TY - JOUR
T1 - Heterozygous IDH1 R132H/WT created by “single base editing” inhibits human astroglial cell growth by downregulating YAP
AU - Wei, Shuang
AU - Wang, Jie
AU - Oyinlade, Olutobi
AU - Ma, Ding
AU - Wang, Shuyan
AU - Kratz, Lisa
AU - Lal, Bachchu
AU - Xu, Qingfu
AU - Liu, Senquan
AU - Shah, Sagar R.
AU - Zhang, Hao
AU - Li, Yunqing
AU - Quiñones-Hinojosa, Alfredo
AU - Zhu, Heng
AU - Huang, Zhi yong
AU - Cheng, Linzhao
AU - Qian, Jiang
AU - Xia, Shuli
N1 - Publisher Copyright:
© 2018, Macmillan Publishers Limited, part of Springer Nature.
PY - 2018/9/20
Y1 - 2018/9/20
N2 - Mutations in the isocitrate dehydrogenase 1 (IDH1) gene have been identified in a number of cancer types, including brain cancer. The Cancer Genome Atlas project has revealed that IDH1 mutations occur in 70–80% of grade II and grade III gliomas. Until recently, most of the functional studies of IDH1 mutations in cellular models have been conducted in overexpression systems with the IDH1 wild type background. In this study, we employed a modified CRISPR/Cas9 genome editing technique called “single base editing”, and efficiently introduced heterozygous IDH1 R132H mutation (IDH1 R132H/WT ) in human astroglial cells. Global DNA methylation profiling revealed hypermethylation as well as hypomethylation induced by IDH1 R132H/WT . Global gene expression analysis identified molecular targets and pathways altered by IDH1 R132H/WT , including cell proliferation, extracellular matrix (ECM), and cell migration. Our phenotype analysis indicated that compared with IDH1 wild type cells, IDH1 R132H/WT promoted cell migration by upregulating integrin β4 (ITGB4); and significantly inhibited cell proliferation. Using our mutated IDH1 models generated by “single base editing”, we identified novel molecular targets of IDH1 R132H/WT , namely Yes-associated protein (YAP) and its downstream signaling pathway Notch, to mediate the cell growth-inhibiting effect of IDH1 R132H/WT . In summary, the “single base editing” strategy has successfully created heterozygous IDH1 R132H mutation that recapitulates the naturally occurring IDH1 mutation. Our isogenic cellular systems that differ in a single nucleotide in one allele of the IDH1 gene provide a valuable model for novel discoveries of IDH1 R132H/WT -driven biological events.
AB - Mutations in the isocitrate dehydrogenase 1 (IDH1) gene have been identified in a number of cancer types, including brain cancer. The Cancer Genome Atlas project has revealed that IDH1 mutations occur in 70–80% of grade II and grade III gliomas. Until recently, most of the functional studies of IDH1 mutations in cellular models have been conducted in overexpression systems with the IDH1 wild type background. In this study, we employed a modified CRISPR/Cas9 genome editing technique called “single base editing”, and efficiently introduced heterozygous IDH1 R132H mutation (IDH1 R132H/WT ) in human astroglial cells. Global DNA methylation profiling revealed hypermethylation as well as hypomethylation induced by IDH1 R132H/WT . Global gene expression analysis identified molecular targets and pathways altered by IDH1 R132H/WT , including cell proliferation, extracellular matrix (ECM), and cell migration. Our phenotype analysis indicated that compared with IDH1 wild type cells, IDH1 R132H/WT promoted cell migration by upregulating integrin β4 (ITGB4); and significantly inhibited cell proliferation. Using our mutated IDH1 models generated by “single base editing”, we identified novel molecular targets of IDH1 R132H/WT , namely Yes-associated protein (YAP) and its downstream signaling pathway Notch, to mediate the cell growth-inhibiting effect of IDH1 R132H/WT . In summary, the “single base editing” strategy has successfully created heterozygous IDH1 R132H mutation that recapitulates the naturally occurring IDH1 mutation. Our isogenic cellular systems that differ in a single nucleotide in one allele of the IDH1 gene provide a valuable model for novel discoveries of IDH1 R132H/WT -driven biological events.
UR - http://www.scopus.com/inward/record.url?scp=85047816917&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85047816917&partnerID=8YFLogxK
U2 - 10.1038/s41388-018-0334-9
DO - 10.1038/s41388-018-0334-9
M3 - Article
C2 - 29849122
AN - SCOPUS:85047816917
SN - 0950-9232
VL - 37
SP - 5160
EP - 5174
JO - Oncogene
JF - Oncogene
IS - 38
ER -