bims-crepig 2021-05-30 papers

bims-crepig

Biomed News

on Chromatin regulation and epigenetics in cell fate and cancer

Issue of 2021‒05‒30
thirty-one papers selected by
Connor Rogerson
University of Cambridge, MRC Cancer Unit

Muscle regeneration controlled by a designated DNA dioxygenase.
Pancreatic cancer prognosis is predicted by an ATAC-array technology for assessing chromatin accessibility.
Epigenomic profiling of primate lymphoblastoid cell lines reveals the evolutionary patterns of epigenetic activities in gene regulatory architectures.
Enhancer release and retargeting activates disease-susceptibility genes.
Live cell dynamics of the NF-Y transcription factor.
Ectopic targeting of CG DNA methylation in Arabidopsis with the bacterial SssI methyltransferase.
RSC primes the quiescent genome for hypertranscription upon cell cycle re-entry.
LncRNA HBL1 is required for genome-wide PRC2 occupancy and function in cardiogenesis from human pluripotent stem cells.
Spreading and epigenetic inheritance of heterochromatin require a critical density of histone H3 lysine 9 tri-methylation.
Impact of DNA methylation on 3D genome structure.
Genome information processing by the INO80 chromatin remodeler positions nucleosomes.
Ruler elements in chromatin remodelers set nucleosome array spacing and phasing.
scDetect: a rank-based ensemble learning algorithm for cell type identification of single-cell RNA sequencing in cancer.
DeepFun: a deep learning sequence-based model to decipher non-coding variant effect in a tissue- and cell type-specific manner.
Yeast cell fate control by temporal redundancy modulation of transcription factor paralogs.
BRD4 methylation by the methyltransferase SETD6 regulates selective transcription to control mRNA translation.
The nuclear periphery is a scaffold for tissue-specific enhancers.
GEPIA2021: integrating multiple deconvolution-based analysis into GEPIA.
Phase separation of OCT4 controls TAD reorganization to promote cell fate transitions.
Multiplexed single-cell profiling of chromatin states at genomic loci by expansion microscopy.
Antagonistic epistasis of Hnf4α and FoxO1 metabolic networks through enhancer interactions in β-cell function.
GRiNCH: simultaneous smoothing and detection of topological units of genome organization from sparse chromatin contact count matrices with matrix factorization.
A 3D system to model human pancreas development and its reference single-cell transcriptome atlas identify signaling pathways required for progenitor expansion.
PTMViz: a tool for analyzing and visualizing histone post translational modification data.
TFcancer: a manually curated database of transcription factors associated with human cancer.
Recognition of the inherently unstable H2A nucleosome by Swc2 is a major determinant for unidirectional H2A.Z exchange.
High Yap and Mll1 promote a persistent regenerative cell state induced by Notch signaling and loss of p53.
CpGmotifs: a tool to discover DNA motifs associated to CpG methylation events.
Mergeomics 2.0: a web server for multi-omics data integration to elucidate disease networks and predict therapeutics.
Multi Locus View: an extensible web-based tool for the analysis of genomic data.
Epigenetic induction of mitochondrial fission is required for maintenance of liver cancer initiating cells.

Cell Death Dis. 2021 May 25. 12(6): 535

Muscle regeneration controlled by a designated DNA dioxygenase.

Hongye Wang, Yile Huang, Ming Yu, Yang Yu, Sheng Li, Huating Wang, Hao Sun, Bing Li, Guoliang Xu, Ping Hu.

Tet dioxygenases are responsible for the active DNA demethylation. The functions of Tet proteins in muscle regeneration have not been well characterized. Here we find that Tet2, but not Tet1 and Tet3, is specifically required for muscle regeneration in vivo. Loss of Tet2 leads to severe muscle regeneration defects. Further analysis indicates that Tet2 regulates myoblast differentiation and fusion. Tet2 activates transcription of the key differentiation modulator Myogenin (MyoG) by actively demethylating its enhancer region. Re-expressing of MyoG in Tet2 KO myoblasts rescues the differentiation and fusion defects. Further mechanistic analysis reveals that Tet2 enhances MyoD binding by demethylating the flanking CpG sites of E boxes to facilitate the recruitment of active histone modifications and increase chromatin accessibility and activate its transcription. These findings shed new lights on DNA methylation and pioneer transcription factor activity regulation.

DOI: https://doi.org/10.1038/s41419-021-03817-2
Nat Commun. 2021 May 24. 12(1): 3044

Pancreatic cancer prognosis is predicted by an ATAC-array technology for assessing chromatin accessibility.

S Dhara, S Chhangawala, H Chintalapudi, G Askan, V Aveson, A L Massa, L Zhang, D Torres, A P Makohon-Moore, N Lecomte, J P Melchor, J Bermeo, A Cardenas, S Sinha, D Glassman, R Nicolle, R Moffitt, K H Yu, S Leppanen, S Laderman, B Curry, J Gui, V P Balachandran, C Iacobuzio-Donahue, R Chandwani, C S Leslie, S D Leach.

Unlike other malignancies, therapeutic options in pancreatic ductal adenocarcinoma (PDAC) are largely limited to cytotoxic chemotherapy without the benefit of molecular markers predicting response. Here we report tumor-cell-intrinsic chromatin accessibility patterns of treatment-naïve surgically resected PDAC tumors that were subsequently treated with (Gem)/Abraxane adjuvant chemotherapy. By ATAC-seq analyses of EpCAM+ PDAC malignant epithelial cells sorted from 54 freshly resected human tumors, we show here the discovery of a signature of 1092 chromatin loci displaying differential accessibility between patients with disease free survival (DFS) < 1 year and patients with DFS > 1 year. Analyzing transcription factor (TF) binding motifs within these loci, we identify two TFs (ZKSCAN1 and HNF1b) displaying differential nuclear localization between patients with short vs. long DFS. We further develop a chromatin accessibility microarray methodology termed "ATAC-array", an easy-to-use platform obviating the time and cost of next generation sequencing. Applying this methodology to the original ATAC-seq libraries as well as independent libraries generated from patient-derived organoids, we validate ATAC-array technology in both the original ATAC-seq cohort as well as in an independent validation cohort. We conclude that PDAC prognosis can be predicted by ATAC-array, which represents a low-cost, clinically feasible technology for assessing chromatin accessibility profiles.

DOI: https://doi.org/10.1038/s41467-021-23237-2
Nat Commun. 2021 May 25. 12(1): 3116

Epigenomic profiling of primate lymphoblastoid cell lines reveals the evolutionary patterns of epigenetic activities in gene regulatory architectures.

Raquel García-Pérez, Paula Esteller-Cucala, Glòria Mas, Irene Lobón, Valerio Di Carlo, Meritxell Riera, Martin Kuhlwilm, Arcadi Navarro, Antoine Blancher, Luciano Di Croce, José Luis Gómez-Skarmeta, David Juan, Tomàs Marquès-Bonet.

Changes in the epigenetic regulation of gene expression have a central role in evolution. Here, we extensively profiled a panel of human, chimpanzee, gorilla, orangutan, and macaque lymphoblastoid cell lines (LCLs), using ChIP-seq for five histone marks, ATAC-seq and RNA-seq, further complemented with whole genome sequencing (WGS) and whole genome bisulfite sequencing (WGBS). We annotated regulatory elements (RE) and integrated chromatin contact maps to define gene regulatory architectures, creating the largest catalog of RE in primates to date. We report that epigenetic conservation and its correlation with sequence conservation in primates depends on the activity state of the regulatory element. Our gene regulatory architectures reveal the coordination of different types of components and highlight the role of promoters and intragenic enhancers (gE) in the regulation of gene expression. We observe that most regulatory changes occur in weakly active gE. Remarkably, novel human-specific gE with weak activities are enriched in human-specific nucleotide changes. These elements appear in genes with signals of positive selection and human acceleration, tissue-specific expression, and particular functional enrichments, suggesting that the regulatory evolution of these genes may have contributed to human adaptation.

DOI: https://doi.org/10.1038/s41467-021-23397-1
Nature. 2021 May 26.

Enhancer release and retargeting activates disease-susceptibility genes.

Soohwan Oh, Jiaofang Shao, Joydeep Mitra, Feng Xiong, Matteo D'Antonio, Ruoyu Wang, Ivan Garcia-Bassets, Qi Ma, Xiaoyu Zhu, Joo-Hyung Lee, Sreejith J Nair, Feng Yang, Kenneth Ohgi, Kelly A Frazer, Zhengdong D Zhang, Wenbo Li, Michael G Rosenfeld.

The functional engagement between an enhancer and its target promoter ensures precise gene transcription1. Understanding the basis of promoter choice by enhancers has important implications for health and disease. Here we report that functional loss of a preferred promoter can release its partner enhancer to loop to and activate an alternative promoter (or alternative promoters) in the neighbourhood. We refer to this target-switching process as 'enhancer release and retargeting'. Genetic deletion, motif perturbation or mutation, and dCas9-mediated CTCF tethering reveal that promoter choice by an enhancer can be determined by the binding of CTCF at promoters, in a cohesin-dependent manner-consistent with a model of 'enhancer scanning' inside the contact domain. Promoter-associated CTCF shows a lower affinity than that at chromatin domain boundaries and often lacks a preferred motif orientation or a partnering CTCF at the cognate enhancer, suggesting properties distinct from boundary CTCF. Analyses of cancer mutations, data from the GTEx project and risk loci from genome-wide association studies, together with a focused CRISPR interference screen, reveal that enhancer release and retargeting represents an overlooked mechanism that underlies the activation of disease-susceptibility genes, as exemplified by a risk locus for Parkinson's disease (NUCKS1-RAB7L1) and three loci associated with cancer (CLPTM1L-TERT, ZCCHC7-PAX5 and PVT1-MYC).

DOI: https://doi.org/10.1038/s41586-021-03577-1
Sci Rep. 2021 May 26. 11(1): 10992

Live cell dynamics of the NF-Y transcription factor.

David G Priest, Andrea Bernardini, Jieqiong Lou, Roberto Mantovani, Elizabeth Hinde.

Transcription factors (TFs) are core players in the control of gene expression, evolutionarily selected to recognise a subset of specific DNA sequences and nucleate the recruitment of the transcriptional machinery. How TFs assemble and move in the nucleus to locate and bind their DNA targets and cause a transcriptional response, remains mostly unclear. NF-Y is a highly conserved, heterotrimeric TF with important roles in both housekeeping and lineage-specific gene expression, functioning as a promoter organiser. Despite a large number of biochemical, structural and genomic studies of NF-Y, there is a lack of experiments in single living cells; therefore, basic assumptions of NF-Y biology remain unproven in vivo. Here we employ a series of dynamic fluorescence microscopy methods (FLIM-FRET, NB, RICS and FRAP) to study NF-Y dynamics and complex formation in live cells. Specifically, we provide quantitative measurement of NF-Y subunit association and diffusion kinetics in the nucleus that collectively suggest NF-Y to move and bind chromatin as a trimeric complex in vivo.

DOI: https://doi.org/10.1038/s41598-021-90081-1
Nat Commun. 2021 May 25. 12(1): 3130

Ectopic targeting of CG DNA methylation in Arabidopsis with the bacterial SssI methyltransferase.

Wanlu Liu, Javier Gallego-Bartolomé, Yuxing Zhou, Zhenhui Zhong, Ming Wang, Somsakul Pop Wongpalee, Jason Gardiner, Suhua Feng, Peggy Hsuanyu Kuo, Steven E Jacobsen.

The ability to target epigenetic marks like DNA methylation to specific loci is important in both basic research and in crop plant engineering. However, heritability of targeted DNA methylation, how it impacts gene expression, and which epigenetic features are required for proper establishment are mostly unknown. Here, we show that targeting the CG-specific methyltransferase M.SssI with an artificial zinc finger protein can establish heritable CG methylation and silencing of a targeted locus in Arabidopsis. In addition, we observe highly heritable widespread ectopic CG methylation mainly over euchromatic regions. This hypermethylation shows little effect on transcription while it triggers a mild but significant reduction in the accumulation of H2A.Z and H3K27me3. Moreover, ectopic methylation occurs preferentially at less open chromatin that lacks positive histone marks. These results outline general principles of the heritability and interaction of CG methylation with other epigenomic features that should help guide future efforts to engineer epigenomes.

DOI: https://doi.org/10.1038/s41467-021-23346-y
Elife. 2021 May 27. pii: e67033. [Epub ahead of print]10

RSC primes the quiescent genome for hypertranscription upon cell cycle re-entry.

Christine E Cucinotta, Rachel H Dell, Keean Ca Braceros, Toshio Tsukiyama.

  Quiescence is a reversible G0 state essential for differentiation, regeneration, stem cell renewal, and immune cell activation. Necessary for long-term survival, quiescent chromatin is compact, hypoacetylated, and transcriptionally inactive. How transcription activates upon cell-cycle re-entry is undefined. Here we report robust, widespread transcription within the first minutes of quiescence exit. During quiescence, the chromatin-remodeling enzyme RSC was already bound to the genes induced upon quiescence exit. RSC depletion caused severe quiescence exit defects: a global decrease in RNA polymerase II (Pol II) loading, Pol II accumulation at transcription start sites, initiation from ectopic upstream loci, and aberrant antisense transcription. These phenomena were due to a combination of highly robust Pol II transcription and severe chromatin defects in the promoter regions and gene bodies. Together, these results uncovered multiple mechanisms by which RSC facilitates initiation and maintenance of large-scale, rapid gene expression despite a globally repressive chromatin state.

Keywords:  S. cerevisiae; chromosomes; gene expression

DOI:  https://doi.org/10.7554/eLife.67033
Development. 2021 May 24. pii: dev.199628. [Epub ahead of print]

LncRNA HBL1 is required for genome-wide PRC2 occupancy and function in cardiogenesis from human pluripotent stem cells.

Juli Liu, Sheng Liu, Lei Han, Yi Sheng, Yucheng Zhang, Il-Man Kim, Jun Wan, Lei Yang.

  Polycomb Repressive Complex 2 (PRC2) deposits H3K27me3 on chromatin to silence transcription. PRC2 broadly interacts with RNAs. Currently, the role of RNA- PRC2 interaction in human cardiogenesis remains elusive. Here, we found human-specific Heart Brake LncRNA 1 (HBL1) interacted with two PRC2 subunits, JARID2 and EED, in human pluripotent stem cells (hPSCs). Loss-of-JARID2, EED or HBL1 significantly enhanced cardiac differentiation from hPSCs. HBL1 depletion disrupted genome-wide PRC2 occupancy and H3K27me3 chromatin modification on essential cardiogenic genes, and broadly enhanced cardiogenic gene transcription in undifferentiated hPSCs and later-on differentiation. Additionally, ChIP-seq revealed reduced EED-occupancy on 62 overlapped cardiogenic genes in HBL1-/- and JARID2-/- hPSCs, indicating the epigenetic state of cardiogenic genes was determined by HBL1 and JARID2 at pluripotency stage. Furthermore, after cardiac development occurred, the cytosolic and nuclear fractions of HBL1 could crosstalk via a conserved "microRNA-1-JARID2" axis to modulate cardiogenic gene transcription. Overall, our findings delineate the indispensable role of HBL1 in guiding PRC2 function during early human cardiogenesis, and expand the mechanistic scope of lncRNA(s) that cytosolic and nuclear portions of HBL1 could coordinate to orchestrate human cardiogenesis.

Keywords:  Cardiac Differentiation; Heart Development; Long Non-Coding RNA; Pluripotent Stem Cells; Polycomb Repressive Complex 2

DOI:  https://doi.org/10.1242/dev.199628
Proc Natl Acad Sci U S A. 2021 Jun 01. pii: e2100699118. [Epub ahead of print]118(22):

Spreading and epigenetic inheritance of heterochromatin require a critical density of histone H3 lysine 9 tri-methylation.

Amber R Cutter DiPiazza, Nitika Taneja, Jothy Dhakshnamoorthy, David Wheeler, Sahana Holla, Shiv I S Grewal.

  Heterochromatin assembly requires methylation of histone H3 lysine 9 (H3K9me) and serves as a paradigm for understanding the importance of histone modifications in epigenetic genome control. Heterochromatin is nucleated at specific genomic sites and spreads across extended chromosomal domains to promote gene silencing. Moreover, heterochromatic structures can be epigenetically inherited in a self-templating manner, which is critical for stable gene repression. The spreading and inheritance of heterochromatin are believed to be dependent on preexisting H3K9 tri-methylation (H3K9me3), which is recognized by the histone methyltransferase Clr4/Suv39h via its chromodomain, to promote further deposition of H3K9me. However, the process involving the coupling of the "read" and "write" capabilities of histone methyltransferases is poorly understood. From an unbiased genetic screen, we characterize a dominant-negative mutation in histone H3 (H3G13D) that impairs the propagation of endogenous and ectopic heterochromatin domains in the fission yeast genome. H3G13D blocks methylation of H3K9 by the Clr4/Suv39h methyltransferase and acts in a dosage-dependent manner to interfere with the spreading and maintenance of heterochromatin. Our analyses show that the incorporation of unmethylatable histone H3G13D into chromatin decreases H3K9me3 density and thereby compromises the read-write capability of Clr4/Suv39h. Consistently, enhancing the affinity of Clr4/Suv39h for methylated H3K9 is sufficient to overcome the defects in heterochromatin assembly caused by H3G13D Our work directly implicates methylated histones in the transmission of epigenetic memory and shows that a critical density threshold of H3K9me3 is required to promote epigenetic inheritance of heterochromatin through the read-write mechanism.

Keywords:  epigenetic; gene silencing; heterochromatin; histone methylation

DOI:  https://doi.org/10.1073/pnas.2100699118
Nat Commun. 2021 May 28. 12(1): 3243

Impact of DNA methylation on 3D genome structure.

Diana Buitrago, Mireia Labrador, Juan Pablo Arcon, Rafael Lema, Oscar Flores, Anna Esteve-Codina, Julie Blanc, Nuria Villegas, David Bellido, Marta Gut, Pablo D Dans, Simon C Heath, Ivo G Gut, Isabelle Brun Heath, Modesto Orozco.

Determining the effect of DNA methylation on chromatin structure and function in higher organisms is challenging due to the extreme complexity of epigenetic regulation. We studied a simpler model system, budding yeast, that lacks DNA methylation machinery making it a perfect model system to study the intrinsic role of DNA methylation in chromatin structure and function. We expressed the murine DNA methyltransferases in Saccharomyces cerevisiae and analyzed the correlation between DNA methylation, nucleosome positioning, gene expression and 3D genome organization. Despite lacking the machinery for positioning and reading methylation marks, induced DNA methylation follows a conserved pattern with low methylation levels at the 5' end of the gene increasing gradually toward the 3' end, with concentration of methylated DNA in linkers and nucleosome free regions, and with actively expressed genes showing low and high levels of methylation at transcription start and terminating sites respectively, mimicking the patterns seen in mammals. We also see that DNA methylation increases chromatin condensation in peri-centromeric regions, decreases overall DNA flexibility, and favors the heterochromatin state. Taken together, these results demonstrate that methylation intrinsically modulates chromatin structure and function even in the absence of cellular machinery evolved to recognize and process the methylation signal.

DOI: https://doi.org/10.1038/s41467-021-23142-8
Nat Commun. 2021 May 28. 12(1): 3231

Genome information processing by the INO80 chromatin remodeler positions nucleosomes.

Elisa Oberbeckmann, Nils Krietenstein, Vanessa Niebauer, Yingfei Wang, Kevin Schall, Manuela Moldt, Tobias Straub, Remo Rohs, Karl-Peter Hopfner, Philipp Korber, Sebastian Eustermann.

The fundamental molecular determinants by which ATP-dependent chromatin remodelers organize nucleosomes across eukaryotic genomes remain largely elusive. Here, chromatin reconstitutions on physiological, whole-genome templates reveal how remodelers read and translate genomic information into nucleosome positions. Using the yeast genome and the multi-subunit INO80 remodeler as a paradigm, we identify DNA shape/mechanics encoded signature motifs as sufficient for nucleosome positioning and distinct from known DNA sequence preferences of histones. INO80 processes such information through an allosteric interplay between its core- and Arp8-modules that probes mechanical properties of nucleosomal and linker DNA. At promoters, INO80 integrates this readout of DNA shape/mechanics with a readout of co-evolved sequence motifs via interaction with general regulatory factors bound to these motifs. Our findings establish a molecular mechanism for robust and yet adjustable +1 nucleosome positioning and, more generally, remodelers as information processing hubs that enable active organization and allosteric regulation of the first level of chromatin.

DOI: https://doi.org/10.1038/s41467-021-23016-z
Nat Commun. 2021 May 28. 12(1): 3232

Ruler elements in chromatin remodelers set nucleosome array spacing and phasing.

Elisa Oberbeckmann, Vanessa Niebauer, Shinya Watanabe, Lucas Farnung, Manuela Moldt, Andrea Schmid, Patrick Cramer, Craig L Peterson, Sebastian Eustermann, Karl-Peter Hopfner, Philipp Korber.

Arrays of regularly spaced nucleosomes dominate chromatin and are often phased by alignment to reference sites like active promoters. How the distances between nucleosomes (spacing), and between phasing sites and nucleosomes are determined remains unclear, and specifically, how ATP-dependent chromatin remodelers impact these features. Here, we used genome-wide reconstitution to probe how Saccharomyces cerevisiae ATP-dependent remodelers generate phased arrays of regularly spaced nucleosomes. We find that remodelers bear a functional element named the 'ruler' that determines spacing and phasing in a remodeler-specific way. We use structure-based mutagenesis to identify and tune the ruler element residing in the Nhp10 and Arp8 modules of the INO80 remodeler complex. Generally, we propose that a remodeler ruler regulates nucleosome sliding direction bias in response to (epi)genetic information. This finally conceptualizes how remodeler-mediated nucleosome dynamics determine stable steady-state nucleosome positioning relative to other nucleosomes, DNA bound factors, DNA ends and DNA sequence elements.

DOI: https://doi.org/10.1038/s41467-021-23015-0
Bioinformatics. 2021 May 28. pii: btab410. [Epub ahead of print]

scDetect: a rank-based ensemble learning algorithm for cell type identification of single-cell RNA sequencing in cancer.

Yifei Shen, Qinjie Chu, Michael P Timko, Longjiang Fan.

MOTIVATION: Single-cell RNA sequencing (scRNA-seq) has enabled the characterization of different cell types in many tissues and tumor samples. Cell type identification is essential for single-cell RNA profiling, currently transforming the life sciences. Often, this is achieved by searching for combinations of genes that have previously been implicated as being cell-type specific, an approach that is not quantitative and does not explicitly take advantage of other scRNA-seq studies. Batch effects and different data platforms greatly decrease the predictive performance in inter-laboratory and different data type validation.RESULTS: Here, we present a new ensemble learning method named as "scDetect" that combines gene expression rank-based analysis and a majority vote ensemble machine-learning probability-based prediction method capable of highly accurate classification of cells based on scRNA-seq data by different sequencing platforms. Because of tumor heterogeneity, in order to accurately predict tumor cells in the single cell RNA-seq data, we have also incorporated cell copy number variation consensus clustering and epithelial score in the classification. We applied scDetect to scRNA-seq data from pancreatic tissue, mononuclear cells, and tumor biopsies cells and show that scDetect classified individual cells with high accuracy and better than other publicly available tools.
AVAILABILITY: scDetect is an open source software. Source code and test data is freely available from Github (https://github.com/IVDgenomicslab/scDetect/) and Zenodo (https://zenodo.org/record/4764132#.YKCOlrH5AYN). The examples and tutorial page is at https://ivdgenomicslab.github.io/scDetect-Introduction/.
SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

DOI: https://doi.org/10.1093/bioinformatics/btab410
Nucleic Acids Res. 2021 May 28. pii: gkab429. [Epub ahead of print]

DeepFun: a deep learning sequence-based model to decipher non-coding variant effect in a tissue- and cell type-specific manner.

Guangsheng Pei, Ruifeng Hu, Peilin Jia, Zhongming Zhao.

More than 90% of the genetic variants identified from genome-wide association studies (GWAS) are located in non-coding regions of the human genome. Here, we present a user-friendly web server, DeepFun (https://bioinfo.uth.edu/deepfun/), to assess the functional activity of non-coding genetic variants. This new server is built on a convolutional neural network (CNN) framework that has been extensively evaluated. Specifically, we collected chromatin profiles from ENCODE and Roadmap projects to construct the feature space, including 1548 DNase I accessibility, 1536 histone mark, and 4795 transcription factor binding profiles covering 225 tissues or cell types. With such comprehensive epigenomics annotations, DeepFun expands the functionality of existing non-coding variant prioritizing tools to provide a more specific functional assessment on non-coding variants in a tissue- and cell type-specific manner. By using the datasets from various GWAS studies, we conducted independent validations and demonstrated the functions of the DeepFun web server in predicting the effect of a non-coding variant in a specific tissue or cell type, as well as visualizing the potential motifs in the region around variants. We expect our server will be widely used in genetics, functional genomics, and disease studies.

DOI: https://doi.org/10.1093/nar/gkab429
Nat Commun. 2021 May 25. 12(1): 3145

Yeast cell fate control by temporal redundancy modulation of transcription factor paralogs.

Yan Wu, Jiaqi Wu, Minghua Deng, Yihan Lin.

Recent single-cell studies have revealed that yeast stress response involves transcription factors that are activated in pulses. However, it remains unclear whether and how these dynamic transcription factors temporally interact to regulate stress survival. Here we show that budding yeast cells can exploit the temporal relationship between paralogous general stress regulators, Msn2 and Msn4, during stress response. We find that individual pulses of Msn2 and Msn4 are largely redundant, and cells can enhance the expression of their shared targets by increasing their temporal divergence. Thus, functional redundancy between these two paralogs is modulated in a dynamic manner to confer fitness advantages for yeast cells, which might feed back to promote the preservation of their redundancy. This evolutionary implication is supported by evidence from Msn2/Msn4 orthologs and analyses of other transcription factor paralogs. Together, we show a cell fate control mechanism through temporal redundancy modulation in yeast, which may represent an evolutionarily important strategy for maintaining functional redundancy between gene duplicates.

DOI: https://doi.org/10.1038/s41467-021-23425-0
Sci Adv. 2021 May;pii: eabf5374. [Epub ahead of print]7(22):

BRD4 methylation by the methyltransferase SETD6 regulates selective transcription to control mRNA translation.

Zlata Vershinin, Michal Feldman, Thilo Werner, Lital Estrella Weil, Margarita Kublanovsky, Elina Abaev-Schneiderman, Menachem Sklarz, Enid Y N Lam, Khawla Alasad, Sarah Picaud, Barak Rotblat, Ruth A McAdam, Vered Chalifa-Caspi, Marcus Bantscheff, Trevor Chapman, Huw D Lewis, Panagis Filippakopoulos, Mark A Dawson, Paola Grandi, Rab K Prinjha, Dan Levy.

The transcriptional coactivator BRD4 has a fundamental role in transcription regulation and thus became a promising epigenetic therapeutic candidate to target diverse pathologies. However, the regulation of BRD4 by posttranslational modifications has been largely unexplored. Here, we show that BRD4 is methylated on chromatin at lysine-99 by the protein lysine methyltransferase SETD6. BRD4 methylation negatively regulates the expression of genes that are involved in translation and inhibits total mRNA translation in cells. Mechanistically, we provide evidence that supports a model where BRD4 methylation by SETD6 does not have a direct role in the association with acetylated histone H4 at chromatin. However, this methylation specifically determines the recruitment of the transcription factor E2F1 to selected target genes that are involved in mRNA translation. Together, our findings reveal a previously unknown molecular mechanism for BRD4 methylation-dependent gene-specific targeting, which may serve as a new direction for the development of therapeutic applications.

DOI: https://doi.org/10.1126/sciadv.abf5374
Nucleic Acids Res. 2021 May 22. pii: gkab392. [Epub ahead of print]

The nuclear periphery is a scaffold for tissue-specific enhancers.

Cheryl L Smith, Andrey Poleshko, Jonathan A Epstein.

Nuclear architecture influences gene regulation and cell identity by controlling the three-dimensional organization of genes and their distal regulatory sequences, which may be far apart in linear space. The genome is functionally and spatially segregated in the eukaryotic nucleus with transcriptionally active regions in the nuclear interior separated from repressive regions, including those at the nuclear periphery. Here, we describe the identification of a novel type of nuclear peripheral chromatin domain that is enriched for tissue-specific transcriptional enhancers. Like other chromatin at the nuclear periphery, these regions are marked by H3K9me2. But unlike the nuclear peripheral Lamina-Associated Domains (LADs), these novel, enhancer-rich domains have limited Lamin B interaction. We therefore refer to them as H3K9me2-Only Domains (KODs). In mouse embryonic stem cells, KODs are found in Hi-C-defined A compartments and feature relatively accessible chromatin. KODs are characterized by low gene expression and enhancers located in these domains bear the histone marks of an inactive or poised state. These results indicate that KODs organize a subset of inactive, tissue-specific enhancers at the nuclear periphery. We hypothesize that KODs may play a role in facilitating and perhaps constraining the enhancer-promoter interactions underlying spatiotemporal regulation of gene expression programs in differentiation and development.

DOI: https://doi.org/10.1093/nar/gkab392
Nucleic Acids Res. 2021 May 29. pii: gkab418. [Epub ahead of print]

GEPIA2021: integrating multiple deconvolution-based analysis into GEPIA.

Chenwei Li, Zefang Tang, Wenjie Zhang, Zhaochen Ye, Fenglin Liu.

In 2017, we released GEPIA (Gene Expression Profiling Interactive Analysis) webserver to facilitate the widely used analyses based on the bulk gene expression datasets in the TCGA and the GTEx projects, providing the biologists and clinicians with a handy tool to perform comprehensive and complex data mining tasks. Recently, the deconvolution tools have led to revolutionary trends to resolve bulk RNA datasets at cell type-level resolution, interrogating the characteristics of different cell types in cancer and controlled cohorts became an important strategy to investigate the biological questions. Thus, we present GEPIA2021, a standalone extension of GEPIA, allowing users to perform multiple interactive analysis based on the deconvolution results, including cell type-level proportion comparison, correlation analysis, differential expression, and survival analysis. With GEPIA2021, experimental biologists could easily explore the large TCGA and GTEx datasets and validate their hypotheses in an enhanced resolution. GEPIA2021 is publicly accessible at http://gepia2021.cancer-pku.cn/.

DOI: https://doi.org/10.1093/nar/gkab418
Cell Stem Cell. 2021 May 20. pii: S1934-5909(21)00181-8. [Epub ahead of print]

Phase separation of OCT4 controls TAD reorganization to promote cell fate transitions.

Jia Wang, Haopeng Yu, Qian Ma, Pengguihang Zeng, Danya Wu, Yingping Hou, Xinyi Liu, Lumeng Jia, Jun Sun, Yilong Chen, Diana Guallar, Miguel Fidalgo, Jiahao Chen, Yangyinhui Yu, Shaoshuai Jiang, Fenjie Li, Cai Zhao, Xianglin Huang, Jianlong Wang, Cheng Li, Yujie Sun, Xiaoxi Zeng, Wei Zhang, Yiliang Miao, Junjun Ding.

  Topological-associated domains (TADs) are thought to be relatively stable across cell types, although some TAD reorganization has been observed during cellular differentiation. However, little is known about the mechanisms through which TAD reorganization affects cell fate or how master transcription factors affect TAD structures during cell fate transitions. Here, we show extensive TAD reorganization during somatic cell reprogramming, which is correlated with gene transcription and changes in cellular identity. Manipulating TAD reorganization promotes reprogramming, and the dynamics of concentrated chromatin loops in OCT4 phase separated condensates contribute to TAD reorganization. Disrupting OCT4 phase separation attenuates TAD reorganization and reprogramming, which can be rescued by fusing an intrinsically disordered region (IDR) to OCT4. We developed an approach termed TAD reorganization-based multiomics analysis (TADMAN), which identified reprogramming regulators. Together, these findings elucidate a role and mechanism of TAD reorganization, regulated by OCT4 phase separation, in cellular reprogramming.

Keywords:  OCT4; TAD reorganization; TADMAN; phase separation; reprogramming

DOI:  https://doi.org/10.1016/j.stem.2021.04.023
Nucleic Acids Res. 2021 May 28. pii: gkab423. [Epub ahead of print]

Multiplexed single-cell profiling of chromatin states at genomic loci by expansion microscopy.

Marcus A Woodworth, Kenneth K H Ng, Aaron R Halpern, Nicholas A Pease, Phuc H B Nguyen, Hao Yuan Kueh, Joshua C Vaughan.

Proper regulation of genome architecture and activity is essential for the development and function of multicellular organisms. Histone modifications, acting in combination, specify these activity states at individual genomic loci. However, the methods used to study these modifications often require either a large number of cells or are limited to targeting one histone mark at a time. Here, we developed a new method called Single Cell Evaluation of Post-TRanslational Epigenetic Encoding (SCEPTRE) that uses Expansion Microscopy (ExM) to visualize and quantify multiple histone modifications at non-repetitive genomic regions in single cells at a spatial resolution of ∼75 nm. Using SCEPTRE, we distinguished multiple histone modifications at a single housekeeping gene, quantified histone modification levels at multiple developmentally-regulated genes in individual cells, and evaluated the relationship between histone modifications and RNA polymerase II loading at individual loci. We find extensive variability in epigenetic states between individual gene loci hidden from current population-averaged measurements. These findings establish SCEPTRE as a new technique for multiplexed detection of combinatorial chromatin states at single genomic loci in single cells.

DOI: https://doi.org/10.1093/nar/gkab423
Mol Metab. 2021 May 25. pii: S2212-8778(21)00101-0. [Epub ahead of print] 101256

Antagonistic epistasis of Hnf4α and FoxO1 metabolic networks through enhancer interactions in β-cell function.

Taiyi Kuo, Wen Du, Yasutaka Miyachi, Prasanna K Dadi, David A Jacobson, Daniel Segrè, Domenico Accili.

  Genetic and acquired abnormalities contribute to pancreatic β-cell failure in diabetes. Transcription factors Hnf4α (MODY1) and FoxO1 are respective examples of these two components, and act through β-cell-specific enhancers. However, their relationship is unclear. Here we show by genome-wide interrogation of chromatin modifications that ablation of FoxO1 in mature β-cells leads to an enrichment of active Hnf4α enhancers according to a HOMER analysis. To model the functional significance of this predicted unusual enhancer utilization we generated single and compound knockouts of FoxO1 and Hnf4α in β-cells. Single knockout of either gene impaired insulin secretion in mechanistically distinct fashions, as indicated by their responses to sulfonylurea and calcium fluxes. Surprisingly, the defective β-cell secretory function of either single mutant in hyperglycemic clamps and isolated islets treated with various secretagogues, was completely reversed in double mutants lacking FoxO1 and Hnf4α. Gene expression analyses revealed distinct epistatic modalities by which the two transcription factors regulate networks associated with reversal of β-cell dysfunction. An antagonistic network regulating glycolysis, including β-cell "disallowed" genes; and a synergistic network regulating protocadherins emerged as likely mediators of the functional restoration of insulin secretion. The findings provide evidence of antagonistic epistasis as a model of gene/environment interactions in the pathogenesis of β-cell dysfunction.

Keywords:  Foxo1; Hnf4a; beta cell; enhancer; epistasis; insulin

DOI:  https://doi.org/10.1016/j.molmet.2021.101256
Genome Biol. 2021 May 25. 22(1): 164

GRiNCH: simultaneous smoothing and detection of topological units of genome organization from sparse chromatin contact count matrices with matrix factorization.

Da-Inn Lee, Sushmita Roy.

  High-throughput chromosome conformation capture assays, such as Hi-C, have shown that the genome is organized into organizational units such as topologically associating domains (TADs), which can impact gene regulatory processes. The sparsity of Hi-C matrices poses a challenge for reliable detection of these units. We present GRiNCH, a constrained matrix-factorization-based approach for simultaneous smoothing and discovery of TADs from sparse contact count matrices. GRiNCH shows superior performance against seven TAD-calling methods and three smoothing methods. GRiNCH is applicable to multiple platforms including SPRITE and HiChIP and can predict novel boundary factors with potential roles in genome organization.

Keywords:  High-throughput chromosomal conformation capture (Hi-C); Matrix factorization; Three-dimensional (3D) genome organization; Topologically associating domains (TADs)

DOI:  https://doi.org/10.1186/s13059-021-02378-z
Nat Commun. 2021 May 25. 12(1): 3144

A 3D system to model human pancreas development and its reference single-cell transcriptome atlas identify signaling pathways required for progenitor expansion.

Carla A Gonçalves, Michael Larsen, Sascha Jung, Johannes Stratmann, Akiko Nakamura, Marit Leuschner, Lena Hersemann, Rashmiparvathi Keshara, Signe Perlman, Lene Lundvall, Lea Langhoff Thuesen, Kristine Juul Hare, Ido Amit, Anne Jørgensen, Yung Hae Kim, Antonio Del Sol, Anne Grapin-Botton.

Human organogenesis remains relatively unexplored for ethical and practical reasons. Here, we report the establishment of a single-cell transcriptome atlas of the human fetal pancreas between 7 and 10 post-conceptional weeks of development. To interrogate cell-cell interactions, we describe InterCom, an R-Package we developed for identifying receptor-ligand pairs and their downstream effects. We further report the establishment of a human pancreas culture system starting from fetal tissue or human pluripotent stem cells, enabling the long-term maintenance of pancreas progenitors in a minimal, defined medium in three-dimensions. Benchmarking the cells produced in 2-dimensions and those expanded in 3-dimensions to fetal tissue identifies that progenitors expanded in 3-dimensions are transcriptionally closer to the fetal pancreas. We further demonstrate the potential of this system as a screening platform and identify the importance of the EGF and FGF pathways controlling human pancreas progenitor expansion.

DOI: https://doi.org/10.1038/s41467-021-23295-6
BMC Bioinformatics. 2021 May 26. 22(1): 275

PTMViz: a tool for analyzing and visualizing histone post translational modification data.

Kevin Chappell, Stefan Graw, Charity L Washam, Aaron J Storey, Chris Bolden, Eric C Peterson, Stephanie D Byrum.

  BACKGROUND: Histone post-translational modifications (PTMs) play an important role in our system by regulating the structure of chromatin and therefore contribute to the regulation of gene and protein expression. Irregularities in histone PTMs can lead to a variety of different diseases including various forms of cancer. Histone modifications are analyzed using high resolution mass spectrometry, which generate large amounts of data that requires sophisticated bioinformatics tools for analysis and visualization. PTMViz is designed for downstream differential abundance analysis and visualization of both protein and/or histone modifications.RESULTS: PTMViz provides users with data tables and visualization plots of significantly differentiated proteins and histone PTMs between two sample groups. All the data is packaged into interactive data tables and graphs using the Shiny platform to help the user explore the results in a fast and efficient manner to assess if changes in the system are due to protein abundance changes or epigenetic changes. In the example data provided, we identified several proteins differentially regulated in the dopaminergic pathway between mice treated with methamphetamine compared to a saline control. We also identified histone post-translational modifications including histone H3K9me, H3K27me3, H4K16ac, and that were regulated due to drug exposure.
CONCLUSIONS: Histone modifications play an integral role in the regulation of gene expression. PTMViz provides an interactive platform for analyzing proteins and histone post-translational modifications from mass spectrometry data in order to quickly identify differentially expressed proteins and PTMs.

Keywords:  Differential abundance; Histone post-translational modifications; Proteomics; Shiny

DOI:  https://doi.org/10.1186/s12859-021-04166-9
Bioinformatics. 2021 May 26. pii: btab405. [Epub ahead of print]

TFcancer: a manually curated database of transcription factors associated with human cancer.

Qingqing Huang, Zhengtang Tan, Yanjing Li, Wenzhu Wang, Mei Lang, Changying Li, Zhiyun Guo.

SUMMARY: Transcription factors (TFs) are critical regulation elements and its dysregulation can lead to a variety of cancers. However, currently, there are no such online resources for large-scale collection, storage and analysis of TF-cancer associations in those cancers. To fill this gap, we present a database called TFcancer (http://lcbb.swjtu.edu.cn/tfcancer/), which contains 3,136 experimentally supported associations between 364 TFs and 33 TCGA cancers by manually curating more than 1,800 literature. TFcancer mainly concentrates on four aspects: TF expression, molecular alteration, regulatory relationships between TFs and target genes, and biological processes and signaling pathways of TFs in cancers. TFcancer not only provides a user-friendly interface for browsing and searching but also allows flexible data downloading and user data submitting. It is believed that TFcancer is a helpful and valuable resource for researchers who seek to understand the functions and molecular mechanisms of TFs involved in human cancers.AVAILABILITY: The TFcancer are freely available at http://lcbb.swjtu.edu.cn/tfcancer/.
SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

DOI: https://doi.org/10.1093/bioinformatics/btab405
Cell Rep. 2021 May 25. pii: S2211-1247(21)00528-3. [Epub ahead of print]35(8): 109183

Recognition of the inherently unstable H2A nucleosome by Swc2 is a major determinant for unidirectional H2A.Z exchange.

Linchang Dai, Xue Xiao, Lu Pan, Liuxin Shi, Ning Xu, Zhuqiang Zhang, Xiaoli Feng, Lu Ma, Shuoxing Dou, Pengye Wang, Bing Zhu, Wei Li, Zheng Zhou.

  The multisubunit chromatin remodeler SWR1/SRCAP/p400 replaces the nucleosomal H2A-H2B dimer with the free-form H2A.Z-H2B dimer, but the mechanism governing the unidirectional H2A-to-H2A.Z exchange remains elusive. Here, we perform single-molecule force spectroscopy to dissect the disassembly/reassembly processes of the H2A nucleosome and H2A.Z nucleosome. We find that the N-terminal 1-135 residues of yeast SWR1 complex protein 2 (previously termed Swc2-Z) facilitate the disassembly of nucleosomes containing H2A but not H2A.Z. The Swc2-mediated nucleosome disassembly/reassembly requires the inherently unstable H2A nucleosome, whose instability is conferred by three H2A α2-helical residues, Gly47, Pro49, and Ile63, as they selectively weaken the structural rigidity of the H2A-H2B dimer. It also requires Swc2-ZN (residues 1-37) that directly anchors to the H2A nucleosome and functions in the SWR1-catalyzed H2A.Z replacement in vitro and yeast H2A.Z deposition in vivo. Our findings provide mechanistic insights into how the SWR1 complex discriminates between the H2A nucleosome and H2A.Z nucleosome, establishing a simple paradigm for the governance of unidirectional H2A.Z exchange.

Keywords:  H2A.Z exchange; SWR1 complex; Swc2; nucleosome dynamics; single-molecule force spectroscopy

DOI:  https://doi.org/10.1016/j.celrep.2021.109183
Proc Natl Acad Sci U S A. 2021 Jun 01. pii: e2019699118. [Epub ahead of print]118(22):

High Yap and Mll1 promote a persistent regenerative cell state induced by Notch signaling and loss of p53.

Julian Heuberger, Johanna Grinat, Frauke Kosel, Lichao Liu, Séverine Kunz, Ramon Oliveira Vidal, Marlen Keil, Johannes Haybaeck, Sylvie Robine, Daniel Louvard, Christian Regenbrecht, Anje Sporbert, Sascha Sauer, Björn von Eyss, Michael Sigal, Walter Birchmeier.

  Specified intestinal epithelial cells reprogram and contribute to the regeneration and renewal of the epithelium upon injury. Mutations that deregulate such renewal processes may contribute to tumorigenesis. Using intestinal organoids, we show that concomitant activation of Notch signaling and ablation of p53 induce a highly proliferative and regenerative cell state, which is associated with increased levels of Yap and the histone methyltransferase Mll1. The induced signaling system orchestrates high proliferation, self-renewal, and niche-factor-independent growth, and elevates the trimethylation of histone 3 at lysine 4 (H3K4me3). We demonstrate that Yap and Mll1 are also elevated in patient-derived colorectal cancer (CRC) organoids and control growth and viability. Our data suggest that Notch activation and p53 ablation induce a signaling circuitry involving Yap and the epigenetic regulator Mll1, which locks cells in a proliferative and regenerative state that renders them susceptible for tumorigenesis.

Keywords:  Kmt2a; Notch; Yap; cancer; regeneration

DOI:  https://doi.org/10.1073/pnas.2019699118
BMC Bioinformatics. 2021 May 26. 22(1): 278

CpGmotifs: a tool to discover DNA motifs associated to CpG methylation events.

Giovanni Scala, Antonio Federico, Dario Greco.

  BACKGROUND: The investigation of molecular alterations associated with the conservation and variation of DNA methylation in eukaryotes is gaining interest in the biomedical research community. Among the different determinants of methylation stability, the DNA composition of the CpG surrounding regions has been shown to have a crucial role in the maintenance and establishment of methylation statuses. This aspect has been previously characterized in a quantitative manner by inspecting the nucleotidic composition in the region. Research in this field still lacks a qualitative perspective, linked to the identification of certain sequences (or DNA motifs) related to particular DNA methylation phenomena.RESULTS: Here we present a novel computational strategy based on short DNA motif discovery in order to characterize sequence patterns related to aberrant CpG methylation events. We provide our framework as a user-friendly, shiny-based application, CpGmotifs, to easily retrieve and characterize DNA patterns related to CpG methylation in the human genome. Our tool supports the functional interpretation of deregulated methylation events by predicting transcription factors binding sites (TFBS) encompassing the identified motifs.
CONCLUSIONS: CpGmotifs is an open source software. Its source code is available on GitHub https://github.com/Greco-Lab/CpGmotifs and a ready-to-use docker image is provided on DockerHub at https://hub.docker.com/r/grecolab/cpgmotifs .

Keywords:  DNA methylation; DNA methylation signature; DNA motifs; R-Shiny; Transcription factors

DOI:  https://doi.org/10.1186/s12859-021-04191-8
Nucleic Acids Res. 2021 May 28. pii: gkab405. [Epub ahead of print]

Mergeomics 2.0: a web server for multi-omics data integration to elucidate disease networks and predict therapeutics.

Jessica Ding, Montgomery Blencowe, Thien Nghiem, Sung-Min Ha, Yen-Wei Chen, Gaoyan Li, Xia Yang.

The Mergeomics web server is a flexible online tool for multi-omics data integration to derive biological pathways, networks, and key drivers important to disease pathogenesis and is based on the open source Mergeomics R package. The web server takes summary statistics of multi-omics disease association studies (GWAS, EWAS, TWAS, PWAS, etc.) as input and features four functions: Marker Dependency Filtering (MDF) to correct for known dependency between omics markers, Marker Set Enrichment Analysis (MSEA) to detect disease relevant biological processes, Meta-MSEA to examine the consistency of biological processes informed by various omics datasets, and Key Driver Analysis (KDA) to identify essential regulators of disease-associated pathways and networks. The web server has been extensively updated and streamlined in version 2.0 including an overhauled user interface, improved tutorials and results interpretation for each analytical step, inclusion of numerous disease GWAS, functional genomics datasets, and molecular networks to allow for comprehensive omics integrations, increased functionality to decrease user workload, and increased flexibility to cater to user-specific needs. Finally, we have incorporated our newly developed drug repositioning pipeline PharmOmics for prediction of potential drugs targeting disease processes that were identified by Mergeomics. Mergeomics is freely accessible at http://mergeomics.research.idre.ucla.edu and does not require login.

DOI: https://doi.org/10.1093/nar/gkab405
Commun Biol. 2021 May 25. 4(1): 623

Multi Locus View: an extensible web-based tool for the analysis of genomic data.

Martin J Sergeant, Jim R Hughes, Lance Hentges, Gerton Lunter, Damien J Downes, Stephen Taylor.

Tracking and understanding data quality, analysis and reproducibility are critical concerns in the biological sciences. This is especially true in genomics where next generation sequencing (NGS) based technologies such as ChIP-seq, RNA-seq and ATAC-seq are generating a flood of genome-scale data. However, such data are usually processed with automated tools and pipelines, generating tabular outputs and static visualisations. Interpretation is normally made at a high level without the ability to visualise the underlying data in detail. Conventional genome browsers are limited to browsing single locations and do not allow for interactions with the dataset as a whole. Multi Locus View (MLV), a web-based tool, has been developed to allow users to fluidly interact with genomics datasets at multiple scales. The user is able to browse the raw data, cluster, and combine the data with other analysis and annotate the data. User datasets can then be shared with other users or made public for quick assessment from the academic community. MLV is publically available at https://mlv.molbiol.ox.ac.uk .

DOI: https://doi.org/10.1038/s42003-021-02097-y
Cancer Res. 2021 May 28. pii: canres.0436.2021. [Epub ahead of print]

Epigenetic induction of mitochondrial fission is required for maintenance of liver cancer initiating cells.

Miaoling Tang, Meisongzhu Yang, Geyan Wu, Shuang Mo, Xingui Wu, Shuxia Zhang, Ruyuan Yu, Yameng Hu, Yingru Xu, Ziwen Li, Xinyi Liao, Jun Li, Libing Song.

Mitochondrial dynamics play vital roles in the tumorigenicity and malignancy of various types of cancers by promoting the tumor-initiating potential of cancer cells, suggesting that targeting crucial factors that drive mitochondrial dynamics may lead to promising anticancer therapies. In the current study, we report that overexpression of mitochondrial fission factor (MFF), which is upregulated significantly in liver cancer initiating cells (LCIC), promotes mitochondrial fission and enhances stemness and tumor-initiating capability in non-LCICs. MFF-induced mitochondrial fission evoked mitophagy and asymmetric stem cell division and promoted a metabolic shift from oxidative phosphorylation to glycolysis that decreased mitochondrial reactive oxygen species (ROS) production, which prevented ROS-mediated degradation of the pluripotency transcription factor OCT4. CRISPR affinity purification in situ of regulatory elements (CAPTURE) showed that T-Box transcription factor 19 (TBX19), which is overexpressed uniquely in LCICs compared to non-LCICs and liver progenitor cells, forms a complex with PRMT1 on the MFF promoter in LCICs, eliciting epigenetic histone H4R3me2a/H3K9ac-mediated transactivation of MFF. Targeting PRMT1 using furamidine, a selective pharmacological inhibitor, suppressed TBX19-induced mitochondrial fission, leading to a profound loss of self-renewal potential and tumor-initiating capacity of LCICs. These findings unveil a novel mechanism underlying mitochondrial fission-mediated cancer stemness and suggest that regulation of mitochondrial fission via inhibition of PRMT1 may be an attractive therapeutic option for liver cancer treatment.

DOI: https://doi.org/10.1158/0008-5472.CAN-21-0436