NSD3 Protein Methylation and Stabilization Transforms Human ES Cells into Variant State

Overview

Journal Life Sci Alliance

Specialties Biochemistry
Biology
Cell Biology
Molecular Biology

Date 2024 Dec 31

PMID 39741006

Authors

Vignesh K Krishnamoorthy

Fariha Hamdani

Pooja Shukla

Radhika Arasala Rao

Shaikh Anaitullah

Kriti Kestur Biligiri

Rajashekar Varma Kadumuri

Purushotham Reddy Pothula

Sreenivas Chavali

Shravanti Rampalli

Affiliations

Soon will be listed here.

Abstract

Cultured human embryonic stem cells (hESCs) can develop genetic anomalies that increase their susceptibility to transformation. In this study, we characterized a variant hESC (vhESC) line and investigated the molecular mechanisms leading to the drift towards a transformed state. Our findings revealed that vhESCs up-regulate EMT-specific markers, accelerate wound healing, exhibit compromised lineage differentiation, and retain pluripotency gene expression in teratomas. Furthermore, we discovered an altered epigenomic landscape and overexpression of the lysine methyltransferases EHMT1, EHMT2, and NSD group of proteins in vhESCs. Remarkably, depleting NSD3 oncogene reversed the molecular and phenotypic changes in vhESCs. We identified a detailed mechanism where EHMT2 interacts and methylates NSD3 at lysine 477, stabilizing its protein levels in vhESCs. In addition, we showed that NSD3 levels are regulated by protein degradation in hESCs, and its stabilization leads to the emergence of the variant state. Overall, our study identify that misregulation of NSD3 in pluripotent stem cells, through methylation-mediated abrogation of its protein degradation, drives hESCs towards oncogenic transformation.

References

Bennett R, Swaroop A, Troche C, Licht J . The Role of Nuclear Receptor-Binding SET Domain Family Histone Lysine Methyltransferases in Cancer. Cold Spring Harb Perspect Med. 2017; 7(6). PMC: 5453381. DOI: 10.1101/cshperspect.a026708. View

Arasala R, Jayaram M, Chattai J, Kumarasamy T, Sambasivan R, Rampalli S . Characterization of new variant human ES line VH9 hESC (INSTEMe001-a): a tool for human stem cell and cancer research. Stem Cell Res. 2019; 37:101444. DOI: 10.1016/j.scr.2019.101444. View

Nachiyappan A, Gupta N, Taneja R . EHMT1/EHMT2 in EMT, cancer stemness and drug resistance: emerging evidence and mechanisms. FEBS J. 2021; 289(5):1329-1351. DOI: 10.1111/febs.16334. View

Sansom S, Griffiths D, Faedo A, Kleinjan D, Ruan Y, Smith J . The level of the transcription factor Pax6 is essential for controlling the balance between neural stem cell self-renewal and neurogenesis. PLoS Genet. 2009; 5(6):e1000511. PMC: 2686252. DOI: 10.1371/journal.pgen.1000511. View

Da Silva L, Parry S, Reid L, Keith P, Waddell N, Kossai M . Aberrant expression of E-cadherin in lobular carcinomas of the breast. Am J Surg Pathol. 2008; 32(5):773-83. DOI: 10.1097/PAS.0b013e318158d6c5. View

Tchieu J, Calder E, Guttikonda S, Gutzwiller E, Aromolaran K, Steinbeck J . NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nat Biotechnol. 2019; 37(3):267-275. PMC: 6591152. DOI: 10.1038/s41587-019-0035-0. View

Zhang L, Zha X . Recent advances in nuclear receptor-binding SET domain 2 (NSD2) inhibitors: An update and perspectives. Eur J Med Chem. 2023; 250:115232. DOI: 10.1016/j.ejmech.2023.115232. View

Cao Q, Dhanasekaran S, Kim J, Mani R, Tomlins S, Mehra R . Repression of E-cadherin by the polycomb group protein EZH2 in cancer. Oncogene. 2008; 27(58):7274-84. PMC: 2690514. DOI: 10.1038/onc.2008.333. View

Amps K, Andrews P, Anyfantis G, Armstrong L, Avery S, Baharvand H . Screening ethnically diverse human embryonic stem cells identifies a chromosome 20 minimal amplicon conferring growth advantage. Nat Biotechnol. 2011; 29(12):1132-44. PMC: 3454460. DOI: 10.1038/nbt.2051. View

10.

Li X, Tao Y, Bradley R, Du Z, Tao Y, Kong L . Fast Generation of Functional Subtype Astrocytes from Human Pluripotent Stem Cells. Stem Cell Reports. 2018; 11(4):998-1008. PMC: 6178885. DOI: 10.1016/j.stemcr.2018.08.019. View

11.

Cho H, Hayami S, Toyokawa G, Maejima K, Yamane Y, Suzuki T . RB1 methylation by SMYD2 enhances cell cycle progression through an increase of RB1 phosphorylation. Neoplasia. 2012; 14(6):476-86. PMC: 3394190. DOI: 10.1593/neo.12656. View

12.

Maeda T, Hobbs R, Merghoub T, Guernah I, Zelent A, Cordon-Cardo C . Role of the proto-oncogene Pokemon in cellular transformation and ARF repression. Nature. 2005; 433(7023):278-85. DOI: 10.1038/nature03203. View

13.

Huang J, Dorsey J, Chuikov S, Zhang X, Jenuwein T, Reinberg D . G9a and Glp methylate lysine 373 in the tumor suppressor p53. J Biol Chem. 2010; 285(13):9636-9641. PMC: 2843213. DOI: 10.1074/jbc.M109.062588. View

14.

Fujii S, Ito K, Ito Y, Ochiai A . Enhancer of zeste homologue 2 (EZH2) down-regulates RUNX3 by increasing histone H3 methylation. J Biol Chem. 2008; 283(25):17324-32. PMC: 2427338. DOI: 10.1074/jbc.M800224200. View

15.

Cowan C, Atienza J, Melton D, Eggan K . Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells. Science. 2005; 309(5739):1369-73. DOI: 10.1126/science.1116447. View

16.

Merkle F, Ghosh S, Kamitaki N, Mitchell J, Avior Y, Mello C . Human pluripotent stem cells recurrently acquire and expand dominant negative P53 mutations. Nature. 2017; 545(7653):229-233. PMC: 5427175. DOI: 10.1038/nature22312. View

17.

Melo S, Figueiredo J, Fernandes M, Goncalves M, Morais-de-Sa E, Sanches J . Predicting the Functional Impact of CDH1 Missense Mutations in Hereditary Diffuse Gastric Cancer. Int J Mol Sci. 2017; 18(12). PMC: 5751289. DOI: 10.3390/ijms18122687. View

18.

Feinberg A, Koldobskiy M, Gondor A . Epigenetic modulators, modifiers and mediators in cancer aetiology and progression. Nat Rev Genet. 2016; 17(5):284-99. PMC: 4888057. DOI: 10.1038/nrg.2016.13. View

19.

Topchu I, Pangeni R, Bychkov I, Miller S, Izumchenko E, Yu J . The role of NSD1, NSD2, and NSD3 histone methyltransferases in solid tumors. Cell Mol Life Sci. 2022; 79(6):285. PMC: 9520630. DOI: 10.1007/s00018-022-04321-2. View

20.

Davies A, Zoubeidi A, Beltran H, Selth L . The Transcriptional and Epigenetic Landscape of Cancer Cell Lineage Plasticity. Cancer Discov. 2023; 13(8):1771-1788. PMC: 10527883. DOI: 10.1158/2159-8290.CD-23-0225. View