TY - JOUR
T1 - A high-throughput screening to identify small molecules that suppress huntingtin promoter activity or activate huntingtin-antisense promoter activity
AU - Khaled, Houda G.
AU - Feng, Hongxuan
AU - Hu, Xin
AU - Sun, Xin
AU - Zheng, Wang
AU - Li, Pan P.
AU - Rudnicki, Dobrila
AU - Ye, Wenjuan
AU - Chen, Yu Chi
AU - Southall, Noel
AU - Marugan, Juan
AU - Ross, Christopher A.
AU - Ferrer, Marc
AU - Henderson, Mark J.
AU - Margolis, Russell L.
N1 - Funding Information:
We thank Dr. Samarjit Patnaik for inspiration, Dr. James Inglese for providing the luminescent reporter vector, Qi Sun for assistance with manuscript preparation, the staff of Evotec and Charles River Laboratories for performing hESC and iPSC cell studies, and CHDI, including Drs. Ramee Li, Douglas MacDonald, and Liz Doherty, for supporting this effort. This work was supported by the intramural research program of the National Center for Advancing Translational Sciences, NIH, and by NIH NS08532, NIH NS100783, the Hereditary Disease Foundation, and the ABCD Charitable Trust.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - 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.
AB - 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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U2 - 10.1038/s41598-021-85279-2
DO - 10.1038/s41598-021-85279-2
M3 - Article
C2 - 33731741
AN - SCOPUS:85102751871
VL - 11
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 6157
ER -