genes ◽  
Cell Transcriptome ◽  
Single Cell Transcriptome ◽  
Upregulated Genes

AbstractAutism spectrum disorder (ASD) is highly heritable but genetically heterogeneous. The affected neural circuits and cell types remain unclear and may vary at different developmental stages. By analyzing multiple sets of human single cell transcriptome profiles, we found that ASD candidates showed enriched gene expression in neurons, especially in inhibitory neurons. ASD candidates were also more likely to be the hubs of the co-expressed module that is highly expressed in inhibitory neurons, a feature not detected for excitatory neurons. In addition, we found that upregulated genes in multiple ASD cortex samples were also enriched with genes highly expressed in inhibitory neurons, suggesting a potential increase of inhibitory neurons and an imbalance in the ratio between excitatory and inhibitory neurons. Furthermore, the downstream targets of several ASD candidates, such as CHD8, EHMT1 and SATB2, also displayed enriched expression in inhibitory neurons. Taken together, our analysis of single cell transcriptomic data suggest that inhibitory neurons may be the major neuron subtype affected by the disruption of ASD gene networks, providing single cell functional evidence to support the excitatory/inhibitory (E/I) imbalance hypothesis.

scholarly journals Single cell transcriptome atlas of mouse mammary epithelial cells across development

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Bhupinder Pal ◽  
Yunshun Chen ◽  
Michael J. G. Milevskiy ◽  
François Vaillant ◽  
Lexie Prokopuk ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Developmental Stages ◽  
Mammary Epithelial Cells ◽  
Expression Profiles ◽  
Single Cells ◽  
Chromatin Accessibility ◽  
Mammary Epithelial ◽  
Cell Transcriptome ◽  
Single Cell Transcriptome

Abstract Background Heterogeneity within the mouse mammary epithelium and potential lineage relationships have been recently explored by single-cell RNA profiling. To further understand how cellular diversity changes during mammary ontogeny, we profiled single cells from nine different developmental stages spanning late embryogenesis, early postnatal, prepuberty, adult, mid-pregnancy, late-pregnancy, and post-involution, as well as the transcriptomes of micro-dissected terminal end buds (TEBs) and subtending ducts during puberty. Methods The single cell transcriptomes of 132,599 mammary epithelial cells from 9 different developmental stages were determined on the 10x Genomics Chromium platform, and integrative analyses were performed to compare specific time points. Results The mammary rudiment at E18.5 closely aligned with the basal lineage, while prepubertal epithelial cells exhibited lineage segregation but to a less differentiated state than their adult counterparts. Comparison of micro-dissected TEBs versus ducts showed that luminal cells within TEBs harbored intermediate expression profiles. Ductal basal cells exhibited increased chromatin accessibility of luminal genes compared to their TEB counterparts suggesting that lineage-specific chromatin is established within the subtending ducts during puberty. An integrative analysis of five stages spanning the pregnancy cycle revealed distinct stage-specific profiles and the presence of cycling basal, mixed-lineage, and 'late' alveolar intermediates in pregnancy. Moreover, a number of intermediates were uncovered along the basal-luminal progenitor cell axis, suggesting a continuum of alveolar-restricted progenitor states. Conclusions This extended single cell transcriptome atlas of mouse mammary epithelial cells provides the most complete coverage for mammary epithelial cells during morphogenesis to date. Together with chromatin accessibility analysis of TEB structures, it represents a valuable framework for understanding developmental decisions within the mouse mammary gland.

scholarly journals Single-Cell Transcriptome Analyses Reveal Signals to Activate Dormant Neural Stem Cells

Cell ◽  
2015 ◽  
Vol 161 (5) ◽  
pp. 1175-1186 ◽  
Author(s):  
Yuping Luo ◽  
Volkan Coskun ◽  
Aibing Liang ◽  
Juehua Yu ◽  
Liming Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
Single Cell ◽  
Transcriptome Analyses ◽  
Cell Transcriptome ◽  
Single Cell Transcriptome
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