DOCSLIB.ORG

HOXB4's Road Map to Stem Cell Expansion

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
HOXB4's Road Map to Stem Cell Expansion HOXB4’s road map to stem cell expansion Bernhard Schiedlmeier*†, Ana Cristina Santos‡§, Ana Ribeiro‡, Natalia Moncaut‡, Dietrich Lesinski*, Herbert Auer¶, Karl Kornackerʈ, Wolfram Ostertag*, Christopher Baum*, Moises Mallo†‡, and Hannes Klump* *Department of Experimental Hematology, Hannover Medical School, Carl-Neuberg-Strasse1, 30625 Hannover, Germany; ‡Instituto Gulbenkian de Ciencia, 2780-156 Oeiras, Portugal; ¶Columbus Children’s Research Institute, Columbus, OH 43210; and ʈDivision of Sensory Biophysics, Ohio State University, Columbus, OH 43205 Edited by Tasuku Honjo, Kyoto University, Kyoto, Japan, and approved September 10, 2007 (received for review April 3, 2007) Homeodomain-containing transcription factors are important reg- BM cells. These HSCs fully replenish the stem cell pool of lethally ulators of stem cell behavior. HOXB4 mediates expansion of adult irradiated mice and maintain a normal supply of HSCs and mature and embryo-derived hematopoietic stem cells (HSCs) when ex- blood cells for the duration of life (2, 3). Ectopic HOXB4 expression pressed ectopically. To define the underlying molecular mecha- also enhances the in vitro development of definitive HSCs from nisms, we performed gene expression profiling in combination differentiating mouse ES cells that are capable of reconstituting with subsequent functional analysis with enriched adult HSCs and recipient mice (4, 5). However, ectopic expresssion of HOXB4 may embryonic derivatives expressing inducible HOXB4. Thereby, we eventually transform hematopoietic cells. Our group and others identified a set of overlapping genes that likely represent ‘‘uni- have described that the amount of ectopically expressed HOXB4 versal’’ targets of HOXB4. A substantial number of loci are involved has an impact on myeloid, lymphoid, and erythroid differentiation in signaling pathways important for controlling self-renewal, of adult HSCs of mice and humans in vitro and in vivo and on ES maintenance, and differentiation of stem cells. Functional assays cell-derived hematopoietic differentiation (6–8). performed on selected pathways confirmed the biological coher- The molecular mechanism behind how this transcription factor ence of the array results. HOXB4 activity protected adult HSCs from acts in immature blood cells is poorly understood. Homeodomain- the detrimental effects mediated by the proinflammatory cytokine mediated DNA binding is essential for the increased self-renewal of TNF-␣. This protection likely contributes to the competitive re- HOXB4-expressing HSCs (9). However, only a few HOXB4 target population advantage of HOXB4-expressing HSCs observed in genes have been identified so far. HOX proteins can form high- ␣ vivo. The concept of TNF- inhibition may also prove beneficial for affinity DNA-binding complexes with other homeodomain- patients undergoing bone marrow transplantation. Furthermore, containing proteins such as PBX1 (10). However, direct HOXB4– we demonstrate that HOXB4 activity and FGF signaling are inter- PBX1 protein interactions are not required for stem cell expansion twined. HOXB4-mediated expansion of adult and ES cell-derived (9). In contrast, PBX1 limits HOXB4-induced HSC expansion in HSCs was enhanced by specific and complete inhibition of FGF vivo (11). receptors. In contrast, the expanding activity of HOXB4 on hema- To further characterize the molecular mechanisms underlying topoietic progenitors in day 4–6 embryoid bodies was blunted in the HOXB4-induced expansion of HSCs, we analyzed the gene the presence of basic FGF (FGF2), indicating a dominant negative expression profiles of adult HSC/hematopoietic progenitor cells effect of FGF signaling on the earliest hematopoietic cells. In (HPCs) and differentiating ES cells expressing inducible forms of summary, our results strongly suggest that HOXB4 modulates the HOXB4. By identification of its target genes and through functional response of HSCs to multiple extrinsic signals in a concerted manner, tests, in vivo, we demonstrate that HOXB4 changes the cellular thereby shifting the balance toward stem cell self-renewal. response of stem cells to conserved signaling pathways known to affect cell fate decisions of HSCs in adult and differentiating ES hematopoiesis ͉ HOX genes ͉ microarray ͉ self-renewal ͉ embryoid bodies cells. These results may also reflect general pathways of homeobox transcription factor activities that are relevant for developmental he lifelong production of blood cells by the hematopoietic decisions. Tsystem strictly depends on the maintenance of its central organizer, the hematopoietic stem cell (HSC). Because of its unique Results and characteristic ability to undergo self-renewal divisions and/or Identification of HOXB4 Target Genes in Primary Murine HSCs and differentiate into mature blood cells, strategies to expand HSCs ex HPCs. To understand the mechanisms of HOXB4 activity, we vivo to increase their clinical potential in cell and gene therapy have wanted to identify target genes of HOXB4 in adult HSCs and become of key interest. HPCs. We thus transduced murine HSC/HPCs with a retroviral Expansion of HSCs naturally occurs in the fetal liver during vector that coexpresses EGFP and a 4-hydroxytamoxifen (TMX)- embryonic development and in the bone marrow (BM) of the adult organism as a response to extrinsic cues. The signaling pathways controlling this process and the molecular mechanisms specifying Author contributions: B.S., H.A., M.M., and H.K. designed research; B.S., A.C.S., A.R., N.M., whether the self-renewal potential is lost or retained during HSC D.L., H.A., and H.K. performed research; H.A. and K.K. contributed new reagents/analytic cell divisions remain largely ill-defined. However, accumulative tools; B.S., H.A., K.K., M.M., and H.K. analyzed data; and B.S., H.A., W.O., C.B., M.M., and evidence shows that HSC fate decisions are controlled by a delicate H.K. wrote the paper. balance of the intrinsic genetic program of HSCs and extrinsic The authors declare no conflict of interest. signals provided by their microenvironment, the stem cell niche. This article is a PNAS Direct Submission. Some of the intrinsic regulatory genes (Bmi1, Cdkn1a, Pten, Etv6, Abbreviations: BM, bone marrow; HSC, hematopoietic stem cell; HPC, hematopoietic Ϫ ϩ ϩ Mcl1, Hoxb4) that control stem cell maintenance, expansion, and progenitor cell; LSK, Lin Sca1 c-kit ; EB, embryoid body; TMX, 4-hydroxytamoxifen; qRT-PCR, quantitative RT-PCR; CHX, cycloheximide; Dox, doxycycline; ES-HC, ES cell-derived differentiation and some extrinsic signals (Wnt, Ang-1, Notch hematopoietic cell; GEMM, granulocyte, erythroid, macrophage, megakaryocyte. ligands, Gp130 ligands) that are exchanged between HSCs and †To whom correspondence may be addressed. E-mail: schiedlmeier.bernhard@ niche cells have been identified recently (1). mh-hannover.de or [email protected]. Numerous members of the homeobox gene family are involved §Present address: Molecular Hematology Laboratory, John Radcliffe Hospital, Oxford, OX3 in the regulation of normal and leukemic hematopoiesis at various 9DS, United Kingdom. stages. In fact, the human homeodomain transcription factor This article contains supporting information online at www.pnas.org/cgi/content/full/ HOXB4 was the first gene shown to lead to profound HSC 0703082104/DC1. expansion in vitro and in vivo when ectopically expressed in murine © 2007 by The National Academy of Sciences of the USA 16952–16957 ͉ PNAS ͉ October 23, 2007 ͉ vol. 104 ͉ no. 43 www.pnas.org͞cgi͞doi͞10.1073͞pnas.0703082104 Downloaded by guest on September 27, 2021 inducible form of HOXB4 (HOXB4-ER). Upon addition of TMX, the HOXB4-ER fusion protein translocates from the cytoplasm to the nucleus, consequently being capable of modulating gene ex- pression. Transduced cell populations were expanded for 14 days in the presence of TMX. Thereafter, HOXB4-ERϩLSK (where LSK is LinϪ, Sca1ϩ, c-kitϩ) cells were flow cytometrically isolated and cultivated either with or without TMX for 1 or 4 h. Inactivation of HOXB4 activity by TMX withdrawal was intended to mimic the naturally occurring down-regulation of HOXB4 in differentiating stem cells (12). RNA was prepared after the aforementioned times, and the transcriptional profiles of HOXB4-ERϩLSK Ϯ TMX were analyzed with the Affimetrix platform. As a control, profiling was also performed with LSK cells expressing unmodified constitutively active HOXB4 (HOXB4const) Ϯ TMX, to exclude changes in gene expression caused by unknown effects of TMX itself [supporting information (SI) Fig. 5]. Genes whose level of expression consistently and significantly (i) changed Ͼ2.0fold4hafterwithdrawal of TMX in one of the two independent experiments and (ii) changed Ͻ 1.5-fold after with- drawal of TMX in both HOXB4const LSK control group samples were considered differentially and specifically regulated by HOXB4. This analysis identified 156 HOXB4-regulated genes in proliferating LSK cells, of which 23 were uncharacterized tran- scripts (SI Table 1). All 23 uncharacterized target genes, and 103 of the 133 characterized genes, were also differentially regulated Ͼ1.5-fold in the second independent experiment. The 133 charac- terized genes identified by the above criteria belonged to different BIOLOGY functional groups (SI Table 2). A comparison of our HOXB4 target DEVELOPMENTAL gene list with molecular signatures shown to be specific for naturally (in vivo) proliferating fetal liver HSCs and their quiescent adult BM-derived counterparts revealed significant overlaps (SI Table 3).
Load more

Recommended publications
  • Genetic Variability in the Italian Heavy Draught Horse from Pedigree Data and Genomic Information
    Supplementary material for manuscript: Genetic variability in the Italian Heavy Draught Horse from pedigree data and genomic information. Enrico Mancin†, Michela Ablondi†, Roberto Mantovani*, Giuseppe Pigozzi, Alberto Sabbioni and Cristina Sartori ** Correspondence: [email protected] † These two Authors equally contributed to the work Supplementary Figure S1. Mares and foal of Italian Heavy Draught Horse (IHDH; courtesy of Cinzia Stoppa) Supplementary Figure S2. Number of Equivalent Generations (EqGen; above) and pedigree completeness (PC; below) over years in Italian Heavy Draught Horse population. Supplementary Table S1. Descriptive statistics of homozygosity (observed: Ho_obs; expected: Ho_exp; total: Ho_tot) in 267 genotyped individuals of Italian Heavy Draught Horse based on the number of homozygous genotypes. Parameter Mean SD Min Max Ho_obs 35,630.3 500.7 34,291 38,013 Ho_exp 35,707.8 64.0 35,010 35,740 Ho_tot 50,674.5 93.8 49,638 50,714 1 Definitions of the methods for inbreeding are in the text. Supplementary Figure S3. Values of BIC obtained by analyzing values of K from 1 to 10, corresponding on the same amount of clusters defining the proportion of ancestry in the 267 genotyped individuals. Supplementary Table S2. Estimation of genomic effective population size (Ne) traced back to 18 generations ago (Gen. ago). The linkage disequilibrium estimation, adjusted for sampling bias was also included (LD_r2), as well as the relative standard deviation (SD(LD_r2)). Gen. ago Ne LD_r2 SD(LD_r2) 1 100 0.009 0.014 2 108 0.011 0.018 3 118 0.015 0.024 4 126 0.017 0.028 5 134 0.019 0.031 6 143 0.021 0.034 7 156 0.023 0.038 9 173 0.026 0.041 11 189 0.029 0.046 14 213 0.032 0.052 18 241 0.036 0.058 Supplementary Table S3.
    [Show full text]
  • Homeobox Gene Expression Profile in Human Hematopoietic Multipotent
    Leukemia (2003) 17, 1157–1163 & 2003 Nature Publishing Group All rights reserved 0887-6924/03 $25.00 www.nature.com/leu Homeobox gene expression profile in human hematopoietic multipotent stem cells and T-cell progenitors: implications for human T-cell development T Taghon1, K Thys1, M De Smedt1, F Weerkamp2, FJT Staal2, J Plum1 and G Leclercq1 1Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium; and 2Department of Immunology, Erasmus Medical Center, Rotterdam, The Netherlands Class I homeobox (HOX) genes comprise a large family of implicated in this transformation proces.14 The HOX-C locus transcription factors that have been implicated in normal and has been primarily implicated in lymphomas.15 malignant hematopoiesis. However, data on their expression or function during T-cell development is limited. Using degener- Hematopoietic cells are derived from stem cells that reside in ated RT-PCR and Affymetrix microarray analysis, we analyzed fetal liver (FL) in the embryo and in the adult bone marrow the expression pattern of this gene family in human multipotent (ABM), which have the unique ability to self-renew and thereby stem cells from fetal liver (FL) and adult bone marrow (ABM), provide a life-long supply of blood cells. T lymphocytes are a and in T-cell progenitors from child thymus. We show that FL specific type of hematopoietic cells that play a major role in the and ABM stem cells are similar in terms of HOX gene immune system. They develop through a well-defined order of expression, but significant differences were observed between differentiation steps in the thymus.16 Several transcription these two cell types and child thymocytes.
    [Show full text]
  • Genome-Wide DNA Methylation Analysis of KRAS Mutant Cell Lines Ben Yi Tew1,5, Joel K
    www.nature.com/scientificreports OPEN Genome-wide DNA methylation analysis of KRAS mutant cell lines Ben Yi Tew1,5, Joel K. Durand2,5, Kirsten L. Bryant2, Tikvah K. Hayes2, Sen Peng3, Nhan L. Tran4, Gerald C. Gooden1, David N. Buckley1, Channing J. Der2, Albert S. Baldwin2 ✉ & Bodour Salhia1 ✉ Oncogenic RAS mutations are associated with DNA methylation changes that alter gene expression to drive cancer. Recent studies suggest that DNA methylation changes may be stochastic in nature, while other groups propose distinct signaling pathways responsible for aberrant methylation. Better understanding of DNA methylation events associated with oncogenic KRAS expression could enhance therapeutic approaches. Here we analyzed the basal CpG methylation of 11 KRAS-mutant and dependent pancreatic cancer cell lines and observed strikingly similar methylation patterns. KRAS knockdown resulted in unique methylation changes with limited overlap between each cell line. In KRAS-mutant Pa16C pancreatic cancer cells, while KRAS knockdown resulted in over 8,000 diferentially methylated (DM) CpGs, treatment with the ERK1/2-selective inhibitor SCH772984 showed less than 40 DM CpGs, suggesting that ERK is not a broadly active driver of KRAS-associated DNA methylation. KRAS G12V overexpression in an isogenic lung model reveals >50,600 DM CpGs compared to non-transformed controls. In lung and pancreatic cells, gene ontology analyses of DM promoters show an enrichment for genes involved in diferentiation and development. Taken all together, KRAS-mediated DNA methylation are stochastic and independent of canonical downstream efector signaling. These epigenetically altered genes associated with KRAS expression could represent potential therapeutic targets in KRAS-driven cancer. Activating KRAS mutations can be found in nearly 25 percent of all cancers1.
    [Show full text]
  • Homeobox A10 Promotes the Proliferation and Invasion of Bladder Cancer Cells Via Regulation of Matrix Metalloproteinase‑3
    ONCOLOGY LETTERS 18: 49-56, 2019 Homeobox A10 promotes the proliferation and invasion of bladder cancer cells via regulation of matrix metalloproteinase‑3 CHUNLEI LIU1*, MINGZHU GE2*, JUN MA1*, YANHUI ZHANG1, YANHUI ZHAO1 and TAO CUI1 Departments of 1Urology and 2Ultrasound, Qingdao Central Hospital, Qingdao, Shandong 266042, P.R. China Received February 9, 2018; Accepted January 31, 2019 DOI: 10.3892/ol.2019.10312 Abstract. Homeobox A10 (HOXA10) belongs to the family Smoking and obesity are risk factors for BC (2), and genetic of HOX genes, which are closely connected with embryonic mutations and abnormal protein expression serve important development and serve important roles in various tumors. roles in the genesis, development and progression of BC (4). However, the role of HOXA10 in bladder cancer (BC) remains Therefore, exploring new anomalous molecules involved in unclear. In the present study, the role of HOXA10 in BC and the development of BC may advance the understanding of the underlying mechanisms by which it promotes the disease the mechanisms behind this disease and contribute to the progression were investigated. Immunohistochemical analysis improvement of treatment strategies. demonstrated that the expression of the HOXA10 protein Homeobox A10 (HOXA10) belongs to the family of HOX was significantly higher in BC tissues as compared with that genes, which are classified into four subgroups, namely HOX in adjacent normal tissues. Subsequent statistical analysis A-D (5), and are closely connected with embryonic develop- revealed that upregulation of HOXA10 was significantly ment (6). HOXA10 encodes a DNA-binding transcription factor associated with the pathological grade and clinical stage of that serves vital roles in regulating gene expression, viability BC patients.
    [Show full text]
  • Noelia Díaz Blanco
    Effects of environmental factors on the gonadal transcriptome of European sea bass (Dicentrarchus labrax), juvenile growth and sex ratios Noelia Díaz Blanco Ph.D. thesis 2014 Submitted in partial fulfillment of the requirements for the Ph.D. degree from the Universitat Pompeu Fabra (UPF). This work has been carried out at the Group of Biology of Reproduction (GBR), at the Department of Renewable Marine Resources of the Institute of Marine Sciences (ICM-CSIC). Thesis supervisor: Dr. Francesc Piferrer Professor d’Investigació Institut de Ciències del Mar (ICM-CSIC) i ii A mis padres A Xavi iii iv Acknowledgements This thesis has been made possible by the support of many people who in one way or another, many times unknowingly, gave me the strength to overcome this "long and winding road". First of all, I would like to thank my supervisor, Dr. Francesc Piferrer, for his patience, guidance and wise advice throughout all this Ph.D. experience. But above all, for the trust he placed on me almost seven years ago when he offered me the opportunity to be part of his team. Thanks also for teaching me how to question always everything, for sharing with me your enthusiasm for science and for giving me the opportunity of learning from you by participating in many projects, collaborations and scientific meetings. I am also thankful to my colleagues (former and present Group of Biology of Reproduction members) for your support and encouragement throughout this journey. To the “exGBRs”, thanks for helping me with my first steps into this world. Working as an undergrad with you Dr.
    [Show full text]
  • SUPPLEMENTARY MATERIAL Bone Morphogenetic Protein 4 Promotes
    www.intjdevbiol.com doi: 10.1387/ijdb.160040mk SUPPLEMENTARY MATERIAL corresponding to: Bone morphogenetic protein 4 promotes craniofacial neural crest induction from human pluripotent stem cells SUMIYO MIMURA, MIKA SUGA, KAORI OKADA, MASAKI KINEHARA, HIROKI NIKAWA and MIHO K. FURUE* *Address correspondence to: Miho Kusuda Furue. Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, 7-6-8, Saito-Asagi, Ibaraki, Osaka 567-0085, Japan. Tel: 81-72-641-9819. Fax: 81-72-641-9812. E-mail: [email protected] Full text for this paper is available at: http://dx.doi.org/10.1387/ijdb.160040mk TABLE S1 PRIMER LIST FOR QRT-PCR Gene forward reverse AP2α AATTTCTCAACCGACAACATT ATCTGTTTTGTAGCCAGGAGC CDX2 CTGGAGCTGGAGAAGGAGTTTC ATTTTAACCTGCCTCTCAGAGAGC DLX1 AGTTTGCAGTTGCAGGCTTT CCCTGCTTCATCAGCTTCTT FOXD3 CAGCGGTTCGGCGGGAGG TGAGTGAGAGGTTGTGGCGGATG GAPDH CAAAGTTGTCATGGATGACC CCATGGAGAAGGCTGGGG MSX1 GGATCAGACTTCGGAGAGTGAACT GCCTTCCCTTTAACCCTCACA NANOG TGAACCTCAGCTACAAACAG TGGTGGTAGGAAGAGTAAAG OCT4 GACAGGGGGAGGGGAGGAGCTAGG CTTCCCTCCAACCAGTTGCCCCAAA PAX3 TTGCAATGGCCTCTCAC AGGGGAGAGCGCGTAATC PAX6 GTCCATCTTTGCTTGGGAAA TAGCCAGGTTGCGAAGAACT p75 TCATCCCTGTCTATTGCTCCA TGTTCTGCTTGCAGCTGTTC SOX9 AATGGAGCAGCGAAATCAAC CAGAGAGATTTAGCACACTGATC SOX10 GACCAGTACCCGCACCTG CGCTTGTCACTTTCGTTCAG Suppl. Fig. S1. Comparison of the gene expression profiles of the ES cells and the cells induced by NC and NC-B condition. Scatter plots compares the normalized expression of every gene on the array (refer to Table S3). The central line
    [Show full text]
  • NF-Ya Activates Multiple Hematopoietic Stem Cell (HSC) Regulatory Genes and Promotes HSC Self-Renewal
    NF-Ya activates multiple hematopoietic stem cell (HSC) regulatory genes and promotes HSC self-renewal Jiang Zhu, Yi Zhang, Gerard J. Joe, Richard Pompetti, and Stephen G. Emerson* Departments of Medicine and Pediatrics, and Abramson Cancer Center, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 Communicated by Zhu Chen, Shanghai Second Medical University, Shanghai, People’s Republic of China, April 25, 2005 (received for review December 15, 2004) Hematopoietic stem cell (HSC) self-renewal and differentiation are chased from The Jackson Laboratory and maintained in the animal influenced through multiple pathways, including homeobox tran- facilities of the University of Pennsylvania with sterile water or with scription factors, signaling through ␤-catenin and Notch-1, telom- the water supplemented with neomycin sulfate and polymyxin B erase, and p27. How these multiple pathways interact and are (Sigma) within 3–4 weeks after BMT. orchestrated is currently unknown. We now report that NF-Ya, the regulatory and DNA-binding subunit of the trimeric transcription NF-Ya cDNA, MigR1 Retroviral Vector, and Preparation of Retrovi- factor NF-Y, plays a central, integrating role in several of these HSC ruses. cDNA for NF-Ya was amplified from human normal bone pathways. NF-Ya is preferentially expressed in HSC-enriched bone marrow (BM) cell RNA by using Pfu DNA polymerase (Strat- marrow subpopulations, and NF-Ya mRNA rapidly declines with agene) (7). MigR1 retroviral vector was obtained from Warren HSC differentiation. Overexpression of NF-Ya in primitive hema- Pear (Department of Pathology and Laboratory Medicine, Uni- topoietic cells activates the transcription of multiple HOX4 paral- versity of Pennsylvania).
    [Show full text]
  • Coordination of Hox Identity Between Germ Layers Along the Anterior-To-Posterior Axis of the Vertebrate Embryo
    Coordination of Hox identity between germ layers along the anterior-to-posterior axis of the vertebrate embryo Ferran Lloret Vilaspasa PhD Developmental Biology Department of Anatomy and Developmental Biology University College of London (UCL) London, United Kingdom 2009 1 Coordination of Hox identity between germ layers along the anterior-to- posterior axis of the vertebrate embryo ‘ I, Ferran Lloret Vilaspasa confirm that the work presented in this thesis is my own. Where information has been derived from other sources, I confirm that this has been indicated in the thesis. ' Thesis for the obtainment of a PhD in Development Biology at the University College of London under the supervision of Prof. dr. Claudio D. Stern and Prof. dr. Antony J. Durston (Leiden University). To be defended by Ferran Lloret Vilaspasa A la meva família… Born in Barcelona, 05-08-1977 2 Abstract During early embryonic development, a relatively undifferentiated mass of cells is shaped into a complex and morphologically differentiated embryo. This is achieved by a series of coordinated cell movements that end up in the formation of the three germ layers of most metazoans and the establishment of the body plan. Hox genes are among the main determinants in this process and they have a prominent role in granting identity to different regions of the embryo. The particular arrangement of their expression domains in early development corresponds to and characterises several future structures of the older embryo and adult animal. Getting to know the molecular and cellular phenomena underlying the correct Hox pattern will help us understand how the complexity of a fully-formed organism can arise from its raw materials, a relatively undifferentiated fertilised egg cell (zygote) and a large but apparently limited repertoire of molecular agents.
    [Show full text]
  • Expression Pattern of the Class I Homeobox Genes in Ovarian Carcinoma
    J Gynecol Oncol Vol. 21, No. 1:29-37, March 2010 DOI:10.3802/jgo.2010.21.1.29 Original Article Expression pattern of the class I homeobox genes in ovarian carcinoma Jin Hwa Hong1, Jae Kwan Lee1, Joong Jean Park2, Nak Woo Lee1, Kyu Wan Lee1, Jung Yeol Na1 Departments of 1Obstetrics and Gynecology, 2Physiology, Korea University College of Medicine, Seoul, Korea Objective: Although some sporadic reports reveal the link between the homeobox (HOX) genes and ovarian carcinoma, there is no comprehensive analysis of the expression pattern of the class I homeobox genes in ovarian carcinoma that determines the candidate genes involved in ovarian carcinogenesis. Methods: The different patterns of expression of 36 HOX genes were analyzed, including 4 ovarian cancer cell lines and 4 normal ovarian tissues. Using a reverse transcription-polymerase chain reaction (RT-PCR) and quantification analysis, the specific gene that showed a significantly higher expression in ovarian cancer cell lines than in normal ovaries was selected, and western blot analysis was performed adding 7 ovarian cancer tissue specimens. Finally, immunohistochemical and immunocytochemical analyses were performed to compare the pattern of expression of the specific HOX gene between ovarian cancer tissue and normal ovaries. Results: Among 36 genes, 11 genes had a different level of mRNA expression between the cancer cell lines and the normal ovarian tissues. Of the 11 genes, only HOXB4 had a significantly higher level of expression in ovarian cancer cell lines than in normal ovaries (p=0.029). Based on western blot, immunohistochemical, and immunocytochemical analyses, HOXB4 was expressed exclusively in the ovarian cancer cell lines or cancer tissue specimens, but not in the normal ovaries.
    [Show full text]
  • Downregulation of Prdm16 Is Critical for HOXB4-Mediated Benign HSC Expansion in Vivo Hui Yu University of Tennessee Health Science Center
    University of Tennessee Health Science Center UTHSC Digital Commons Theses and Dissertations (ETD) College of Graduate Health Sciences 12-2014 Downregulation of Prdm16 Is Critical for HOXB4-mediated Benign HSC Expansion In Vivo Hui Yu University of Tennessee Health Science Center Follow this and additional works at: https://dc.uthsc.edu/dissertations Part of the Medical Genetics Commons, and the Medical Molecular Biology Commons Recommended Citation Yu, Hui , "Downregulation of Prdm16 Is Critical for HOXB4-mediated Benign HSC Expansion In Vivo" (2014). Theses and Dissertations (ETD). Paper 307. http://dx.doi.org/10.21007/etd.cghs.2014.0367. This Dissertation is brought to you for free and open access by the College of Graduate Health Sciences at UTHSC Digital Commons. It has been accepted for inclusion in Theses and Dissertations (ETD) by an authorized administrator of UTHSC Digital Commons. For more information, please contact [email protected]. Downregulation of Prdm16 Is Critical for HOXB4-mediated Benign HSC Expansion In Vivo Document Type Dissertation Degree Name Doctor of Philosophy (PhD) Program Biochemistry Track Therapeutics and Cell Signaling Research Advisor Brian P. Sorrentino, MD Committee Suzanne J. Baker, PhD Wing H. Leung, MD, PhD Janet F. Partridge, PhD Lawrence M. Pfeffer, PhD DOI 10.21007/etd.cghs.2014.0367 This dissertation is available at UTHSC Digital Commons: https://dc.uthsc.edu/dissertations/307 DOWNREGULATION OF PRDM16 IS CRITICAL FOR HOXB4-MEDIATED BENIGN HSC EXPANSION IN VIVO A Dissertation Presented for The Graduate Studies Council The University of Tennessee Health Science Center In Partial Fulfillment Of the Requirements for the Degree Doctor of Philosophy From The University of Tennessee By Hui Yu December 2014 Copyright © 2014 by Hui Yu All rights reserved ii DEDICATION This dissertation is dedicated to my parents, Guoquan Yu and Yan Xu, my husband, Satish Kumar Nandakumar and my precious daughter Emma Satish for all their love and support.
    [Show full text]
  • PRDM16 Isoforms Differentially Regulate Normal and Leukemic Hematopoiesis and Inflammatory Gene Signature
    RESEARCH ARTICLE The Journal of Clinical Investigation PRDM16 isoforms differentially regulate normal and leukemic hematopoiesis and inflammatory gene signature David J. Corrigan,1,2 Larry L. Luchsinger,1,3 Mariana Justino de Almeida,1,2 Linda J. Williams,1,3 Alexandros Strikoudis,1,3 and Hans-Willem Snoeck1,2,3,4 1Columbia Center of Human Development, 2Department of Microbiology and Immunology, 3Department of Medicine, and 4Columbia Center for Translational Immunology, Columbia University Medical Center, New York, New York, USA. PRDM16 is a transcriptional coregulator involved in translocations in acute myeloblastic leukemia (AML), myelodysplastic syndromes, and T acute lymphoblastic leukemia that is highly expressed in and required for the maintenance of hematopoietic stem cells (HSCs), and can be aberrantly expressed in AML. Prdm16 is expressed as full-length (fPrdm16) and short (sPrdm16) isoforms, the latter lacking the N-terminal PR domain. The role of both isoforms in normal and malignant hematopoiesis is unclear. We show here that fPrdm16 was critical for HSC maintenance, induced multiple genes involved in GTPase signaling, and repressed inflammation, while sPrdm16 supported B cell development biased toward marginal zone B cells and induced an inflammatory signature. In a mouse model of human MLL-AF9 leukemia, fPrdm16 extended latency, while sPrdm16 shortened latency and induced a strong inflammatory signature, including several cytokines and chemokines that are associated with myelodysplasia and with a worse prognosis in human AML. Finally, in human NPM1-mutant and in MLL-translocated AML, high expression of PRDM16, which negatively impacts outcome, was associated with inflammatory gene expression, thus corroborating the mouse data. Our observations demonstrate distinct roles for Prdm16 isoforms in normal HSCs and AML, and identify sPrdm16 as one of the drivers of prognostically adverse inflammation in leukemia.
    [Show full text]
  • Mammalian Polycomb-Mediated Repression of Hox Genes Requires the Essential Spliceosomal Protein Sf3b1
    Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press RESEARCH COMMUNICATION complex, which contains Eed/Ezh2, is associated with Mammalian histone deacetylase and methyltransferase activity. The Polycomb-mediated repression PRC1 or Class II complex consists of, for example, Zfp144 (Mel18), Rnf2 (Ring1B), Cbx2/M33, and Phc2/ of Hox genes requires the Edr2. The Class II complex, which characteristically in- cludes the products from highly related pairs of genes, essential spliceosomal has been shown to inhibit nucleosome remodeling by protein Sf3b1 the SWI/SNF complex in vitro (Shao et al. 1999). How- ever, as the inhibition requires preincubation of the Kyoichi Isono,1 Yoko Mizutani-Koseki,1 Class II with the nucleosomal template (it does not occur Toshihisa Komori,2 when Class II and SWI/SNF products are added together), Marion S. Schmidt-Zachmann,3 and this suggests that the Class II complex does not interact 1,4 directly with SWI/SNF, but instead competes for the Haruhiko Koseki nucleosome template. Possibly, it is this binding of the 1Developmental Genetics Group, RIKEN Research Center for Class II complex, which prevents nucleosome remodel- Allergy and Immunology, Tsurumi-ku, Yokohama 230-0045, ing, that silences the genes by blocking the access of Japan; 2Department of Medicine III, Osaka University Medical transcription activators to cis-regulatory elements such School, Suita, Osaka 565-0871, Japan; 3Division of Cell as promoters and enhancers (Francis et al. 2001). Inter- Biology, German Cancer Research Center, estingly, recent studies have proposed an alternative D-69120 Heidelberg, Germany mechanism for maintaining gene silence (Breiling et al.
    [Show full text]