ΔNp73/ETS2 complex drives glioblastoma pathogenesis— targeting downstream mediators by rebastinib prolongs survival in preclinical models of glioblastoma

Abstract Background Glioblastoma (GBM) remains one of the least successfully treated cancers. It is essential to understand the basic biology of this lethal disease and investigate novel pharmacological targets to treat GBM. The aims of this study were to determine the biological consequences of elevated expression of ΔNp73, an N-terminal truncated isoform of TP73, and to evaluate targeting of its downstream mediators, the angiopoietin 1 (ANGPT1)/tunica interna endothelial cell kinase 2 (Tie2) axis, by using a highly potent, orally available small-molecule inhibitor (rebastinib) in GBM. Methods ΔNp73 expression was assessed in glioma sphere cultures, xenograft glioblastoma tumors, and glioblastoma patients by western blot, quantitative reverse transcription PCR, and immunohistochemistry. Immunoprecipitation, chromatin immunoprecipitation (ChiP) and sequential ChIP were performed to determine the interaction between ΔNp73 and E26 transformation-specific (ETS) proto-oncogene 2 (ETS2) proteins. The oncogenic consequences of ΔNp73 expression in glioblastomas were examined by in vitro and in vivo experiments, including orthotopic zebrafish and mouse intracranial-injection models. Effects of rebastinib on growth of established tumors and survival were examined in an intracranial-injection mouse model. Results ΔNp73 upregulates both ANGPT1 and Tie2 transcriptionally through ETS conserved binding sites on the promoters by interacting with ETS2. Elevated expression of ΔNp73 promotes tumor progression by mediating angiogenesis and survival. Therapeutic targeting of downstream ΔNp73 signaling pathways by rebastinib inhibits growth of established tumors and extends survival in preclinical models of glioblastoma. Conclusion Aberrant expression of ΔNp73 in GBM promotes tumor progression through autocrine and paracrine signaling dependent on Tie2 activation by ANGPT1. Disruption of this signaling by rebastinib improves tumor response to treatment in glioblastoma.

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Non-Coding RNAs as Mediators of Epigenetic Changes in Malignancies

Cancers ◽

10.3390/cancers12123657 ◽

2020 ◽

Vol 12 (12) ◽

pp. 3657

Author(s):

Subhasree Kumar ◽

Edward A. Gonzalez ◽

Pranela Rameshwar ◽

Jean-Pierre Etchegaray

Keyword(s):

Cellular Proliferation ◽

Circular Rnas ◽

Cancer Type ◽

Epigenetic Alterations ◽

Gene Expressions ◽

Aberrant Expression ◽

In Vivo Experiments ◽

Non Coding Rnas

Non-coding RNAs (ncRNAs) are untranslated RNA molecules that regulate gene expressions. NcRNAs include small nuclear RNAs (snRNAs), small nucleolar RNAs (snoRNAs), ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), circular RNAs (cRNAs) and piwi-interacting RNAs (piRNAs). This review focuses on two types of ncRNAs: microRNAs (miRNAs) or short interfering RNAs (siRNAs) and long non-coding RNAs (lncRNAs). We highlight the mechanisms by which miRNAs and lncRNAs impact the epigenome in the context of cancer. Both miRNAs and lncRNAs have the ability to interact with numerous epigenetic modifiers and transcription factors to influence gene expression. The aberrant expression of these ncRNAs is associated with the development and progression of tumors. The primary reason for their deregulated expression can be attributed to epigenetic alterations. Epigenetic alterations can cause the misregulation of ncRNAs. The experimental evidence indicated that most abnormally expressed ncRNAs impact cellular proliferation and apoptotic pathways, and such changes are cancer-dependent. In vitro and in vivo experiments show that, depending on the cancer type, either the upregulation or downregulation of ncRNAs can prevent the proliferation and progression of cancer. Therefore, a better understanding on how ncRNAs impact tumorigenesis could serve to develop new therapeutic treatments. Here, we review the involvement of ncRNAs in cancer epigenetics and highlight their use in clinical therapy.

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Elevated Expression of lncRNA MEG3 Induces Endothelial Dysfunction on HUVECs of IVF Born Offspring via Epigenetic Regulation

10.21203/rs.3.rs-69983/v1 ◽

2020 ◽

Author(s):

Ying Jiang ◽

Hong Zhu ◽

Hong Chen ◽

Meng-Meng Yang ◽

Yi-Chen Yu ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Umbilical Vein ◽

Methylation Status ◽

Vitro Fertilization ◽

In Vivo Experiments ◽

Elevated Expression ◽

Nitrite Nitrate ◽

Umbilical Vein Endothelial Cells

Abstract Background: The cardiovascular dysfunction in children born after in vitro fertilization (IVF) has been of great concern, in our study, we aim to explore potential molecular mechanism for such long-term outcomes. Methods：Real-time qPCR was used to test long non-coding RNA MEG3 and endothelium-derived factors, endothelial nitric oxide synthase (eNOS), endothelin-1(ET1), vascular endothelial growth factor (VEGF). Primary HUVEC after caesarean section was treated with different estradiol concentrations in vitro. Besides, knockdown of MEG3 on HUVEC provided further evidence between MEG3 expression and alteration of NO, ET1, VEGF. Then, by using pyrosequencing, we detected MEG3 promoter methylation status.Results: We found that the expression level of MEG3 was higher in human umbilical vein endothelial cells (HUVECs) of IVF offspring than that in spontaneously born offspring. Furthermore, we found decreased expression of eNOS, VEGF, elevated expression of ET1 in HUVECs from IVF offspring compared to spontaneously born offspring. We further confirmed the results from in-vivo experiments by demonstrating that high-estradiol intrauterine environments lead to abnormal expression of MEG3 and endothelium-derived factors. Meanwhile, silencing MEG3 expression decreased ET1 expression, and increased nitrite, nitrate, VEGF secretion, which could correct the effect we observed in-vivo. With pyrosequencing technology, we found that elevated expression of MEG3 in IVF offspring derived HUVECs was the result of hypomethylation of the MEG3 promoter. Conclusions: Our results demonstrated that higher expression of MEG3 in IVF-born HUVECs, accompanied by lower secretion of eNOS, VEGF, and higher secretion of ET1, which is closely related with endothelial dysfunction, which together provide a potential mechanism addressing high-risk of hypertension in IVF offspring.

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TGFβ-induced expression of long noncoding lincRNA Platr18 controls breast cancer axonogenesis

Life Science Alliance ◽

10.26508/lsa.202101261 ◽

2021 ◽

Vol 5 (2) ◽

pp. e202101261

Author(s):

Simon Grelet ◽

Cécile Fréreux ◽

Clémence Obellianne ◽

Ken Noguchi ◽

Breege V Howley ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Cell ◽

Tumor Progression ◽

Primary Tumor ◽

Epithelial Mesenchymal Transition ◽

Metastatic Progression ◽

Mesenchymal Transition ◽

Elevated Expression

Metastasis is the leading driver of cancer-related death. Tumor cell plasticity associated with the epithelial–mesenchymal transition (EMT), an embryonic program also observed in carcinomas, has been proposed to explain the colonization of distant organs by the primary tumor cells. Many studies have established correlations between EMT marker expression in the primary tumor and metastasis in vivo. However, the longstanding model of EMT-transitioned cells disseminating to secondary sites is still actively debated and hybrid states are presently considered as more relevant during tumor progression and metastasis. Here, we describe an unexplored role of EMT on the tumor microenvironment by controlling tumor innervation. Using in vitro and in vivo breast tumor progression models, we demonstrate that TGFβ-mediated tumor cell EMT triggers the expression of the embryonic LincRNA Platr18 those elevated expression controls the expression of the axon guidance protein semaphorin-4F and other neuron-related molecules such as IGSF11/VSIG-3. Platr18/Sema4F axis silencing abrogates axonogenesis and attenuates metastasis. Our observations suggest that EMT-transitioned cells are also locally required in the primary tumor to support distant dissemination by promoting axonogenesis, a biological process known for its role in metastatic progression of breast cancer.

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LINC00152 promotes Ovarian tumor progression and Predicts Poor Prognosis by inhibiting the ubiquitination of BCL6

10.21203/rs.3.rs-17686/v1 ◽

2020 ◽

Author(s):

Shunni Wang ◽

Weiwei Weng ◽

Tingting Chen ◽

Midie Xu ◽

Ping Wei ◽

...

Keyword(s):

Ovarian Cancer ◽

Tumor Progression ◽

Protein Interactions ◽

Ovarian Tumor ◽

Molecular Mechanisms ◽

Tumor Proliferation ◽

In Vivo Experiments ◽

Rna Immunoprecipitation

Abstract Background : Long non-coding RNA 00152 (LINC00152) has been found oncogenic in multiple somatic malignancies. But its roles in the ovarian cancer remain elusive. We aim to investigate its functions and mechanism in the ovarian tumor progression. Methods: We used RNAscope and RT-qPCR assays to detect the LINC00152 levels in 152 pairs of ovarian tumor and paratumorous tissues, conducted in vitro and in vivo experiments to evaluate its biological functions, and performed RNA immunoprecipitation, RNA-pulldown, MS/LS analysis, mutant construction and ubiquitination assays to explore the LINC00152-BCL6 binding and regulation. Results: LINC00152 was abnormally increased in the ovarian tumor tissues and its high expression predicted poorer survivals of patients; overexpression of LINC00152 promoted ovarian tumor proliferation and metastasis both in vitro and in vivo ; LINC00152 bond to Serine333 and Serine343 of BCL6 and then suppressed its ubiquitination; Finally, LINC00152 facilitated its oncogenic functions in a BCL6-mediated manner in ovarian cancer. Conclusions: LINC00152 is the independent prognostic predictor of ovarian cancer; and the abnormal expression of LINC00152 facilitates ovarian tumor proliferation and invasion by suppress the ubiquitination of BCL6. Our data might be helpful in exploring the molecular mechanisms of lncRNA-protein interactions, and might provide the potential target for the tumor pharmacology of ovarian cancer.

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Elevated expression of lncRNA MEG3 induces endothelial dysfunction on HUVECs of IVF born offspring via epigenetic regulation

10.21203/rs.3.rs-69983/v2 ◽

2020 ◽

Author(s):

Ying Jiang ◽

Hong Zhu ◽

Hong Chen ◽

Meng-Meng Yang ◽

Yi-Chen Yu ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Umbilical Vein ◽

Methylation Status ◽

Vitro Fertilization ◽

In Vivo Experiments ◽

Elevated Expression ◽

Nitrite Nitrate ◽

Umbilical Vein Endothelial Cells

Abstract Background: The cardiovascular dysfunction in children born after in vitro fertilization (IVF) has been of great concern, in our study, we aim to explore potential molecular mechanism for such long-term outcomes. Methods： Real-time qPCR was used to test long non-coding RNAMEG3and endothelium-derived factors, endothelial nitric oxide synthase (eNOS), endothelin-1(ET1), vascular endothelial growth factor (VEGF). Primary HUVEC after caesarean section was treated with different estradiol concentrations in vitro. Besides, knockdown ofMEG3on HUVEC provided further evidence between MEG3 expression and alteration of NO, ET1, VEGF. Then, by using pyrosequencing, we detectedMEG3promoter methylation status. Results: We found that the expression level of MEG3was higher in human umbilical vein endothelial cells (HUVECs) of IVF offspring than that in spontaneously born offspring. Furthermore, we found decreased expression ofeNOS, VEGF, elevated expression of ET1in HUVECs from IVF offspring compared to spontaneously born offspring. We further confirmed the results from in-vivo experiments by demonstrating that high-estradiol intrauterine environments lead to abnormal expression of MEG3 and endothelium-derived factors. Meanwhile, silencing MEG3expression decreased ET1expression, and increased nitrite, nitrate, VEGFsecretion, which could correct the effect we observed in-vivo. With pyrosequencing technology, we found that elevated expression of MEG3in IVF offspring derived HUVECs was the result of hypomethylation of the MEG3promoter. Conclusions: Our results demonstrated that higher expression ofMEG3in IVF-born HUVECs, accompanied by lower secretion of eNOS, VEGF, and higher secretion of ET1, which is closely related with endothelial dysfunction, which togetherprovide a potential mechanism addressing high-risk of hypertension in IVF offspring.

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SOX9 in prostate cancer is upregulated by cancer-associated fibroblasts to promote tumor progression through HGF/c-Met-FRA1 signaling

10.21203/rs.3.rs-32822/v1 ◽

2020 ◽

Author(s):

Haixiang Qin ◽

Yang Yang ◽

Bo Jiang ◽

Chun Pan ◽

Wei Chen ◽

...

Keyword(s):

Prostate Cancer ◽

Tumor Progression ◽

Molecular Mechanism ◽

The Cancer Genome Atlas ◽

Cancer Associated Fibroblasts ◽

Sox9 Expression ◽

In Vivo Experiments ◽

Vitro Model

Abstract Background Previous studies have demonstrated that transcription factor SOX9 which was reactivated in prostate cancer (Pca) and promoted tumor growth was a poor prognostic biomarker for Pca. Nevertheless, the regulatory mechanism underlying SOX9 upregulation in Pca still remains unclear. Several cytokines widely distributed in the tumor microenvironment (TME) have been reported to be involved in the regulation of SOX9, suggesting that cancer-associated fibroblasts (CAFs), one of the main sources of secreted factors in TME, may play a role in regulating SOX9 expression. Methods Herein, an in vitro model of paracrine interaction between primary CAFs and Pca cells (both AR-positive and AR-negative Pca cells), was applied to investigate the molecular mechanism of SOX9 upregulation during Pca progression. The regulatory axis was validated by in vivo experiments and The Cancer Genome Atlas (TCGA) data. Results Conditional medium from Pca CAFs (CAF-CM) upregulated the expression of SOX9, which was also proved to be essential for CAF-induced tumor progression. Further analysis showed that it was hepatocyte growth factor (HGF) secreted by CAFs that was responsible for the SOX9 elevation in Pca cells via activating c-Met signaling. Mechanistically, HGF/c-Met signaling specifically activated MEK1/2-ERK1/2 pathway which then induced phosphorylated status and protein upregulation of FRA1. Furthermore, ChIP assay demonstrated that FRA1 transcriptionally upregulated SOX9 expression by binding to the TPA-responsive element (TRE) sequence in the promoter of SOX9 gene. We also found that HGF/c-Met-ERK1/2-FRA1-SOX9 axis was relatively conserved in human and mouse species by validating in mouse Pca cells (RM-1). Conclusions Our results revealed a novel insight into the molecular mechanism that SOX9 expression in Pca cells is promoted by CAFs, through the HGF/cMet-ERK1/2-FRA1 axis. Besides, SOX9 may serve as an alternative marker for the activated HGF/c-Met signaling to enroll the optimal Pca patients for HGF/c-Met inhibition treatment, since it is much more stable and easier to detect.

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Effect of BPM31510 on radiosensitivity of temozolomide-resistant glioblastoma cell model and survival in in vivo C6 glioma rat model supporting phase I clinical investigation in GBM.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.e13509 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. e13509-e13509

Author(s):

Seema Nagpal ◽

Tulin Dadali ◽

Taichang Jang ◽

Milton Merchant ◽

Anne R. Diers ◽

...

Keyword(s):

Phase I ◽

Rat Model ◽

Clinical Investigation ◽

C6 Glioma ◽

Response To Treatment ◽

Preclinical Models ◽

Anti Cancer ◽

Cancer Activity

e13509 Background: Glioblastoma (GB) is characterized by dysregulated metabolism, utilizing glycolysis for energy production to support unrestricted growth. BPM 31510, an ubidecarenone containing lipid nanodispersion effectuates a switch in cancer energy sourcing from glycolysis towards mitochondrial OXPHOS, i.e. reverses Warburg effect, providing rationale for its potential utility in treatment of GB. The current study investigated utility of BPM31510 alone and in combination with temozolomide. Methods: In vitro (U251-MG human GB cell line) and in vivo (C6 glioma rat model) preclinical models of GB were used to assess the anti-cancer activity of BPM 31510 alone (100 mg/kg/d) and combination with TMZ/bevacizumab (BEV). In addition, an in vitro model of acquired TMZ chemo-resistance was established by progressive adaptation of parental U251-MG cells to increasing doses of TMZ. Parental and resistant subclones (TMZ-R) were used to define activity of BPM31510 in the TMZ-refractory setting. Results: In vitro results demonstrated that BPM 31510 has anti-cancer activity in both parental and TMZ-R U251-MG cells with EC50 values of ~400 µM and 800 µM, respectively. Importantly, BPM 31510 treatment also resensitized TMZ-R cell lines to TMZ. In vivo, BPM 31510 treatment was associated with increasing duration of survival; one fifth of the rats treated achieved survival greater than 15 days post implantation, a response not observed in the control or irradiation arms of the study. Assessment of the combination of BPM 31510 with TMZ or BEV in the in vivoC6 glioma rat model is ongoing. A phase I open-label, non-randomized clinical trial to evaluate the safety and tolerability of BPM31510 in patients with recurrent BEV-refractory GB, as well as the changes in GB metabolism by SUV-PET imaging in response to treatment is under investigation. Conclusions: Preclinical data demonstrate that BPM 31510 has potential anti-cancer activity alone and in combination with standard therapy regimens and alleviates TMZ chemo-resistance in preclinical models of GB. These results provide support of a Phase 1 clinical study of BPM31510 in GB; this study is actively enrolling.

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miR-155 as a Biomarker in B-Cell Malignancies

BioMed Research International ◽

10.1155/2016/9513037 ◽

2016 ◽

Vol 2016 ◽

pp. 1-14 ◽

Cited By ~ 22

Author(s):

Hanne Due ◽

Pernille Svendsen ◽

Julie Støve Bødker ◽

Alexander Schmitz ◽

Martin Bøgsted ◽

...

Keyword(s):

B Cell ◽

Relevant Literature ◽

B Cell Lymphoma ◽

Predictive Biomarker ◽

Lymphocytic Leukemia ◽

Response To Treatment ◽

B Cell Malignancies ◽

Elevated Expression

MicroRNAs have the potential to be useful biomarkers in the development of individualized treatment since they are easy to detect, are relatively stable during sample handling, and are important determinants of cellular processes controlling pathogenesis, progression, and response to treatment of several types of cancers including B-cell malignancies. miR-155 is an oncomiR with a crucial role in tumor initiation and development of several B-cell malignancies. The present review elucidates the potential of miR-155 as a diagnostic, prognostic, or predictive biomarker in B-cell malignancies using a systematic search strategy to identify relevant literature. miR-155 was upregulated in several malignancies compared to nonmalignant controls and overexpression of miR-155 was further associated with poor prognosis. Elevated expression of miR-155 shows potential as a diagnostic and prognostic biomarker in diffuse large B-cell lymphoma and chronic lymphocytic leukemia. Additionally,in vitroandin vivostudies suggest miR-155 as an efficient therapeutic target, supporting its oncogenic function. The use of inhibiting anti-miR structures indicates promising potential as novel anticancer therapeutics. Reports from 53 studies prove that miR-155 has the potential to be a molecular tool in personalized medicine.

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Berberine Inhibits FoxM1 Dependent Transcriptional Regulation of POLE2 and Interferes with the Survival of Lung Adenocarcinoma

10.21203/rs.3.rs-845748/v1 ◽

2021 ◽

Author(s):

Lulu Ni ◽

Ping Sun ◽

Xiaochun Fan ◽

Zhongjie Li ◽

Hongli Ren ◽

...

Keyword(s):

Dna Replication ◽

Lung Adenocarcinoma ◽

Molecular Mechanisms ◽

Cellular Functions ◽

In Vivo Experiments ◽

Elevated Expression ◽

Tcga Dataset ◽

Plasmid Transfection

Abstract Background：Berberine is one of the most interesting and promising natural anticancer drugs. POLE2 is involved in many cellular functions such as DNA replication and is highly expressed in a variety of cancers. However, the specific molecular mechanism of berberine interfering with POLE2 expression in lung adenocarcinoma (LUAD) is still unknown to a great extent.Method:The KEGG database (Release 91.0) and Gene Ontology (GO) category database were used for functional annotation of differentially expressed genes after berberine treatment. Reproducibility assessment using TCGA dataset. The biological functions of berberine in LUAD were investigated by a series of in vitro and in vivo experiments: MTT, colony formation, mouse xenograft and plasmid transfection. The molecular mechanisms of berberine were demonstrated by plasmid transfection, quantitative RT-PCR and Western blotting.Result:The elevated expression of FoxM1 and the high enrichment of DNA replication pathway were confirmed in LUAD by microarray and TCGA analysis, and were positively correlated with poor prognosis. Functionally, berberine inhibited the proliferation and survival of LUAD cell lines in vitro and in vivo. Mechanistically, berberine treatment down regulated the expression of FoxM1which closely related to survival, survival related genes in cell cycle and DNA replication pathway, and significantly down regulated the expression of survival related POLE2. Interestingly, we found that the transcription factor FoxM1 could act as a bridge between berberine and POLE2.Conclusion: Berberine significantly inhibited LUAD progression via the FoxM1/POLE2, and FoxM1/POLE2 may act as a clinical prognostic factor and a therapeutic target for LUAD. Berberine may be used as a promising therapeutic candidate for LUAD patients.

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The BCL9 Oncogene Promotes Tumor Progression by Conferring Enhanced Proliferative, Metastatic and Angiogenic Properties to Myeloma Cells

Blood ◽

10.1182/blood.v112.11.648.648 ◽

2008 ◽

Vol 112 (11) ◽

pp. 648-648

Author(s):

Mala Mani ◽

Jui Dutta ◽

Yunyu Zhang ◽

Daniel E Carrasco ◽

Yiming Zhou ◽

...

Keyword(s):

Wnt Signaling ◽

Tumor Progression ◽

Cyclin D1 ◽

Wnt Pathway ◽

Target Genes ◽

Metastatic Potential ◽

Luciferase Reporter ◽

Aberrant Expression

Abstract Wnt signaling plays an important role in tissue development and maintenance during embryogenesis, cell differentiation, and stem cell growth. Several components of the Wnt signaling cascade have been shown to function as either tumor suppressor proteins or as oncogenes in multiple human cancers, underscoring the relevance of this pathway in oncogenesis. Deregulation of the canonical Wnt/b-catenin pathway has been implicated in numerous human epithelial malignancies as well as hematologic malignancies including multiple myeloma (MM), generating immense interest in these molecules as targets for cancer therapy. Activation of Wnt/b-catenin in cancer has been associated with mutations that enable b-catenin to escape degradation by the proteasome, thereby allowing its accumulation in the nucleus where it functions as a transcriptional regulator in conjunction with coactivators by constitutively activating target genes such as c-Myc and Cyclin D1. To date, however, no mutations in Wnt pathway have been documented in MM, suggesting that mechanisms other than gene mutation may contribute to Wnt pathway deregulation. BCL9, a key component of the Wnt pathway, is required for b-catenin transcriptional activity and resides on chromosome 1q21, a region frequently involved in secondary chromosomal aberrations associated with MM tumor progression. Here we provide evidence that dysregulation of BCL9 expression is a novel oncogenic mechanism of Wnt pathway activation in MM. Using in vitro and in vivo functional analyses, we demonstrate that BCL9 is a bonafide oncogene that is aberrantly expressed in MM and associated with survival. Using the TCF- specific luciferase reporter, we show that enforced expression of BCL9 in MM cells enhanced b-catenin mediated transcription by >12 fold, suggesting a possible role of BCL9 overexpression in the pathogenesis of MM. BCL9 enhanced proliferation (1.5 fold, P<0.02), migration (3.5 fold, P<0.0001) and the metastatic potential of MM cells. We also showed that BCL9 plays an important role in tumor progression by regulating Cyclin D1 and c-Myc mediated cell proliferation, CD44 mediated tumor metastasis, as well as VEGF mediated host angiogenesis. Importantly, BCL9 knockdown significantly increased the survival in a xenograft mouse model of human MM (P=0.001), associated with decreased tumor burden and host angiogenesis. In summary, we have demonstrated that BCL9 is a novel and potent oncogene of the Wnt pathway in MM, playing fundamental roles in tumor progression by regulating proliferation, migration, invasion, angiogenesis and the metastatic potential of tumor cells. The pleiotropic roles of BCL9 and its aberrant expression highlight its importance as an attractive and novel therapeutic target in the treatment of MM.

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