LIN28B Underlies the Pathogenesis of a Subclass of Ewing Sarcoma

Keskin, Tuğba; Bakaric, Arnaud; Waszyk, Patricia; Boulay, Gaylor; Torsello, Matteo; Cornaz-Buros, Sandrine; Chevalier, Nadja; Geiser, Thibaud; Martin, Patricia; Volorio, Angela; Iyer, Sowmya; Kulkarni, Anupriya S.; Letovanec, Igor; Cherix, Stéphane; Coté, Gregory M.; Choy, Edwin; Digklia, Antonia; Montemurro, Michael; Chebib, Ivan; Nielsen, Petur; Carcaboso, Ángel Montero; Mora, Jaume; Renella, Raffaele; Suvà, Mario L.; Fusco, Carlo; Provero, Paolo; Rivera, Miguel N.; Riggi, Nicolò; Stamenkovic, Ivan

doi:10.1016/j.celrep.2020.107539

Cited by 6 publications

(13 citation statements)

References 54 publications

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“…S4A) does not support the notion of variable EWS-FLI-1 expression between GFP + and GFP − cells. In addition, expression of 106 previously described EWS-FLI-1 direct target genes (23) was not found to be significantly different between GFP + and GFP − cells (fig. S4B), indicating that their distinct behavior was not related to differences in EWSR1-FLI1 expression or function.…”

Section: Identification Of Candidate Genes That Underlie Ews Aggressi...mentioning

confidence: 79%

“…The coding sequence of enhanced GFP (EGFP) was amplified by PCR using pcDNA3-EGFP (Addgene, #13031) as template and EGFP-F and EGFP-R primers. The amplified EGFP cDNA sequence, which included a 3´UTR containing three miR-145 recognition sites, was cloned into the Bst BI and Mlu I restriction sites of the pLIV lentiviral vector (23). An EGFP cDNA containing five miR-145 recognition sites was subsequently generated by inserting two more miR-145 target sequences (oligo DNA A and oligo DNA B; table S4) into the Mlu I and Bam HI cloning site in the pLIV-EGFP vector containing the three miR-145 target sequence repeats.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…We then assessed mirRep145 function in two EwS 3D cultures, EwS1 and EwS2 (21)(22)(23), derived, respectively, from a metastatic lung lesion removed after chemotherapy and an untreated primary tumor (table S1). Both 3D cultures, which appeared as spheroids, retained the hierarchical cellular organization of the primary tumors from which they were derived (21,22).…”

Section: Detection Of Mir-145 Activity In Primary Ews Cellsmentioning

confidence: 99%

“…Using these parameters, we found 55 and 29 significantly up-and down-regulated genes, respectively, in GFP + compared to GFP − cells (tables S2 and S3). To address a possible link between miR-145 and EWS-FLI-1 expression levels, we compared differentially expressed genes between GFP + and GFP − EwS1 cells to those that displayed a change in expression upon EWSR1-FLI1 depletion by short hairpin RNA (shRNA) in the same model (23). However, the observation that the same EWS-FLI-1-dependent gene expression signature was enriched in both GFP + and GFP − populations (fig.…”

Section: Identification Of Candidate Genes That Underlie Ews Aggressi...mentioning

confidence: 99%

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A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

et al. 2021

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Targeting of the most aggressive tumor cell subpopulations is key for effective management of most solid malignancies. However, the metastable nature of tumor heterogeneity, which allows cells to transition between strong and weak tumorigenic phenotypes, and the lack of reliable markers of tumor-promoting properties hamper identification of the most relevant cells. To overcome these obstacles, we designed a functional microRNA (miR)–based live-cell reporter assay to identify highly tumorigenic cells in xenotransplants of primary Ewing sarcoma (EwS) 3D cultures. Leveraging the inverse relationship between cell pluripotency and miR-145 expression, we successfully separated highly tumorigenic, metastasis-prone (miR-145low) cells from poorly tumorigenic, nonmetastatic (miR-145high) counterparts. Gene expression and functional studies of the two cell populations identified the EPHB2 receptor as a prognostic biomarker in patients with EwS and a major promoter of metastasis. Our study provides a simple and powerful means to identify and isolate tumor cells that display aggressive behavior.

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Section: Identification Of Candidate Genes That Underlie Ews Aggressi...mentioning

confidence: 79%

Section: Plasmid Constructionmentioning

confidence: 99%

Section: Detection Of Mir-145 Activity In Primary Ews Cellsmentioning

confidence: 99%

Section: Identification Of Candidate Genes That Underlie Ews Aggressi...mentioning

confidence: 99%

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A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

et al. 2021

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“…Increased transcription levels of mirRNA-145 and Let-7 family members reduces the transcription levels of EWSR1-FLI1 (Ban et al, 2011;Keskin et al, 2020).…”

Section: Ewsr1 Promotermentioning

confidence: 99%

HDAC6 regulates expression of the oncogenic driver EWSR1-FLI1 through theEWSR1promoter in Ewing sarcoma

García-Domínguez

Hajji

Sánchez-Molina

et al. 2021

Preprint

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Ewing sarcoma (EWS) is an aggressive developmental sarcoma driven by a fusion gene, EWSR1-FLI1. However, little is known about the regulation of EWSR1-FLI1 chimeric fusion gene expression. Here, we demonstrate that active nuclear HDAC6 in EWS modulates acetylation status of specificity protein 1 (SP1), consequently regulating SP1/P300 activator complex binding to EWSR1 and EWSR1-FLI1 promoters. Selective inhibition of HDAC6 impairs binding of activator complex SP1/P300, thereby inducing EWSR1-FLI1 downregulation and significantly reducing its oncogenic functions. In addition, sensitivity of EWS cell lines to HDAC6 inhibition is higher than other tumor or non-tumor cell lines. Overexpression of HDAC6 in primary EWS tumor clinical samples correlates with a poor prognosis. Notably, a combination treatment of a selective HDAC6 inhibitor and doxorubicin (standard of care in EWS) dramatically inhibits tumor growth in two EWS murine xenograft models. These results could lead to suitable and promising therapeutic alternatives for EWS patients.

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The LIN28B–let‐7–PBK pathway is essential for group 3 medulloblastoma tumor growth and survival

Shahab,

Roggeveen,

Sun

et al. 2023

Molecular Oncology

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Children with Group 3 medulloblastoma (G3 MB) have a very poor prognosis, and many do not survive beyond 5 years after diagnosis. A factor that may contribute to this is the lack of available targeted therapy. Expression of protein lin‐28 homolog B (LIN28B), a regulator of developmental timing, is upregulated in several cancers, including G3 MB, and is associated with worse survival in this disease. Here, we investigate the role of the LIN28B pathway in G3 MB and demonstrate that the LIN28B–lethal‐7 (let‐7; a microRNA that is a tumor suppressor)–lymphokine‐activated killer T‐cell‐originated protein kinase (PBK; also known as PDZ‐binding kinase) axis promotes G3 MB proliferation. LIN28B knockdown in G3‐MB‐patient‐derived cell lines leads to a significant reduction in cell viability and proliferation in vitro and in prolonged survival of mice with orthotopic tumors. The LIN28 inhibitor N‐methyl‐N‐[3‐(3‐methyl‐1,2,4‐triazolo[4,3‐b]pyridazin‐6‐yl)phenyl]acetamide (1632) significantly reduces G3 MB cell growth and demonstrates efficacy in reducing tumor growth in mouse xenograft models. Inhibiting PBK using HI‐TOPK‐032 also results in a significant reduction in G3 MB cell viability and proliferation. Together, these results highlight a critical role for the LIN28B–let‐7–PBK pathway in G3 MB and provide preliminary preclinical results for drugs targeting this pathway.

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LIN28B Underlies the Pathogenesis of a Subclass of Ewing Sarcoma

Cited by 6 publications

References 54 publications

A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

HDAC6 regulates expression of the oncogenic driver EWSR1-FLI1 through theEWSR1promoter in Ewing sarcoma

The LIN28B–let‐7–PBK pathway is essential for group 3 medulloblastoma tumor growth and survival

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