Interaction of Quindoline derivative with telomeric repeat–containing RNA induces telomeric DNA-damage response in cancer cells through inhibition of telomeric repeat factor 2

• Published in: Biochimica et biophysica acta. General subjects Vol. 1861; no. 12; pp. 3246 - 3256
• Main Authors: Zhang, Yan, Zeng, Deying, Cao, Jiaojiao, Wang, Mingxue, Shu, Bing, Kuang, Guotao, Ou, Tian-Miao, Tan, Jia-Heng, Gu, Lian-Quan, Huang, Zhi-Shu, Li, Ding
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2017
• Subjects: Alkaloids - metabolism; Alkaloids - pharmacology; Allosteric Regulation; apoptosis; Apoptosis - drug effects; binding proteins; cell cycle checkpoints; Cell Cycle Checkpoints - drug effects; Cell Line, Tumor; cell proliferation; dissociation; DNA; DNA Damage; enzyme-linked immunosorbent assay; fluorescent antibody technique; G-quadruplex; G-Quadruplexes; heterochromatin; Humans; Indoles - metabolism; Indoles - pharmacology; ligands; mammals; neoplasm cells; neoplasms; Neoplasms - drug therapy; Neoplasms - genetics; Neoplasms - pathology; non-coding RNA; precipitin tests; Quindoline; Quinolines - metabolism; Quinolines - pharmacology; RNA, Long Noncoding - metabolism; screening; Telomere; telomeres; Telomeric Repeat Binding Protein 2 - antagonists & inhibitors; TERRA; TRF2; Western blotting; DSBs; PBS; CD; DMSO; RTCA; G-quadruplex; ALT; DAPI; ChIP; MTT; TRF2; Telomere; TRF1; MST; TERRA; TAMRA; DDR; FAM; SPR; OD; Quindoline; PI; ATM; ELISA
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.09.015

Telomeric repeat–containing RNA (TERRA) is a large non-coding RNA in mammalian cells, which forms an integral component of telomeric heterochromatin. TERRA can bind to an allosteric site of telomeric repeat factor 2 (TRF2), a key component of Shelterin that protect chromosome termini. Both TERRA and TRF2 have been recognized as promising new therapeutic targets for cancer treatment. Our methods include FRET assay, SPR, CD, microscale thermophoresis (MST), enzyme-linked immunosorbent assay (ELISA), chromatin immunoprecipitation (ChIP), colony formation assays, Western blot, immunofluorescence, cell cycle arrest and apoptosis detection, and xCELLigence real-time cell analysis (RTCA). In our routine screening of small molecule libraries, we found that a Quindoline derivative, CK1-14 could bind to and stabilize TERRA G-quadruplex structure, which could bind more tightly with an allosteric site of a telomeric binding protein TRF2, resulting in dissociation of TRF2 from telomeric DNA. Further in cellular studies indicated that the above effect of CK1-14 on TERRA G-quadruplex could activate DNA-damage response and cause cell cycle arrest, resulting in inhibition of U2OS cell proliferation and causing cell apoptosis. Our mechanistic studies indicated that interaction of CK1-14 with TERRA induces telomeric DNA-damage response in U2OS cancer cells through inhibition of TRF2. CK1-14 could be further developed as a promising lead compound targeting telomere for cancer treatment. Our present study provides the first evidence that allosteric modulation of TRF2 by TERRA G-quadruplex with a binding ligand could become a promising new strategy for cancer treatment especially for ALT tumor cells. [Display omitted] •Telomeric repeat–containing RNA (TERRA) is a component of telomeric heterochromatin.•Telomeric repeat factor 2 (TRF2) is a component of Shelterin protecting chromosome.•Quindoline derivative CK1-14 could bind to and stabilize TERRA G-quadruplex.•The above complex could inhibit TRF2, resulting in its dissociation from telomere.•CK1-14 could activate DNA-damage response and induce tumor cell apoptosis.