Anthracyclines have been a cornerstone in the cure of diffuse large B-cell lymphoma (DLBCL) and other hematological cancers. The ability of anthracyclines to eliminate DLBCL depends on the presence of topoisomerase-II-alpha (TopIIA), a DNA repair enzyme complex. We identified nucleolin as a novel binding partner of TopIIA. Abrogation of nucleolin sensitized DLBCL cells to TopIIA targeting agents (doxorubicin/etoposide). Silencing nucleolin and challenging DLBCL cells with doxorubicin enhanced the phosphorylation of H2AX (γH2AX-marker of DNA damage) and allowed DNA fragmentation. Reconstitution of nucleolin expression in nucleolin-knockdown DLBCL cells prevented TopIIA targeting agent-induced apoptosis. Nucleolin binding to TopIIA was mapped to RNA-binding domain 3 of nucleolin, and this interaction was essential for blocking DNA damage and apoptosis. Nucleolin silencing decreased TopIIA decatenation activity, but enhanced formation of TopIIA–DNA cleavable complexes in the presence of etoposide. Moreover, combining nucleolin inhibitors: aptamer AS1411 or nucant N6L with doxorubicin reduced DLBCL cell survival. These findings are of clinical importance because low nucleolin levels versus high nucleolin levels in DLBCL predicted 90-month estimated survival of 70% versus 12% (P<0.0001) of patients treated with R-CHOP-based therapy.
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This work was supported by National Cancer Institute grant (CA 1206173), National Institute of Diabetes and Digestive and Kidney Diseases grant (DK091490), the Richard Spencer Lewis Memorial Foundation, and patients’ families. The MD Anderson Flow Cytometry and Cellular Imaging Facility are supported by the NIH/NCI under award number P30CAQ16672.
NJ, HFZ LS and FS designed the research studies, analyzed the data and wrote the manuscript. NJ, HFZ, BG, TK, QY, RM, RKS, FKB, XW, ZB, JW, XYXM and KHY performed experiments, analyzed the data and contributed to clinical resources under approved IRB. QY and KHY analyzed the tissue staining, quantified protein expression and identified correlations with patient clinical outcome. LS and FS contributed to the research design, writing and finalized the manuscript.
The authors declare no conflict of interest.
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Jain, N., Zhu, H., Khashab, T. et al. Targeting nucleolin for better survival in diffuse large B-cell lymphoma. Leukemia 32, 663–674 (2018). https://doi.org/10.1038/leu.2017.215
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