Mapping Human Pluripotent Stem Cell-derived Erythroid Differentiation by Single-cell Transcriptome Analysis

AbstractThere is currently an imbalance between the supply and demand of functional red blood cells (RBCs) in clinical applications, and this imbalance can be addressed by regenerating RBCs with a variety of in vitro methods. Induced pluripotent stem cells (iPSCs) can address the low supply of cord blood and the ethical issues in embryonic stem cell research and provide a promising strategy to eliminate immune rejection. However, no complete single-cell level differentiation pathway exists for the iPSC-derived RBC differentiation system. In this study, we used iPSC line BC1 to establish an RBC regeneration system and used the 10× Genomics single-cell transcriptome platform to map the cell lineage and differentiation trajectories on day 14 (D14) of the regeneration system. We found iPSC differentiation was not synchronized during embryoid body (EB) culture, and the D14 cells in the system mainly consisted of mesodermal and various blood cells, similar to yolk sac hematopoiesis. During asynchronous EB differentiation, iPSCs undergo three bifurcations before they enter erythroid differentiation, and the driver genes of each bifurcation were identified. The key roles of cell adhesion and estradiol in RBC regeneration were observed. This study provides systematically theoretical guidance for the optimization of the iPSC-derived RBC differentiation system.

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TAMI-26. SINGLE-CELL TRANSCRIPTOME ANALYSIS REVEALS THAT ENDOTHELIAL TRANSFORMATION DRIVES GLIOBLASTOMA CHEMORESISTANCE VIA Wnt/β-CATENIN/MRP-1

Neuro-Oncology ◽

10.1093/neuonc/noaa215.914 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii218-ii218

Author(s):

Menggui Huang ◽

Duo Zhang ◽

Yi Fan

Keyword(s):

Stem Cell ◽

Single Cell ◽

Transcriptome Analysis ◽

Therapeutic Resistance ◽

Tumor Resistance ◽

Genetic Ablation ◽

Cytotoxic Treatment ◽

Cell Transcriptome ◽

Mesenchymal Transformation ◽

Single Cell Transcriptome

Abstract Therapeutic resistance remains a persistent challenge for patients with glioblastoma (GBM). Here, we report that endothelial cells (ECs) acquire transformation into mesenchymal stem cell (MSC)-like cells in GBM, driving tumor resistance to cytotoxic treatment. Single-cell transcriptome analysis identified an EC-derived cell population that robustly expresses mesenchymal genes and stem cell-associated markers, suggesting that ECs undergo mesenchymal transformation and stemness-like activation in GBM microenvironment. Furthermore, we identified a c-Met-mediated axis that induces β-catenin phosphorylation at Ser675 and Wnt signaling activation, inducing multidrug resistance-associated protein-1 (MRP-1) expression and leading to EC stemness-like activation and chemoresistance. Last, genetic ablation of β-catenin in ECs overcome GBM tumor resistance to temozolomide (TMZ) chemotherapy in vivo. Combination of Wnt inhibition and TMZ chemotherapy eliminated tumor-associated ECs, inhibited GBM growth, and increased mouse survival. These findings identified a cell plasticity-based, microenvironment-dependent mechanism that controls tumor chemoresistance, and suggest that targeting Wnt/β-catenin-mediated EC transformation and stemness activation may overcome therapeutic resistance in GBM.

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