Gene essentiality landscape and druggable oncogenic dependencies in herpesviral primary effusion lymphoma

Manzano, Mark; Patil, Ajinkya; Waldrop, Alex; Davé, Sandeep S.; Behdad, Amir; Gottwein, Eva

doi:10.1038/s41467-018-05506-9

Cited by 51 publications

(122 citation statements)

References 70 publications

(109 reference statements)

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“…Recently, CMs have been shown to induce efficient cell death in PEL cell lines 6,7 and LEN is currently part of a clinical trial in PEL (ClinicalTrials.gov Identifier: NCT02911142). As in multiple myeloma 28 , the transcription factor IRF4 is a highly essential survival gene in PEL cells and its expression is reduced upon treatment with CMs 6,7,29 . Surprisingly, however, in this setting, downregulation of IRF4 and CM toxicity were independent of IKZF1 and/or IKZF3 7 .…”

Section: Primary Effusion Lymphoma (Pel) Is a Non-hodgkin B Cell Lympmentioning

confidence: 99%

“…All parental cell lines were maintained as described recently 29 . Lenalidomide was from Cayman Chemicals (Ann Arbor, MI) and pomalidomide and CC-122 were from Selleck Chemicals (Houston, TX).…”

Section: Cell Lines and Reagentsmentioning

confidence: 99%

“…Lentiviruses were prepared as described previously and functionally titrated 29 . Generation and validation of clonal Cas9-expressing BC-3 and BCBL-1 cells were described 29 . Clonal BC-3 CRBN KO cells were described previously 7 .…”

Section: Cell Lines and Reagentsmentioning

confidence: 99%

“…Clonal Cas9 expressing BC-3 cells were transduced with the Brunello library at ~500X coverage and an MOI of ~0.3 and selected with puromycin as described 29 . Cells were passaged for two weeks to allow drop-out of sgRNAs targeting essential genes from the cell pool 29 . Then, cells were treated with 5µM LEN, 1µM POM, 1µM CC-122 or DMSO, in two replicates per condition.…”

Section: Genome-wide Crispr Screensmentioning

confidence: 99%

“…A second sample for all conditions was taken on day 21. Preparation of genomic DNA and next generation sequencing libraries was performed as described 29 . Trimmed reads were aligned to the Brunello sgRNA library 32 using Bowtie and raw read counts were generated for each sgRNA as previously described 29 (Table S1).…”

Section: Genome-wide Crispr Screensmentioning

confidence: 99%

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Genome-wide CRISPR Screens Reveal Genetic Mediators of Cereblon Modulator Toxicity in Primary Effusion Lymphoma

Patil¹,

Manzano²,

Gottwein³

2019

Preprint

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Genome-wide CRISPR/Cas9 screens represent a powerful approach to study mechanisms of drug action and resistance. Cereblon modulating agents (CMs) have recently emerged as candidates for therapeutic intervention in primary effusion lymphoma (PEL), a highly aggressive cancer caused by Kaposi's sarcoma-associated herpesvirus. CMs bind to cereblon (CRBN), the substrate receptor of the cullin-RING type E3 ubiquitin ligase CRL4 CRBN , and thereby trigger the acquisition and proteasomal degradation of neosubstrates. Downstream mechanisms of CM toxicity are incompletely understood, however. To identify novel CM effectors and mechanisms of CM resistance, we performed positive selection CRISPR screens using three CMs with increasing toxicity in PEL: lenalidomide (LEN), pomalidomide (POM), and CC-122.Results identified several novel modulators of the activity of CRL4 CRBN . The number of genes whose inactivation confers resistance decreases with increasing CM efficacy.Only inactivation of CRBN conferred complete resistance to CC-122. Inactivation of the E2 ubiquitin conjugating enzyme UBE2G1 also conferred robust resistance against LEN and POM. Inactivation of additional genes, including the Nedd8-specific protease SENP8, conferred resistance to only LEN. SENP8 inactivation indirectly increased levels of unneddylated CUL4A/B, which limits CRL4 CRBN activity in a dominant negative manner. Accordingly, sensitivity of SENP8-inactivated cells to LEN is restored by overexpression of CRBN. In sum, our screens identify several novel players in CRL4 CRBN function and define pathways to CM resistance in PEL. These results provide rationale for increasing CM efficacy upon patient relapse from a less efficient CM.Identified genes could finally be developed as biomarkers to predict CM efficacy in PEL and other cancers. Key Points1. Genome-wide CRISPR/Cas9 screens identify novel mediators of resistance to lenalidomide, pomalidomide and CC-122 in PEL cells.2. UBE2G1 and SENP8 are modulators of CRL4 CRBN and their inactivation drives resistance to CMs in PEL-derived cell lines.

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Section: Primary Effusion Lymphoma (Pel) Is a Non-hodgkin B Cell Lympmentioning

confidence: 99%

Section: Cell Lines and Reagentsmentioning

confidence: 99%

Section: Cell Lines and Reagentsmentioning

confidence: 99%

Section: Genome-wide Crispr Screensmentioning

confidence: 99%

Section: Genome-wide Crispr Screensmentioning

confidence: 99%

See 3 more Smart Citations

Genome-wide CRISPR Screens Reveal Genetic Mediators of Cereblon Modulator Toxicity in Primary Effusion Lymphoma

Patil¹,

Manzano²,

Gottwein³

2019

Preprint

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Antitumor effect of acanthoic acid against primary effusion lymphoma via inhibition of c‐FLIP

Sittithumcharee

Kariya

Kasemsuk

et al. 2021

Phytotherapy Research

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Acanthoic acid (AA) is an active substance that is extracted from Croton oblongifolius Roxb., a traditional plant in Thailand. The antiinflammatory effect of AA on NF-κB pathway has been exclusively reported, however, its anticancer effect is still lacking. PEL is a B cell lymphoma that is mostly found in HIV patients. The prognosis and progression of PEL patients are terribly poor with a median survival time less than 6 months, so the new effective treatment is urgently needed. In this study, we found that AA effectively inhibited PEL cell proliferation with IC50s at 120-130 μM in well-representative cells, while the IC50s of AA in PBMC were higher (>200 μM). AA increased percentages of Annexin V/PI positive cells, whereas adding of caspase inhibitor (Q-VD-OPh) prevented AA-induced cell death. The antiapoptotic protein, c-FLIP, was downregulated by AA which leading to the activation of caspase-8 and -3. Combination of AA and TRAIL dramatically enhanced apoptotic cell death. In PEL xenograft model, AA at the dose of 250 mg/kg effectively

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Update: The molecular spectrum of virus-associated high-grade B-cell non-Hodgkin lymphomas

Witte,

Künstner,

Gebauer

2024

Blood Reviews

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Gene essentiality landscape and druggable oncogenic dependencies in herpesviral primary effusion lymphoma

Cited by 51 publications

References 70 publications

Genome-wide CRISPR Screens Reveal Genetic Mediators of Cereblon Modulator Toxicity in Primary Effusion Lymphoma

Genome-wide CRISPR Screens Reveal Genetic Mediators of Cereblon Modulator Toxicity in Primary Effusion Lymphoma

Antitumor effect of acanthoic acid against primary effusion lymphoma via inhibition of c‐FLIP

Update: The molecular spectrum of virus-associated high-grade B-cell non-Hodgkin lymphomas

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