EXO1 protects BRCA1-deficient cells against toxic DNA lesions

Molecular Cell, Год журнала: 2024, Номер 84(4), С. 659 - 674.e7

Опубликована: Янв. 23, 2024

Inactivating mutations in the BRCA1 and BRCA2 genes impair DNA double-strand break (DSB) repair by homologous recombination (HR), leading to chromosomal instability cancer. Importantly, BRCA1/2 deficiency also causes therapeutically targetable vulnerabilities. Here, we identify dependency on end resection factor EXO1 as a key vulnerability of BRCA1-deficient cells. generates poly(ADP-ribose)-decorated lesions during S phase that associate with unresolved DSBs genomic but not wild-type or BRCA2-deficient Our data indicate BRCA1/EXO1 double-deficient cells accumulate due impaired single-strand annealing (SSA) top their HR defect. In contrast, retain SSA activity absence hence tolerate loss. Consistent EXO1-mediated SSA, find BRCA1-mutated tumors show elevated expression increased SSA-associated scars compared BRCA1-proficient tumors. Overall, our findings uncover promising therapeutic target for

Язык: Английский

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DNA Damage Response Mechanisms in Head and Neck Cancer: Significant Implications for Therapy and Survival

International Journal of Molecular Sciences, Год журнала: 2023, Номер 24(3), С. 2760 - 2760

Опубликована: Фев. 1, 2023

Head and neck cancer (HNC) is a term collectively used to describe heterogeneous group of tumors that arise in the oral cavity, larynx, nasopharynx, oropharynx, hypopharynx, represents sixth most common type malignancy worldwide. Despite advances multimodality treatment, disease has recurrence rate around 50%, prognosis metastatic patients remains poor. HNCs are characterized by high degree genomic instability, which involves vicious circle accumulating DNA damage, defective damage repair (DDR), replication stress. Nonetheless, induced on tumor cells chemo radiotherapy relies DDR processes for successful response may play an important role development novel more effective therapies. This review summarizes current knowledge genes proteins appear be deregulated pathways, their implication HNC pathogenesis, rationale behind targeting these pathways new We give particular emphasis therapeutic targets have shown promising results at pre-clinical stage those so far been associated with advantage clinical setting.

Язык: Английский

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DNA Repair Deficiency Regulates Immunity Response in Cancers: Molecular Mechanism and Approaches for Combining Immunotherapy

Cancers, Год журнала: 2023, Номер 15(5), С. 1619 - 1619

Опубликована: Март 6, 2023

Defects in DNA repair pathways can lead to genomic instability multiple tumor types, which contributes immunogenicity. Inhibition of damage response (DDR) has been reported increase susceptibility anticancer immunotherapy. However, the interplay between DDR and immune signaling remains unclear. In this review, we will discuss how a deficiency affects anti-tumor immunity, highlighting cGAS-STING axis as an important link. We also review clinical trials that combine inhibition immune-oncology treatments. A better understanding these help exploit cancer immunotherapy improve treatment outcomes for various cancers.

Язык: Английский

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Unveiling a pathogenic FANCA gene variant in a Mexican family with Fanconi anemia through next‑generation sequencing

Experimental and Therapeutic Medicine, Год журнала: 2025, Номер 29(3)

Опубликована: Янв. 9, 2025

Fanconi anemia (FA) is the most common hereditary bone marrow failure syndrome, with an incidence of 1 in 5,000,000. This disease caused by alteration one 23 genes associated FA/BRCA DNA repair pathway, which responsible for repairing interstrand bridges generated during homologous recombination. FA has been a predisposition to other types neoplasm. The current study aimed present pathogenic variant FANCA observed three Mexican siblings, as detected next‑generation sequencing (NGS). results induced chromosomal breakage test showed breaks and radial figures, were compatible FA, normal karyotype. NGS TruSight Hereditary Cancer Panel analysis resulted FANCA:c.3931_3932delAG being classified according bioinformatics analysis. reports that was found family siblings exhibited suggestive mucosa‑assisted lymphoid tissue lymphoma, atypical presentation neoplasia FA.

Язык: Английский

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Cell and Gene Therapy for Anemia: Hematopoietic Stem Cells and Gene Editing

International Journal of Molecular Sciences, Год журнала: 2021, Номер 22(12), С. 6275 - 6275

Опубликована: Июнь 10, 2021

Hereditary anemia has various manifestations, such as sickle cell disease (SCD), Fanconi anemia, glucose-6-phosphate dehydrogenase deficiency (G6PDD), and thalassemia. The available management strategies for these disorders are still unsatisfactory do not eliminate the main causes. As genetic aberrations causes of all forms hereditary optimal approach involves repairing defective gene, possibly through transplantation normal hematopoietic stem cells (HSCs) from a matching donor or gene therapy approaches (either in vivo ex vivo) to correct patient's HSCs. To clearly illustrate importance this paper provides review aberration, epidemiology, clinical features, current management, endeavors related SCD, thalassemia, G6PDD. Moreover, we expound future research direction HSC derivation induced pluripotent (iPSCs), edit HSCs, risk mitigation, their perspectives. In conclusion, gene-corrected promising outcomes it may overcome limitation source allogenic bone marrow transplantation.

Язык: Английский

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AmNA‐Modified Antisense Oligonucleotide Targeting MCM8 as a Cancer‐Specific Chemosensitizer for Platinum Compounds

Cancer Science, Год журнала: 2025, Номер unknown

Опубликована: Март 17, 2025

ABSTRACT MCM8 and MCM9 participate in homologous recombination with long‐tract gene conversion to repair double‐strand breaks caused by replication stress, which is generally higher cancer cells than normal cells. highly expressed certain cells, where it necessary for maintaining cell growth, migration, invasion, although the molecular mechanisms remain unclear. Knockdown siRNAs or knockout of selectively sensitizes cisplatin. Thus, drugs inhibiting could serve as novel anti‐neoplastic agents and/or chemosensitizers that sensitize platinum compounds. The present study describes development an amido‐bridged nucleic acid (AmNA)‐modified gapmer antisense oligonucleotide (ASO) targeting , called ASO 8–3419. In vitro, 8–3419 inhibited expression several human lines sensitized Moreover, modestly suppressed growth whose proliferation has been reported depend on MCM8. vivo, xenografted tumors colon cancer‐derived HCT116 nude mice increased tumor sensitivity cisplatin minimal toxicity. These findings suggest AmNA‐modified, MCM8‐specific ASOs hold promise anti‐cancer agents.

Язык: Английский

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BRCA2-DSS1 interaction is dispensable for RAD51 recruitment at replication-induced and meiotic DNA double strand breaks

Nature Communications, Год журнала: 2022, Номер 13(1)

Опубликована: Апрель 1, 2022

The interaction between tumor suppressor BRCA2 and DSS1 is essential for RAD51 recruitment repair of DNA double stand breaks (DSBs) by homologous recombination (HR). We have generated mice with a leucine to proline substitution at position 2431 BRCA2, which disrupts this interaction. Although significant number mutant die during embryogenesis, some homozygous hemizygous undergo normal postnatal development. Despite lack radiation induced foci formation severe HR defect in somatic cells, are fertile exhibit meiosis. hypothesize that the presence chromosomes close proximity early prophase I may compensate BRCA2-DSS1 show restoration cells when Topoisomerase inhibitor-induced single strand converted into DSBs replication. also partially rescue tethering donor site using streptavidin-fused Cas9. Our findings demonstrate complex dispensable loading DSB.

Язык: Английский

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SCAI promotes error‐free repair of DNA interstrand crosslinks via the Fanconi anemia pathway

EMBO Reports, Год журнала: 2022, Номер 23(4)

Опубликована: Фев. 14, 2022

Article14 February 2022Open Access Transparent process SCAI promotes error-free repair of DNA interstrand crosslinks via the Fanconi anemia pathway Lisa Schubert Protein Signaling Program, Novo Nordisk Foundation Center for Research, University Copenhagen, Denmark Contribution: Conceptualization, Data curation, Funding acquisition, ​Investigation, Methodology, Project administration Search more papers by this author Ivo A Hendriks orcid.org/0000-0002-1439-3701 Proteomics Methodology Emil P T Hertz Formal analysis, Wei Wu Chromosome Stability, Department Cellular and Molecular Medicine, ​Investigation Selene Sellés-Baiget Saskia Hoffmann Keerthana Stine Viswalingam orcid.org/0000-0002-2179-1804 Biology, Irene Gallina orcid.org/0000-0002-3741-9038 Satyakrishna Pentakota Bente Benedict orcid.org/0000-0002-7503-8527 Joachim Johansen Disease Systems Katja Apelt Human Genetics, Leiden Medical Center, Leiden, The Netherlands Martijn S Luijsterburg Supervision Simon Rasmussen orcid.org/0000-0001-6323-9041 Michael Lisby orcid.org/0000-0002-4830-5247 Supervision, Writing - original draft, administration, review & editing Ying Liu L Nielsen Niels Mailand Corresponding Author [email protected] orcid.org/0000-0002-6623-709X Julien Duxin orcid.org/0000-0001-9389-4186 Information Schubert1, Hendriks2,†, Hertz1,†, Wu3,†, Sellés-Baiget1, Hoffmann1, Viswalingam4, Gallina1, Pentakota1, Benedict1, Johansen5, Apelt6, Luijsterburg6, Rasmussen5, Lisby3,4, Liu3, Nielsen2, *,1,3 *,1 1Protein 2Proteomics 3Center 4Department 5Disease 6Department † These authors contributed equally to work *Corresponding author. Tel: +45 35325023; E-mail: ***Corresponding 93565571; EMBO Reports (2022)23:e53639https://doi.org/10.15252/embr.202153639 PDFDownload PDF article text main figures. Peer ReviewDownload a summary editorial decision including letters, reviewer comments responses feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures Info Abstract (ICLs) are cytotoxic lesions that threaten genome integrity. (FA) orchestrates ICL during replication, with ubiquitylated FANCI-FANCD2 (ID2) marking activation step triggers incisions on unhook ICL. Restoration intact requires coordinated actions polymerase ζ (Polζ)-mediated translesion synthesis (TLS) homologous recombination (HR). While proteins mediating FA have been well characterized, effectors regulating choice promote resolution remain poorly defined. Here, we uncover an indispensable role in ensuring upon pathway. We show forms complex Polζ localizes ICLs replication. SCAI-deficient cells exquisitely sensitive ICL-inducing drugs display major hallmarks gene inactivation. In absence SCAI, HR-mediated is defective, breaks instead re-ligated θ-dependent microhomology-mediated end-joining, generating deletions spanning site radial chromosomes. Our establishes as integral component, acting at interface between TLS HR repair. Synopsis This critical regulator crosslink (ICL) Loss manifests deficiency ensures faithful replication-coupled interacts direct binding REV3 prevents end joining Introduction pathways counteract wide spectrum protect from detrimental mutations, breaks, chromosome rearrangements. some directly removed dedicated enzymes (e.g., oxidized bases readily restored base excision repair), other deleterious such require precise coordination multiple enzymatic activities re-establish DNA. highly lesions, whose tightly coupled replication (Akkari et al, 2000; Räschle 2008). large number participate repair, defects least 22 which causative (FA), rare, inherited disease characterized developmental birth defects, bone marrow failure, cancer predisposition (Rageul Kim, 2020). factors can be functionally subdivided according where they operate multistep (Wang, 2007; Ceccaldi 2016; Rageul First, core (comprising FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, FANCL, FANCM, FAAP20, FAAP100) associates damaged chromatin monoubiquitylates heterodimer UBE2T (FANCT) (Garcia-Higuera 2001; Smogorzewska Hira 2015; Rickman 2015). Monoubiquitylated ID2 marks downstream effector (Knipscheer 2009; Douwel 2014). include nucleases nick one strand both sides it (SLX4 (FANCP)-XPF (FANCQ)-ERCC1 complex), bypasses adduct (REV1-Polζ), (HR) two-ended double-strand break (DSB) final DSB distinct processes, prior restoration template TLS. Thus, must ensure but these processes largely unknown. demonstrate protein (suppressor cell invasion) has pathway, operating steps prevent erroneous intermediates Polθ-dependent (MMEJ), thereby resolution. Results principal To identify novel involved carried out genome-scale CRISPR-Cas9 dropout screen genes required survival presence low dose drug mitomycin C (MMC), corresponding 20% lethality (LD20). end, human RPE-1 targeted knockout (KO) p53 were transfected lentiviral sgRNA library propagated or MMC 12 days (Fig 1A). Validating our screening approach, many FA-associated scored among top hits (Figs 1B EV1A; Dataset EV1). fact, KO 20 known manifested hypersensitivity 1B). Unexpectedly, also revealed sgRNAs targeting selectively depleted exposed 1B; was originally identified transcriptional suppressor migration (Brandt 2009) later found play interaction 53BP1 (Hansen Isobe 2017). However, unlike did not significant MMC-treated (Dataset thus suggested previously unrecognized function set further explore. accordance screen, cisplatin, another inducer, could fully rescued stable re-expression 1C–E EV1B C). As observed established genes, displayed strong accumulation G2 phase accompanied increased level damage foci following treatment 1F–H EV1D–H). Likewise, metaphase spreads exhibited marked increase breaks/gaps formation treatment, characteristic feature patient 1I–K; García-de-Teresa Again, all phenotypes complemented ectopic 1F–K). conclude loss hypersensitizes agents phenocopies defective resulting Figure 1. dysfunction Schematic outline sensitizes MMC. LD20, lethal dose; NGS, next-generation sequencing. DrugZ analysis depletion CRISPR (A) low-dose (n = 2 technical replicates). highlighted blue; red. Immunoblot U2OS WT, U2OS/SCAI KO, stably reconstituted Strep-HA-SCAI (U2OS/SCAI KO/Strep-HA-SCAI). Clonogenic KO/Strep-HA-SCAI subjected indicated doses 24 h (mean ± SEM; n 3 independent experiments). (D), except treated Cisplatin (9 nM) 48 h, fixed, co-stained PCNA antibody DAPI. Cell cycle distribution analyzed quantitative image-based cytometry (QIBC) (≥ 2,000 per condition). representative experiment shown. (90 1 fixed later, RPA2 quantified QIBC 3,000 condition; mean SD; experiments; *P < 0.05; ns, significant, two-tailed paired t-test). (G), γH2AX DAPI **P 0.01; Experimental workflow morphology (top) images lines (bottom). stained Scale bars, 10 µm. Quantification chromosomes (I) 180 0.01, chromosomal 99 each condition pooled three ****P 0.0001, Mann–Whitney U test). Download figure PowerPoint Click here expand figure. EV1. (related Figs 2) GO term (NormZ −3) Fig 1A, using Reactome PANTHER16.0. WT (B), cisplatin Scatter plot showing 1F. Light grey, G1 phase; dark red: G2/M phase. Proportion indicated. (E), RAD51 Representative experiments 1G H. bar, (E) (F). FANCD2 siRNA knockdown efficiency cells. non-targeting control (CTRL) siRNAs harvested times after exposure (0.5 µM). Preventing alleviates better define emerging complementary suppresses aim, transduced near-lethal (LD80) 2A). Strikingly, ontology (GO) components most enriched class inactivation conferred significantly resistance 2B comprised associated (FANCA, FAAP100), complex, its deubiquitylation (USP1 WDR48), ordinarily roles activating against toxicity C; EV2; By contrast, functioning ubiquitylation score EV2). Genes encoding remodeling histone acetylation regulators proliferation far less prominently represented than 2C; Verifying results, FANCA suppressed DSBs 2D–G). Knockdown similarly alleviated EV1I J). Notably, much stronger background 2D E, exclude possibility consequence residual due incomplete siRNAs, knocked 2H). confirmed indeed cells, strongly 2I Importantly, whereas monoubiquitylation abolished expected, had no impact 2K EV1K). Collectively, data suggest essential proper once their processing initiated, se. 2. LD80, 80% MaGECK enrichment replicates; false discovery rate (FDR) 0.2 dotted line). blue. (FDR 0.2) (A), (F), lines. U2OS/FANCA U2OS, U2OS/FANCA+SCAI double (DKO) DKO elucidate how functions used Xenopus egg extracts, efficiently recapitulate plasmid containing site-specific (pICLpt) (Räschle system, two replisomes quickly converge lesion 3A, i). Upon collision, CMG first TRAIP unloaded p97 ii) (Fullbright 2019). One leading strands then extended within nucleotide (nt) (−1 position) 2008), point forks undergoes reversal produce substrate suitable iii) (Amunugama 2018). Ubiquitylated SLX4-XPF-ERCC1 iv), daughter molecules while leaving gap adducted molecule two-step involves insertion across unknown polymerase, extension past REV1-Polζ v) (Budzowska Finally, repaired utilizing vi–vii; Long 2011). 3. Current model ICLpt extracts pICLpt isolated pull down incubation extract CDC7i (100 µM) immunoblotting. replicated [α-32P]dATP times, reactions native agarose gel electrophoresis. inhibitor NMS-873 (p97i; 200 μM) supplemented reaction Note activity, CMGs longer plasmid, intermediate products (RI) 2016). OC, open circular; SC, supercoiled. Samples

Язык: Английский

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Fanconi anemia and dyskeratosis congenita/telomere biology disorders: Two inherited bone marrow failure syndromes with genomic instability

Frontiers in Oncology, Год журнала: 2022, Номер 12

Опубликована: Авг. 25, 2022

Inherited bone marrow failure syndromes (IBMFS) are a complex and heterogeneous group of genetic diseases. To date, at least 13 IBMFS have been characterized. Their pathophysiology is associated with germline pathogenic variants in genes that affect hematopoiesis. A couple these diseases also genomic instability, Fanconi anemia due to DNA damage repair deficiency dyskeratosis congenita/telomere biology disorders as result an alteration telomere maintenance. Patients can extramedullary manifestations, including cancer functional or structural physical abnormalities. Furthermore, the phenotypic spectrum varies from cryptic features patients significantly evident manifestations. These require high index suspicion should be considered any patient abnormal hematopoiesis, even if manifestations not evident. This review describes disrupted cellular processes lead affected maintenance genome structure, contrasting dysmorphological oncological phenotypes disorders. Through analysis, we describe allow make differential diagnosis early identification patients, before onset hematological From perspective, analyzed risks cancers carriers.

Язык: Английский

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Fanconi Anemia Pathway in Colorectal Cancer: A Novel Opportunity for Diagnosis, Prognosis and Therapy

Journal of Personalized Medicine, Год журнала: 2022, Номер 12(3), С. 396 - 396

Опубликована: Март 4, 2022

Colorectal cancer (CRC) is the third most commonly diagnosed malignancy and has second highest mortality rate globally. Thanks to advent of next-generation sequencing technologies, several novel candidate genes have been proposed for CRC susceptibility. Germline biallelic mutations in one or more 22 currently recognized Fanconi anemia (FA) associated with disease, while germline monoallelic mutations, somatic promoter hypermethylation some

Язык: Английский

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