Use of AD Informer Set compounds to explore validity of novel targets in Alzheimer's disease pathology.
Use of AD Informer Set compounds to explore validity of novel targets in Alzheimer's disease pathology.
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DOI：
10.1002/trc2.12253
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发表时间：
2022
影响因子：
4.8
通讯作者：
Axtman, Alison D
中科院分区：
文献类型：
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作者：
Potjewyd, Frances M;Annor-Gyamfi, Joel K;Aube, Jeffrey;Chu, Shaoyou;Conlon, Ivie L;Frankowski, Kevin J;Guduru, Shiva K R;Hardy, Brian P;Hopkins, Megan D;Kinoshita, Chizuru;Kireev, Dmitri B;Mason, Emily R;Moerk, Charles T;Nwogbo, Felix;Pearce, Kenneth H Jr;Richardson, Timothy I;Rogers, David A;Soni, Disha M;Stashko, Michael;Wang, Xiaodong;Wells, Carrow;Willson, Timothy M;Frye, Stephen V;Young, Jessica E;Axtman, Alison D
最近开发了一组小分子的化学基因组学，其具有注释的活性和在阿尔茨海默病（AD）中的相关作用，称为AD Informer Set，并可供AD研究社区使用：https：//treatad.org/data-tools/ad-informer-set/。选择AD Informer Set化合物的小子集进行AD相关分析。靶向由加速阿尔茨海默病治疗开发的靶向启动（TREAT-AD）团队优先研究的六种AD相关基因表达的蛋白质的九种化合物被选择用于G蛋白偶联受体（GPCR）、淀粉样蛋白β（Aβ）和tau蛋白以及药代动力学（PK）研究。在小胶质细胞毒性和吞噬作用测定中分析了四种非重叠化合物。靶向CAPN 2、EPHX 2、MDK、MerTK/FLT 3或SYK蛋白的9种化合物在46至47个主要GPCR结合测定中进行了分析。用相同的9种化合物处理人诱导多能干细胞（iPSC）衍生的神经元，并在两种浓度和两个时间点测量Aβ肽（Aβ40和Aβ42）的分泌以及磷酸化tau（p-tau，Thr 231）和总tau（t-tau）肽的水平。最后，对CD 1小鼠进行静脉给药，以确定这些化合物的初步PK和/或脑特异性半衰期值。作为最终的基于细胞的研究，基于单浓度筛选选择四种化合物的非重叠子集，用于分析鼠和人小胶质细胞中的细胞毒性和吞噬作用。我们已经证明了AD Informer Set在使用生物化学、细胞（原发性和永生化）和体内研究验证新型AD假设中的实用性。对于我们的化合物，确定了其主要靶标相对于脑中必需GPCR的选择性。观察到tau、p-tau、Aβ40和/或Aβ42和血脑屏障通透性的统计学变化，巩固了特定化合物对AD的效用。经验证，单浓度吞噬结果可预测剂量-反应结果。这些研究建立了工作流程，验证了检测方法，并阐明了蛋白质靶点和化合物的下一步。
A chemogenomic set of small molecules with annotated activities and implicated roles in Alzheimer's disease (AD) called the AD Informer Set was recently developed and made available to the AD research community: https://treatad.org/data‐tools/ad‐informer‐set/. Small subsets of AD Informer Set compounds were selected for AD‐relevant profiling. Nine compounds targeting proteins expressed by six AD‐implicated genes prioritized for study by Target Enablement to Accelerate Therapy Development for Alzheimer's Disease (TREAT‐AD) teams were selected for G‐protein coupled receptor (GPCR), amyloid beta (Aβ) and tau, and pharmacokinetic (PK) studies. Four non‐overlapping compounds were analyzed in microglial cytotoxicity and phagocytosis assays. The nine compounds targeting CAPN2, EPHX2, MDK, MerTK/FLT3, or SYK proteins were profiled in 46 to 47 primary GPCR binding assays. Human induced pluripotent stem cell (iPSC)‐derived neurons were treated with the same nine compounds and secretion of Aβ peptides (Aβ40 and Aβ42) as well as levels of phosphophorylated tau (p‐tau, Thr231) and total tau (t‐tau) peptides measured at two concentrations and two timepoints. Finally, CD1 mice were dosed intravenously to determine preliminary PK and/or brain‐specific penetrance values for these compounds. As a final cell‐based study, a non‐overlapping subset of four compounds was selected based on single‐concentration screening for analysis of both cytotoxicity and phagocytosis in murine and human microglia cells. We have demonstrated the utility of the AD Informer Set in the validation of novel AD hypotheses using biochemical, cellular (primary and immortalized), and in vivo studies. The selectivity for their primary targets versus essential GPCRs in the brain was established for our compounds. Statistical changes in tau, p‐tau, Aβ40, and/or Aβ42 and blood–brain barrier penetrance were observed, solidifying the utility of specific compounds for AD. Single‐concentration phagocytosis results were validated as predictive of dose–response findings. These studies established workflows, validated assays, and illuminated next steps for protein targets and compounds.