Stroke target identification guided by astrocyte transcriptome analysis

Rakers C,Schleif M,Blank N,Matu?ková H,2,Ulas T,H?ndler K,Torres SV,Schumacher T,Tai K,Schultze JL,,Jackson WS,Petzold GC,2

1.German Center for Neurodegenerative Diseases(DZNE),Bonn,Germany

2.Department of Neurology,University Hospital Bonn,Bonn,Germany

3.Genomics and Immunoregulation,LIMES-Institute,University of Bonn,Bonn,Germany

Abstract Astrocytes support normal brain function,but may also contribute to neurodegeneration when they become reactive under pathological conditions such as stroke.However,the molecular underpinnings of this context-dependent interplay between beneficial and detrimental properties in reactive astrogliosis have remained incompletely understood.Therefore,using the RiboTag technique,we immunopurified translating mRNAs specifically from astrocytes 72 hr after transient middle cerebral artery occlusion in mice(tMCAO),thereby generating a stroke-specific astroglial translatome database.We found that compared to control brains,reactive astrocytes after tMCAO show an enrichment of transcripts linked to the A2 phenotype,which has been associated with neuroprotection.However,we found that astrocytes also upregulate a large number of potentially neurotoxic genes.In total,we identified the differential expression of 1,003 genes and 38 transcription factors,of which Stat3,Sp1,and Spi1 were the most prominent.To further explore the effects of Stat3-mediated pathways on stroke pathogenesis,we subjected mice with an astrocyte-specific conditional deletion of Stat3 to tMCAO,and found that these mice have reduced stroke volume and improved motor outcome 72 hr after focal ischemia.Taken together,our study extends the emerging database of novel astrocyte-specific targets for stroke therapy,and supports the role of astrocytes as critical safeguards of brain function in health and disease.

Key words astroglia;ischemia;next generation sequencing

通過星形膠質(zhì)細胞轉(zhuǎn)錄組分析尋找腦卒中治療靶標

摘要星形膠質(zhì)細胞支持正常的腦功能，但當(dāng)它們在卒中等病理條件下變得具有反應(yīng)性時也可能促成神經(jīng)退行性病變。然而，反應(yīng)性星形膠質(zhì)細胞增生的有益和有害特性之間依賴于背景的相互作用的分子基礎(chǔ)仍然未被完全理解。我們使用RiboTag技術(shù)，在短暫性大腦中動脈閉塞（tMCAO）72 h后的小鼠星形膠質(zhì)細胞中特異性地純化翻譯mRNA，從而產(chǎn)生卒中特異性星形膠質(zhì)細胞翻譯組數(shù)據(jù)庫。與對照組大腦相比，tMCAO后的反應(yīng)性星形膠質(zhì)細胞顯示與A2表型相關(guān)的轉(zhuǎn)錄物富集，這與神經(jīng)保護相關(guān)。我們還發(fā)現(xiàn)，星形膠質(zhì)細胞也會上調(diào)大量潛在的神經(jīng)毒性基因。本研究共鑒定了1,003個基因和38個轉(zhuǎn)錄因子的差異表達，其中Stat3，Sp1和Spi1是最顯著的。為了進一步探索Stat3介導(dǎo)的途徑對卒中發(fā)病機制的影響，我們對星形膠質(zhì)細胞特異性條件性缺失Stat3的小鼠進行tMCAO處理，發(fā)現(xiàn)局灶性缺血72 h后的小鼠腦梗死體積減小，運動功能改善。綜上所述，本研究擴展了新興的星形膠質(zhì)細胞特異性靶向卒中治療數(shù)據(jù)庫，支持星形膠質(zhì)細胞在生理和病理條件下均對腦功能提供了關(guān)鍵保障作用的觀點。

關(guān)鍵詞星形膠質(zhì)細胞；缺血；下一代測序

中圖分類號R741；R741.02文獻標識碼ADOI10.16780/j.cnki.sjssgncj.2019.02.017