Recurrent Somatic<i>DICER1</i>Mutations in Nonepithelial Ovarian Cancers

Heravi-Moussavi, Alireza; Anglesio, Michael S.; Cheng, S-W Grace; Senz, Janine; Yang, Winnie; Prentice, Leah M; Fejes, Anthony P.; Chow, Christine; Tone, Alicia A.; Kalloger, Steve E.; Hamel, Nancy; Roth, Andrew; Ha, Gavin; Wan, Adrian; Maines-Bandiera, Sarah; Salamanca, Clara; Pasini, Barbara; Clarke, Blaise A.; Lee, Anna F.; Lee, Cheng‐Han; Zhao, Chengquan; Young, Robert H.; Aparicio, Samuel; Sorensen, Poul H.; Woo, Michelle; Boyd, Niki; Jones, Steven J.M.; Hirst, Martin; Marra, Marco A.; Gilks, Blake; Shah, Sohrab P.; Foulkes, William D.; Morin, Gregg B.; Huntsman, David G.

doi:10.1056/nejmoa1102903

Cited by 426 publications

(449 citation statements)

References 38 publications

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“…Both G1809R and D1709N DICER1 mutants failed to cleave the 5 ′ -labelled pre-let-7c miRNA ( Figure 4A) at the RNase IIIb processing site in vitro, as shown by the nearly absent amount of the 22 nt 5p let-7c miRNA and the accumulation of the 45 nt let-7c intermediate RNA, whereas WT DICER1 produced the expected 22 nt 5p let-7c miRNA ( Figure 4B). These data suggested that the G1809R mutation adversely affects 5p miRNA production, similar to the D1709N and other mutations at metal-binding sites [4,11].…”

Section: Characterization Of the Functional Impact Of G1809r On Mirnamentioning

confidence: 69%

“…We recently identified recurrent somatic DICER1 hotspot mutations in rare non-epithelial ovarian tumours [11]. Subsequent studies have shown that hotspot mutations at the four metal-binding sites in the RNase IIIb domain (E1705, D1709, D1810 and E1813) impair DICER1's ability to generate mature 5p miRNAs [4,11,12]. More recently, DICER1 hotspot mutations were identified in tumours from PPB and PPB-FTDS cases possessing germline DICER1 mutations, suggesting a two-hit hypothesis of tumourigenesis [11,[13][14][15][16].…”

Section: J Chen Et Almentioning

confidence: 99%

“…The mutational analyses of DICER1 have previously focused on rare paediatric cancers [6][7][8][9][10][11][13][14][15][16]. In this study, we investigated the prevalence of DICER1 mutations in a spectrum of more common malignancies.…”

Section: J Chen Et Almentioning

confidence: 99%

“…All the patient tumour and blood samples were collected from the Ovarian Cancer Research Programme (OvCaRe) tissue bank (Vancouver, British Columbia, Canada) under ethical approvals from the University of British Columbia and BC Cancer Agency Research Ethics Board [11].…”

Section: Patient Samples and Cell Culturementioning

confidence: 99%

“…The RNA transcript was gel-purified and 5 ′ -32 P end-labelled. The preparation of DICER1 enzymes and in vitro DICER cleavage assays were performed as previously described [11]. The resulting products were visualized on a storage phosphor screen, using a FLA-7000 instrument (GE Healthcare).…”

Section: Western Blottingmentioning

confidence: 99%

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Recurrent DICER1 hotspot mutations in endometrial tumours and their impact on microRNA biogenesis

Chen

Wang

McMonechy

et al. 2015

The Journal of Pathology

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DICER1 plays a critical role in microRNA (miRNA) biogenesis. Recurrent somatic 'hotspot' mutations at the four metal-binding sites within the RNase IIIb domain of DICER1 were identified in ovarian sex cord-stromal tumours and have since been described in other paediatric tumours. In this study, we screened the RNase IIIb domain of DICER1 in 290 endometrial tumours and identified six cases with hotspot mutations, including two cases affected by an atypical G1809R mutation directly adjacent to a metal-binding site. Using Illumina and Sanger targeted resequencing, we observed and validated biallelic DICER1 mutations in several cases with hotspot mutations. Through in vitro DICER1 cleavage assays, small RNA deep sequencing and real-time PCR, we demonstrated that mutations adding a positively charged side chain to residue 1809 have similar detrimental effects on 5p miRNA production to mutations at the metal-binding sites. As expected, 5p miRNAs were globally reduced in tumours and cell lines with hotspot mutations. Pathway analysis of gene expression profiles indicated that genes de-repressed due to loss of 5p miRNAs are strongly associated with pathways regulating the cell cycle. Using a Dicer1-null mouse cell line model, we found that expression of DICER1 hotspot mutants promoted cell proliferation, whereas wild-type (WT) DICER1 inhibited cell proliferation. Furthermore, targets of let-7 family miRNAs are enriched among the up-regulated genes, suggesting that loss of let-7 may be impacting downstream pathways. Our results reveal that DICER1 hotspot mutations are implicated in common malignancies and may constitute a unique oncogenic pathway.

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Section: Characterization Of the Functional Impact Of G1809r On Mirnamentioning

confidence: 69%

Section: J Chen Et Almentioning

confidence: 99%

Section: J Chen Et Almentioning

confidence: 99%

Section: Patient Samples and Cell Culturementioning

confidence: 99%

Section: Western Blottingmentioning

confidence: 99%

See 3 more Smart Citations

Recurrent DICER1 hotspot mutations in endometrial tumours and their impact on microRNA biogenesis

Chen

Wang

McMonechy

et al. 2015

The Journal of Pathology

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A Genome-First Approach to Characterize DICER1 Pathogenic Variant Prevalence, Penetrance, and Phenotype

et al. 2021

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Key Points Question What are the prevalence, risk, and phenotypic spectrum of individuals with a germline putative loss-of-function (pLOF) variant in DICER1 according to a genome-first approach in a population-scale cohort? Findings In this cohort study, DICER1 pLOF variants were more than twice as common (even after adjustment for relatedness) than previously observed. Malignant tumors were observed in 16% of participants with a DICER1 pLOF variant, which is comparable to the frequency of neoplasms in the largest phenotype-first DICER1 studies published to date. Meaning The genome-first approach complements more traditional approaches to syndrome delineation and may be an efficient approach for risk estimation in monogenic disorders.

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Replenishing co‐downregulated miR‐100‐5p and miR‐125b‐5p in malignant germ cell tumors causes growth inhibition through cell cycle disruption

Ferraresso,

Bailey,

Alonso‐Crisostomo

et al. 2024

Molecular Oncology

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MicroRNAs (miRNAs) are short, nonprotein‐coding RNAs, and their expression is dysregulated in malignant germ cell tumors (GCTs). Here, we investigated the causes and consequences of downregulated miR‐99a‐5p/miR‐100‐5p (functionally identical) and miR‐125b‐5p levels in malignant GCTs regardless of age, site, or subtype. Quantitative RT‐PCR was used to assess miR‐99a‐5p/miR‐100‐5p, miR‐125b‐5p, and associated gene expression in malignant GCT tissues/cell lines [seminoma (Sem), yolk sac tumor (YST), embryonal carcinoma (EC)]. Cells were treated with demethylating 5‐azacytidine and pyrosequencing was performed. Combination miR‐100‐5p/miR‐125b‐5p mimic replenishment was used to treat malignant GCT cells. Global messenger RNA (mRNA) targets of the replenished miRNAs were identified and Metascape used to study pathway effects. We found that expression levels of miR‐99a‐5p/miR‐100‐5p and miR‐125b‐5p, their respective pri‐miRNAs, and associated genes from chromosomes 11 and 21 (chr11/chr21) were downregulated and highly correlated in malignant GCT cells. Treatment with 5‐azacytidine caused upregulation of these miRNAs, with pyrosequencing revealing hypermethylation of their chr11/chr21 loci, likely contributing to miR‐100‐5p/miR‐125b‐5p downregulation. Combination miR‐100‐5p/miR‐125b‐5p mimic replenishment resulted in growth inhibition in Sem/YST cells, with miR‐100‐5p/miR‐125b‐5p mRNA targets enriched in downregulated genes, which were involved in cell cycle (confirmed by flow cytometry) and signaling pathways. Knockdown of the miR‐100‐5p/miR‐125b‐5p target tripartite motif containing 71 (TRIM71kd) recapitulated miR‐100‐5p/miR‐125b‐5p replenishment, with growth inhibition and cell cycle disruption of Sem/YST/EC cells. Further, replenishment led to reduced lin‐28 homolog A (LIN28A) levels and concomitant increases in let‐7 (MIRLET7B) tumor suppressor miRNAs, creating a sustained reversion of cell phenotype. In summary, combination miR‐100‐5p/miR‐125b‐5p mimic replenishment or TRIM71kd caused growth inhibition in malignant GCT cells via cell cycle disruption. Further studies are now warranted, including mimic treatment alongside conventional platinum‐based chemotherapy.

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Recurrent SomaticDICER1Mutations in Nonepithelial Ovarian Cancers

Cited by 426 publications

References 38 publications

Recurrent DICER1 hotspot mutations in endometrial tumours and their impact on microRNA biogenesis

Recurrent DICER1 hotspot mutations in endometrial tumours and their impact on microRNA biogenesis

A Genome-First Approach to Characterize DICER1 Pathogenic Variant Prevalence, Penetrance, and Phenotype

Replenishing co‐downregulated miR‐100‐5p and miR‐125b‐5p in malignant germ cell tumors causes growth inhibition through cell cycle disruption

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