Pluripotent stem cell-derived epithelium misidentified as brain microvascular endothelium requires ETS factors to acquire vascular fate.
Pluripotent stem cell-derived epithelium misidentified as brain microvascular endothelium requires ETS factors to acquire vascular fate.
复制标题
DOI：
10.1073/pnas.2016950118
复制
发表时间：
2021-02-23
影响因子：
11.1
通讯作者：
Lis R
中科院分区：
文献类型：
--
作者：
Lu TM;Houghton S;Magdeldin T;Durán JGB;Minotti AP;Snead A;Sproul A;Nguyen DT;Xiang J;Fine HA;Rosenwaks Z;Studer L;Rafii S;Agalliu D;Redmond D;Lis R
人PSC来源的iBMEC已被用于研究疾病机制和治疗神经疾病的药物开发。然而，它们的完整转录特征尚不清楚，这可能导致不准确的生理研究和治疗方法的开发，从而导致无效的临床结果。利用生理学研究证实的全面的转录荟萃分析，我们发现目前许多用于产生iBMEC的方案产生了同质的上皮细胞群。ETS转录因子的过度表达使这些细胞重新编程为表型内皮细胞(RECs)，这些细胞概括了某些血管功能，尽管在体外缺乏一些器质性转运蛋白基因的表达和高电阻。然而，它们代表着朝着产生适合于血脑屏障的生理和药物研究的体外模型迈出了关键的一步。体外多能来源的细胞必须在转录和功能水平上与体内发现的细胞尽可能地相似，才能成为研究疾病和开发治疗方法的有用工具。最近，有报道将人多能干细胞(HPSCs)分化为具有血脑屏障(BBB)样特性的脑微血管内皮细胞(ECs)。自那以后，这些细胞被用作一个强大的体外血脑屏障模型，用于药物输送和对神经疾病的机制理解。然而，这些诱导的脑微血管内皮细胞(IBMECs)的确切细胞特性还没有得到很好的描述。通过对先前发表的hPSC来源的细胞进行全面的转录荟萃分析并经生理学分析验证，我们证明iBMEC缺乏内皮细胞的功能属性，因为它们缺乏血管谱系基因，而表达与神经外胚层上皮谱系相关的基因簇(Epi-Ibmec)。关键的内皮ETS转录因子(ETV2、ERG和FLI1)的过表达将Epi-iBMECs重新编程为真正的内皮细胞，这些细胞在转录水平和某些功能水平上与真正的内皮细胞一致。这种方法最终可能被用于开发一种强大的体外人类血脑屏障模型，该模型类似于体内人脑EC，用于功能研究和药物开发。
Human PSC-derived iBMECs have been generated to study disease mechanisms and drug development for neurological disorders. However, their full transcriptomic characterization is unclear, which could result in inaccurate physiological studies and development of treatments with ineffective clinical outcomes. Utilizing a comprehensive transcriptomic metaanalysis validated by physiological studies, we find that many current protocols used to generate iBMECs produce a homogenous epithelial cell population. Overexpression of ETS transcription factors reprogram these cells into phenotypic endothelial cells (rECs) which recapitulate certain vascular functions, albeit lacking expression of some organotypic transporter genes and high electrical resistance in vitro. Nevertheless, they represent a crucial step toward the generation of an in vitromodel suitable for physiological and pharmaceutical studies of the blood–brain barrier. Cells derived from pluripotent sources in vitro must resemble those found in vivo as closely as possible at both transcriptional and functional levels in order to be a useful tool for studying diseases and developing therapeutics. Recently, differentiation of human pluripotent stem cells (hPSCs) into brain microvascular endothelial cells (ECs) with blood–brain barrier (BBB)-like properties has been reported. These cells have since been used as a robust in vitro BBB model for drug delivery and mechanistic understanding of neurological diseases. However, the precise cellular identity of these induced brain microvascular endothelial cells (iBMECs) has not been well described. Employing a comprehensive transcriptomic metaanalysis of previously published hPSC-derived cells validated by physiological assays, we demonstrate that iBMECs lack functional attributes of ECs since they are deficient in vascular lineage genes while expressing clusters of genes related to the neuroectodermal epithelial lineage (Epi-iBMEC). Overexpression of key endothelial ETS transcription factors (ETV2, ERG, and FLI1) reprograms Epi-iBMECs into authentic endothelial cells that are congruent with bona fide endothelium at both transcriptomic as well as some functional levels. This approach could eventually be used to develop a robust human BBB model in vitro that resembles the human brain EC in vivo for functional studies and drug discovery.
登录
查看更多内容
影响因子：
64.5
作者：
Ginsberg M;James D;Ding BS;Nolan D;Geng F;Butler JM;Schachterle W;Pulijaal VR;Mathew S;Chasen ST;Xiang J;Rosenwaks Z;Shido K;Elemento O;Rabbany SY;Rafii S
通讯作者：
Rafii S
影响因子：
4.6
作者：
Grifno, Gabrielle N.;Farrell, Alanna M.;Searson, Peter C.
通讯作者：
Searson, Peter C.
影响因子：
20.3
作者：
Hu, Kejin;Yu, Junying;Slukvin, Igor I.
通讯作者：
Slukvin, Igor I.
影响因子：
5.9
作者：
Faley, Shannon L.;Neal, Emma H.;Bellan, Leon M.
通讯作者：
Bellan, Leon M.
影响因子：
5.9
作者：
Appelt-Menzel A;Cubukova A;Günther K;Edenhofer F;Piontek J;Krause G;Stüber T;Walles H;Neuhaus W;Metzger M
通讯作者：
Metzger M