Cell Division Differentiation Research Articles

Gene expression dynamics during temperature-dependent sex determination in a sea turtle

Fifty years ago, researchers discovered a link between ambient temperature and the sex of turtle embryos. More recently, significant progress has been made in understanding the influence of temperature on freshwater turtles. However, our understanding of the key genetic factors in other turtle groups, such as sea turtles, remains limited. To address this gap, we conducted RNA-seq analyses on embryonic tissues from the sea olive ridley turtle during the thermosensitive period (stages 21–26) at temperatures known to produce males (26 °C) and females (33 °C). Our findings revealed that incubation temperatures primarily influence genes with broad expression across tissues due to differential cell division rates and later have an effect regulating gonad-specific transcripts. This effect is mostly related to gene activation rather than transcription repression. We performed transcriptome analyses following shifts in incubation temperatures of bi-potential gonads. This approach allowed us to identify genes that respond rapidly and may be closer to the beginning of the temperature-sensing pathway. Notably, we observed swift adaptations in the expression levels of chromatin modifiers JARID2 and KDM6B, as well as the splicing factor SRSF5, and transcription regulators THOC2, DDX3X and CBX3, but little impact in the overall gonad-specific pathways, indicating that temperature-sensing genes may change rapidly but the rewiring of the gonad's developmental fate is complex and resilient. Author summarySea turtles, one of the most iconic creatures of our oceans, confront a troubling reality of endangerment, a peril magnified by the looming specter of climate change. This climatic shift is gradually increasing the temperature of the nesting beaches thus causing dramatic male/female population biases. Conservation efforts will need genetic and molecular information to reverse the negative effects of climate change on the populations. In this study, we conducted the first transcriptomic analysis of embryonic tissues, including gonads, brain, liver, and mesonephros, in the olive ridley sea turtle during the critical thermosensitive period spanning stages 21–26. We examined both male-producing (26 °C) and female-producing (33 °C) temperatures and found that incubation temperatures influence temperature-sensitive genes that are either expressed globally or specifically associated with the gonads. These findings indicate that incubation temperatures predominantly sway genes with broad expression patterns due to differential cell division rates. This natural process was opted in the gonads to drive sex determination. We also identified genes that are rapidly capable of sensing temperature changes and that could play a role in the activation of the sex determination pathway. Overall, our study sheds light on the intricate interplay between temperature and gene expression during sea turtle development, revealing dynamic changes in the transcriptome and highlighting the involvement of key genetic players in sex determination.

StkP- and PhpP-Mediated Posttranslational Modifications Modulate the S. pneumoniae Metabolism, Polysaccharide Capsule, and Virulence.

Pneumococcal Ser/Thr kinase (StkP) and its cognate phosphatase (PhpP) play a crucial role in bacterial cytokinesis. However, their individual and reciprocal metabolic and virulence regulation-related functions have yet to be adequately investigated in encapsulated pneumococci. Here, we demonstrate that the encapsulated pneumococcal strain D39-derived D39ΔPhpP and D39ΔStkP mutants displayed differential cell division defects and growth patterns when grown in chemically defined media supplemented with glucose or nonglucose sugars as the sole carbon source. Microscopic and biochemical analyses supported by RNA-seq-based global transcriptomic analyses of these mutants revealed significantly down- and upregulated polysaccharide capsule formation and cps2 genes in D39ΔPhpP and D39ΔStkP mutants, respectively. While StkP and PhpP individually regulated several unique genes, they also participated in sharing the regulation of the same set of differentially regulated genes. Cps2 genes were reciprocally regulated in part by the StkP/PhpP-mediated reversible phosphorylation but independent of the MapZ-regulated cell division process. StkP-mediated dose-dependent phosphorylation of CcpA proportionately inhibited CcpA-binding to Pcps2A, supporting increased cps2 gene expression and capsule formation in D39ΔStkP. While the attenuation of the D39ΔPhpP mutant in two mouse infection models corroborated with several downregulated capsules-, virulence-, and phosphotransferase systems (PTS)-related genes, the D39ΔStkP mutant with increased amounts of polysaccharide capsules displayed significantly decreased virulence in mice compared to the D39 wild-type, but more virulence compared to D39ΔPhpP. NanoString technology-based inflammation-related gene expression and Meso Scale Discovery-based multiplex chemokine analysis of human lung cells cocultured with these mutants confirmed their distinct virulence phenotypes. StkP and PhpP may, therefore, serve as critical therapeutic targets.

Author Correction: Distinct strengths of mTORC1 control T-cell memory via transcriptional FOXO1 and metabolic AMPKα1 pathways in linear cell differentiation and asymmetric cell division models

Author Correction: Distinct strengths of mTORC1 control T-cell memory via transcriptional FOXO1 and metabolic AMPKα1 pathways in linear cell differentiation and asymmetric cell division models

Complex developmental and transcriptional dynamics underlie pollinator-driven evolutionary transitions in nectar spur morphology in Aquilegia (columbine).

Determining the developmental programs underlying morphological variation is key to elucidating the evolutionary processes that generated the stunning biodiversity of the angiosperms. Here, we characterized the developmental and transcriptional dynamics of the elaborate petal nectar spur of Aquilegia (columbine) in species with contrasting pollination syndromes and spur morphologies. We collected petal epidermal cell number and length data across four Aquilegia species, two with short, curved nectar spurs of the bee-pollination syndrome and two with long, straight spurs of the hummingbird-pollination syndrome. We also performed RNA-seq on A. brevistyla (bee) and A. canadensis (hummingbird) distal and proximal spur compartments at multiple developmental stages. Finally, we intersected these data sets with a previous QTL mapping study on spur length and shape to identify new candidate loci. The differential growth between the proximal and distal surfaces of curved spurs is primarily driven by differential cell division. However, independent transitions to straight spurs in the hummingbird syndrome have evolved by increasing differential cell elongation between spur surfaces. The RNA-seq data reveal these tissues to be transcriptionally distinct and point to auxin signaling as being involved with the differential cell elongation responsible for the evolution of straight spurs. We identify several promising candidate genes for future study. Our study, taken together with previous work in Aquilegia, reveals the complexity of the developmental mechanisms underlying trait variation in this system. The framework we established here will lead to exciting future work examining candidate genes and processes involved in the rapid radiation of the genus.

Distinct strengths of mTORC1 control T-cell memory via transcriptional FOXO1 and metabolic AMPKα1 pathways in linear cell differentiation and asymmetric cell division models.

Distinct strengths of mTORC1 control T-cell memory via transcriptional FOXO1 and metabolic AMPKα1 pathways in linear cell differentiation and asymmetric cell division models.

Auxin concentration and xylem production of Pinus massoniana in a subtropical forest in south China.

Auxin is involved in various developmental processes of plants, including cell division in cambium and xylem differentiation. However, most studies linking auxin and xylem cell production are performed in environments with a strong seasonality (i.e., temperate and boreal climates). The temporal dynamics of auxin and cambial activity of subtropical trees remain basically unknown. In this study, we sampled four microcores weekly in three individuals of Chinese red pine (Pinus massoniana Lamb.) from February to December 2015-16 to compare xylem formation with auxin concentration in subtropical China. During the entire period of sampling, the number of cambial cells varied from 2 to 7, while the number of cells in the enlarging zone ranged from 1 to 4 and from 1 to 5 in the wall-thickening zone. In 2015, the average auxin concentration was 3.46ngg-1, with 33 xylem cells being produced at the end of the year. In 2016, a lower auxin concentration (2.59ngg-1) corresponded to a reduced annual xylem production (13.7 cells). No significant relationship between auxin concentration and number of xylem cells in differentiation was found at the weekly scale. Unlike in boreal and temperate forests, the lack of wood formation seasonality in subtropical forests makes it more difficult to reveal the relationship between auxin concentration and number of xylem cells in differentiation at the intra-annual scale. The frequent and repeated samplings might have reduced auxin concentration in the developing cambium and xylem, resulting in a lower xylem cell production.

The Staufen1-dependent cell cycle regulon or how a misregulated RNA-binding protein leads to cancer.

In recent years, an increasing number of reports have linked the RNA-binding protein Staufen1 (STAU1) to the control of cell decision making. In non-transformed cells, STAU1 balances the expression of messenger RNA (mRNA) regulons that regulate differentiation and well-ordered cell division. Misregulation of STAU1 expression and/or functions changes the fragile balance in the expression of pro- and anti-proliferative and apoptotic genes and favours a novel equilibrium that supports cell proliferation and cancer development. The misregulation of STAU1 functions causes multiple coordinated modest effects in the post-transcriptional regulation of many RNA targets that code for cell cycle regulators, leading to dramatic consequences at the cellular level. The new tumorigenic equilibrium in STAU1-mediated gene regulation observed in cancer cells can be further altered by a slight increase in STAU1 expression that favours expression of pro-apoptotic genes and cell death. The STAU1-dependent cell cycle regulon is a good model to study how abnormal expression of an RNA-binding protein promotes cell growth and provides an advantageous selection of malignant cells in the first step of cancer development.

Depletion of kinesin motor KIF20A to target cell fate control suppresses medulloblastoma tumour growth

During mammalian brain development, neural progenitor cells proliferate extensively but can ensure the production of correct numbers of various types of mature cells by balancing symmetric proliferative versus asymmetric differentiative cell divisions. This process of cell fate determination may be harnessed for developing cancer therapy. Here, we test this idea by targeting KIF20A, a mitotic kinesin crucial for the control of cell division modes, in a genetic model of medulloblastoma (MB) and human MB cells. Inducible Kif20a knockout in both normal and MB-initiating granule neuron progenitors (GNPs) causes early cell cycle exit and precocious neuronal differentiation without causing cytokinesis failure and suppresses the development of Sonic Hedgehog (SHH)-activated MB. Inducible KIF20A knockdown in human MB cells inhibits proliferation both in cultures and in growing tumors. Our results indicate that targeting the fate specification process of nascent daughter cells presents a novel avenue for developing anti-proliferation treatment for malignant brain tumors.

Auxin-related genes associated with leaf petiole angle at the seedling stage are involved in adaptation to low temperature in Brassica napus

Rapeseed (Brassica napus L.) has winter and semi-winter ecotypes which need vernalization in the development process. Leaf development coincides with vernalization, and the leaf petiole angle (LPA) is associated with cold environmental adaptation during the overwintering period in rapeseed. In the present study, a genome-wide association study (GWAS) using a Brassica 60 K SNP array for LPA identified 45 significantly associated quantitative trait loci (QTLs) and 3525 candidate genes. Moreover, two rapeseed accessions (T193, winter type and T268, semi-winter type) with significantly different LPA degrees were used for RNA sequencing (RNA-seq). And transcriptome analysis revealed that most of the differentially expressed genes (DEGs) were involved in auxin, brassinosteroid (BR) and ethylene signaling pathways as well as the cell division pathway. Using integration of GWAS and RNA-seq analysis, several promising candidate genes, including BZR1, IAA7, IAA15, PIN3 and SPL orthologs were prioritized for further research. Quantitative real time PCR (qRT-PCR) also validated the differential expression patterns of nine candidate genes between T193 and T268. Through regulating differential cell division of the abaxial and adaxial petiole for small LPA, auxin signaling genes could be involved in the adaptation of rapeseed seedlings to low temperature conditions. These findings provide insights into the molecular mechanism of environmental adaptation at the seedling stage and provide valuable information for facilitating marker-based breeding in B. napus.

Leaf development in some ferns with variable dissection patterns (Dryopteridaceae and Lomariopsidaceae)

Although ferns are an important group for understanding leaf evolution, their developmental processes are still few understood. To better understand the organization of meristems and ontogeny of leaves in ferns, we studied their development in Elaphoglossum vagans (Mett.) Hieron., Lastreopsis amplissima (C. Presl) Tindale, Lomariopsis marginata (Schrad.) Kuhn, Mickelia scandens (Aubl.) R.C. Moran et al., Polybotrya cylindrica Kaulf., and Rumohra adiantiformis (G. Forst.) Ching, by using anatomical techniques. In addition, we observed herbarium specimens of those species, plus Elaphoglossum lingua (C. Presl) Brack, searching for unusual morphologies that may occur, that could represent a deviation from developmental processes here described. We propose hypotheses to explain these “natural mutants” based on known mechanisms of development.Shoot apical meristems of these species bear the typical tetrahedral cell with three dividing faces and the leaf apical meristem has two dividing faces. The leaf forms marginal meristems with enlarged initial cells, each of them bearing similarities to the leaf apical cell geometry and size, except in the simple-leaved fern Elaphoglossum vagans. Pinnae and segments primordia have grouped apical cells that are contiguous with their own marginal cells. In multi-pinnate leaves, one of these grouped apical cells in pinnae may enlarge and become similar to the leaf apical cell, indicating a reiterative process. The observed plants with unusual morphologies include plants with mixed reproductive identity, differential marginal growth, compromised apical growth, and reduced determinacy, supporting previous studies for leptosporangiate ferns.Apical and marginal cells of ferns may be homologous between them. Their characteristic geometry may allow specific cell divisions and their interconvertibility may drive the balance between apical and marginal growth. Unusual activity of these cells or changes in transcription factors may generate the observed abnormalities. A reiterative development in fern leaves supports the homology with shoots and the identity-in-parallel of their divisions.

Dimorphic Leaf Development of the Aquatic Plant Callitriche palustris L. Through Differential Cell Division and Expansion.

Heterophylly, or phenotypic plasticity in leaf form, is a remarkable feature of amphibious plants. When the shoots of these plants grow underwater, they often develop surprisingly different leaves from those that emerge in air. Among aquatic plants, it is typical for two or more distinct leaf development processes to be observed in the same individual exposed to different environments. Here, we analyze the developmental processes of heterophylly in the amphibious plant Callitriche palustris L. (Plantaginaceae). First, we reliably cultured this species under laboratory conditions and established a laboratory strain. We also established a framework for molecular-based developmental analyses, such as whole-mount in situ hybridization. We observed several developmental features of aerial and submerged leaves, including changes in form, stomata and vein formation, and transition of the meristematic zone. Then we defined developmental stages for C. palustris leaves. We found that in early stages, aerial and submerged leaf primordia had similar forms, but became discriminable through cell divisions with differential direction, and later became highly distinct via extensive cell elongation in submerged leaf primordia.

The Arabidopsis Receptor Kinase IRK Is Polarized and Represses Specific Cell Divisions in Roots.

Development of multicellular organisms requires coordination of cell division and differentiation across tissues. In plants, directional signaling, and implicitly cell polarity, is proposed to participate in this coordination; however, mechanistic links between intercellular signaling, cell polarity, and cellular organization remain unclear. Here, we investigate the localization and function of INFLORESCENCE AND ROOT APICES RECEPTOR KINASE (IRK) in root development. We find that IRK-GFP localizes to the outer plasma membrane domain in endodermal cells but localizes to different domains in other cell types. Our results suggest that IRK localization is informed locally by adjacent cell types. irk mutants have excess cell divisions in the ground tissue stem cells and endodermis, indicating IRK functions to maintain tissue organization through inhibition of specific cell divisions. We predict that IRK perceives a directional cue that negatively regulates these cell divisions, thus linking intercellular signaling and cell polarity with the control of oriented cell divisions during development.

Cell division method for mobility analysis of multi-loop mechanisms

This paper presents a combination of biological and screw theories for analyzing mechanisms, namely, cytology of mechanism (CM). In CM, kinematic pairs are considered as cells and they are the most important part of mechanisms. CM comprises four concepts, namely, cell division, cell division–differentiation, cell combination, and cell combination–variation. Consequently, mathematical models based on screw theory are employed to describe those concepts. A cell division method (CDM) is also proposed for mobility analysis of multi-loop mechanisms based on cell division. The basic principle of this method is to transform mobility analysis into constraint analysis between kinematic pairs and mobility analysis of parallel sub-mechanisms. Finally, CDM is used for analyzing the mobility of three typical multi-loop mechanisms to verify its correctness and practicality.

Histone Deacetylase HDA19 Affects Root Cortical Cell Fate by Interacting with SCARECROW.

The Arabidopsis (Arabidopsis thaliana) root epidermis is a simple model for investigating cell fate specification and pattern formation. In addition to regulatory networks consisting of transcription factors, histone deacetylases are also involved in the formation of cellular patterns. Here, we report thatHistone Deacetylase19 (HDA19) affects the root epidermal cellular pattern through regulation of cortical cell fate by interacting with SCARECROW (SCR). HDA19 binds to the DNA sequence upstream of SCR, as well as to those of several of SCR's target genes, and regulates their expression. Mutant lines of several SCR target genes show impaired patterns of epidermal differentiation and cortical cell division, similar to that of hda19 This work presents HDA19 and SCR as two further players in the regulation of cortical and epidermal cell specification and describes an additional function for SCR.

DOF2.1 Controls Cytokinin-Dependent Vascular Cell Proliferation Downstream of TMO5/LHW

SummaryTo create a three-dimensional structure, plants rely on oriented cell divisions and cell elongation. Oriented cell divisions are specifically important in procambium cells of the root to establish the different vascular cell types [1, 2]. These divisions are in part controlled by the auxin-controlled TARGET OF MONOPTEROS5 (TMO5) and LONESOME HIGHWAY (LHW) transcription factor complex [3, 4, 5, 6, 7]. Loss-of-function of tmo5 or lhw clade members results in strongly reduced vascular cell file numbers, whereas ectopic expression of both TMO5 and LHW can ubiquitously induce periclinal and radial cell divisions in all cell types of the root meristem. TMO5 and LHW interact only in young xylem cells, where they promote expression of two direct target genes involved in the final step of cytokinin (CK) biosynthesis, LONELY GUY3 (LOG3) and LOG4 [8, 9] Therefore, CK was hypothesized to act as a mobile signal from the xylem to trigger divisions in the neighboring procambium cells [3, 6]. To unravel how TMO5/LHW-dependent cytokinin regulates cell proliferation, we analyzed the transcriptional responses upon simultaneous induction of both transcription factors. Using inferred network analysis, we identified AT2G28510/DOF2.1 as a cytokinin-dependent downstream target gene. We further showed that DOF2.1 controls specific procambium cell divisions without inducing other cytokinin-dependent effects such as the inhibition of vascular differentiation. In summary, our results suggest that DOF2.1 and its closest homologs control vascular cell proliferation, thus leading to radial expansion of the root.

Single Cell Resolution of Glucocorticoid Effects on Erythroid Progenitor Cells

To generate the billions of new erythrocytes required on a daily basis, erythroid progenitor cells must exponentially increase in number before undergoing terminal differentiation. A limited number of cell divisions occur during erythropoietin (EPO)-regulated erythroid terminal differentiation, but the principal regulation of erythroid transit-amplification occurs earlier in erythropoiesis between the burst forming unit-erythroid (BFU-E) and colony forming unit-erythroid (CFU-E) stages of development. The importance of this EPO-independent early erythropoietic process is highlighted in Diamond-Blackfan Anemia (DBA). DBA is characterized by pure red cell aplasia, loss of BFU-E and CFU-E progenitors in the bone marrow, and severe anemia despite high circulating EPO levels. The only known effective medical therapy for DBA also provides insight into the regulation of erythroid transit-amplification. In patients responsive to glucocorticoid treatment, there are increased numbers of BFU-E and CFU-E progenitors in the bone marrow, and ex vivo culture studies indicate that the synthetic glucocorticoid dexamethasone (Dex) predominantly increases proliferative capacity of BFU-Es, with minimal effect on proliferative capacity of CFU-Es. These findings led to a prevailing model that glucocorticoids increase BFU-E proliferative capacity by stimulating several rounds of self-renewal cell divisions. However, a limitation of this model is that there is no mechanistic explanation for how BFU-Es regulate the fate choice of undergoing a self-renewal cell division versus a differentiation cell division in the presence or absence of glucocorticoids. In this study, we address this question by examining progression of early erythroid progenitor development at single cell resolution, and subsequently elucidate the true mechanistic nature of glucocorticoid-induced erythroid progenitor proliferative capacity amplification. By performing single cell transcriptome profiling (scRNAseq) of primary-isolated mouse fetal liver BFU-Es, CFU-Es, and their developmental intermediates, we identify a continuum of transcriptomic states during erythroid transit-amplification when performing principle component analysis (PCA) on transcriptomes of individual cells. We show that ex vivo culture of primary-isolated BFU-Es in serum free media supplemented with stem cell factor, insulin-like growth factor 1, and EPO results in developmental progression along the transcriptome continuum when performing scRNAseq and PCA on cultured BFU-Es. The addition of Dex into this culture system does not result in self-renewal of BFU-Es at the transcriptome level, but rather still results in developmental progression, albeit to less of a degree per cell division than BFU-Es cultured without Dex. We additionally show that the continuum of transcriptome states in erythroid transit-amplification is reflective of a continuum of functional states, with developmental progression characterized by decreasing proliferative capacity and decreasing glucocorticoid-responsiveness. Lastly, through manual separation of daughter cells resulting from a BFU-E cell division, we demonstrate that BFU-E cell division is a symmetric process at the transcriptome level, both with and without the addition of Dex. Our results clarify the nature of how glucocorticoids amplify BFU-E proliferative capacity. As opposed to stimulating a finite number of BFU-E self-renewal cell divisions, glucocorticoids decrease the extent of progression through the erythroid transit-amplifying developmental continuum per cell division. Thus, a decreased rate of progression through the developmental continuum is associated with an increased number of transit-amplifying cell divisions prior to terminal differentiation. These findings are important not only for the rational development of glucocorticoid-alternatives for treating DBA, but also for all bone marrow failure syndromes characterized by progenitor cell hypoplasia. DisclosuresNo relevant conflicts of interest to declare.

E2F1 Mediates the Retinoic Acid-Induced Transcription of Tshz1 during Neuronal Differentiation in a Cell Division-Dependent Manner.

The involvement of cell division in cellular differentiation has long been accepted. Cell division may be required not only for the expansion of a differentiated cell population but also for the execution of differentiation processes. Nonetheless, knowledge regarding how specific differentiation processes are controlled in a cell division-dependent manner is far from complete. Here, we determined the involvement of cell division in neuronal differentiation. We initially confirmed that cell division is an essential event for the neuronal differentiation of P19 embryonic carcinoma cells. We investigated the induction mechanisms of Tshz1, whose expression is induced by retinoic acid (RA) in a cell division-dependent manner. Promoter analysis of Tshz1 revealed a specific region required for RA-dependent transcription. A series of experiments was used to identify E2F1 as the induction factor for the RA-dependent transcription of Tshz1 We propose that E2F1 mediates neuronal differentiation in a cell division-dependent manner.

Lgl1 controls NG2 endocytic pathway to regulate oligodendrocyte differentiation and asymmetric cell division and gliomagenesis

Oligodendrocyte progenitor cells (OPC) undergo asymmetric cell division (ACD) to generate one OPC and one differentiating oligodendrocyte (OL) progeny. Loss of pro-mitotic proteoglycan and OPC marker NG2 in the OL progeny is the earliest immunophenotypic change of unknown mechanism that indicates differentiation commitment. Here, we report that expression of the mouse homolog of Drosophila tumor suppressor Lethal giant larvae 1 (Lgl1) is induced during OL differentiation. Lgl1 conditional knockout OPC progeny retain NG2 and show reduced OL differentiation, while undergoing more symmetric self-renewing divisions at the expense of asymmetric divisions. Moreover, Lgl1 and hemizygous Ink4a/Arf knockouts in OPC synergistically induce gliomagenesis. Time lapse and total internal reflection microscopy reveals a critical role for Lgl1 in NG2 endocytic routing and links aberrant NG2 recycling to failed differentiation. These data establish Lgl1 as a suppressor of gliomagenesis and positive regulator of asymmetric division and differentiation in the healthy and demyelinated murine brain.

Analysis of KRT1, KRT10, KRT19, TP53 and MMP9 expression in pediatric and adult cholesteatoma.

Cholesteatoma is an epidermal cyst with still unknown pathomechanism. The aim of the current study was to investigate molecular differences in the background of the hyperproliferative property and aggressive behavior typical of the cholesteatoma epithelium. The expression of three cytokeratin genes (KRT1, KRT10 and KRT19), the matrix metalloproteinase 9 gene (MMP9) and the tumor suppressor TP53 gene was measured by qRT-PCR in surgical samples of pediatric and adult cholesteatoma cases and their expression level was compared to that of normal skin samples from the retroauricular region of control individuals. Cholesteatoma samples were stratified according to the age of onset and recurrence for more detailed analysis. Our results showed identical expression pattern for KRT1 and KRT10, their expression was higher in pediatric cases than in adults, especially in pediatric recurrent samples. The expression level of KRT19 was inversely proportional to that of KRT1/KRT10, it was lower in the more invasive recurrent cases both in our pediatric and adult groups. As it was expected from the bone destructive behavior of cholesteatoma, a significantly elevated expression of MMP9 was measured in cholesteatoma samples, the highest level was found in adult recurrent cases. Low expression levels characterize the TP53 gene without significant differences in our samples. These findings demonstrate that cytokeratin expression distinguishes between pediatric/adult, nonrecurrent/recurrent cases, suggesting that distinct differentiation state and cell division potential characterize these cholesteatoma cases. KRT19 with a tumor suppressor potential might restrict the recurrence of cholesteatoma. The differences observed in gene expression profiles between cholesteatoma and control samples support the notion that cholesteatoma is a cystic lesion with tumor-like behavior because it is characterized by invasive, destructive growth and high tendency for recurrence.

Plasma cell differentiation is controlled by multiple cell division-coupled epigenetic programs

The genomic loci associated with B cell differentiation that are subject to transcriptional and epigenetic regulation in vivo are not well defined, leaving a gap in our understanding of the development of humoral immune responses. Here, using an in vivo T cell independent B cell differentiation model, we define a cellular division-dependent cis-regulatory element road map using ATAC-seq. Chromatin accessibility changes correlate with gene expression and reveal the reprogramming of transcriptional networks and the genes they regulate at specific cell divisions. A subset of genes in naive B cells display accessible promoters in the absence of transcription and are marked by H3K27me3, an EZH2 catalyzed repressive modification. Such genes encode regulators of cell division and metabolism and include the essential plasma cell transcription factor Blimp-1. Chemical inhibition of EZH2 results in enhanced plasma cell formation, increased expression of the above gene set, and premature expression of Blimp-1 ex vivo. These data provide insights into cell-division coupled epigenetic and transcriptional processes that program plasma cells.