MV1035 Overcomes Temozolomide Resistance in Patient-Derived Glioblastoma Stem Cell Lines

Glioblastoma (GBM, grade IV glioma) represents the most aggressive brain tumor and patients with GBM have a poor prognosis. Until now surgical resection followed by radiotherapy and temozolomide (TMZ) treatment represents the standard strategy for GBM. We showed that the imidazobenzoxazin-5-thione MV1035 is able to significantly reduce GBM U87-MG cells migration and invasiveness through inhibition of the RNA demethylase ALKBH5. In this work, we focus on the DNA repair protein ALKBH2, a further MV1035 target resulting from SPILLO-PBSS proteome-wide scale in silico analysis. Our data demonstrate that MV1035 inhibits the activity of ALKBH2, known to be involved in GBM TMZ resistance. MV1035 was used on both U87-MG and two patient-derived (PD) glioma stem cells (GSCs): in combination with TMZ, it has a significant synergistic effect in reducing cell viability and sphere formation. Moreover, MV1035 induces a reduction in MGMT expression in PD-GSCs cell lines most likely through a mechanism that acts on MGMT promoter methylation. Taken together our data show that MV1035 could act as an inhibitor potentially helpful to overcome TMZ resistance and able to reduce GBM migration and invasiveness.

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Innovative 3D proteome-wide scale identification of ALKBH5 target for MV1035 small molecule able to reduce migration and invasiveness in U87 glioblastoma cell lines by SPILLO-PBSS

Biomedical Science and Engineering ◽

10.4081/bse.2021.178 ◽

2021 ◽

Vol 4 (s1) ◽

Author(s):

Alessio Malacrida ◽

Mirko Rivara ◽

Omar Ben Mariem ◽

Alessandro Di Domizio ◽

Giacomo Cislaghi ◽

...

Keyword(s):

Cell Lines ◽

Small Molecule ◽

Binding Sites ◽

Research Projects ◽

Animal Testing ◽

Repair Protein ◽

Potential Binding ◽

Dna Repair Protein ◽

Wide Scale ◽

Glioblastoma Cell Lines

The innovative in silico technologies developed at SPILLOproject,1 e.g., the SPILLO potential binding sites searcher (SPILLO-PBSS) software,2,3 allow to identify targets and off-targets of any small molecule on a multiple-organism proteomewide scale, and to perform an accurate multilevel cross-organism transferability analysis (MCOTA) aimed at rationalising animal testing. SPILLO-PBSS has been successfully used in several research projects, such as a study in which a compound (MV1035) was found to reduce migration and invasiveness in U87 glioblastoma (GBM) cell lines: the human structural proteome was analyzed and the RNA demethylase ALKBH5 has been identified as a target responsible for the observed effects (target experimentally validated). Another top-ranked target identified by SPILLO-PBSS, the DNA repair protein AlkB homolog 2 (ALKBH2), abundantly expressed in GBM cell lines, resulted particularly interesting for its pivotal role in the onset of resistance to Temozolomide (TMZ), the standard firstline treatment for GBM.2

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The effects of 41°C hyperthermia on the DNA repair protein, MRE11, correlate with radiosensitization in four human tumor cell lines

International Journal of Hyperthermia ◽

10.1080/02656730701383007 ◽

2007 ◽

Vol 23 (4) ◽

pp. 343-351 ◽

Cited By ~ 17

Author(s):

M. Xu ◽

R. J. Myerson ◽

Y. Xia ◽

T. Whitehead ◽

E. G. Moros ◽

...

Keyword(s):

Dna Repair ◽

Tumor Cell ◽

Cell Lines ◽

Human Tumor ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines ◽

Repair Protein ◽

Dna Repair Protein

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CSIG-02. VAL-083 INHIBITS PROLIFERATION OF A PANEL OF EIGHT GLIOBLASTOMA STEM CELL LINES: DOWNREGULATION OF BDR4 AS A NOVEL ANTI-NEOPLASTIC MECHANISM

Neuro-Oncology ◽

10.1093/neuonc/noy148.168 ◽

2018 ◽

Vol 20 (suppl_6) ◽

pp. vi43-vi43

Author(s):

Elise Fernandez ◽

Anne Steino ◽

Glenn Lesser ◽

Jeffrey Bacha ◽

Dennis Brown ◽

...

Keyword(s):

Stem Cell ◽

Cell Lines ◽

Glioblastoma Stem Cell ◽

Stem Cell Lines

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A Novel Amino Acid Substitution, Fibrinogen Bβp.Pro234Leu, Associated with Hypofibrinogenemia Causing Impairment of Fibrinogen Assembly and Secretion

International Journal of Molecular Sciences ◽

10.3390/ijms21249422 ◽

2020 ◽

Vol 21 (24) ◽

pp. 9422

Author(s):

Takahiro Kaido ◽

Masahiro Yoda ◽

Tomu Kamijo ◽

Shinpei Arai ◽

Chiaki Taira ◽

...

Keyword(s):

Cell Lines ◽

In Silico ◽

Contact Region ◽

Chinese Hamster ◽

In Silico Analysis ◽

Enzyme Linked Immunosorbent Assay ◽

Immunoblotting Analysis ◽

Fibrin Polymerization ◽

The Impact ◽

Silico Analysis

We identified a novel heterozygous variant, Bβp.Pro234Leu (fibrinogen Tokorozawa), which was suspected to be associated with hypofibrinogenemia. Therefore, we analyzed the assembly and secretion of this fibrinogen using Chinese hamster ovary (CHO) cells. To determine the impact on the synthesis and secretion of fibrinogen of the Bβp.P234L and γp.G242E substitutions, we established recombinant variant fibrinogen-producing CHO cell lines. Synthesis and secretion analyses were performed using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting analysis with the established cell lines. In addition, we performed fibrin polymerization using purified plasma fibrinogen and in-silico analysis. Both Bβp.P234L and γp.G242E impaired the secretion and synthesis of fibrinogen. Moreover, immunoblotting analysis elucidated the mobility migration of the Bβγ complex in Bβp.P234L. On the other hand, the fibrin polymerization of fibrinogen Tokorozawa was similar to that of normal fibrinogen. In-silico analysis revealed that the Bβp.P234 residue is located in the contact region between the Bβ and γ chains and contacts γp.G242 residue. The present study demonstrated that the Bβp.P234L substitution resulted in hypofibrinogenemia by decreasing the assembly and secretion of fibrinogen. Therefore, there is a possibility that substitutions in the contact region between the Bβ and γ chains impact the assembly and secretion of fibrinogen.

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MBRS-13. MiR-1253 POTENTIATES CISPLATIN RESPONSE IN PEDIATRIC MEDULLOBLASTOMA BY REGULATING FERROPTOSIS

Neuro-Oncology ◽

10.1093/neuonc/noaa222.530 ◽

2020 ◽

Vol 22 (Supplement_3) ◽

pp. iii400-iii400

Author(s):

Ranjana K Kanchan ◽

Naveenkumar Perumal ◽

Pranita Atri ◽

Ramakanth Chirravuri Venkata ◽

Ishwor Thapa ◽

...

Keyword(s):

High Risk ◽

Cell Lines ◽

In Silico Analysis ◽

Poor Overall Survival ◽

Cancer Cell Death ◽

Pediatric Medulloblastoma ◽

Mitochondrial Ros ◽

Group 3 ◽

Silico Analysis

Abstract Despite improvements in targeted therapies, few group 3 medulloblastoma patients survive long-term. Haploinsufficiency of 17p13.3 is a hallmark of these high-risk tumors; included within this locus is miR-1253, which has tumor suppressive properties in medulloblastoma. Therapeutic strategies capitalizing on the anti-neoplastic properties of miRNAs can provide promising adjuncts to chemotherapy. In this study, we explored the potentiation of miR-1253 on cisplatin cytotoxicity in group 3 MB. Overexpression of miR-1253 sensitized group 3 MB cell lines to cisplatin, leading to a pronounced downregulation in cell viability and induction of apoptosis. Cisplatin is reported as an inducer of both apoptosis and ferroptosis-mediated cancer cell death. In silico analysis revealed an upregulation of several ABC transporters in high-risk MB tumors. When compared to cell lines overexpressing miR-1253, the ABC transporter ABCB7, which regulates both apoptosis and ferroptosis, was revealed as a putative target of miR-1253 with poor survival that may facilitate its chemosensitizing effects by modulating mitochondrial ROS and HIF1α-driven NFκB signaling. We observed high expression of ABCB7 and GPX4, ferroptosis regulators, in MB patients with poor overall survival. MiR-1253 negatively regulated the expression of ABCB7 in Group 3 MB cell lines and induced cytoplasmic ROS and mitochondrial O2-, suggesting ROS-mediated induction of ferroptosis through regulation of ABCB7 and GPX4. Treatment with ROS and ferroptosis inhibitors rescued miR-1253 transfected cells treated with cisplatin. We conclude that miR-1253 induced ROS and potentiated the ferroptotic effects of cisplatin via targeting miR-1253/ABCB7/GPX4/mtROS axis.

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