PU.1 regulates Alzheimer's disease-associated genes in primary human microglia.
PU.1 regulates Alzheimer's disease-associated genes in primary human microglia.
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DOI：
10.1186/s13024-018-0277-1
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发表时间：
2018-08-20
影响因子：
15.1
通讯作者：
Dragunow M
中科院分区：
文献类型：
--
作者：
Rustenhoven J;Smith AM;Smyth LC;Jansson D;Scotter EL;Swanson MEV;Aalderink M;Coppieters N;Narayan P;Handley R;Overall C;Park TIH;Schweder P;Heppner P;Curtis MA;Faull RLM;Dragunow M
小胶质细胞在大脑内稳态和病理状态中起着关键作用。了解支持小胶质细胞功能和激活状态的分子事件将进一步使我们能够靶向这些细胞治疗神经系统疾病。转录因子PU.1在髓细胞的发育中起关键作用，是小胶质细胞基因表达的主要调节因子。在大脑中，PU.1在小胶质细胞中特异性表达，最近来自全基因组关联研究的证据表明，PU.1的减少有助于阿尔茨海默病（AD）的延迟发病，可能是通过限制神经炎症反应。为了研究PU.1如何促进人小胶质细胞的免疫激活，我们对sirna介导的PU.1敲低的人混合胶质细胞进行了微阵列分析。在混合胶质细胞培养和分离的小胶质细胞中，通过qRT-PCR和免疫细胞化学证实了PU.1敲除后的微阵列击中。为了鉴定PU.1在小胶质细胞中的表达衰减剂，使用含有fda批准药物的化合物文库进行了高通量药物筛选。利用NanoString和免疫组化技术研究了神经正常和临床病理证实的AD患者原代人脑组织中PU.1自身和PU.1调节介质的表达。对混合胶质培养中PU.1沉默后基因表达的生物信息学分析显示，ad相关的修饰小胶质基因网络参与先天和适应性免疫系统，特别是那些参与抗原呈递和吞噬的基因。这些基因变化在分离的小胶质细胞培养中得到证实。利用高通量筛选fda批准的混合胶质细胞培养物中的化合物，我们发现组蛋白去乙酰化酶抑制剂伏立诺他是人类小胶质细胞中PU.1表达的有效衰减剂。vorinostat在分离的小胶质细胞培养中的进一步表征显示，基因和蛋白质的变化部分再现了sirna介导的PU.1敲除后所见的变化。最后，我们证明了这些pu .1调节基因中的几个在人类AD大脑中的小胶质细胞原位表达。总之，这些结果表明，减弱PU.1可能是一种有效的治疗方法，以限制阿尔茨海默病中小胶质细胞介导的炎症反应，并证明伏立诺他在这方面的效用。本文的在线版本（10.1186/s13024-018-0277-1）包含补充资料，仅供授权用户使用。
Microglia play critical roles in the brain during homeostasis and pathological conditions. Understanding the molecular events underpinning microglial functions and activation states will further enable us to target these cells for the treatment of neurological disorders. The transcription factor PU.1 is critical in the development of myeloid cells and a major regulator of microglial gene expression. In the brain, PU.1 is specifically expressed in microglia and recent evidence from genome-wide association studies suggests that reductions in PU.1 contribute to a delayed onset of Alzheimer’s disease (AD), possibly through limiting neuroinflammatory responses. To investigate how PU.1 contributes to immune activation in human microglia, microarray analysis was performed on primary human mixed glial cultures subjected to siRNA-mediated knockdown of PU.1. Microarray hits were confirmed by qRT-PCR and immunocytochemistry in both mixed glial cultures and isolated microglia following PU.1 knockdown. To identify attenuators of PU.1 expression in microglia, high throughput drug screening was undertaken using a compound library containing FDA-approved drugs. NanoString and immunohistochemistry was utilised to investigate the expression of PU.1 itself and PU.1-regulated mediators in primary human brain tissue derived from neurologically normal and clinically and pathologically confirmed cases of AD. Bioinformatic analysis of gene expression upon PU.1 silencing in mixed glial cultures revealed a network of modified AD-associated microglial genes involved in the innate and adaptive immune systems, particularly those involved in antigen presentation and phagocytosis. These gene changes were confirmed using isolated microglial cultures. Utilising high throughput screening of FDA-approved compounds in mixed glial cultures we identified the histone deacetylase inhibitor vorinostat as an effective attenuator of PU.1 expression in human microglia. Further characterisation of vorinostat in isolated microglial cultures revealed gene and protein changes partially recapitulating those seen following siRNA-mediated PU.1 knockdown. Lastly, we demonstrate that several of these PU.1-regulated genes are expressed by microglia in the human AD brain in situ. Collectively, these results suggest that attenuating PU.1 may be a valid therapeutic approach to limit microglial-mediated inflammatory responses in AD and demonstrate utility of vorinostat for this purpose. The online version of this article (10.1186/s13024-018-0277-1) contains supplementary material, which is available to authorized users.
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DOI：
10.1056/nejmoa1211851
发表时间：
2013-01-10
期刊：
The New England journal of medicine
影响因子：
--
作者：
Guerreiro R;Wojtas A;Bras J;Carrasquillo M;Rogaeva E;Majounie E;Cruchaga C;Sassi C;Kauwe JS;Younkin S;Hazrati L;Collinge J;Pocock J;Lashley T;Williams J;Lambert JC;Amouyel P;Goate A;Rademakers R;Morgan K;Powell J;St George-Hyslop P;Singleton A;Hardy J;Alzheimer Genetic Analysis Group
通讯作者：
Alzheimer Genetic Analysis Group
影响因子：
5.1
作者：
Fu, Ruying;Shen, Qingyu;Xu, Pengfei;Luo, Jin Jun;Tang, Yamei
通讯作者：
Tang, Yamei
DOI：
10.4049/jimmunol.1301758
发表时间：
2014-03-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Gross TJ;Kremens K;Powers LS;Brink B;Knutson T;Domann FE;Philibert RA;Milhem MM;Monick MM
通讯作者：
Monick MM
影响因子：
16.8
作者：
Crotti A;Glass CK
通讯作者：
Glass CK
影响因子：
30.8
作者：
Harold, Denise;Abraham, Richard;Hollingworth, Paul;Sims, Rebecca;Gerrish, Amy;Hamshere, Marian L.;Pahwa, Jaspreet Singh;Moskvina, Valentina;Dowzell, Kimberley;Williams, Amy;Jones, Nicola;Thomas, Charlene;Stretton, Alexandra;Morgan, Angharad R.;Lovestone, Simon;Powell, John;Proitsi, Petroula;Lupton, Michelle K.;Brayne, Carol;Rubinsztein, David C.;Gill, Michael;Lawlor, Brian;Lynch, Aoibhinn;Morgan, Kevin;Brown, Kristelle S.;Passmore, Peter A.;Craig, David;McGuinness, Bernadette;Todd, Stephen;Holmes, Clive;Mann, David;Smith, A. David;Love, Seth;Kehoe, Patrick G.;Hardy, John;Mead, Simon;Fox, Nick;Rossor, Martin;Collinge, John;Maier, Wolfgang;Jessen, Frank;Schuermann, Britta;van den Bussche, Hendrik;Heuser, Isabella;Kornhuber, Johannes;Wiltfang, Jens;Dichgans, Martin;Froelich, Lutz;Hampel, Harald;Huell, Michael;Rujescu, Dan;Goate, Alison M.;Kauwe, John S. K.;Cruchaga, Carlos;Nowotny, Petra;Morris, John C.;Mayo, Kevin;Sleegers, Kristel;Bettens, Karolien;Engelborghs, Sebastiaan;De Deyn, Peter P.;Van Broeckhoven, Christine;Livingston, Gill;Bass, Nicholas J.;Gurling, Hugh;McQuillin, Andrew;Gwilliam, Rhian;Deloukas, Panagiotis;Al-Chalabi, Ammar;Shaw, Christopher E.;Tsolaki, Magda;Singleton, Andrew B.;Guerreiro, Rita;Muehleisen, Thomas W.;Noethen, Markus M.;Moebus, Susanne;Joeckel, Karl-Heinz;Klopp, Norman;Wichmann, H-Erich;Carrasquillo, Minerva M.;Pankratz, V. Shane;Younkin, Steven G.;Holmans, Peter A.;O'Donovan, Michael;Owen, Michael J.;Williams, Julie
通讯作者：
Williams, Julie