Inhibition of the Spectraplakin Protein Microtubule Actin Crosslinking Factor 1 Sensitizes Glioblastomas to Radiation

Kala Bonner,Danielle Borlay,Orica Kutten,Quincy A. Quick 대한뇌종양학회 2020 Brain Tumor Research and Treatment Vol.8 No.1

Background: Microtubule actin crosslinking factor 1 (MACF1) is a spectraplakin cytoskeletal crosslinking protein whose function and role in cancer biology has lacked investigation. Recent studies have identified MACF1 as a novel target in glioblastomas expressed in tissue from tumor patient explants but not normal brain tissue and when silenced has an antitumorigenic impact on these tumors. Radiation as a single agent therapy to treat glioblastomas has been used for decades and has done little to improve survival of individuals diagnosed with this disease. However, contemporary clinical radiotherapy protocols have provided evidence that combinatorial radiotherapy approaches confer a therapeutic benefit in glioblastoma patients. In this study MACF1 was investigated as a radiosensitization target in glioblastomas. Methods: To provide context of MACF1 in glioblastomas, The Cancer Genome Atlas expression analyses were performed in conjunction with genes associated with glioblastoma evolution, while a genetic inhibitory approach, cell migratory assays, and immunofluorescence procedures were used to evaluate responses to MACF1 suppression with radiation. Additionally, expression analyses were conducted to assess co-expression of mTOR signaling pathway regulators and MACF1 in glioblastoma patient samples Results: Our amalgamation approach demonstrated that negative regulation of MACF1, which was positively correlated with epidermal growth factor receptor and p70s6k expression, enhanced the sensitivity of glioblastoma cells to radiation as a consequence of reducing glioblastoma cell viability and migration. Mechanistically, the antitumorigenic effects on glioblastoma cell behaviors after radiation and impairing MACF1 function were associated with decreased expression of ribosomal protein S6, a downstream effector of p70s6k. Conclusion: MACF1 represents a diagnostic marker with target specificity in glioblastomas that can enhance the efficacy of radiation while minimizing normal tissue toxicity. This approach could potentially expand combinatorial radiation strategies for glioblastoma treatments via impairment of translational regulatory processes that contribute to poor patient survival.

Differentiation between Glioblastoma and Solitary Metastasis: Morphologic Assessment by Conventional Brain MR Imaging and Diffusion-Weighted Imaging

Jung, Bo Young,Lee, Eun Ja,Bae, Jong Myon,Choi, Young Jae,Lee, Eun Kyoung,Kim, Dae Bong Korean Society of Magnetic Resonance in Medicine 2021 Investigative Magnetic Resonance Imaging Vol.25 No.1

Purpose: Differentiating between glioblastoma and solitary metastasis is very important for the planning of further workup and treatment. We assessed the ability of various morphological parameters using conventional MRI and diffusion-based techniques to distinguish between glioblastomas and solitary metastases in tumoral and peritumoral regions. Materials and Methods: We included 38 patients with solitary brain tumors (21 glioblastomas, 17 solitary metastases). To find out if there were differences in the morphologic parameters of enhancing tumors, we analyzed their shape, margins, and enhancement patterns on postcontrast T1-weighted images. During analyses of peritumoral regions, we assessed the extent of peritumoral non-enhancing lesion on T2- and postcontrast T1-weighted images. We also aimed to detect peritumoral neoplastic cell infiltration by visual assessment of T2-weighted and diffusion-based images, including DWI, ADC maps, and exponential DWI, and evaluated which sequence depicted peritumoral neoplastic cell infiltration most clearly. Results: The shapes, margins, and enhancement patterns of tumors all significantly differentiated glioblastomas from metastases. Glioblastomas had an irregular shape, ill-defined margins, and a heterogeneous enhancement pattern; on the other hand, metastases had an ovoid or round shape, well-defined margins, and homogeneous enhancement. Metastases had significantly more extensive peritumoral T2 high signal intensity than glioblastomas had. In visual assessment of peritumoral neoplastic cell infiltration using T2-weighted and diffusion-based images, all sequences differed significantly between the two groups. Exponential DWI had the highest sensitivity for the diagnosis of both glioblastoma (100%) and metastasis (70.6%). A combination of exponential DWI and ADC maps was optimal for the depiction of peritumoral neoplastic cell infiltration in glioblastoma. Conclusion: In the differentiation of glioblastoma from solitary metastatic lesions, visual morphologic assessment of tumoral and peritumoral regions using conventional MRI and diffusion-based techniques can also offer diagnostic information.

Nanoarchitectured Conjugates Targeting Angiogenesis: Investigating Heparin-Taurocholate Acid Conjugates (LHT7) as an Advanced Anti-Angiogenic Therapy for Brain Tumor Treatment

김형식,설재학,황해현,이동윤 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00

Background Glioblastoma is a highly malignant brain tumor associated with poor prognosis. Conventional therapeutic approaches have limitations due to their toxic effects on normal tissue and the development of tumor cell resistance. This study aimed to explore alternative mechanisms for glioblastoma treatment by targeting angiogenesis. Methods The study investigated the anti-angiogenic properties of heparin in glioblastoma treatment. To overcome the limitations of heparin, a heparin-taurocholate conjugate (LHT7) was synthesized by conjugating heparin to taurocholic acid. The study utilized the U87MG human glioblastoma cell line and human umbilical vein endothelial cells (HUVEC) as experimental models. Cell viability assays and sprouting assays were performed to assess the effects of LHT7. Additionally, phosphorylation of angiogenesis-related proteins, such as phospho-ERK and phospho-VEGFR2, was measured. The anti-angiogenic effects of LHT7 were further evaluated using a glioblastoma orthotopic mouse model. Results Treatment with LHT7 resulted in a dose-dependent reduction in cell viability in U87MG human glioblastoma cells. The sprouting of HUVEC cells was significantly decreased upon LHT7 treatment. Furthermore, LHT7 treatment led to a decrease in the phosphorylation of angiogenesis-related proteins, including phospho-ERK and phospho- VEGFR2. In the glioblastoma orthotopic mouse model, LHT7 exhibited anti-angiogenic effects, supporting its potential as a therapeutic agent. Conclusions The conjugation of heparin and taurocholic acid to create LHT7 offers several advantages over conventional therapeutic approaches for glioblastoma. LHT7 demonstrated anti-angiogenic properties, as evidenced by the reduction in cell viability and inhibition of endothelial cell sprouting. Moreover, LHT7 modulated the phosphorylation of angiogenesis-related proteins. These findings suggest that LHT7 holds promise as a medication for glioblastoma treatment, offering potential implications for improving patient outcomes.

Immunohistochemical Classification of Primary and Secondary Glioblastomas

이규상,남경한,서안나,윤수미,김경주,조화진,박성혜,최기영 대한병리학회 2013 Journal of Pathology and Translational Medicine Vol.47 No.6

Background: Glioblastomas may develop de novo (primary glioblastomas, P-GBLs) or through progression from lower-grade astrocytomas (secondary glioblastomas, S-GBLs). The aim of this study was to compare the immunohistochemical classification of glioblastomas with clinically determined P-GBLs and S-GBLs to identify the best combination of antibodies for immunohistochemical classification. Methods: We evaluated the immunohistochemical expression of epidermal growth factor receptor (EGFR), p53, and isocitrate dehydrogenase 1 (IDH-1) in 150 glioblastoma cases. Results: According to clinical history, the glioblastomas analyzed in this study consisted of 146 P-GBLs and 4 S-GBLs. Immunohistochemical expression of EGFR, p53, and IDH-1 was observed in 62.6%, 49.3%, and 11.1%, respectively. Immunohistochemical profiles of EGFR(+)/ p53(–), IDH-1(–)/EGFR(+)/p53(–), and EGFR(–)/p53(+) were noted in 41.3%, 40.2%, and 28.7%, respectively. Expression of IDH-1 and EGFR(–)/p53(+) was positively correlated with young age. The typical immunohistochemical features of S-GBLs comprised IDH-1(+)/EGFR(–)/p53(+), and were noted in 3.6% of clinically P-GBLs. The combination of IDH-1(–) or EGFR(+) was the best set of immunohistochemical stains for identifying P-GBLs, whereas the combination of IDH-1(+) and EGFR(–) was best for identifying S-GBLs. Conclusions: We recommend a combination of IDH-1 and EGFR for immunohistochemical classification of glioblastomas. We expect our results to be useful for determining treatment strategies for glioblastoma patients.

Rapid Progression of Gliomatosis Cerebri to Secondary Glioblastoma, Factors That Affect the Progression Rate: A Case Report

김희경,유인규,김승민,김주헌,이승훈,이승연 대한영상의학회 2017 대한영상의학회지 Vol.76 No.3

Glioblastomas may develop de novo or through progression from low-grade or anaplastic astrocytomas. The term ‘primary glioblastoma’ refers to a glioblastoma that lacks a precursor lesion and has a clinical history of less than three months. On the other hand, the term ‘secondary glioblastoma’ indicates that the glioblastoma has progressed from a low-grade tumor after a long latency period and often manifests in younger patients. These subtypes of glioblastoma develop via different genetic pathways, and they differ in prognosis and response to therapy. Thus, differential diagnosis of these subtypes and prediction of the factors that affect the progression from low-grade diffuse astrocytoma to secondary glioblastoma would be clinically very important. We present a rare case of secondary glioblastoma, which developed only three months after the follow up imaging evaluations, with a history of low grade glioma, and present the factors that cause rapid progression.

Delivery of Hypoxia and Glioma Dual-Specific Suicide Gene Using Dexamethasone Conjugated Polyethylenimine for Glioblastoma-Specific Gene Therapy

Kim, Hyun Ah,Park, Jin Hyeong,Yi, Na,Lee, Minhyung American Chemical Society 2014 MOLECULAR PHARMACEUTICS Vol.11 No.3

<P>Gene therapy has been considered a promising approach for glioblastoma therapy. To avoid side effects and increase the specificity of gene expression, gene expression should be tightly regulated. In this study, glioma and hypoxia dual-specific plasmids (pEpo-NI2-SV-Luc and pEpo-NI2-SV-HSVtk) were developed by combining the erythropoietin (Epo) enhancer and nestin intron 2 (NI2). In the <I>in vitro</I> studies, pEpo-NI2-SV-Luc showed higher gene expression under hypoxia than normoxia in a glioblastoma-specific manner. The MTT and caspase assays demonstrated that pEpo-NI2-SV-HSVtk specifically induced caspase activity and cell death in hypoxic glioblastoma cells. For <I>in vivo</I> evaluation, subcutaneous and intracranial glioblastoma models were established. Dexamethasone-conjugated-polyethylenimine (PEI-Dexa) was used as a gene carrier, since PEI-Dexa efficiently delivers plasmid to glioblastoma cells and also has an antitumor effect due to the effect of dexamethasone. In the <I>in vivo</I> study in the subcutaneous and intracranial glioblastoma models, the tumor size was reduced more effectively in the pEpo-NI2-SV-HSVtk group than in the control and pSV-HSVtk groups. In addition, higher levels of HSVtk gene expression and TUNEL-positive cells were observed in the pEpo-NI2-SV-HSVtk group compared with the control and pSV-HSVtk groups, suggesting that pEpo-NI2-SV-HSVtk increased the therapeutic efficacy in hypoxic glioblastoma. Therefore, pEpo-NI2-SV-HSVtk/PEI-Dexa complex may be useful for glioblastoma-specific gene therapy.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/mpohbp/2014/mpohbp.2014.11.issue-3/mp4006003/production/images/medium/mp-2013-006003_0014.gif'></P>

Repositioning of the antipsychotic trifluoperazine: Synthesis, biological evaluation and <i>in silico</i> study of trifluoperazine analogs as anti-glioblastoma agents

Kang, Seokmin,Lee, Jung Moo,Jeon, Borami,Elkamhawy, Ahmed,Paik, Sora,Hong, Jinpyo,Oh, Soo-Jin,Paek, Sun Ha,Lee, C. Justin,Hassan, Ahmed H.E.,Kang, Sang Soo,Roh, Eun Joo Elsevier 2018 European journal of medicinal chemistry Vol.151 No.-

<P><B>Abstract</B></P> <P>Repositioning of the antipsychotic drug trifluoperazine for treatment of glioblastoma, an aggressive brain tumor, has been previously suggested. However, trifluoperazine did not increase the survival time in mice models of glioblastoma. In attempt to identify an effective trifluoperazine analog, fourteen compounds have been synthesized and biologically <I>in vitro</I> and <I>in vivo</I> assessed. Using MTT assay, compounds <B>3dc</B> and <B>3dd</B> elicited 4–5 times more potent inhibitory activity than trifluoperazine with IC<SUB>50</SUB> = 2.3 and 2.2 μM against U87MG glioblastoma cells, as well as, IC<SUB>50</SUB> = 2.2 and 2.1 μM against GBL28 human glioblastoma patient derived primary cells, respectively. Furthermore, they have shown a reasonable selectivity for glioblastoma cells over NSC normal neural cell. <I>In vivo</I> evaluation of analog <B>3dc</B> confirmed its advantageous effect on reduction of tumor size and increasing the survival time in brain xenograft mouse model of glioblastoma. Molecular modeling simulation provided a reasonable explanation for the observed variation in the capability of the synthesized analogs to increase the intracellular Ca<SUP>2+</SUP> levels. In summary, this study presents compound <B>3dc</B> as a proposed new tool for the adjuvant chemotherapy of glioblastoma.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fourteen TFP derivatives were synthesized and biologically assessed. </LI> <LI> <B>3dc</B> and <B>3dd</B> elicited promising inhibitory activities against U87MG and GBL28 cells. </LI> <LI> <B>3dc</B> was assessed in brain xenograft mouse model of glioblastoma. </LI> <LI> Molecular modeling provided explanation for observed activity variations. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Functional Biological Activity of Sorafenib as a Tumor-Treating Field Sensitizer for Glioblastoma Therapy

Jo, Yunhui,Kim, Eun Ho,Sai, Sei,Kim, Jin Su,Cho, Jae-Min,Kim, Hyeongi,Baek, Jeong-Hwa,Kim, Jeong-Yub,Hwang, Sang-Gu,Yoon, Myonggeun MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.11

<P>Glioblastoma, the most common primary brain tumor in adults, is an incurable malignancy with poor short-term survival and is typically treated with radiotherapy along with temozolomide. While the development of tumor-treating fields (TTFields), electric fields with alternating low and intermediate intensity has facilitated glioblastoma treatment, clinical outcomes of TTFields are reportedly inconsistent. However, combinatorial administration of chemotherapy with TTFields has proven effective for glioblastoma patients. Sorafenib, an anti-proliferative and apoptogenic agent, is used as first-line treatment for glioblastoma. This study aimed to investigate the effect of sorafenib on TTFields-induced anti-tumor and anti-angiogenesis responses in glioblastoma cells in vitro and in vivo. Sorafenib sensitized glioblastoma cells to TTFields, as evident from significantly decreased post-TTFields cell viability (<I>p</I> < 0.05), and combinatorial treatment with sorafenib and TTFields accelerated apoptosis via reactive oxygen species (ROS) generation, as evident from Poly (ADP-ribose) polymerase (PARP) cleavage. Furthermore, use of sorafenib plus TTFields increased autophagy, as evident from LC3 upregulation and autophagic vacuole formation. Cell cycle markers accumulated, and cells underwent a G2/M arrest, with an increased G0/G1 cell ratio. In addition, the combinatorial treatment significantly inhibited tumor cell motility and invasiveness, and angiogenesis. Our results suggest that combination therapy with sorafenib and TTFields is slightly better than each individual therapy and could potentially be used to treat glioblastoma in clinic, which requires further studies.</P>

Propranolol Inhibits the Proliferation of Human Glioblastoma Cell Lines through Notch1 and Hes1 Signaling System

Kim, Hyun Sik,Park, Young Han,Lee, Heui Seung,Kwon, Mi Jung,Song, Joon Ho,Chang, In Bok The Korean Neurosurgical Society 2021 Journal of Korean neurosurgical society Vol.64 No.5

Objective : The anti-tumor effect of the beta-adrenergic receptor antagonist propranolol in breast cancer is well known; however, its activity in glioblastoma is not well-evaluated. The Notch-Hes pathway is known to regulate cell differentiation, proliferation, and apoptosis. We investigated the effect of propranolol to human glioblastoma cell lines, and the role of Notch and Hes signaling in this process. Methods : We performed immunohistochemical staining on 31 surgically resected primary human glioblastoma tissues. We also used glioblastoma cell lines of U87-MG, LN229, and neuroblastoma cell line of SH-SY5Y in this study. The effect of propranolol and isoproterenol on cell proliferation was evaluated using the MTT assay (absorbance 570 nm). The impact of propranolol on gene expression (Notch and Hes) was evaluated using real-time polymerase chain reaction (RT-PCR, whereas protein levels of Notch1 and Hes1 were measured using Western blotting (WB), simultaneously. Small interfering RNA (siRNA) was used to suppress the Notch gene to investigate its role in the proliferation of glioblastoma. Results : Propranolol and isoproterenol caused a dose-dependent decrease in cell proliferation (MTT assay). RT-PCR showed an increase in Notch1 and Hes1 expression by propranolol, whereas WB demonstrated increase in Notch1 protein, but a decrease in Hes1 by propranolol. The proliferation of U87-MG and LN229 was not significantly suppressed after transfection with Notch siRNA. Conclusion : These results demonstrated that propranolol suppressed the proliferation of glioblastoma cell lines and neuroblastoma cell line, and Hes1 was more closely involved than Notch1 was in glioblastoma proliferation.

기초 : Subtractive Hybridization법에 의한 교모세포종에서 특이적으로 발현하는 신규 유전자의 기능 연구

안정용 ( Jung Yong Ahn ),유나미 ( Na Mi Yu ),한인보 ( In Bo Han ),김진경 ( Jin Kyeoung Kim ) 대한뇌종양학회 2005 대한뇌종양학회지 Vol.4 No.2

A variety of genetic alterations in human glioblastomas comprises signal transduction and cell cycle arrest control of cellular processes. In an attempt to identify novel genes implicated in glioblastoma development, we performed experiments using suppression subtraction hybridization(SSH) with human glioblastoma tissue, as well as with cloned genes, that are expressed more than in normal brain tissues. After the identification of a novel gene, RT-PCR was performed to determine its mRNA expression in human glioblastoma tissues. In order to learn more about the expression profile of this gene, PCR was performed using various commercially available normal or carcinoma cell lines. The novel gene was found to be strongly expressed in glioblastoma tissues and other carcinoma cell lines. To determine whether this novel gene was associated with cell cycle regulation, a serum stimulation study with normal WI-38 fibroblast cells treated medium containing 0.1% FBS for 48hr was used for its examination. The novel clone was identified as being expressed in several glioblastoma tissues and carcinoma cell lines. The time-course expression of this novel gene in the WI-38(8PDL) normal lung cell line indicated a significant increase for G1-phase arrest. We have used suppression subtractive hybridization(SSH) to identify cDNA of a gene “Clone 15” overexpressed in human glioblastoma. When the expression of “Clone 15” was screened in numerous cell types, we could confirm that this novel gene was overexpressed in tumor compared to matched normal tissue.