Oncogenic ETS fusions promote DNA damage and proinflammatory responses via pericentromeric RNAs in extracellular vesicles
Peter Ruzanov, … , Lincoln D. Stein, Poul H. Sorensen
Peter Ruzanov, … , Lincoln D. Stein, Poul H. Sorensen
Published March 26, 2024
Citation Information: J Clin Invest. 2024;134(9):e169470. https://doi.org/10.1172/JCI169470.
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Research Article Inflammation

Oncogenic ETS fusions promote DNA damage and proinflammatory responses via pericentromeric RNAs in extracellular vesicles

Abstract

Aberrant expression of the E26 transformation-specific (ETS) transcription factors characterizes numerous human malignancies. Many of these proteins, including EWS:FLI1 and EWS:ERG fusions in Ewing sarcoma (EwS) and TMPRSS2:ERG in prostate cancer (PCa), drive oncogenic programs via binding to GGAA repeats. We report here that both EWS:FLI1 and ERG bind and transcriptionally activate GGAA-rich pericentromeric heterochromatin. The respective pathogen-like HSAT2 and HSAT3 RNAs, together with LINE, SINE, ERV, and other repeat transcripts, are expressed in EwS and PCa tumors, secreted in extracellular vesicles (EVs), and are highly elevated in plasma of patients with EwS with metastatic disease. High human satellite 2 and 3 (HSAT2,3) levels in EWS:FLI1- or ERG-expressing cells and tumors were associated with induction of G2/M checkpoint, mitotic spindle, and DNA damage programs. These programs were also activated in EwS EV-treated fibroblasts, coincident with accumulation of HSAT2,3 RNAs, proinflammatory responses, mitotic defects, and senescence. Mechanistically, HSAT2,3-enriched cancer EVs induced cGAS-TBK1 innate immune signaling and formation of cytosolic granules positive for double-strand RNAs, RNA-DNA, and cGAS. Hence, aberrantly expressed ETS proteins derepress pericentromeric heterochromatin, yielding pathogenic RNAs that transmit genotoxic stress and inflammation to local and distant sites. Monitoring HSAT2,3 plasma levels and preventing their dissemination may thus improve therapeutic strategies and blood-based diagnostics.

Authors

Peter Ruzanov, Valentina Evdokimova, Manideep C. Pachva, Alon Minkovich, Zhenbo Zhang, Sofya Langman, Hendrik Gassmann, Uwe Thiel, Marija Orlic-Milacic, Syed H. Zaidi, Vanya Peltekova, Lawrence E. Heisler, Manju Sharma, Michael E. Cox, Trevor D. McKee, Mark Zaidi, Eve Lapouble, John D. McPherson, Olivier Delattre, Laszlo Radvanyi, Stefan E.G. Burdach, Lincoln D. Stein, Poul H. Sorensen

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Figure 2

Repeat RNAs are the most abundant species in blood plasma of patients with EwS.

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Repeat RNAs are the most abundant species in blood plasma of patients wi...
(A) RNA-Seq of plasma EV RNAs from patients with EwS (EW cohort; n = 12) and age-matched healthy donors (n = 7). Pie charts represent averaged values for each repeat and nonrepeat category. Comparisons made between patients with EwS versus healthy donors; P = 0.012, Wilcoxon signed-rank test. (B) RepeatMasker-annotated families from A shown as mean RPM ± SEM; patients with EwS versus healthy donors, *P < 0.05, **P < 0.01, Wilcoxon signed-rank test. (C) Violin plots of representative RNAs from B; patients with EwS versus healthy donors, *P < 1.36×10–11, **P < 2.00×10–41. Note that ACRO1 was not detected in healthy donors and patients with localized disease. (D) RT-ddPCR of indicated RNAs in plasma of patients with EwS (TUM cohort, n = 20) versus healthy donors (n = 49); *P < 0.05, **P = 0.0002, ***P < 2.00×10–8. Variance between groups was analyzed with the Welch F test, P = 4.27×10–12 (ACRO1), P = 0.0038 (HSAT2) and P = 0.0158 (HERV-Kenv). P values in C and D were adjusted for multiple comparisons using BH method.