An RNAi screen in human cell lines reveals conserved DNA damage repair pathways that mitigate formaldehyde sensitivity

Juarez, Eleonora; Chambwe, Nyasha; Tang, Weiliang; Mitchell, Asia; Owen, Nichole; Kumari, Anuradha; Monnat, Raymond J.; McCullough, Amanda K.

doi:10.1101/310730

Cited by 3 publications

(4 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The loss of FA pathway activity did not consistently sensitize FA patient-derived or sporadic, FANCA -mutant HNSCC lines to formaldehyde dose-dependent cell killing over a 4 day/continuous exposure time course (Figure S4). This may reflect cell lineage-specific or other differences that collectively determine formaldehyde toxicity (38,39). In contrast, loss of FA pathway function sensitized cells regardless of origin to platinum-based chemotherapeutic drugs with cis -Pt displaying the strongest - and oxaliplatin the least – dose-dependent reductions in survival.…”

Section: Resultsmentioning

confidence: 99%

Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource

Nguyen

Tang

Webster

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Fanconi anemia (FA) is a heritable malformation, bone marrow failure and cancer predisposition syndrome that confers an exceptionally high risk of developing carcinomas arising in squamous mucosal epithelia lining the mouth, proximal esophagus, vulva and anus. The origin of these cancers is not understood, and no effective way has been identified to prevent or delay their appearance. FA-associated carcinomas are also therapeutically challenging, as they may be multi-focal and stage-advanced at diagnosis making surgical control challenging. Moreover, individuals with FA have systemic DNA damage hypersensitivity and thus an elevated risk of toxicity when treated with standard-of-care therapies such as DNA cross-linking drugs and ionizing radiation. We developed the Fanconi Anemia Cancer Cell Line Resource (FA-CCLR) in order to foster new research on the origins, treatment, and prevention of FA-associated cancers. The FA-CCLR consists of FANC-isogenic head and neck squamous cell carcinoma (HNSCC) cell line pairs from cancers arising in individuals with FA, or newly engineered from sporadic HNSCC cell lines. Molecular, cellular, and biochemical analyses were used to demonstrate the causal dependence of key FA-associated phenotypes on FANC genotype, expression and pathway activity. These FANC-isogenic cell line pairs are available to academic and non-profit investigators, with ordering information available at the Fanconi Anemia Research Materials Resource and Repository at Oregon Health & Sciences University, Portland OR.

show abstract

Section: Resultsmentioning

confidence: 99%

Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource

Nguyen

Tang

Webster

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…To further characterize STING effect on replication, we asked if we can reproduce it in a BRCA1-proficient cell line. A derivative of the SV40-transformed fibroblast cell line GM639 that we have previously used in our studies and that carries a wild type BRCA1 and a benign SNP variant of BRCA2, Q2384K (Juarez et al, 2018), displays some NDD upon HU arrest. We depleted STING by siRNA in the Frontiers in Molecular Biosciences frontiersin.org GM639 background (Figure 2E).…”

Section: Sting Is Found In the Nuclear Envelope And Chromatin Fractionsmentioning

confidence: 99%

“…The SV40-transformed human fibroblast GM639 (GM00639, Cellosaurus ID CVCL_7299) has been used by us and others previously (Swanson et al, 2004;Sidorova et al, 2008) and was obtained from the NIGMS Human Genetic Mutant Cell Repository. Exome sequencing of this cell line was reported in (Juarez et al, 2018). The U2OS line was acquired from ATCC (ATCC HTB-96).…”

Section: Cells and Culturementioning

confidence: 99%

Innate immunity mediator STING modulates nascent DNA metabolism at stalled forks in human cells

Lazarchuk

Nguyen

Brunon

et al. 2023

Front. Mol. Biosci.

View full text Add to dashboard Cite

Background: The cGAS/STING pathway, part of the innate immune response to foreign DNA, can be activated by cell’s own DNA arising from the processing of the genome, including the degradation of nascent DNA at arrested replication forks, which can be upregulated in cancer cells. Recent evidence raises a possibility that the cGAS/STING pathway may also modulate the very processes that trigger it, e.g., DNA damage repair or processing of stalled forks.Methods: We manipulated STING levels in human cells by depleting or re-expressing it, and assessed the effects of STING on replication using microfluidics-assisted replication track analysis, or maRTA, a DNA fiber assay, as well as immuno-precipitation of nascent DNA, or iPOND. We also assessed STING subcellular distribution and its ability to activate.Results: Depletion of STING suppressed and its re-expression in STING-deficient cancer cells upregulated the degradation of nascent DNA at arrested replication forks. Replication fork arrest was accompanied by the STING pathway activation, and a STING mutant that does not activate the pathway failed to upregulate nascent DNA degradation. cGAS was required for STING’s effect on degradation, but this requirement could be bypassed by treating cells with a STING agonist. Cells expressing inactive STING had a reduced level of RPA on parental and nascent DNA of arrested forks and a reduced CHK1 activation compared to cells with the wild type STING. STING also affected unperturbed fork progression in a subset of cell lines. STING fractionated to the nuclear fractions enriched for structural components of chromatin and nuclear envelope, and furthermore, it associated with the chromatin of arrested replication forks as well as post-replicative chromatin.Conclusion: Our data highlight STING as a determinant of stalled replication fork integrity, thus revealing a novel connection between the replication stress and innate immune responses.

show abstract

“…Next to chemical screens, genetic screens are a powerful tool to identify novel gene functions. At first restricted to the use of small interference RNA (siRNA), short hairpin RNA (shRNA) and insertional mutagenesis, recent years have seen a rapid development of the toolbox available for such screens, especially with the advent of CRISPR-Cas9 [ 207 , 208 , 209 ]. CRISPR screens can be performed both in a pooled manner, where NGS is used to identify enrichment or depletion of sgRNAs, or in an arrayed manner.…”

Section: Outlook: Approaches To Identify Interactions Between Metamentioning

confidence: 99%

Interplay between Cellular Metabolism and the DNA Damage Response in Cancer

Moretton

Loizou

2020

Cancers

View full text Add to dashboard Cite

Metabolism is a fundamental cellular process that can become harmful for cells by leading to DNA damage, for instance by an increase in oxidative stress or through the generation of toxic byproducts. To deal with such insults, cells have evolved sophisticated DNA damage response (DDR) pathways that allow for the maintenance of genome integrity. Recent years have seen remarkable progress in our understanding of the diverse DDR mechanisms, and, through such work, it has emerged that cellular metabolic regulation not only generates DNA damage but also impacts on DNA repair. Cancer cells show an alteration of the DDR coupled with modifications in cellular metabolism, further emphasizing links between these two fundamental processes. Taken together, these compelling findings indicate that metabolic enzymes and metabolites represent a key group of factors within the DDR. Here, we will compile the current knowledge on the dynamic interplay between metabolic factors and the DDR, with a specific focus on cancer. We will also discuss how recently developed high-throughput technologies allow for the identification of novel crosstalk between the DDR and metabolism, which is of crucial importance to better design efficient cancer treatments.

show abstract

An RNAi screen in human cell lines reveals conserved DNA damage repair pathways that mitigate formaldehyde sensitivity

Cited by 3 publications

References 62 publications

Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource

Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource

Innate immunity mediator STING modulates nascent DNA metabolism at stalled forks in human cells

Interplay between Cellular Metabolism and the DNA Damage Response in Cancer

Contact Info

Product

Resources

About