scholarly journals Time-Sequential Change in Immune-Related Gene Expression After Irradiation in Glioblastoma: Next-Generation Sequencing Analysis

2020 ◽  
Author(s):  
Yi-Jun Kim ◽  
Kwangsoo Kim ◽  
Soo Yeon Seo ◽  
Juyeon Yu ◽  
Il Han Kim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Next Generation Sequencing ◽  
T Cell ◽  
Cell Line ◽  
Interferon Γ ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Related Gene ◽  
Immune Related Gene ◽  
Generation Sequencing

Abstract The time-sequential change in immune-related gene expression of the glioblastoma cell line after irradiation was evaluated to speculate the effect of combined immunotherapy with radiotherapy. The U373 MG glioblastoma cell line was irradiated with 6 Gy single dose. Next-generation sequencing (NGS) transcriptome data was generated before irradiation (control), and at 6, 24, and 48 hours post-irradiation. Immune-related pathways were analyzed at each time period. The same analyses were also performed for A549 lung cancer and U87 MG glioblastoma cell lines. Western blotting confirmed the programmed death-ligand 1 (PD-L1) expression levels over time. In the U373 MG cell line, neutrophil-mediated immunity, type I interferon signaling, antigen cross-presentation to T cell, and interferon-γ signals began to increase significantly at 24 hours and were upregulated until 48 hours after irradiation. The results were similar to those of the A549 and U87 MG cell lines. Without T cell infiltration, PD-L1 did not increase even with upregulated interferon-γ signaling in cancer cells. In conclusions, In the glioblastoma cell line, immune-related signals were significantly upregulated at 24 hours after irradiation. Therefore, the time interval between daily radiotherapy might not be enough to expect full immune responses by combined immune checkpoint inhibitors and newly infiltrating immune cells after irradiation.

scholarly journals WWOX modulates the gene expression profile in the T98G glioblastoma cell line rendering its phenotype less malignant

2014 ◽  
Vol 32 (4) ◽  
pp. 1362-1368 ◽  
Author(s):  
KATARZYNA KOŚLA ◽  
MAGDALENA NOWAKOWSKA ◽  
KAROLINA POSPIECH ◽  
ANDRZEJ K. BEDNAREK
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell

scholarly journals Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis

Oncogene ◽  
2004 ◽  
Vol 23 (27) ◽  
pp. 4681-4689 ◽  
Author(s):  
Sajani S Lakka ◽  
Christopher S Gondi ◽  
Niranjan Yanamandra ◽  
William C Olivero ◽  
Dzung H Dinh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Tumor Growth ◽  
Tumor Cell ◽  
Cell Line ◽  
Cell Invasion ◽  
Cathepsin B ◽  
Tumor Cell Invasion ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell

Next-Generation Sequencing Suggests Complex, Heterogeneous Pathogenesis In Peripheral T-Cell Lymphoma Unspecified

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 843-843 ◽  
Author(s):  
Jonathan H. Schatz ◽  
Steven M. Horwitz ◽  
Matthew A. Lunning ◽  
Igor Dolgalev ◽  
Kety Huberman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Next Generation Sequencing ◽  
T Cell ◽  
Candidate Genes ◽  
Research Funding ◽  
Cell Lymphoma ◽  
Next Generation ◽  
Peripheral T Cell Lymphoma ◽  
Tumor Dna ◽  
Generation Sequencing

Abstract Peripheral T-cell lymphoma (PTCL) makes up about 12 percent of non-Hodgkin lymphoma, comprising 18 diseases that are poorly understood and carry a generally worse prognosis than B-lymphomas. PTCL not otherwise specified (PTCL-NOS), a diagnosis of exclusion, is most common, making up 25-30 percent. Gene-expression studies suggest a heterogeneous origin of this diagnosis, with overlap to other PTCL types, but the genetic factors underlying its pathogenesis are undefined. Current therapy for PTCL-NOS is empiric and ultimately ineffective for most patients. Identification of specific therapeutic targets is therefore a high priority. We have sought better understanding of pathogenesis through next-generation sequencing of PTCL-NOS tumor DNA. Whole-exome sequencing revealed candidate genes but low availability of fresh-frozen samples limited our ability to draw conclusions by this method alone. We therefore sequenced the coding regions of 237 candidate genes in a collection formalin-fixed paraffin-embedded samples. We used Nimblegen Sequence Capture for PCR amplification of exons and Illumina hiSeq for raw sequence generation. Results were aligned to hg19 and compared to dbSNP and the 1,000 genomes data to exclude germline variants. Analysis, including comparison to the COSMIC database of cancer-specific mutations, revealed high-confidence mutations affecting more than 60 known cancer-related genes in 25 PTCL-NOS cases. Recurrent mutations pointed to frequent activation of three key signaling pathways: NF-kB (TNFAIP3), WNT/B-Catenin (APC, CHD8, CELSR2), and NOTCH (NOTCH1, FBXW7). Recurrent deregulation of epigenetic processes was indicated by mutations in genes affecting histone acetylation (EP300, CREBBP), histone methylation (MLL2, KDM6A), and DNA methylation (TET2, DNMT3A). In addition, components of core tumor suppressor pathways showed evidence of frequent inactivation (TP53, ATM, RB1, CUL9, PRKDC). In all, 22 of 25 cases had mutations in at least one of these 17 recurrently mutated genes. Multiple additional candidate disease mechanisms also were suggested by lower-confidence mutations but require confirmation studies, which are under way. In sum, analysis of the coding region of PTCL-NOS tumor DNA suggests a complex and heterogeneous pathogenesis, in line with gene-expression profiling. This work provides an opportunity to better sub-classify entities within the diagnosis of PTCL-NOS and identify specific therapeutic targets and their associated biomarkers. Disclosures: Horwitz: Seattle Genetics, Inc.: Consultancy, Research Funding; Millennium: Consultancy, Research Funding.

578 Tumor selective immune responses of STA551, a novel anti-CD137 agonist antibody activated by extracellular ATP

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A612-A612
Author(s):  
Yoshinori Narita ◽  
Mika Kamata-Sakurai
Keyword(s):  
Gene Expression ◽  
T Cell ◽  
Immune Responses ◽  
Tumor Growth ◽  
Tumor Tissue ◽  
Antigen Expression ◽  
Related Gene ◽  
Immune Related Gene ◽  
Ifn Γ ◽  
Agonist Antibody

BackgroundAgonistic antibodies targeting CD137 in clinic have failed due to severe hepatotoxicity, leading to the development of bispecific approaches that must rely on high tumor-associated antigen expression to crosslink CD137. Here we report on STA551, a novel anti-CD137 agonist antibody which binds to CD137 only in the presence of ATP. Extracellular ATP concentration is well-known to be elevated in tumor tissue while remaining tightly regulated in non-tumor tissue, suggesting that STA551 can activate immune cells only in tumor tissue and not elsewhere. Thus, STA551 has great potential to overcome the limitations of conventional CD137-targeted antibodies.MethodsWe evaluated in vitro STA551’s effect on IFN-γ production from human CD8+ T cells. We also evaluated in vivo STA551’s effect on tumor growth, RNAseq-based immune-related gene expression, immunohistochemistry, and T cell activation in tumor and non-tumor tissues in human CD137 knock-in mice treated with mouse surrogate STA551 (Sta-MB) or urelumab (Ure-MB) in combination with anti-PD-L1 antibody.ResultsIn a human T cell assay, STA551 induced IFN-γ only in the presence of ATP. In contrast, urelumab induced IFN-γ regardless of ATP concentration. In mice with Colon 38 tumors, Sta-MB inhibited tumor growth at least as strongly as Ure-MB, but whereas Ure-MB elicited systemic immune responses in draining lymph node, spleen, and liver, Sta-MB appeared to evade such responses. To confirm immune responses in tumors, we evaluated immune-related gene expression and found changes after treatment with Sta-MB or Ure-MB. These results suggest that STA551 works only in tumor tissue. Furthermore, Sta-MB with anti-PD-L1 antibody synergistically inhibited tumor growth and dramatically changed immune-related gene expression, CD8+ T cell infiltration, and PD-L1 expression without systemic immune responses. Also, it was well-tolerated in cynomolgus monkey in a repeated-dose toxicity study*.ConclusionsSTA551 is a novel anti-CD137 agonist antibody that exerts agonistic activity selectively in tumors without on-target toxicity in non-tumor tissues, regardless of tumor-associated antigen expression. These results strongly support the clinical testing of STA551 for the treatment of solid tumors. STA551 is currently being tested in a phase 1 clinical study.Ethics ApprovalAll animal studies were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC).

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