Spatial and transcriptional heterogeneity of pancreatic beta cell neogenesis revealed by a time-resolved reporter system
Spatial and transcriptional heterogeneity of pancreatic beta cell neogenesis revealed by a time-resolved reporter system
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DOI：
10.1007/s00125-022-05662-0
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发表时间：
2022-03-03
期刊：
影响因子：
8.2
通讯作者：
Miyatsuka, Takeshi
中科院分区：
文献类型：
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作者：
Sasaki, Shugo;Lee, Michelle Y. Y.;Miyatsuka, Takeshi
目的/假说虽然胰腺β细胞已被证明起源于导管区域的内分泌前体细胞，但仍不清楚β细胞确切地从哪里产生，以及哪些转录物定义了新生儿的β细胞。方法建立小鼠模型‘Ins1-GFP；Timer’，以高时间分辨率提供β细胞新生过程中的空间信息。单细胞RNA测序(scRNA-seq)是对荧光报告分选的小鼠β细胞进行的，以揭示新生β细胞的转录图谱。对人胚胎干细胞(HESC)来源的β样细胞进行scRNA-seq分析，比较小鼠和人新生β细胞的特征。结果Ins1-GFP荧光成像；Timer小鼠胰腺成功地将新生成的β细胞切割成绿色荧光占优势的细胞。这一报告系统显示，不出所料，一些新生β细胞出现在靠近导管(β(Ducts))的地方；出乎意料的是，另一些细胞出现在远离导管的地方，靠近血管(beta(血管))。单细胞转录组分析显示，新生β细胞中有5个不同的群体，证实了β细胞新生的空间异质性，如高概率的胰升糖素阳性的β细胞(DUTE)、肌肉腱膜纤维肉瘤癌基因家族B(MafB)阳性的β细胞(DUT)和肌肉膜神经性纤维肉瘤癌基因家族A(MAFA)阳性的β细胞(血管)。通过与小鼠新生β细胞和人胚胎干细胞来源的类β细胞的scRNA-seq数据的比较分析，发现小鼠和人的β细胞新生之间在转录上的相似性，包括微粒体谷胱甘肽S转移酶1和突触素13(SYT13)的高表达状态。结论/解释时间分辨组织成像和单细胞转录图谱相结合显示了β细胞新生的空间和转录异质性的新特征，这将有助于更好地理解未来细胞治疗中的β细胞分化。
Aims/hypothesis While pancreatic beta cells have been shown to originate from endocrine progenitors in ductal regions, it remains unclear precisely where beta cells emerge from and which transcripts define newborn beta cells. We therefore investigated characteristics of newborn beta cells extracted by a time-resolved reporter system.Methods We established a mouse model, 'Ins1-GFP; Timer', which provides spatial information during beta cell neogenesis with high temporal resolution. Single-cell RNA-sequencing (scRNA-seq) was performed on mouse beta cells sorted by fluorescent reporter to uncover transcriptomic profiles of newborn beta cells. scRNA-seq of human embryonic stem cell (hESC)-derived beta-like cells was also performed to compare newborn beta cell features between mouse and human.Results Fluorescence imaging of Ins1-GFP; Timer mouse pancreas successfully dissected newly generated beta cells as green fluorescence-dominant cells. This reporter system revealed that, as expected, some newborn beta cells arise close to the ducts (beta(duct)); unexpectedly, the others arise away from the ducts and adjacent to blood vessels (beta(vessel)). Single-cell transcriptomic analyses demonstrated five distinct populations among newborn beta cells, confirming spatial heterogeneity of beta cell neogenesis such as high probability of glucagon-positive beta(duct), musculoaponeurotic fibrosarcoma oncogene family B (MafB)-positive beta(duct) and musculoaponeurotic fibrosarcoma oncogene family A (MafA)-positive beta(vessel) cells. Comparative analysis with scRNA-seq data of mouse newborn beta cells and hESC-derived beta-like cells uncovered transcriptional similarity between mouse and human beta cell neogenesis including microsomal glutathione S-transferase 1 (MGST1)- and synaptotagmin 13 (SYT13)-highly-expressing state.Conclusions/interpretation The combination of time-resolved histological imaging with single-cell transcriptional mapping demonstrated novel features of spatial and transcriptional heterogeneity in beta cell neogenesis, which will lead to a better understanding of beta cell differentiation for future cell therapy.Data availability Raw and processed single-cell RNA-sequencing data for this study has been deposited in the Gene Expression Omnibus under accession number GSE155742.