Identification of Cell Fate Determining Transcription Factors for Generating Brain Endothelial Cells.

IF 4.5 3区医学 Q2 CELL & TISSUE ENGINEERING

Stem Cell Reviews and Reports Pub Date : 2025-01-24 DOI:10.1007/s12015-025-10842-7

Roya Ramezankhani, Jonathan De Smedt, Burak Toprakhisar, Bernard K van der Veer, Tine Tricot, Gert Vanmarcke, Bradley Balaton, Leo van Grunsven, Massoud Vosough, Yoke Chin Chai, Catherine Verfaillie

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Abstract

Reliable models of the blood-brain barrier (BBB), wherein brain microvascular endothelial cells (BMECs) play a key role in maintenance of barrier function, are essential tools for developing therapeutics and disease modeling. Recent studies explored generating BMEC-like cells from human pluripotent stem cells (hPSCs) by mimicking brain-microenvironment signals or genetic reprogramming. However, due to the lack of comprehensive transcriptional studies, the exact cellular identity of most of these cells remains poorly defined. In this study we aimed to identify the most likely master transcription factors (TFs) for inducing brain endothelial cell (EC) fate and assess the transcriptomic changes following their introduction into immature ECs. Therefore, we first generated PSC-derived immature ECs by transient overexpression of the TF, ETV2. Subsequently, by performing an extensive meta-analysis of transcriptome studies of brain and non-brain ECs, 12 candidate TFs were identified, which might fate immature ECs towards cells with brain EC features. Following combinatorial overexpression of these 12 TFs tagged with unique barcodes, single cell transcriptomics identified a subset of transduced cells that resembled mid-gestational human brain ECs. Assessment of the TF barcodes present in these cells revealed significant enrichment of the TFs ZIC3, TFAP2C, TFAP2A, and DLX2. These TFs might be useful to fate PSC-EC to BMEC-like cells, which could be incorporated in human in vitro BBB models.

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脑微血管内皮细胞（BMECs）在维持血脑屏障（BBB）功能方面发挥着关键作用，而可靠的血脑屏障（BBB）模型是开发治疗药物和建立疾病模型的重要工具。最近的研究探索了通过模拟脑微环境信号或基因重编程，从人多能干细胞（hPSC）中生成类脑微血管内皮细胞（BMEC）。然而，由于缺乏全面的转录研究，大多数此类细胞的确切细胞身份仍未明确。在这项研究中，我们旨在确定最有可能诱导脑内皮细胞（EC）命运的主转录因子（TFs），并评估将它们引入未成熟EC后的转录组变化。因此，我们首先通过瞬时过表达 TF ETV2 生成了源自 PSC 的未成熟 EC。随后，我们对脑和非脑EC的转录组研究进行了广泛的荟萃分析，确定了12个候选TF，它们可能会使未成熟EC趋向于具有脑EC特征的细胞。在以独特条形码标记的这12个TFs进行组合过表达后，单细胞转录组学鉴定出了一个转导细胞亚群，该亚群类似于妊娠中期的人类脑ECs。对这些细胞中存在的 TF 条形码进行评估后发现，ZIC3、TFAP2C、TFAP2A 和 DLX2 等 TF 显著富集。这些TFs可能有助于将PSC-EC转化为BMEC样细胞，并将其纳入人类体外BBB模型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Stem Cell Reviews and Reports 医学-细胞生物学

CiteScore

9.30

自引率

4.20%

发文量

审稿时长

3 months

期刊介绍： The purpose of Stem Cell Reviews and Reports is to cover contemporary and emerging areas in stem cell research and regenerative medicine. The journal will consider for publication: i) solicited or unsolicited reviews of topical areas of stem cell biology that highlight, critique and synthesize recent important findings in the field. ii) full length and short reports presenting original experimental work. iii) translational stem cell studies describing results of clinical trials using stem cells as therapeutics. iv) papers focused on diseases of stem cells. v) hypothesis and commentary articles as opinion-based pieces in which authors can propose a new theory, interpretation of a controversial area in stem cell biology, or a stem cell biology question or paradigm. These articles contain more speculation than reviews, but they should be based on solid rationale. vi) protocols as peer-reviewed procedures that provide step-by-step descriptions, outlined in sufficient detail, so that both experts and novices can apply them to their own research. vii) letters to the editor and correspondence. In order to facilitate this exchange of scientific information and exciting novel ideas, the journal has created five thematic sections, focusing on: i) the role of adult stem cells in tissue regeneration; ii) progress in research on induced pluripotent stem cells, embryonic stem cells and mechanism governing embryogenesis and tissue development; iii) the role of microenvironment and extracellular microvesicles in directing the fate of stem cells; iv) mechanisms of stem cell trafficking, stem cell mobilization and homing with special emphasis on hematopoiesis; v) the role of stem cells in aging processes and cancerogenesis.