Discovery of small molecules that normalize the transcriptome and enhance cysteine cathepsin activity in progranulin-deficient microglia.
Discovery of small molecules that normalize the transcriptome and enhance cysteine cathepsin activity in progranulin-deficient microglia.
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发现了使颗粒体蛋白前体缺陷的小胶质细胞转录组正常化并增强半胱氨酸组织蛋白酶活性的小分子。
DOI：
10.1038/s41598-020-70534-9
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发表时间：
2020
影响因子：
4.6
通讯作者：
Gan,Li
中科院分区：
文献类型：
--
作者：
Telpoukhovskaia,MariaA;Liu,Kai;Sayed,FatenA;Etchegaray,JonIker;Xie,Min;Zhan,Lihong;Li,Yaqiao;Zhou,Yungui;Le,David;Bahr,BenA;Bogyo,Matthew;Ding,Sheng;Gan,Li
由颗粒蛋白（GRN）单倍不足引起的额颞叶痴呆（FTD）患者颗粒蛋白原水平降低，并表现出炎症和溶酶体网络的失调。小胶质细胞产生高水平的颗粒蛋白前体，并且仅小胶质细胞中颗粒蛋白前体的减少足以以细胞自主的方式再现炎症、溶酶体功能障碍和过度增殖。因此，靶向治疗颗粒蛋白前体不足引起的小胶质细胞功能障碍代表了一种管理FTD神经退行性变的潜在治疗策略。目前的progranulin增强策略的局限性，需要发现新的目标。为了鉴定可以逆转Grn缺陷小鼠小胶质细胞中小胶质细胞缺陷的化合物，我们进行了化合物筛选，并结合高通量测序来评估炎症和溶酶体途径中的关键转录变化。然后基于其拯救组织蛋白酶活性的能力，进一步缩小来自该初始筛选的阳性命中，这是溶酶体能力的关键生化读数。该筛选鉴定了去甲-binaltorphimine dihydrochloride（nor-BNI）和二丁酰-cAMP钠盐（DB-cAMP）作为颗粒蛋白前体缺乏的两种表型调节剂。此外，nor-BNI和DB-cAMP也挽救了前颗粒蛋白缺陷细胞的细胞周期异常。这些数据强调了基于转录的药物筛选平台的潜力，并提出了两种新的FTD先导化合物。
Patients with frontotemporal dementia (FTD) resulting from granulin (GRN)haploinsufficiency have reduced levels of progranulin and exhibit dysregulation in inflammatory and lysosomal networks. Microglia produce high levels of progranulin, and reduction of progranulin in microglia alone is sufficient to recapitulate inflammation, lysosomal dysfunction, and hyperproliferation in a cell-autonomous manner. Therefore, targeting microglial dysfunction caused by progranulin insufficiency represents a potential therapeutic strategy to manage neurodegeneration in FTD. Limitations of current progranulin-enhancing strategies necessitate the discovery of new targets. To identify compounds that can reverse microglial defects inGrn-deficient mouse microglia, we performed a compound screen coupled with high throughput sequencing to assess key transcriptional changes in inflammatory and lysosomal pathways. Positive hits from this initial screen were then further narrowed down based on their ability to rescue cathepsin activity, a critical biochemical readout of lysosomal capacity. The screen identified nor-binaltorphimine dihydrochloride (nor-BNI) and dibutyryl-cAMP, sodium salt (DB-cAMP) as two phenotypic modulators of progranulin deficiency. In addition,nor-BNI and DB-cAMP also rescued cell cycle abnormalities in progranulin-deficient cells. These data highlight the potential of a transcription-based platform for drug screening, and advance two novel lead compounds for FTD.