Cell-free DNA promotes malignant transformation in non-tumor cells

Souza, Aline Gomes de; Bastos, Víctor Alexandre Félix; Fujimura, Patrícia Tiemi; Ferreira, Izabella C C; Leal, Letícia Ferro; Silva, Luciane Sussuchi da; Laus, Ana Carolina; Reis, Rui Manuel; Martins, Mário Machado; Santos, Paula de Souza; Corrêa, Natássia Caroline Resende; Marangoni, Karina; Thomé, Carolina Hassibe; Colli, Leandro M.; Goulart, Luíz Ricardo; Alonso-Goulart, Vivian

doi:10.1038/s41598-020-78766-5

Cited by 14 publications

(10 citation statements)

References 50 publications

(45 reference statements)

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“…A portion of cfDNA is produced by tumor cells through apoptosis, necrosis, or active secretion [ 34 , 35 ]. In addition to its role in the field of cancer diagnostics, cfDNA could influence the immune response, or promote tumorigenesis and “genometastasis” [ 36 ]. These biological effects can be triggered by signaling pathways activated by the interplay of cfDNA with certain cell receptors (including TLRs) or by increasing the transcriptional levels of several genes in an interaction similar to that observed with DNA aptamers [ 37 ].…”

Section: Discussionmentioning

confidence: 99%

Survival of HT29 Cancer Cells Is Affected by IGF1R Inhibition via Modulation of Self-DNA-Triggered TLR9 Signaling and the Autophagy Response

Sípos¹,

Barta²,

Simon³

et al. 2022

Pathol. Oncol. Res.

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Purpose: In HT29 colon cancer cells, a close interplay between self-DNA-induced TLR9 signaling and autophagy response was found, with remarkable effects on cell survival and differentiation. IGF1R activation drives the development and malignant progression of colorectal cancer. IGF1R inhibition displays a controversial effect on autophagy. The interrelated roles of IGF1R inhibition and TLR9/autophagy signaling in HT29 cancer cells have not yet been clarified. In our study, we aimed to investigate the complex interplay of IGF1R inhibition and TLR9/autophagy signaling in HT29 cells.Methods: HT29 cells were incubated with tumor-originated self-DNA with or without inhibitors of IGF1R (picropodophyllin), autophagy (chloroquine), and TLR9 (ODN2088), respectively. Cell proliferation and metabolic activity measurements, direct cell counting, NanoString and Taqman gene expression analyses, immunocytochemistry, WES Simple Western blot, and transmission electron microscopy investigations were performed.Results: The concomitant use of tumor-derived self-DNA and IGF1R inhibitors displays anti-proliferative potential, which can be reversed by parallel TLR9 signaling inhibition. The distinct effects of picropodophyllin, ODN2088, and chloroquine per se or in combination on HT29 cell proliferation and autophagy suggest that either the IGF1R-associated or non-associated autophagy machinery is “Janus-faced” regarding its actions on cell proliferation. Autophagy, induced by different combinations of self-DNA and inhibitors is not sufficient to rescue HT29 cells from death but results in the survival of some CD133-positive stem-like HT29 cells.Conclusion: The creation of new types of combined IGF1R, autophagy, and/or TLR9 signaling inhibitors would play a significant role in the development of more personalized anti-tumor therapies for colorectal cancer.

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Section: Discussionmentioning

confidence: 99%

Survival of HT29 Cancer Cells Is Affected by IGF1R Inhibition via Modulation of Self-DNA-Triggered TLR9 Signaling and the Autophagy Response

Sípos¹,

Barta²,

Simon³

et al. 2022

Pathol. Oncol. Res.

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“…If this were to occur, this could possibly contribute to tumor progression through effects on cellular microenvironment such as initiation of extracellular DNA trap formation to promote inflammation [35]. Extracellular DNA has also been reported to promote tumor survival after chemotherapy [36] and even transformation of non-tumor cells [37]. Thus, cancers bearing an extra-cellular DNA shedding phenotype may be more aggressive than others, suggesting that ATR-I774Yfs*5 may be a biomarker of disease progression.…”

Section: Discussionmentioning

confidence: 99%

ATR-I774Yfs*5 promotes genomic instability through micronuclei formation

D’Orazio

Holcomb

Hong

et al. 2022

Preprint

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Although mismatch repair (MMR) defects are associated with high risk of malignancy, the specific oncogenic drivers pertinent to MMR-affected cancers are poorly characterized. The heterozygous ATR-I774Yfs*5 mutation, the result of strand slippage in a poly-A tract of the Ataxia Telangiectasia and Rad3 related (ATR) gene, is overexpressed in MMR-defective malignancies including colorectal carcinoma (CRC) and is the most common ATR mutation in cancer. Here, we explore the contribution of ATR-I774Yfs*5 to genomic integrity. Using heterozygous ATR-I774Yfs*5 HCT-116 cells to mimic the native mutation, we found this mutation reduced ATR activity as measured by damage-induced Chk1 phosphorylation at S317 and ATR autophosphorylation ATR at T1989. ATR-I774Yfs*5 expression impaired genomic stability as visualized by the appearance of micronuclei in two stable expression models as well as in cell lines transfected with ATR-I774Yfs*5. Micronucleus development was dependent on replication and independent of ATR copy number. ATR-I774Yfs*5 expression did not alter cellular viability, cell cycle progression, or replicative rate, suggesting this mutation is well-tolerated despite its destabilizing effect on the genome. Taken together, these data suggest that the ATR-I774Yfs*5, whose development is favored in the context of MMR deficiency, may represent an important driver of a mutator phenotype by promoting genomic instability.

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“…In line with this, there is evidence that cfDNA released by dead or dying tumor cells can activate TLR9 signaling, which in turn inhibits apoptosis and enhances autophagy, thereby promoting tumor growth [ 371 ]. Furthermore, it is possible that cfDNA promotes metastasis in recipient cells by inducing the overexpression of several pro-metastatic genes, which for example enhance cell invasion and migration, through the TLR9/MYD88 independent pathway [ 365 , 372 , 373 ], while cfDNA may also promote metastasis by altering the expression levels of the inflammatory chemokine CXCL8 [ 374 ], or other genes such as MMP9 and CD44 and miRNAs [ 375 ]. Other reports suggest that the malignant phenotype of tumor cells may be transferred to neighboring normal cells via the assimilation and transfection of cfDNA and other nucleic acids complexed with EVs, such as apoptotic bodies [ 326 ] and exosomes [ 291 , 376 ].…”

Section: Potential Pathological Effects and Biological Functions Of C...mentioning

confidence: 99%

New Perspectives on the Importance of Cell-Free DNA Biology

et al. 2022

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Body fluids are constantly replenished with a population of genetically diverse cell-free DNA (cfDNA) fragments, representing a vast reservoir of information reflecting real-time changes in the host and metagenome. As many body fluids can be collected non-invasively in a one-off and serial fashion, this reservoir can be tapped to develop assays for the diagnosis, prognosis, and monitoring of wide-ranging pathologies, such as solid tumors, fetal genetic abnormalities, rejected organ transplants, infections, and potentially many others. The translation of cfDNA research into useful clinical tests is gaining momentum, with recent progress being driven by rapidly evolving preanalytical and analytical procedures, integrated bioinformatics, and machine learning algorithms. Yet, despite these spectacular advances, cfDNA remains a very challenging analyte due to its immense heterogeneity and fluctuation in vivo. It is increasingly recognized that high-fidelity reconstruction of the information stored in cfDNA, and in turn the development of tests that are fit for clinical roll-out, requires a much deeper understanding of both the physico-chemical features of cfDNA and the biological, physiological, lifestyle, and environmental factors that modulate it. This is a daunting task, but with significant upsides. In this review we showed how expanded knowledge on cfDNA biology and faithful reverse-engineering of cfDNA samples promises to (i) augment the sensitivity and specificity of existing cfDNA assays; (ii) expand the repertoire of disease-specific cfDNA markers, thereby leading to the development of increasingly powerful assays; (iii) reshape personal molecular medicine; and (iv) have an unprecedented impact on genetics research.

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Cell-free DNA promotes malignant transformation in non-tumor cells

Cited by 14 publications

References 50 publications

Survival of HT29 Cancer Cells Is Affected by IGF1R Inhibition via Modulation of Self-DNA-Triggered TLR9 Signaling and the Autophagy Response

Survival of HT29 Cancer Cells Is Affected by IGF1R Inhibition via Modulation of Self-DNA-Triggered TLR9 Signaling and the Autophagy Response

ATR-I774Yfs*5 promotes genomic instability through micronuclei formation

New Perspectives on the Importance of Cell-Free DNA Biology

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