Anguera Laboratory

XIST FISH, Anguera Lab, Penn VetOur laboratory investigates X-chromosome Inactivation, and how this epigenetic process contributes to female-biased autoimmunity.

We are investigating how female lymphocytes maintain X-chromosome Inactivation, which is an epigenetic process responsible for equalizing gene expression between sexes.  X-chromosome Inactivation silences one X-chromosome in female cells, and this process is initiated and maintained by the long noncoding RNA Xist.

Lab Mission:

  • To understand the female bias underlying autoimmune disorders such as lupus
  • To investigate how X-Chromosome Inactivation is maintained in female lymphocytes and becomes mis-regulated in female-biased autoimmune disorders
  • To investigate the function and mechanisms for novel X-linked long noncoding RNAs important for early human development
  • To identify therapeutic opportunities for correcting X-linked dosage imbalances in autoimmune disorders 

Interested in Working With Us?

We are always seeking highly motivated students and post-doctoral fellows with an interest in:

  • Epigenetics, X-chromosome Inactivation, Imprinting
  • Immunology and female-biased autoimmune disorders
  • Long noncoding RNAs
  • Genetics
  • Bioinformatics

Interested post-doctoral candidates should inquire by sending e-mail to anguera@vet.upenn.edu

Interested graduate students should visit the Department of Cellular and Molecular Biology (CAMB) at UPenn or inquire by sending e-mail to: anguera@vet.upenn.edu

Contact Information

The Anguera Lab
University of Pennsylvania
School of Veterinary Medicine
Department of Biomedical Studies
Rosenthal Building, Room 304
Philadelphia, PA 19104

3800 Spruce St., Rm 390EB
Office Phone: 215.898.0567
Lab Phone: 215.898.9541

FAX: 215.573.6810

anguera@vet.upenn.edu

X-Chromosome Inactivation (XCI) is one of theCalico cats demonstrate x-mosaicism best-characterized epigenetic phenomena where long noncoding RNAs are key players that regulate gene expression. Female mammals (XX) have two X-chromosomes, and one X is randomly chosen for transcriptional silencing in order to equalize the expression of X-linked genes compared to males (XY).  Thus females are mosaic for X-chromosome expression, where a cell will express the maternal or paternal X.  One great example of female X-mosaicism are calico cats.

The X-chromosome contains about 1000 genes with roles in development, reproduction, metabolism, and immunity. Mammalian females (including humans) are healthier and have longer life-spans than males. Females have better survival rates than males from illnesses caused by infectious diseases, injury, or sepsis. The X-chromosome plays an important role in the hyper-responsive female immune system because this chromosome has the highest density of immune-related genes. Females are also more susceptible to a number of autoimmune diseases, suggesting that the enhanced immune function may be related to the loss of self-tolerance.

The X-chromosome contains about 1000 genes with roles in development, reproduction, metabolism, and immunity. Mammalian females (including humans) are healthier and have longer life-spans than males. Females have better survival rates than males from illnesses caused by infectious diseases, injury, or sepsis. The X-chromosome plays an important role in the hyper-responsive female immune system because this chromosome has the highest density of immune-related genes. Females are also more susceptible to a number of autoimmune diseases, suggesting that the enhanced immune function may be related to the loss of self-tolerance.

XCI occurs during early female embryonic development, and is initiated by expression of Xist RNA which recruits heterochromatic complexes for transcriptional silencing.  Xist RNA and heterochromatin modifications associate with the inactive X chromosome with each cell division, and can be visualized using fluorescence microscopy. Xist RNA also functions to maintain XCI in somatic cells.

We recently discovered that XCI is maintained differently in female lymphocytes. Naïve T and B cells, which are quiescent, lack Xist RNA and heterochromatin marks on the inactive X. After lymphocyte stimulation, these epigenetic modifications re-localize back to the inactive X, as the cells re-enter the cell cycle and actively proliferate. This dynamic mechanism for XCI maintenance is unique to immune cells, and our lab investigates the molecular details of this process. The major areas of investigation in our lab are: (1) to understand how XCI is maintained in female lymphocytes, and how this contributes to female-biased autoimmunity; (2) to investigate epigenetic mechanisms involving long noncoding RNAs during early human development and placental progenitors.

Penn Vet banner MCA 2016 PNAS

Research Techniques

  • Microscopy: RNA and DNA fluorescence in situ hybridization (FISH), immunofluorescence detection of proteins and chromatin modifications
  • Cell culture of human and mouse immune cells
  • Mouse models of autoimmunity
  • Genome editing using TALENs and CRISPRs for introducing mutations of coding and noncoding genes
  • Molecular Biology: qPCR and next-generation sequencing technologies (RNAseq, CHIRPseq, CUT&RUN)


Mouse Sic region contains chromatin modifications

XCI maintenance is different in mammalian female lymphocytes

We have discovered that the inactive X chromosome lacks the typical heterochromatic modifications in female mature naïve T and B cells. This is the first example of physiologically-relevant female somatic cells that don’t have XIST/Xist RNA, H3K27me3, macroH2A, H2A-ubiquitin, and H4K20me modifications enriched on the inactive X. We also observed that in vitro activation of T and B cells stimulates the return of XIST/Xist RNA and some heterochromatic modifications back to the inactive X.

naive vs activated human T Cells

Using loss of function assays, we identified protein factors responsible for localizing XIST/Xist RNA back to the inactive X during lymphocyte activation.  YY1 and hnRNP U, known XIST RNA binding proteins, return the XIST/Xist transcripts back to the inactive X. Elucidating the molecular mechanism of XIST/Xist RNA relocalization back to the inactive X, and how this transcript initiates the return of heterochromatin modifications, will reveal how the memory of transcriptional silencing is maintained after cell division in lymphocytes.

 XIST/Xist RNA localization on the inactive X is perturbed in lymphocytes from female lupus patients and lupus mouse models

Remarkably, we have recently found that female lupus patients exhibit differences with XCI compared to healthy controls, which may explain the female bias of autoimmune disorders and why female lupus patients express higher levels of X-linked autoimmunity related genes compared to male patients. SLE-patient B cells have fewer typical XIST RNA clouds and more cells with dispersed patterns of XIST RNA or completely lacking XIST localization. Abnormal Xist RNA localization suggests partial reactivation of the inactive X-chromosome, which could increase gene expression. Using RNA FISH, we demonstrated that female lupus T and B cells frequently exhibit biallelic expression of CD40LG, CXCR3 and TLR7, which correlates with elevated expression of these genes. Moving forward, we are investigating causality of partial X-reactivation for lupus pathogenesis using mouse models. These will be the first experiments that directly connect transcription from the inactive X to lupus pathology.

healthy female activated T cells SLE female activated T cells
  

Diversity of XCI maintenance mechanisms in immune cells

Our lab recently found that other immune cell populations, including dendritic cells, NK cells, and monocytes, have different mechanisms to maintain XCI.  Using single-cell microscopy to image Xist RNA and heterochromatin modifications, we observed that monocytes and NK cells have faint Xist RNA ‘clouds’ and H3K27me3 foci, yet plasmacytoid dentritic cells (p-DCs) completely lack Xist RNA signals and enrichment of this histone mark. Unlike in humans, mouse p-DCs do not exhibit sex differences with interferon alpha production, and interferon signature gene expression in p-DCs is similar between males and females.  Thus, immune cells use diverse (& unknown) mechanisms to maintain XCI, and we propose that these mechanisms are susceptible to error, thereby contributing to sex-linked autoimmune diseases. Our lab is actively investigating the molecular mechanisms that maintain XCI in a variety of immune cells.

Anguera Lab - Immune Cells
DC cells - anguera lab
Hantsoo Liisa, Kornfield Sara, Anguera Montserrat C, Epperson C Neill: Inflammation: A Proposed Intermediary Between Maternal Stress and Offspring Neuropsychiatric Risk. [PMID 30314641] Biological psychiatry 85(2): 97-106, Jan 2019.

Sierra Isabel, Anguera Montserrat C: Enjoy the silence: X-chromosome inactivation diversity in somatic cells.[PMID 31108425Current opinion in genetics & development 55: 26-31, May 2019.

Syrett Camille M, Anguera Montserrat C: When the balance is broken: X-linked gene dosage from two X chromosomes and female-biased autoimmunity. [PMID 31125996Journal of leukocyte biology May 2019.

Kotzin Jonathan J, Iseka Fany, Wright Jasmine, Basavappa Megha G, Clark Megan L, Ali Mohammed-Alkhatim, Abdel-Hakeem Mohamed S, Robertson Tanner F, Mowel Walter K, Joannas Leonel, Neal Vanessa D, Spencer Sean P, Syrett Camille M, Anguera Montserrat C, Williams Adam, Wherry E John, Henao-Mejia Jorge: The long noncoding RNA regulates CD8 T cells in response to viral infection.[PMID 31138702Proceedings of the National Academy of Sciences of the United States of America 116(24): 11916-11925, Jun 2019.

Syrett Camille M, Paneru Bam, Sandoval-Heglund Donavon, Wang Jianle, Banerjee Sarmistha, Sindhava Vishal, Behrens Edward M, Atchison Michael, Anguera Montserrat C: Altered X-chromosome inactivation in T cells may promote sex-biased autoimmune diseases. [PMID 30944248 JCI insight 4(7), Apr 2019.

Syrett Camille M, Sindhava Vishal, Sierra Isabel, Dubin Aimee H, Atchison Michael, Anguera Montserrat C: Diversity of Epigenetic Features of the Inactive X-Chromosome in NK Cells, Dendritic Cells, and Macrophages. [PMID 30671059Frontiers in immunology 9: 3087, 2018.

Le Coz Carole, Trofa Melissa, Syrett Camille M, Martin Anna, Jyonouchi Harumi, Jyonouchi Soma, Anguera Montserrat C, Romberg Neil: CD40LG duplication-associated autoimmune disease is silenced by nonrandom X-chromosome inactivation.[PMID 29499223] The Journal of allergy and clinical immunology 141(6): 2308-2311.e7, Jun 2018.

Syrett Camille M, Sierra Isabel, Berry Corbett L, Beiting Daniel, Anguera Montserrat C: Sex-Specific Gene Expression Differences Are Evident in Human Embryonic Stem Cells and During In Vitro Differentiation of Human Placental Progenitor Cells. [PMID 29993333] Stem cells and development 27(19): 1360-1375, Oct 2018.

Wang Jianle, Anguera Montserrat C: In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells. [PMID 28362386] Journal of visualized experiments : JoVE(121), Mar 2017.

Syrett Camille M, Sindhava Vishal, Hodawadekar Suchita, Myles Arpita, Liang Guanxiang, Zhang Yue, Nandi Satabdi, Cancro Michael, Atchison Michael, Anguera Montserrat C: Loss of Xist RNA from the inactive X during B cell development is restored in a dynamic YY1-dependent two-step process in activated B cells. [PMID 28991910] PLoS genetics 13(10): e1007050, Oct 2017.

Penkala Ian, Wang Jianle, Syrett Camille M, Goetzl Laura, López Carolina B, Anguera Montserrat C: lncRHOXF1, a Long Noncoding RNA from the X Chromosome That Suppresses Viral Response Genes during Development of the Early Human Placenta. [PMID 27066803] Molecular and cellular biology 36(12): 1764-75, Jun 2016.

Wang Jianle, Syrett Camille M, Kramer Marianne C, Basu Arindam, Atchison Michael L, Anguera Montserrat C: Unusual maintenance of X chromosome inactivation predisposes female lymphocytes for increased expression from the inactive X. [PMID 27001848] Proceedings of the National Academy of Sciences of the United States of America 113(14): E2029-38, Apr 2016.

Luo Mengcheng, Zhou Jian, Leu N Adrian, Abreu Carla M, Wang Jianle, Anguera Montserrat C, de Rooij Dirk G, Jasin Maria, Wang P Jeremy: Polycomb protein SCML2 associates with USP7 and counteracts histone H2A ubiquitination in the XY chromatin during male meiosis. [PMID 25634095] PLoS genetics 11(1): e1004954, Jan 2015.

Lessing D, Anguera MC, Lee JT.: X chromosome inactivation and epigenetic responses to cellular reprogramming. [PMID 23662665] Annu Rev Genomics Hum Genet. 14: 85-110, May 2013.

Anguera Montserrat C, Sadreyev Ruslan, Zhang Zhaoqing, Szanto Attila, Payer Bernhard, Sheridan Steven D, Kwok Showming, Haggarty Stephen J, Sur Mriganka, Alvarez Jason, Gimelbrant Alexander, Mitalipova Maisam, Kirby James E, Lee Jeannie T: Molecular signatures of human induced pluripotent stem cells highlight sex differences and cancer genes. [PMID 22770242] Cell stem cell 11(1): 75-90, Jul 2012.

Anguera Montserrat C, Ma Weiyuan, Clift Danielle, Namekawa Satoshi, Kelleher Raymond J, Lee Jeannie T: Tsx produces a long noncoding RNA and has general functions in the germline, stem cells, and brain. [PMID 21912526] PLoS genetics 7(9): e1002248, Sep 2011.

Kim Daniel H, Jeon Yesu, Anguera Montserrat C, Lee Jeannie T: X-chromosome epigenetic reprogramming in pluripotent stem cells via noncoding genes. [PMID 21376830] Seminars in cell & developmental biology 22(4): 336-42, Jun 2011.

Field Martha S, Anguera Montserrat C, Page Rodney, Stover Patrick J: 5,10-Methenyltetrahydrofolate synthetase activity is increased in tumors and modifies the efficacy of antipurine LY309887. [PMID 19022216] Archives of biochemistry and biophysics 481(2): 145-50, Jan 2009.

Anguera Montserrat C, Liu Matthew, Avruch Joseph, Lee Jeannie T: Characterization of two Mst1-deficient mouse models. [PMID 18942145] Developmental dynamics : an official publication of the American Association of Anatomists 237(11): 3424-34, Nov 2008.

  • Montserrat was promoted to Associate Professor with tenure! (July 2019)
  • Natalie Toothacre (CAMB program, G&E) joined our lab – WELCOME!!  (May 2019)
  • Welcome to Makheni Jean-Pierre, who joined us during the summer of 2019 with the Penn Summer Internship Program (SUIP) (Summer 2019)
  • Welcome to Penn Vet student, Kaitlyn Cassel, who joined us during the summer of 2019 for the Penn Vet BI Summer Research Program (Summer 2019)
  • Camille’s review article (“When the balance is broken”) on X-linked gene dosage and female-biased autoimmunity is accepted! Yet another witty title for a review article from our lab. (May 2019)
  • Zack passed his prelim, and is an official candidate for the PhD program in the Penn CAMB DSRB program!! (May 2019)
  • Anguera lab graduates for 2019: Congratulations to Don, who graduated with a B.S. with honors from Penn; Congrats to Steve Hayes, who graduated from Penn Vet also with honors; Congrats to Anna Martin, who also graduated from Penn Vet! (May 2019)
  • Montserrat & Camille participated in the first Anguera lab hooding ceremony, for the Penn BGS Graduation! (May 2019)
  • Isabel’s review article (“Enjoy the silence”) on XCI diversity in somatic cells is accepted! (April 2019)
  • Camille defended her dissertation with flying colors, and joined Gerd Blobel’s lab at CHOP.  We wish her all the best! (March 2019)
  • Welcome to Cardboard_Camille, our special 2D slightly-larger-than-life replica of Camille. She likes to move around the 3rd floor of Rosenthal and surprise members of other labs. (March 2019)
  • Mr. Sparkles continued to get bigger (& sassier) in the lab, demanding more frequent feedings. (Spring 2019)
  • Our lab’s first paper on XCI in T cells is accepted at JCI Insight!! (March 2019)
  • Camille presented her research in a short talk at the 2019 Keystone RNAs in Immunity meeting (Feb 2019)
  • Camille & Isabel’s paper on XCI features in macrophages, dendritic cells, and NK cells was accepted!! (January 2019)
  • Camille’s manuscript on sex differences in human ESCs and early placental progenitors was accepted!! (October 2018)
  • Camille, Isabel, & Montserrat presented at the 2018 CSHL Epigenetics meeting in Cold Spring Harbor, NY (September 2018)
  • Welcome to the lab, Zachary Beetham! Zack is a CAMB G&E student (June 2018)
  • Welcome Natalia Enid Aponte Borges! Natalia is part of the Penn Summer Undergraduate Internship Program (June 2018)
  • Congratulations to Don who is doing a summer research program in NYC at Sloan Kettering (June 2018)
  • Farewell Anna, we wish you all the best in your final year of Vet School (May 2018)
  • The Anguera lab welcomes Aimee Dubin as our new Research Specialist! (May 2018)
  • Congratulations to Isabel for being appointed to the T-32 in Immune System Development and Regulation (May 2018)
  • Isabel passed her prelim with flying colors and is officially a PhD candidate in DSRB!!! (May 2018)
  • Anna presented her research at the 2018 AAI meeting in Austin, Texas (May 2018)
  • Congratulations to Montserrat for receiving an NIAID R01 and a Lupus Foundation of America D.O.D grant!! Read about it here (May 2018)
  • Anna wins first place for her talk at the Vet Student Research Day! (March 2018)
  • Turns out Sparkles is a boy, and he celebrates his first birthday this month. Happy birthday, Mr. Sparkles! (March 2018)
  • Montserrat and Camille attended the 2018 Midwinter Conference of Immunologists in Monterey, CA (January 2018)
  • Camille was selected for a talk at the 2018 Penn Epigenetics Spatial and Functional Genomics Symposium (January 2018)
  • Camille’s paper was accepted in Plos Genetics.
  • Camille's NRSA predoctoral fellowship (F31) was awarded from the NIH! (August 2017).
  • Summer lab axe throwing competition: Anguera lab against the Lengner lab (Aug 2017).
  • Camille received a travel award to attend the EMBO Workshop on B cells in health and disease in Girona, Spain (August 2017).
  • The Anguera Lab welcomes its newest member, Sparkles, the axolotl (August 2017).
  • Welcome to the Lab, Bam Paneru! (July 2017).
  • Camille receives a Goldie Simon Preceptorship award from the Lupus Foundation of America (June 2017).
  • Welcome to the Lab, Isabel Sierra! Isabel is a CAMB-DSRB PhD Student (June 2017).
  • Welcome Donavon Sandoval-Heglund! Don is a Penn Undergraduate and is a part of the PURM summer research program (May 2017).
  • Erin Achilles (Vet ’19) received an NIH-Merial Summer Research grant to work in the lab (May 2017).
  • Steve received the Morris Animal Foundation summer research fellowship, Congrats Steve! (May 2017).
  • Welcome to the Anguera Lab, Anna Martin! Anna received an HHMI career fellowship grant! She will be taking a year off of Vet school to do research full-time in the Anguera & Atchison labs (April 2017).
  • Jianle’s JOVE methods paper is now in press (April 2017).
  • Camille was appointed to the Developmental Biology Training Grant (T32) at Penn (June 2016).
  • First lab publication to make the cover! Read more...
  • "LNCRHOXF1: a long noncoding RNA from the X-chromosome that suppresses viral response genes during development of the early human placenta" is now in press in Molecular Cell Biology (May 2016).
  • Jianle was selected to give a talk at the 10th annual meeting of the Organization for the Society of Sex Differences (May 2016). 
  • Camile was selected to attend the 2016 Van Andel Research Institute summer course on Epigenomics (May 2016). 
  • Steve Hanes received an NIH-Merial Summer Research grant (2016).
  • First publication from the Anguera Lab now in press at PNAS (March 2016).
  • Montserrat received an NIH-BIRCWH Scholarship (January 2016).
  • Ian Penkala placed 3rd in the 2015 Penn Vet Student Research Day presentations (March 2015).

    camille

  • Camille received a travel award to present her research at the 2016 Keystone Conference on Long noncoding RNAs (October 2015).
  • Jianle was selected to give a seminar on her research at the Penn Postdoctoral Research Symposium (October 2015).
  • Camille received a grant from the American Lupus Foundation (June 2015).

Team Anguera

In the Anguera Lab, life isn't always about work. Here are some examples of how the Anguera team unwind, too.

Anguera Lab at Penn Vet 2019
anguera-party-5_rs960wx608_cprs Sparkles, Anguera Lab Axolotl

Anguera Photo Collage_1

Anguera Photo Collage_2

Anguera Graduations
Anguera Team
Anguera Promotion

 

Anguera Lab Members
  • Associate Professor, Biomedical Sciences
  • Head of Anguera Lab
  • Montserrat grew up in San Diego, CA and attended UC San Diego as an undergraduate, where she studied environmental chemistry. She received her PhD in Biochemistry from Cornell University in Ithaca, NY, where she studied folate metabolism using cell culture and mouse models with Dr. Patrick Stover. She became interested in epigenetics, and joined Jeannie Lee’s lab at the Massachusetts General Hospital and she received a Ruth Kirschstein NRSA. She worked on a variety of projects in the Lee lab investigating long noncoding RNAs from the X-chromosome in mouse and human pluripotent stem cells.
  • Dr. Sarah Pyfrom, Penn VetSarah Pyfrom, PhD
  • Post-Doctoral Fellow
  • Born and raised in the beautiful islands of the Bahamas, Sarah is a postdoctoral researcher in the Anguera lab. She completed her PhD at Washington University in St. Louis in 2019 in the Molecular Genetics and Genomics Program in the lab of Jacqueline Payton. During her time in the Payton lab she worked on several projects from mechanistic studies of lymphoma-associated lncRNAs to development of bioinformatic tools to interrogate genomic level transcriptional regulation (and many others in between). Her goal is to become a “swiss army knife” of science, able to directly tackle any research question of interest that might arise. Outside of lab, Sarah enjoys reading, crochet, and essentially any aerobic exercise class you could imagine.
  • Isabel Sierra, Penn VetIsabel Sierra, BS
  • Graduate Student
  • Isabel, born and raised in Philadelphia, is a PhD student in the Developmental, Stem Cell, and Regenerative Biology graduate group. After graduating from Drexel University in 2013, she took a research assistant position at the Wistar Institute in a lab studying the histone H3.3 deposition pathway and it's involvement in pediatric glioblastoma. After three years, she applied to graduate school with the hopes of pursuing her two favorite topics: developmental biology and epigenetics. Outside of the lab Isabel likes to go to shows and explore the food scene around Philly.
  • Amy Dubin_160x200Aimee Dubin, BS
  • Aimee grew up in the suburbs of Philadelphia and graduated from Temple University in 2018 with a degree in Biology. During her undergraduate studies, she did research in organic chemistry in Dr. Sarah Wengryniuk’s lab. She studied polycationic hypervalent iodine mediated oxidative ring expansion of secondary alcohols. Aimee is a research specialist with aspirations to become an MD or MD/PhD. In her free time, Aimee likes to play soccer, explore the city, and try new restaurants.
  • Zack Beethem_160x200Zachary Beethem, BS
  • Zack grew up in Los Angeles and graduated in 2017 from University of Wisconsin-Madison with degrees in Genetics and Scandinavian Studies. He is a PhD student in the Developmental, Stem Cell and Regenerative Biology graduate group. During undergrad, he did research in Dr. Sean Schoville's lab, where he used phylogenomic techniques to study the evolutionary history of the Grylloblattidae family. Outside of lab, Zack likes to watch movies, eat food, read books, and travel.
  • Natalie Toothacre Anguera LabNatalie Toothacre, BS
  • Natalie is a PhD student in the Genetics and Epigenetics graduate group at Penn. She was born and raised in San Diego, CA and graduated from UC San Diego in 2018 with a degree in General Biology. During her undergraduate career she worked with Dr. Charles C. King to study the differentiation of human embryonic stem cells into glucose-responsive insulin-producing pancreatic islet cells. In her free time Natalie likes to knit, watch movies, try new restaurants, and travel.
  • Amanda Driscoll - Anguera LabAmanda Driscoll, BS
  • Native to the Jersey Shore, Amanda received her BS in Biology from Drexel University, where she worked in a paleontology lab researching titanosaurs. She now is a research specialist with an interest in immunology and microbiology. She enjoys reading genre fiction and tending to her large collection of houseplants.
 Former Anguera Lab Members
Anguera Lab - Bam Picture 160
Bam Dev Paneru, PhD
Don Sandoval-Anguera lab 160 Donald Sandoval, BS
CamilleSyrett Camille Syrett, PhD
Jianle Wang, PhD, Penn Vet            

Jianle Wang, PhD

Anna Jones, Anguera Lab, Penn Vet

Anna Jones, BS

Ian Penkala, Anguera Lab 

Ian Penkala, BS

MarianneKrammer

Marianne Krammer, BS

TaylorBryant

TaylorBryant, BS

 

Anna Martin, BS

 

Steve Hanes, BS