Supplementary Figures 1 through 5 from Identification of the Cell-Intrinsic and -Extrinsic Pathways Downstream of EGFR and IFNγ That Induce PD-L1 Expression in Head and Neck Cancer
Fernando Concha‐BenaventeRaghvendra M. SrivastavaSumita TrivediYu LeiUma ChandranRaja R. SeethalaGordon J. FreemanRobert L. Ferris
0
Citation
0
Reference
10
Related Paper
Abstract:
<p>Supplementary Figure 1. IFNγ-induced PD-L1 upregulation is not PI3K dependent. Supplementary Figure 2. IFNα induces pSTAT1 and HLA class I upregulation. Supplementary figure 3. A. STAT1 and STAT3 siRNA knockdown efficiency. Supplementary figure 4. EGF induces JAK2 phosphorylation. Supplementary figure 5. Knockdown efficiency of STAT1 siRNA in HNC cell lines.</p>Keywords:
STAT1
Sendai virus expresses C protein that blocks interferon (IFN) signaling. We previously reported suppression of IFN‐stimulated tyrosine phosphorylation of signal transducers and activators of transcription (Stats) in infected cells. However this conclusion has remained controversial. To settle it, we re‐examined the effect of C protein expression on phosphorylation of Stat1 in detail. IFN‐stimulated tyrosine phosphorylation of Stat1 was doubtlessly suppressed early in infection, but the suppression was incomplete, suggesting the importance of the unknown blocking mechanism that inactivates the tyrosine‐phosphorylated (pY)‐Stat1 generated as the signaling leak. Interestingly, the dephosphorylation process of pY‐Stat1 was also impaired. These effects on both phosphorylation and dephosphorylation processes were attributable to the function of the C protein.
Dephosphorylation
STAT1
Sendai virus
Cite
Citations (65)
Overexpression of members of the HER/erbB transmembrane tyrosine kinase family like HER2/erbB2/neu is associated with various cancers. Some heterodimers, especially HER2/HER3 heterodimers, are particularly potent inducers of oncogenic signaling. Still, from a clinical viewpoint their inhibition has yielded only moderate success so far, despite promising data from cell cultures. This suggests acquired resistance upon inhibitor therapy as one putative issue, requiring further studies in cell culture also aiming at rational combination therapies. In this paper, we demonstrate in ovarian carcinoma cells that the RNAi-mediated single knockdown of HER2 or HER3 leads to the rapid counter-upregulation of the respective other HER family member, thus providing a rational basis for combinatorial inhibition. Concomitantly, combined knockdown of HER2/HER3 exerts stronger anti-tumor effects as compared to single inhibition. In a tumor cell line xenograft mouse model, therapeutic intervention with nanoscale complexes based on polyethylenimine (PEI) for siRNA delivery, again reveals HER3 upregulation upon HER2 single knockdown and a therapeutic benefit from combination therapy. On the mechanistic side, we demonstrate that HER2 knockdown or inhibition reduces miR-143 levels with subsequent de-repression of HER3 expression, and validates HER3 as a direct target of miR-143. HER3 knockdown or inhibition, in turn, increases HER2 expression through the upregulation of the transcriptional regulator SATB1. These counter-upregulation processes of HER family members are thus based on distinct molecular mechanisms and may provide the basis for the rational combination of inhibitors.
Cite
Citations (9)
The balance between Ang II/AT1R and Ang-(1-7)/Mas plays a pivotal role in the development of lipopolysaccharides (LPS)-induced acute respiratory distress syndrome. However, the mechanisms underlying the balancing process still remain unclear. Here we investigated the roles of nuclear factor (NF)-κB and p53 in regulating AT1R and Mas expression. The results demonstrated that Ang II pretreatment resulted in downregulation of Mas and upregulation of AT1R, phosphorylated p65, and apoptosis in LPS-treated Human pulmonary microvascular endothelial cells (HPMVECs), but had no effect on p53 expression. Lentiviral vector-mediated P65 knockdown, but not a P53 knockdown, reversed all these effects of Ang II. On the other hand, Ang-(1-7) pretreatment lead to an increased in Mas expression and a decrease in AT1R, p53, and phosphorylated p65 expressions with suppressed apoptosis in LPS-treated cells. P65 knockdown promoted the protein expression of both AT1R and Mas while inhibiting p53 expression. P53 knockdown, but not a p65 knockdown, reversed all these effects of Ang-(1-7). Interestingly, p65 overexpression upregulated p53 and AT1R but downregulated Mas. P53 knockdown activated p65. These results suggest that there is a two-way feedback regulation between AT1R and Mas receptor via the NF-kB p65/P53 pathway, which may play a key role in LPS-induced HPMVECs apoptosis.
Cite
Citations (17)
Abstract Approximately 25,000 ovarian cancers are diagnosed in the United States annually, and 75% of cases are in the advanced stage when they are largely incurable. There is a critical need for improved early detection tools and development of novel treatments. Recently, we showed that among 20q13‐amplified genes in ovarian cancer, ADRM1 overexpression was the most highly correlated with amplification and was significantly upregulated with respect to stage, recurrence, and metastasis. In addition, overexpression of ADRM1 correlated significantly with shorter time to recurrence and overall survival. Herein, array‐CGH and microarray expression of ovarian cancer cell lines provides evidence consistent with the primary tumor data that ADRM1 is a 20q13 amplification target. Knockdown of ADRM1 in amplified ovarian cell‐line OAW42 results in downregulation of growth factor GIPC1 and upregulation of tumor‐suppressor RECK RNA and protein. In our dataset of 141 ovarian primary tumors, ADRM1 overexpression significantly correlates with GIPC1 overexpression. In addition, there is a significant anticorrelation between ADRM1 overexpression and RECK expression. Further research is necessary to determine whether targeting knockdown of ADRM1 in 20q13‐amplified ovarian cancers results in growth inhibition and tumor suppression via downstream targets GIPC1 and RECK . © 2011 Wiley‐Liss, Inc.
Ovarian tumor
Cite
Citations (16)
Cite
Citations (17)
Cite
Citations (0)
Sendai virus
Dephosphorylation
STAT1
Cite
Citations (34)
We recently developed a piggyback knockdown method that was used to knockdown genes in adult zebrafish. In this method, a vivo morpholino (VMO) piggybacks an antisense deoxyoligonucleotide (dO) into the somatic cells and reduces the cognate mRNA levels. In this paper, we tested whether we can piggyback more than one dO with one VMO. We designed various hybrids that had more than one dO that could be piggybacked with one VMO. We chose f7, f8, and αIIb genes and tested their knockdown by the appropriate assays. The knockdown with piggybacking either two or three dOs by one VMO yielded > 85% knockdown efficiency. We also performed knockdown of argonautes and rnaseh separately along with f7. We found the knockdown of f7 occurs when knockdown of argonautes happens and not when rnaseh knockdown was performed, suggesting that RNaseH is involved in mRNA degradation. In conclusion, we developed a method where we could knockdown three genes at one time, and by increasing the concentration of VMO by twofold, we could knockdown six genes simultaneously. These multiple gene knockdowns will not only increase the efficiency of the method in whole genome-wide knockdowns but will also be useful to study multifactorial disorders.
Cite
Citations (10)
SNGH5 and TGFBR3 messenger RNA were downregulated while miR-181a-5p was upregulated in osteoarthritis tissues and models. Knockdown of SNGH5 impeded chondrocytes proliferation, while accelerated the apoptosis. However, miR-181a-5p had opposite effects. TGFBR3 was identified as a target gene of miR-181a-5p, which could be indirectly suppressed by SNGH5 knockdown. Taken together, downregulation of SNGH5 could inhibit the proliferation abilities of chondrocytes and facilitate apoptosis via regulating the miR-181a-5p/TGFBR3 axis
Cite
Citations (0)
Event Abstract Back to Event Multi-step regulation of STAT1 phosphorylation in response to double-stranded RNA Tomoh Matsumiya1*, Junichi Dempoya1, Fei Xing1, Ryo Hayakari1, Hidemi Yoshida1 and Tadaatsu Imaizumi1 1 Hirosaki University Graduate School of Medicine, Japan Pattern recognition receptors which sense non-self RNA trigger to produce type I IFN. Type I IFN binds to its cognate receptor, IFNAR, resulting in the activation of STAT1. Therefore, it has long been thought that dsRNA-introduced STAT1 phosphorylation is through transactivation of type I IFN signaling. This study asked whether the phosphorylation of STAT1 is absolutely type I IFN-dependent. A synthetic dsRNA polyinosinic-polycytidylic acid (polyI:C) induces STAT1 phosphorylation in A549 human lung epithelial cells. We found the polyI:C induced STAT1 phosphorylation in a predominantly IFN-dependent manner. However, we also found that polyI:C was able to stimulate STAT1 phosphorylation in type I IFN receptor-deficient U5A cells, suggesting this phosphorylation is type I IFN-independent. Retinoic acid-inducible gene-I (RIG-I) was involved in the initial STAT1 phosphorylation in response to polyI:C; however, little contribution of RIG-I was observed in the late STAT1 phosphorylation. Taken together, our results showed comprehensive regulation in which dsRNA induces STAT1 phosphorylation, indicating importance of STAT1 with such a tight regulation in innate immune system. References Dempoya J, Matsumiya T, Imaizumi T, Hayakari R, Xing F, Yoshida H, Okumura K, Satoh K. Double-Stranded RNA Induces Biphasic STAT1 Phosphorylation by both Type I Interferon (IFN)-Dependent and Type I IFN-Independent Pathways. Journal of Virology 86: 12760-9, 2012 Keywords: RIG-I-like receptors, STAT1, type I IFN, dsRNA, Innate immune system Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Innate immunity Citation: Matsumiya T, Dempoya J, Xing F, Hayakari R, Yoshida H and Imaizumi T (2013). Multi-step regulation of STAT1 phosphorylation in response to double-stranded RNA. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00660 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 12 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Tomoh Matsumiya, Hirosaki University Graduate School of Medicine, Hirosaki, 036-8562, Japan, tomo1027@cc.hirosaki-u.ac.jp Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tomoh Matsumiya Junichi Dempoya Fei Xing Ryo Hayakari Hidemi Yoshida Tadaatsu Imaizumi Google Tomoh Matsumiya Junichi Dempoya Fei Xing Ryo Hayakari Hidemi Yoshida Tadaatsu Imaizumi Google Scholar Tomoh Matsumiya Junichi Dempoya Fei Xing Ryo Hayakari Hidemi Yoshida Tadaatsu Imaizumi PubMed Tomoh Matsumiya Junichi Dempoya Fei Xing Ryo Hayakari Hidemi Yoshida Tadaatsu Imaizumi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.
STAT1
RNA Silencing
Cite
Citations (0)