A high-throughput screening to identify small molecules that suppress huntingtin promoter activity or activate huntingtin-antisense promoter activity.
A high-throughput screening to identify small molecules that suppress huntingtin promoter activity or activate huntingtin-antisense promoter activity.
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DOI：
10.1038/s41598-021-85279-2
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发表时间：
2021-03-17
影响因子：
4.6
通讯作者：
Margolis RL
中科院分区：
文献类型：
--
作者：
Khaled HG;Feng H;Hu X;Sun X;Zheng W;Li PP;Rudnicki DD;Ye W;Chen YC;Southall N;Marugan J;Ross CA;Ferrer M;Henderson MJ;Margolis RL
亨廷顿病 (HD) 是一种神经退行性疾病，由亨廷顿蛋白 (HTT) 外显子 1 的 CAG 重复扩增引起。虽然目前没有针对 HD 的疾病缓解疗法，但最近的努力集中在开发基于核苷酸的疗法来降低 HTT 表达。作为 siRNA 或寡核苷酸方法的替代方法，我们假设抑制 HTT 表达可能是通过小分子来实现的，这些小分子要么（1）通过抑制 HTT 启动子活性直接降低 HTT 表达，要么（2）通过增加 HTT-AS 的启动子活性间接降低 HTT 表达，HTT-AS 是 HTT 的反义基因，似乎抑制 HTT 的表达。我们使用发光报告 HEK293 细胞开发并采用了 HTT 和 HTT-AS 启动子活性修饰剂的高通量筛选；在测试的 52,041 种化合物中，我们鉴定出了 898 种可复制的化合物。我们采用严格的逐步方法来评估化合物毒性以及化合物特异性降低 5 种不同细胞系中亨廷顿蛋白的能力，包括 HEK293 细胞、HD 淋巴母细胞、小鼠原代神经元、分化为皮质样神经元的 HD iPSC 和 HD hESC。在所有测定中，我们发现没有能够降低亨廷顿蛋白而不降低细胞活力的化合物，尽管不能排除一些无毒剂量的化合物的潜在功效。我们的结果表明，更具体的靶标可能有助于采用小分子方法抑制 HTT。
Huntington’s disease (HD) is a neurodegenerative disorder caused by a CAG repeat expansion in exon 1 of huntingtin (HTT). While there are currently no disease-modifying treatments for HD, recent efforts have focused on the development of nucleotide-based therapeutics to lower HTT expression. As an alternative to siRNA or oligonucleotide methods, we hypothesized that suppression of HTT expression might be accomplished by small molecules that either (1) directly decrease HTT expression by suppressing HTT promoter activity or (2) indirectly decrease HTT expression by increasing the promoter activity of HTT-AS, the gene antisense to HTT that appears to inhibit expression of HTT. We developed and employed a high-throughput screen for modifiers of HTT and HTT-AS promoter activity using luminescent reporter HEK293 cells; of the 52,041 compounds tested, we identified 898 replicable hits. We used a rigorous stepwise approach to assess compound toxicity and the capacity of the compounds to specifically lower huntingtin protein in 5 different cell lines, including HEK293 cells, HD lymphoblastoid cells, mouse primary neurons, HD iPSCs differentiated into cortical-like neurons, and HD hESCs. We found no compounds which were able to lower huntingtin without lowering cell viability in all assays, though the potential efficacy of a few compounds at non-toxic doses could not be excluded. Our results suggest that more specific targets may facilitate a small molecule approach to HTT suppression.
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影响因子：
2.6
作者：
Kawabata, Hiroshi;Uchiyama, Tatsuki;Takaori-Kondo, Akifumi
通讯作者：
Takaori-Kondo, Akifumi
DOI：
10.1073/pnas.182426599
发表时间：
2002-12-10
影响因子：
11.1
作者：
Heiser, V;Engernann, S;Wanker, EE
通讯作者：
Wanker, EE
影响因子：
2.2
作者：
De Simone, Uliana;Caloni, Francesca;Coccini, Teresa
通讯作者：
Coccini, Teresa
影响因子：
11.1
作者：
Hsiao J;Yuan TY;Tsai MS;Lu CY;Lin YC;Lee ML;Lin SW;Chang FC;Liu Pimentel H;Olive C;Coito C;Shen G;Young M;Thorne T;Lawrence M;Magistri M;Faghihi MA;Khorkova O;Wahlestedt C
通讯作者：
Wahlestedt C
影响因子：
29
作者：
Kieburtz, Karl;Landwehrmeyer, Georg B.;Miyasaki, Janis
通讯作者：
Miyasaki, Janis