Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2024

Open Access 01-12-2024 | Neuroblastoma | Research

KAP1 stabilizes MYCN mRNA and promotes neuroblastoma tumorigenicity by protecting the RNA m6A reader YTHDC1 protein degradation

Authors: Yi Yang, Yingwen Zhang, Guoyu Chen, Bowen Sun, Fei Luo, Yijin Gao, Haizhong Feng, Yanxin Li

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2024

Login to get access

Abstract

Background

Neuroblastoma (NB) patients with amplified MYCN often face a grim prognosis and are resistant to existing therapies, yet MYCN protein is considered undruggable. KAP1 (also named TRIM28) plays a crucial role in multiple biological activities. This study aimed to investigate the relationship between KAP1 and MYCN in NB.

Methods

Transcriptome analyses and luciferase reporter assay identified that KAP1 was a downstream target of MYCN. The effects of KAP1 on cancer cell proliferation and colony formation were explored using the loss-of-function assays in vitro and in vivo. RNA stability detection was used to examine the influence of KAP1 on MYCN expression. The mechanisms of KAP1 to maintain MYCN mRNA stabilization were mainly investigated by mass spectrum, immunoprecipitation, RIP-qPCR, and western blotting. In addition, a xenograft mouse model was used to reveal the antitumor effect of STM2457 on NB.

Results

Here we identified KAP1 as a critical regulator of MYCN mRNA stability by protecting the RNA N6-methyladenosine (m6A) reader YTHDC1 protein degradation. KAP1 was highly expressed in clinical MYCN-amplified NB and was upregulated by MYCN. Reciprocally, KAP1 knockdown reduced MYCN mRNA stability and inhibited MYCN-amplified NB progression. Mechanistically, KAP1 regulated the stability of MYCN mRNA in an m6A-dependent manner. KAP1 formed a complex with YTHDC1 and RNA m6A writer METTL3 to regulate m6A-modified MYCN mRNA stability. KAP1 depletion decreased YTHDC1 protein stability and promoted MYCN mRNA degradation. Inhibiting MYCN mRNA m6A modification synergized with chemotherapy to restrain tumor progression in MYCN-amplified NB.

Conclusions

Our research demonstrates that KAP1, transcriptionally activated by MYCN, forms a complex with YTHDC1 and METTL3, which in turn maintain the stabilization of MYCN mRNA in an m6A-dependent manner. Targeting m6A modification by STM2457, a small-molecule inhibitor of METTL3, could downregulate MYCN expression and attenuate tumor proliferation. This finding provides a new alternative putative therapeutic strategy for MYCN-amplified NB.
Appendix
Available only for authorised users
Literature
1.
Swift CC, Eklund MJ, Kraveka JM, Alazraki AL. Updates in diagnosis, management, and treatment of neuroblastoma. Radiographics. 2018;38:566–80.PubMedCrossRef
2.
Schmitt-Hoffner F, Van Rijn S, Toprak UH, Mauermann M, Rosemann F, Heit-Mondrzyk A, et al. FOXR2 stabilizes MYCN protein and identifies non-MYCN-amplified neuroblastoma patients with unfavorable outcome. J Clin Oncol. 2021;39:3217–28.PubMedPubMedCentralCrossRef
3.
Otte J, Dyberg C, Pepich A, Johnsen JI. MYCN function in neuroblastoma development. Front Oncol. 2020;10:624079.PubMedCrossRef
4.
Pinto NR, Applebaum MA, Volchenboum SL, Matthay KK, London WB, Ambros PF, et al. Advances in risk classification and treatment strategies for neuroblastoma. J Clin Oncol. 2015;33:3008–17.PubMedPubMedCentralCrossRef
5.
Fletcher JI, Ziegler DS, Trahair TN, Marshall GM, Haber M, Norris MD. Too many targets, not enough patients: rethinking neuroblastoma clinical trials. Nat Rev Cancer. 2018;18:389–400.PubMedCrossRef
6.
Wolpaw AJ, Bayliss R, Büchel G, Dang CV, Eilers M, Gustafson WC, et al. Drugging the “Undruggable” MYCN oncogenic transcription factor: overcoming previous obstacles to impact childhood cancers. Cancer Res. 2021;81:1627–32.PubMedPubMedCentralCrossRef
7.
Otto T, Horn S, Brockmann M, Eilers U, Schüttrumpf L, Popov N, et al. Stabilization of N-Myc is a critical function of Aurora a in human neuroblastoma. Cancer Cell. 2009;15:67–78.PubMedCrossRef
8.
Gustafson WC, Meyerowitz JG, Nekritz EA, Chen J, Benes C, Charron E, et al. Drugging MYCN through an allosteric transition in Aurora kinase a. Cancer Cell. 2014;26:414–27.PubMedPubMedCentralCrossRef
9.
Puissant A, Frumm SM, Alexe G, Bassil CF, Qi J, Chanthery YH, et al. Targeting MYCN in neuroblastoma by BET bromodomain inhibition. Cancer Discov. 2013;3:308–23.PubMedPubMedCentralCrossRef
10.
Rohaan MW, Gomez-Eerland R, Van Den Berg JH, Geukes Foppen MH, Van Zon M, Raud B, et al. MART-1 TCR gene-modified peripheral blood T cells for the treatment of metastatic melanoma: a phase I/IIa clinical trial. Immunooncol Technol. 2022;15:100089.PubMedPubMedCentralCrossRef
11.
Toyoshima M, Howie HL, Imakura M, Walsh RM, Annis JE, Chang AN, et al. Functional genomics identifies therapeutic targets for MYC-driven cancer. Proc Natl Acad Sci USA. 2012;109:9545–50.PubMedPubMedCentralCrossRef
12.
13.
Huang N, Sun X, Li P, Liu X, Zhang X, Chen Q, et al. TRIM family contribute to tumorigenesis, cancer development, and drug resistance. Exp Hematol Oncol. 2022;11:75.PubMedPubMedCentralCrossRef
14.
Pineda CT, Ramanathan S, Fon Tacer K, Weon JL, Potts MB, Ou YH, et al. Degradation of AMPK by a cancer-specific ubiquitin ligase. Cell. 2015;160:715–28.PubMedPubMedCentralCrossRef
15.
Wang C, Ivanov A, Chen L, Fredericks WJ, Seto E, Rauscher FJ 3rd, et al. MDM2 interaction with nuclear corepressor KAP1 contributes to p53 inactivation. EMBO J. 2005;24:3279–90.PubMedPubMedCentralCrossRef
16.
17.
Li M, Xu X, Chang CW, Liu Y. TRIM28 functions as the SUMO E3 ligase for PCNA in prevention of transcription induced DNA breaks. Proc Natl Acad Sci USA. 2020;117:23588–96.PubMedPubMedCentralCrossRef
18.
Diao Z, Ji Q, Wu Z, Zhang W, Cai Y, Wang Z, et al. SIRT3 consolidates heterochromatin and counteracts senescence. Nucleic Acids Res. 2021;49:4203–19.PubMedPubMedCentralCrossRef
19.
Rowe HM, Jakobsson J, Mesnard D, Rougemont J, Reynard S, Aktas T, et al. KAP1 controls endogenous retroviruses in embryonic stem cells. Nature. 2010;463:237–40.PubMedCrossRef
20.
Ecco G, Cassano M, Kauzlaric A, Duc J, Coluccio A, Offner S, et al. Transposable elements and their KRAB-ZFP controllers regulate gene expression in adult tissues. Dev Cell. 2016;36:611–23.PubMedPubMedCentralCrossRef
21.
Bacon CW, Challa A, Hyder U, Shukla A, Borkar AN, Bayo J, et al. KAP1 is a chromatin reader that couples steps of RNA polymerase II transcription to sustain oncogenic programs. Mol Cell. 2020;78:1133–51.PubMedPubMedCentralCrossRef
22.
Mcnamara RP, Reeder JE, Mcmillan EA, Bacon CW, Mccann JL, D’orso I. KAP1 recruitment of the 7SK snRNP complex to promoters enables transcription elongation by RNA polymerase II. Mol Cell. 2016;61:39–53.PubMedCrossRef
23.
Xu W, Li J, He C, Wen J, Ma H, Rong B, et al. METTL3 regulates heterochromatin in mouse embryonic stem cells. Nature. 2021;591:317–21.PubMedCrossRef
24.
Percharde M, Lin CJ, Yin Y, Guan J, Peixoto GA, Bulut-Karslioglu A, et al. A LINE1-Nucleolin partnership regulates early development and ESC identity. Cell. 2018;174:391–405.PubMedPubMedCentralCrossRef
25.
Sang Y, Li Y, Song L, Alvarez AA, Zhang W, Lv D, et al. TRIM59 promotes Gliomagenesis by inhibiting TC45 Dephosphorylation of STAT3. Cancer Res. 2018;78:1792–804.PubMedPubMedCentralCrossRef
26.
Song L, Yu B, Yang Y, Liang J, Zhang Y, Ding L, et al. Identification of functional cooperative mutations of GNAO1 in human acute lymphoblastic leukemia. Blood. 2021;137:1181–91.PubMedCrossRef
27.
Yang Y, Wang S, Cai J, Liang J, Zhang Y, Xie Y, et al. Targeting ARHGEF12 promotes neuroblastoma differentiation, MYCN degradation, and reduces tumorigenicity. Cell Oncol (Dordr). 2023;46:133–43.PubMedCrossRef
28.
29.
30.
Zhang Z, Theler D, Kaminska KH, Hiller M, De La Grange P, Pudimat R, et al. The YTH domain is a novel RNA binding domain. J Biol Chem. 2010;285:14701–10.PubMedPubMedCentralCrossRef
31.
Xu C, Wang X, Liu K, Roundtree IA, Tempel W, Li Y, et al. Structural basis for selective binding of m6A RNA by the YTHDC1 YTH domain. Nat Chem Biol. 2014;10:927–9.PubMedCrossRef
32.
Li F, Yi Y, Miao Y, Long W, Long T, Chen S, et al. N(6)-Methyladenosine modulates nonsense-mediated mRNA decay in human glioblastoma. Cancer Res. 2019;79:5785–98.PubMedPubMedCentralCrossRef
33.
Liu S, Li G, Li Q, Zhang Q, Zhuo L, Chen X, et al. The roles and mechanisms of YTH domain-containing proteins in cancer development and progression. Am J Cancer Res. 2020;10:1068–84.PubMedPubMedCentral
34.
Fong KW, Zhao JC, Song B, Zheng B, Yu J. TRIM28 protects TRIM24 from SPOP-mediated degradation and promotes prostate cancer progression. Nat Commun. 2018;9:5007.PubMedPubMedCentralCrossRef
35.
Zhu K, Gao T, Wang Z, Zhang L, Tan K, Lv Z. RNA N6-methyladenosine reader IGF2BP3 interacts with MYCN and facilitates neuroblastoma cell proliferation. Cell Death Discov. 2023;9:151.PubMedPubMedCentralCrossRef
36.
Cheng J, Xu L, Deng L, Xue L, Meng Q, Wei F, et al. RNA N(6)-methyladenosine modification is required for miR-98/MYCN axis-mediated inhibition of neuroblastoma progression. Sci Rep. 2020;10:13624.PubMedPubMedCentralCrossRef
37.
38.
Yankova E, Blackaby W, Albertella M, Rak J, De Braekeleer E, Tsagkogeorga G, et al. Small-molecule inhibition of METTL3 as a strategy against myeloid leukaemia. Nature. 2021;593:597–601.PubMedPubMedCentralCrossRef
39.
Wang L, Yang Q, Zhou Q, Fang F, Lei K, Liu Z, et al. METTL3-m(6)A-EGFR-axis drives lenvatinib resistance in hepatocellular carcinoma. Cancer Lett. 2023;559:216122.PubMedCrossRef
40.
Zhu J, Wang J, Sun F, Zhen Z, Chen T, Lu S, et al. Vincristine, Irinotecan, and Temozolomide in patients with relapsed/refractory neuroblastoma. Front Oncol. 2022;12:804310.PubMedPubMedCentralCrossRef
41.
Bunch H, Zheng X, Burkholder A, Dillon ST, Motola S, Birrane G, et al. TRIM28 regulates RNA polymerase II promoter-proximal pausing and pause release. Nat Struct Mol Biol. 2014;21:876–83.PubMedPubMedCentralCrossRef
42.
Noon AT, Shibata A, Rief N, Löbrich M, Stewart GS, Jeggo PA, et al. 53BP1-dependent robust localized KAP-1 phosphorylation is essential for heterochromatic DNA double-strand break repair. Nat Cell Biol. 2010;12:177–84.PubMedCrossRef
43.
White D, Rafalska-Metcalf IU, Ivanov AV, Corsinotti A, Peng H, Lee SC, et al. The ATM substrate KAP1 controls DNA repair in heterochromatin: regulation by HP1 proteins and serine 473/824 phosphorylation. Mol Cancer Res. 2012;10:401–14.PubMedCrossRef
44.
Jin JO, Lee GD, Nam SH, Lee TH, Kang DH, Yun JK, et al. Sequential ubiquitination of p53 by TRIM28, RLIM, and MDM2 in lung tumorigenesis. Cell Death Differ. 2021;28:1790–803.PubMedCrossRef
45.
Yu C, Zhan L, Jiang J, Pan Y, Zhang H, Li X, et al. KAP-1 is overexpressed and correlates with increased metastatic ability and tumorigenicity in pancreatic cancer. Med Oncol. 2014;31:25.PubMedCrossRef
46.
Li J, Xi Y, Li W, Mccarthy RL, Stratton SA, Zou W, et al. TRIM28 interacts with EZH2 and SWI/SNF to activate genes that promote mammosphere formation. Oncogene. 2017;36:2991–3001.PubMedPubMedCentralCrossRef
47.
Song T, Lv S, Ma X, Zhao X, Fan L, Zou Q, et al. TRIM28 represses renal cell carcinoma cell proliferation by inhibiting TFE3/KDM6A-regulated autophagy. J Biol Chem. 2023;299:104621.PubMedPubMedCentralCrossRef
48.
Lin LF, Li CF, Wang WJ, Yang WM, Wang DD, Chang WC, et al. Loss of ZBRK1 contributes to the increase of KAP1 and promotes KAP1-mediated metastasis and invasion in cervical cancer. PLoS One. 2013;8:e73033.PubMedPubMedCentralCrossRef
49.
Agarwal N, Rinaldetti S, Cheikh BB, Zhou Q, Hass EP, Jones RT, et al. TRIM28 is a transcriptional activator of the mutant TERT promoter in human bladder cancer. Proc Natl Acad Sci USA. 2021;118:e2102423118.PubMedPubMedCentralCrossRef
50.
Lionnard L, Duc P, Brennan MS, Kueh AJ, Pal M, Guardia F, et al. TRIM17 and TRIM28 antagonistically regulate the ubiquitination and anti-apoptotic activity of BCL2A1. Cell Death Differ. 2019;26:902–17.PubMedCrossRef
51.
Chen L, Chen DT, Kurtyka C, Rawal B, Fulp WJ, Haura EB, et al. Tripartite motif containing 28 (Trim28) can regulate cell proliferation by bridging HDAC1/E2F interactions. J Biol Chem. 2012;287:40106–18.PubMedPubMedCentralCrossRef
52.
Bojkowska K, Aloisio F, Cassano M, Kapopoulou A, Santoni De Sio F, Zangger N, et al. Liver-specific ablation of Krüppel-associated box-associated protein 1 in mice leads to male-predominant hepatosteatosis and development of liver adenoma. Hepatology. 2012;56:1279–90.PubMedCrossRef
53.
54.
Tan B, Zhou K, Liu W, Prince E, Qing Y, Li Y, et al. RNA N(6) -methyladenosine reader YTHDC1 is essential for TGF-beta-mediated metastasis of triple negative breast cancer. Theranostics. 2022;12:5727–43.PubMedPubMedCentralCrossRef
55.
Zhao H, Xu Y, Xie Y, Zhang L, Gao M, Li S, et al. m6A regulators is differently expressed and correlated with immune response of esophageal Cancer. Front Cell Dev Biol. 2021;9:650023.PubMedPubMedCentralCrossRef
56.
Liu J, Sun G, Pan S, Qin M, Ouyang R, Li Z, et al. The Cancer genome atlas (TCGA) based m(6) a methylation-related genes predict prognosis in hepatocellular carcinoma. Bioengineered. 2020;11:759–68.PubMedPubMedCentralCrossRef
57.
Widagdo J, Anggono V, Wong JJ. The multifaceted effects of YTHDC1-mediated nuclear m(6)a recognition. Trends Genet. 2022;38:325–32.PubMedCrossRef
58.
Yan H, Zhang L, Cui X, Zheng S, Li R. Roles and mechanisms of the m(6)a reader YTHDC1 in biological processes and diseases. Cell Death Discov. 2022;8:237.PubMedPubMedCentralCrossRef
59.
Jiang X, Liu B, Nie Z, Duan L, Xiong Q, Jin Z, et al. The role of m6A modification in the biological functions and diseases. Signal Transduct Target Ther. 2021;6:74.PubMedPubMedCentralCrossRef
60.
Zaccara S, Ries RJ, Jaffrey SR. Reading, writing and erasing mRNA methylation. Nat Rev Mol Cell Biol. 2019;20:608–24.PubMedCrossRef
61.
Sun T, Wu R, Ming L. The role of m6A RNA methylation in cancer. Biomed Pharmacother. 2019;112:108613.PubMedCrossRef
62.
63.
64.
Zhang ZW, Teng X, Zhao F, Ma C, Zhang J, Xiao LF, et al. METTL3 regulates m(6)a methylation of PTCH1 and GLI2 in sonic hedgehog signaling to promote tumor progression in SHH-medulloblastoma. Cell Rep. 2022;41:111530.PubMedCrossRef
Metadata
Title
KAP1 stabilizes MYCN mRNA and promotes neuroblastoma tumorigenicity by protecting the RNA m6A reader YTHDC1 protein degradation
Authors
Yi Yang
Yingwen Zhang
Guoyu Chen
Bowen Sun
Fei Luo
Yijin Gao
Haizhong Feng
Yanxin Li
Publication date
01-12-2024
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2024
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-024-03040-9

Other articles of this Issue 1/2024

Journal of Experimental & Clinical Cancer Research 1/2024 Go to the issue

Research

SF3B3-regulated mTOR alternative splicing promotes colorectal cancer progression and metastasis

Review

GMP-manufactured CRISPR/Cas9 technology as an advantageous tool to support cancer immunotherapy

Retraction Note

Retraction Note: MicroRNA-338-3p suppresses ovarian cancer cells growth and metastasis: implication of Wnt/catenin beta and MEK/ ERK signaling pathways

Research

EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening

Review

Tumor immune microenvironment-based therapies in pancreatic ductal adenocarcinoma: time to update the concept

Review

Transcriptional regulation of cancer stem cell: regulatory factors elucidation and cancer treatment strategies
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits