DIRAS3 (ARHI) Blocks RAS/MAPK Signaling by Binding Directly to RAS and Disrupting RAS Clusters

Sutton, Margie N.; Lü, Zhen; Li, Yao Cheng; Zhou, Yong; Huang, Tao; Reger, Albert S.; Hurwitz, Amy M.; Palzkill, Timothy; Logsdon, Craig D.; Liang, Xiaowen; Gray, Joe W.; Nan, Xiaolin; Hancock, John F.; Wahl, Geoffrey M.; Bast, Robert C.

doi:10.1016/j.celrep.2019.11.045

Cited by 51 publications

(37 citation statements)

References 41 publications

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“…Zhang et al [ 10 ] evaluated DIRAS3 expression in paired adjacent normal and cancerous tongue tissue samples from 20 patients with tongue squamous cell carcinoma and found that DIRAS3 expression was significantly downregulated in cancerous tongue tissues. The re-expression of DIRAS3 led to retarded cell growth, angiogenesis, migration and invasion in breast, ovarian and liver cancer [ 11 – 13 ]. However, whether DIRAS3 re-expression can suppress tumor growth in HNSCC has not yet been studied.…”

Section: Introductionmentioning

confidence: 99%

Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

et al. 2020

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Background p53 and DIRAS3 are tumor suppressors that are frequently silenced in tumors. In this study, we sought to determine whether the concurrent re-expression of p53 and DIRAS3 could effectively induce head and neck squamous cell carcinoma (HNSCC) cell death. Methods CAL-27 and SCC-25 cells were treated with Ad-DIRAS3 and rAd-p53 to induce re-expression of DIRAS3 and p53 respectively. The effects of DIRAS3 and p53 re-expression on the growth and apoptosis of HNSCC cells were examined by TUNEL assay, flow cytometric analysis and MTT. The effects of DIRAS3 and p53 re-expression on Akt phosphorylation, oncogene expression, and the interaction of 4E-BP1 with eIF4E were determined by real-time PCR, Western blotting and immunoprecipitation analysis. The ability of DIRAS3 and p53 re-expression to induce autophagy was evaluated by transmission electron microscopy, LC3 fluorescence microscopy and Western blotting. The effects of DIRAS3 and p53 re-expression on HNSCC growth were evaluated by using an orthotopic xenograft mouse model. Results TUNEL assay and flow cytometric analysis showed that the concurrent re-expression of DIRAS3 and p53 significantly induced apoptosis (P < 0.001). MTT and flow cytometric analysis revealed that DIRAS3 and p53 re-expression significantly inhibited proliferation and induced cell cycle arrest (P < 0.001). Mechanistically, the concurrent re-expression of DIRAS3 and p53 down-regulated signal transducer and activation of transcription 3 (STAT3) and up-regulated p21WAF1/CIP1 and Bax (P < 0.001). DIRAS3 and p53 re-expression also inhibited Akt phosphorylation, increased the interaction of eIF4E with 4E-BP1, and reduced the expression of c-Myc, cyclin D1, vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor receptor (EGFR) and Bcl-2 (P < 0.001). Moreover, the concurrent re-expression of DIRAS3 and p53 increased the percentage of cells with GFP-LC3 puncta compared with that in cells treated with control adenovirus (50.00% ± 4.55% vs. 4.67% ± 1.25%, P < 0.001). LC3 fluorescence microscopy and Western blotting further showed that DIRAS3 and p53 re-expression significantly promoted autophagic activity but also inhibited autophagic flux, resulting in overall impaired autophagy. Finally, the concurrent re-expression of DIRAS3 and p53 significantly decreased the tumor volume compared with the control group in a HNSCC xenograft mouse model [(3.12 ± 0.75) mm3 vs. (189.02 ± 17.54) mm3, P < 0.001]. Conclusions The concurrent re-expression of DIRAS3 and p53 is a more effective approach to HNSCC treatment than current treatment strategies.

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Section: Introductionmentioning

confidence: 99%

Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

et al. 2020

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“…This growth promotion might be accomplished through the imprinted gene networks as mentioned above, although PLAGL1 was first discovered with its proapoptotic and antiproliferative molecular properties [ 11 , 12 ] and regarded as a tumor suppressor gene [ 15 , 18 ]. Nevertheless, this hypothetical theorem for imprinted paternal expression of tumor suppressor genes towards growth may not be generally applicable to other tumor suppressor genes, given that Igf2r is a maternally expressed imprinted gene in mice and inhibits embryonic growth [ 45 ] and DIRAS3 is a paternally expressed imprinted gene in humans (and in pigs, submitted for publication by our group) and also hinders growth [ 46 , 47 , 48 ], and both counteract the effects of growth promoting Igf2 and RAS , respectively.…”

Section: Discussionmentioning

confidence: 99%

Genomic Imprinting at the Porcine PLAGL1 Locus and the Orthologous Locus in the Human

Ahn

Hwang

Park

et al. 2021

Genes

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Implementation of genomic imprinting in mammals often results in cis-acting silencing of a gene cluster and monoallelic expression, which are important for mammalian growth and function. Compared with widely documented imprinting status in humans and mice, current understanding of genomic imprinting in pigs is relatively limited. The objectives of this study were to identify DNA methylation status and allelic expression of alternative spliced isoforms at the porcine PLAGL1 locus and assess the conservation of the locus compared to the orthologous human locus. DNA methylome and transcriptome were constructed using porcine parthenogenetic or biparental control embryos. Using methylome, differentially methylated regions between those embryos were identified. Alternative splicing was identified by differential splicing analysis, and monoallelic expression was examined using single nucleotide polymorphism sites. Moreover, topological boundary regions were identified by analyzing CTCF binding sites and compared with the boundary of human orthologous locus. As a result, it was revealed that the monoallelic expression of the PLAGL1 gene in porcine embryos via genomic imprinting was maintained in the adult stage. The porcine PLAGL1 locus was largely conserved in regard to maternal hypermethylation, tissue distribution of mRNA expression, monoallelic expression, and biallelic CTCF-binding, with exceptions on transcript isoforms produced by alternative splicing instead of alternative promoter usage. These findings laid the groundwork for comparative studies on the imprinted PLAGL1 gene and related regulatory mechanisms across species.

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“…Mediated by adaptor proteins SHC (Src homology 2 domain containing) and Grb2 (growth factor receptor-binding protein 2), RTK activates SOS, which acts to substitute guanosine diphosphate (GDP) by GTP at the membrane to activate Ras (Bandaru et al, 2019;Huang et al, 2019;Liao et al, 2018Liao et al, , 2020. Membrane-anchored Ras dimers or nanoclusters bind and activate Raf kinase (Nussinov et al, 2019b(Nussinov et al, , 2019c(Nussinov et al, , 2020Solman et al, 2015;Sutton et al, 2019;Zhou et al, 2018). In the absence of Ras, Raf is autoinhibited.…”

Section: Ras: An Overviewmentioning

confidence: 99%

Inhibition of Nonfunctional Ras

Nussinov

Jang

Gürsoy

et al. 2021

Cell Chemical Biology

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Intuitively, functional states should be targeted; not nonfunctional ones. So why could drugging the inactive K-Ras4B G12C work-but drugging the inactive kinase will likely not? The reason is the distinct oncogenic mechanisms. Kinase driver mutations work by stabilizing the active state and/or destabilizing the inactive state. Either way, oncogenic kinases are mostly in the active state. Ras driver mutations work by quelling its deactivation mechanisms, GTP hydrolysis, and nucleotide exchange. Covalent inhibitors that bind to the inactive GDP-bound K-Ras4B G12C conformation can thus work. By contrast, in kinases, allosteric inhibitors work by altering the active-site conformation to favor orthosteric drugs. From the translational standpoint this distinction is vital: it expedites effective pharmaceutical development and extends the drug classification based on the mechanism of action. Collectively, here we postulate that drug action relates to blocking the mechanism of activation, not to whether the protein is in the active or inactive state. BACKGROUND ll

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DIRAS3 (ARHI) Blocks RAS/MAPK Signaling by Binding Directly to RAS and Disrupting RAS Clusters

Cited by 51 publications

References 41 publications

Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

Genomic Imprinting at the Porcine PLAGL1 Locus and the Orthologous Locus in the Human

Inhibition of Nonfunctional Ras

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