Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors
The CRISPR-associated Cas9 system can modulate disease-causing alleles both in vivo and ex vivo, raising the possibility of therapeutic genome editing. In addition to gene targeting, epigenetic modulation by the catalytically inactive dCas9 may also be a potential form of cancer therapy. Granulin (G...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2018-06-01
|
Series: | Molecular Therapy: Nucleic Acids |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118300039 |
id |
doaj-9dd4267d3f104272b65ca433f7e1d21f |
---|---|
record_format |
Article |
spelling |
doaj-9dd4267d3f104272b65ca433f7e1d21f2020-11-24T20:58:44ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-06-0111C233310.1016/j.omtn.2018.01.002Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressorsHong Wang0Rui Guo1Zhonghua Du2Ling Bai3Lingyu Li4Jiuwei Cui5Wei Li6Andrew R. Hoffman7Ji-Fan Hu8Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaClinical Laboratory, First Affiliated Hospital, Jilin University, Changchun, ChinaStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaVA Palo Alto Health Care System and Stanford University Medical School, Palo Alto, CA 94304, USAStem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, ChinaThe CRISPR-associated Cas9 system can modulate disease-causing alleles both in vivo and ex vivo, raising the possibility of therapeutic genome editing. In addition to gene targeting, epigenetic modulation by the catalytically inactive dCas9 may also be a potential form of cancer therapy. Granulin (GRN), a potent pluripotent mitogen and growth factor that promotes cancer progression by maintaining self-renewal of hepatic stem cancer cells, is upregulated in hepatoma tissues and is associated with decreased tumor survival in patients with hepatoma. We synthesized a group of dCas9 epi-suppressors to target GRN by tethering the C terminus of dCas9 with three epigenetic suppressor genes: DNMT3a (DNA methyltransferase), EZH2 (histone 3 lysine 27 methyltransferase), and KRAB (the Krüppel-associated box transcriptional repression domain). In conjunction with guide RNAs (gRNAs), the dCas9 epi-suppressors caused significant decreases in GRN mRNA abundance in Hep3B hepatoma cells. These dCas9 epi-suppressors initiated de novo CpG DNA methylation in the GRN promoter, and they produced histone codes that favor gene suppression, including decreased H3K4 methylation, increased H3K9 methylation, and enhanced HP1a binding. Epigenetic knockdown of GRN led to the inhibition of cell proliferation, decreased tumor sphere formation, and reduced cell invasion. These changes were achieved at least partially through the MMP/TIMP pathway. This study thus demonstrates the potential utility of using dCas9 epi-suppressors in the development of epigenetic targeting against tumors.http://www.sciencedirect.com/science/article/pii/S2162253118300039hepatomaCRISPR dCas9epigenetic targetingDNA methylationhistone methylationDNMT3aEZH2KRABcancer |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hong Wang Rui Guo Zhonghua Du Ling Bai Lingyu Li Jiuwei Cui Wei Li Andrew R. Hoffman Ji-Fan Hu |
spellingShingle |
Hong Wang Rui Guo Zhonghua Du Ling Bai Lingyu Li Jiuwei Cui Wei Li Andrew R. Hoffman Ji-Fan Hu Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors Molecular Therapy: Nucleic Acids hepatoma CRISPR dCas9 epigenetic targeting DNA methylation histone methylation DNMT3a EZH2 KRAB cancer |
author_facet |
Hong Wang Rui Guo Zhonghua Du Ling Bai Lingyu Li Jiuwei Cui Wei Li Andrew R. Hoffman Ji-Fan Hu |
author_sort |
Hong Wang |
title |
Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors |
title_short |
Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors |
title_full |
Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors |
title_fullStr |
Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors |
title_full_unstemmed |
Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors |
title_sort |
epigenetic targeting of granulin in hepatoma cells by synthetic crispr dcas9 epi-suppressors |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2018-06-01 |
description |
The CRISPR-associated Cas9 system can modulate disease-causing alleles both in vivo and ex vivo, raising the possibility of therapeutic genome editing. In addition to gene targeting, epigenetic modulation by the catalytically inactive dCas9 may also be a potential form of cancer therapy. Granulin (GRN), a potent pluripotent mitogen and growth factor that promotes cancer progression by maintaining self-renewal of hepatic stem cancer cells, is upregulated in hepatoma tissues and is associated with decreased tumor survival in patients with hepatoma. We synthesized a group of dCas9 epi-suppressors to target GRN by tethering the C terminus of dCas9 with three epigenetic suppressor genes: DNMT3a (DNA methyltransferase), EZH2 (histone 3 lysine 27 methyltransferase), and KRAB (the Krüppel-associated box transcriptional repression domain). In conjunction with guide RNAs (gRNAs), the dCas9 epi-suppressors caused significant decreases in GRN mRNA abundance in Hep3B hepatoma cells. These dCas9 epi-suppressors initiated de novo CpG DNA methylation in the GRN promoter, and they produced histone codes that favor gene suppression, including decreased H3K4 methylation, increased H3K9 methylation, and enhanced HP1a binding. Epigenetic knockdown of GRN led to the inhibition of cell proliferation, decreased tumor sphere formation, and reduced cell invasion. These changes were achieved at least partially through the MMP/TIMP pathway. This study thus demonstrates the potential utility of using dCas9 epi-suppressors in the development of epigenetic targeting against tumors. |
topic |
hepatoma CRISPR dCas9 epigenetic targeting DNA methylation histone methylation DNMT3a EZH2 KRAB cancer |
url |
http://www.sciencedirect.com/science/article/pii/S2162253118300039 |
work_keys_str_mv |
AT hongwang epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT ruiguo epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT zhonghuadu epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT lingbai epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT lingyuli epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT jiuweicui epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT weili epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT andrewrhoffman epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors AT jifanhu epigenetictargetingofgranulininhepatomacellsbysyntheticcrisprdcas9episuppressors |
_version_ |
1716784806429196288 |