The three-dimensional structure of Epstein-Barr virus genome varies by latency type and is regulated by PARP1 enzymatic activity.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
17 01 2022
Historique:
received: 23 10 2020
accepted: 21 12 2021
entrez: 18 1 2022
pubmed: 19 1 2022
medline: 9 2 2022
Statut: epublish

Résumé

Epstein-Barr virus (EBV) persists in human B-cells by maintaining its chromatinized episomes within the nucleus. We have previously shown that cellular factor Poly [ADP-ribose] polymerase 1 (PARP1) binds the EBV genome, stabilizes CTCF binding at specific loci, and that PARP1 enzymatic activity correlates with maintaining a transcriptionally active latency program. To better understand PARP1's role in regulating EBV latency, here we functionally characterize the effect of PARP enzymatic inhibition on episomal structure through in situ HiC mapping, generating a complete 3D structure of the EBV genome. We also map intragenomic contact changes after PARP inhibition to global binding of chromatin looping factors CTCF and cohesin across the EBV genome. We find that PARP inhibition leads to fewer total unique intragenomic interactions within the EBV episome, yet new chromatin loops distinct from the untreated episome are also formed. This study also illustrates that PARP inhibition alters gene expression at the regions where chromatin looping is most effected. We observe that PARP1 inhibition does not alter cohesin binding sites but does increase its frequency of binding at those sites. Taken together, these findings demonstrate that PARP has an essential role in regulating global EBV chromatin structure and latent gene expression.

Identifiants

pubmed: 35039491
doi: 10.1038/s41467-021-27894-1
pii: 10.1038/s41467-021-27894-1
pmc: PMC8764100
doi:

Substances chimiques

CCCTC-Binding Factor 0
CTCF protein, human 0
Cell Cycle Proteins 0
Chromatin 0
Chromosomal Proteins, Non-Histone 0
Phthalazines 0
Piperazines 0
PARP1 protein, human EC 2.4.2.30
Poly (ADP-Ribose) Polymerase-1 EC 2.4.2.30
olaparib WOH1JD9AR8

Types de publication

Journal Article Research Support, N.I.H., Extramural

Langues

eng

Sous-ensembles de citation

IM

Pagination

187

Subventions

Organisme : NIAID NIH HHS
ID : R01 AI130209
Pays : United States
Organisme : NIDCR NIH HHS
ID : R01 DE017336
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA093606
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA010815
Pays : United States
Organisme : NCI NIH HHS
ID : R50 CA211199
Pays : United States

Informations de copyright

© 2022. The Author(s).

Références

Price, A. M. & Luftig, M. A. To Be or Not IIb: a multi-step process for Epstein-Barr virus latency establishment and consequences for B cell tumorigenesis. PLOS Pathog. 11, e1004656 (2015).
pubmed: 25790223 pmcid: 4366242 doi: 10.1371/journal.ppat.1004656
Hsu, J. L. & Glaser, S. L. Epstein-Barr virus-associated malignancies: epidemiologic patterns and etiologic implications. Crit. Rev. Oncol. Hematol. 34, 27–53 (2000).
pubmed: 10781747 doi: 10.1016/S1040-8428(00)00046-9
Thorley-Lawson, D. A. & Gross, A. Persistence of the Epstein-Barr virus and the origins of associated lymphomas. N. Engl. J. Med. 350, 1328–1337 (2004).
pubmed: 15044644 doi: 10.1056/NEJMra032015
Parkin, D. M. The global health burden of infection-associated cancers in the year 2002. Int. J. Cancer 118, 3030–3044 (2006).
pubmed: 16404738 doi: 10.1002/ijc.21731
Ferlay, J. et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int. J. Cancer 127, 2893–2917 (2010).
pubmed: 21351269 doi: 10.1002/ijc.25516
Khan, G. & Hashim, M. Global burden of deaths from Epstein-Barr virus attributable malignancies 1990-2010. Infect. Agents Cancer 9, 38–11 (2014).
doi: 10.1186/1750-9378-9-38
Farrell, P. J. Epstein-Barr virus and cancer. Annu. Rev. Pathol. 14, 29–53 (2019).
pubmed: 30125149 doi: 10.1146/annurev-pathmechdis-012418-013023
Gupte, R., Liu, Z. & Kraus, W. L. PARPs and ADP-ribosylation: recent advances linking molecular functions to biological outcomes. Genes Dev. 31, 101–126 (2017).
pubmed: 28202539 pmcid: 5322727 doi: 10.1101/gad.291518.116
Lupey-Green, L. N. et al. PARP1 restricts Epstein Barr Virus lytic reactivation by binding the BZLF1 promoter. Virology 507 IS -, 220–230 (2017).
doi: 10.1016/j.virol.2017.04.006
Lupey-Green, L. N. et al. PARP1 stabilizes CTCF binding and chromatin structure to maintain Epstein-Barr virus latency type. J. Virol. 92, e00755–00718 (2018).
pubmed: 29976663 pmcid: 6146685 doi: 10.1128/JVI.00755-18
Tempera, I., Wiedmer, A., Dheekollu, J. & Lieberman, P. M. CTCF prevents the epigenetic drift of EBV latency promoter Qp. PLoS Pathog. 6, e1001048 (2010).
pubmed: 20730088 pmcid: 2921154 doi: 10.1371/journal.ppat.1001048
Martin, K. A., Lupey, L. N. & Tempera, I. Epstein-Barr virus oncoprotein LMP1 mediates epigenetic changes in host gene expression through PARP1. J. Virol. 90, 8520–8530 (2016).
pubmed: 27440880 pmcid: 5021435 doi: 10.1128/JVI.01180-16
Phillips, J. E. & Corces, V. G. CTCF: master weaver of the genome. Cell 137, 1194–1211 (2009).
pubmed: 19563753 pmcid: 3040116 doi: 10.1016/j.cell.2009.06.001
Rowley, M. J. & Corces, V. G. Organizational principles of 3D genome architecture. Nat. Rev. Genet. 19, 1–12 (2018).
doi: 10.1038/s41576-018-0060-8
Ong, C.-T., Bortle, K. V., Ramos, E. & Corces, V. G. Poly(ADP-ribosyl)ation regulates insulator function and intrachromosomal interactions in Drosophila. Cell 155, 148–159 (2013).
pubmed: 24055367 doi: 10.1016/j.cell.2013.08.052
Farrar, D. et al. Mutational analysis of the poly(ADP-ribosyl)ation sites of the transcription factor CTCF provides an insight into the mechanism of its regulation by poly(ADP-ribosyl)ation. Mol. Cell. Biol. 30, 1199–1216 (2010).
pubmed: 20038529 doi: 10.1128/MCB.00827-09
Zampieri, M. et al. ADP-ribose polymers localized on Ctcf-Parp1-Dnmt1 complex prevent methylation of Ctcf target sites. Biochem. J. 441, 645–652 (2012).
pubmed: 21985173 doi: 10.1042/BJ20111417
Holdorf, M. M., Cooper, S. B., Yamamoto, K. R. & Miranda, J. J. L. Occupancy of chromatin organizers in the Epstein-Barr virus genome. Virology https://doi.org/10.1016/j.virol.2011.04.004 (2011).
Arvey, A. et al. An atlas of the Epstein-Barr virus transcriptome and epigenome reveals host-virus regulatory interactions. Cell Host Microbe 12, 233–245 (2012).
pubmed: 22901543 pmcid: 3424516 doi: 10.1016/j.chom.2012.06.008
Chau, C. M., Zhang, X.-Y., McMahon, S. B. & Lieberman, P. M. Regulation of Epstein-Barr virus latency type by the chromatin boundary factor CTCF. J. Virol. 80, 5723–5732 (2006).
pubmed: 16731911 pmcid: 1472585 doi: 10.1128/JVI.00025-06
Day, L. et al. Chromatin profiling of Epstein-Barr virus latency control region. J. Virol. 81, 6389–6401 (2007).
pubmed: 17409162 pmcid: 1900095 doi: 10.1128/JVI.02172-06
Davidson, I. F. et al. DNA loop extrusion by human cohesin. Science https://doi.org/10.1126/science.aaz3418 (2019).
Kagey, M. H. et al. Mediator and cohesin connect gene expression and chromatin architecture. Nature 467, 430–435 (2010).
pubmed: 20720539 pmcid: 2953795 doi: 10.1038/nature09380
Millau, J.-F. & Gaudreau, L. CTCF, cohesin, and histone variants: connecting the genome. Biochem. Cell Biol. 89, 505–513 (2011).
pubmed: 21970734 doi: 10.1139/o11-052
Chen, H.-S. et al. Epigenetic deregulation of the LMP1/LMP2 locus of Epstein-Barr virus by mutation of a single CTCF-cohesin binding site. J. Virol. 88, 1703–1713 (2014).
pubmed: 24257606 pmcid: 3911611 doi: 10.1128/JVI.02209-13
Langmead, B. & Salzberg, S. L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 9, 357–359 (2012).
pubmed: 22388286 pmcid: 3322381 doi: 10.1038/nmeth.1923
Tempera, I., Klichinsky, M. & Lieberman, P. M. EBV latency types adopt alternative chromatin conformations. PLoS Pathog. 7, e1002180 (2011).
pubmed: 21829357 pmcid: 3145795 doi: 10.1371/journal.ppat.1002180
Rao, S. S. et al. A 3D map of the human genome at kilobase resolution reveals principles of chromatin looping. Cell 159, 1665–1680 (2014).
pubmed: 25497547 pmcid: 5635824 doi: 10.1016/j.cell.2014.11.021
Menear, K. A. et al. 4-[3-(4-cyclopropanecarbonylpiperazine-1-carbonyl)-4-fluorobenzyl]-2H-phthalazin- 1-one: a novel bioavailable inhibitor of poly(ADP-ribose) polymerase-1. J. Med. Chem. 51, 6581–6591 (2008).
pubmed: 18800822 doi: 10.1021/jm8001263
Johannsen, E. et al. Proteins of purified Epstein-Barr virus. Proc. Natl Acad. Sci. USA 101, 16286–16291 (2004).
pubmed: 15534216 pmcid: 528973 doi: 10.1073/pnas.0407320101
Chiu, S. H. et al. Epstein-Barr virus BALF3 has nuclease activity and mediates mature virion production during the lytic cycle. J. Virol. 88, 4962–4975 (2014).
pubmed: 24554665 pmcid: 3993834 doi: 10.1128/JVI.00063-14
Paulsen, S. J., Rosenkilde, M. M., Eugen-Olsen, J. & Kledal, T. N. Epstein-Barr virus-encoded BILF1 is a constitutively active G protein-coupled receptor. J. Virol. 79, 536–546 (2005).
pubmed: 15596846 pmcid: 538743 doi: 10.1128/JVI.79.1.536-546.2005
Beisser, P. S. et al. The Epstein-Barr virus BILF1 gene encodes a G protein-coupled receptor that inhibits phosphorylation of RNA-dependent protein kinase. J. Virol. 79, 441–449 (2005).
pubmed: 15596837 pmcid: 538699 doi: 10.1128/JVI.79.1.441-449.2005
Zuo, J. et al. The Epstein-Barr virus G-protein-coupled receptor contributes to immune evasion by targeting MHC class I molecules for degradation. PLoS Pathog. 5, e1000255 (2009).
pubmed: 19119421 pmcid: 2603334 doi: 10.1371/journal.ppat.1000255
Gill, M. B., Edgar, R., May, J. S. & Stevenson, P. G. A gamma-herpesvirus glycoprotein complex manipulates actin to promote viral spread. PLoS ONE 3, e1808 (2008).
pubmed: 18350146 pmcid: 2262946 doi: 10.1371/journal.pone.0001808
Xiao, J., Palefsky, J. M., Herrera, R., Berline, J. & Tugizov, S. M. The Epstein-Barr virus BMRF-2 protein facilitates virus attachment to oral epithelial cells. Virology 370, 430–442 (2008).
pubmed: 17945327 doi: 10.1016/j.virol.2007.09.012
Matveeva, E. et al. Involvement of PARP1 in the regulation of alternative splicing. Cell Discov. 2, 15046 (2016).
pubmed: 27462443 pmcid: 4860959 doi: 10.1038/celldisc.2015.46
Zhang, K., Li, N., Ainsworth, R. I. & Wang, W. Systematic identification of protein combinations mediating chromatin looping. Nat. Commun. 7, 12249 (2016).
pubmed: 27461729 pmcid: 4974460 doi: 10.1038/ncomms12249
Sonoda, E. et al. Scc1/Rad21/Mcd1 is required for sister chromatid cohesion and kinetochore function in vertebrate cells. Dev. Cell 1, 759–770 (2001).
pubmed: 11740938 doi: 10.1016/S1534-5807(01)00088-0
James, D. I. et al. First-in-class chemical probes against poly(ADP-ribose) glycohydrolase (PARG) inhibit DNA repair with differential pharmacology to olaparib. ACS Chem. Biol. 11, 3179–3190 (2016).
pubmed: 27689388 doi: 10.1021/acschembio.6b00609
Murai, J. et al. Stereospecific PARP trapping by BMN 673 and comparison with olaparib and rucaparib. Mol. Cancer Therap. 13, 433–443 (2014).
doi: 10.1158/1535-7163.MCT-13-0803
Tempera, I. et al. Regulation of Epstein-Barr virus OriP replication by poly(ADP-ribose) polymerase 1. J. Virol. 84, 4988–4997 (2010).
pubmed: 20219917 pmcid: 2863838 doi: 10.1128/JVI.02333-09
Robinson, M. D., McCarthy, D. J. & Smyth, G. K. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140 (2010).
pubmed: 19910308 doi: 10.1093/bioinformatics/btp616
Varoquaux, N., Ay, F., Noble, W. S. & Vert, J. P. A statistical approach for inferring the 3D structure of the genome. Bioinformatics 30, i26–i33 (2014).
pubmed: 24931992 pmcid: 4229903 doi: 10.1093/bioinformatics/btu268
Hughes, D. J. et al. Contributions of CTCF and DNA methyltransferases DNMT1 and DNMT3B to Epstein-Barr virus restricted latency. J. Virol. 86, 1034–1045 (2012).
pubmed: 22072770 pmcid: 3255836 doi: 10.1128/JVI.05923-11
O’Neil, N. J., van Pel, D. M. & Hieter, P. Synthetic lethality and cancer: cohesin and PARP at the replication fork. Trends Genet. 29, 290–297 (2013).
pubmed: 23333522 doi: 10.1016/j.tig.2012.12.004
Busslinger, G. A. et al. Cohesin is positioned in mammalian genomes by transcription, CTCF and Wapl. Nature 544, 503–507 (2017).
pubmed: 28424523 pmcid: 6080695 doi: 10.1038/nature22063
Kojic, A. et al. Distinct roles of cohesin-SA1 and cohesin-SA2 in 3D chromosome organization. Nat. Struct. Mol. Biol. 25, 496–504 (2018).
pubmed: 29867216 pmcid: 6122591 doi: 10.1038/s41594-018-0070-4
McAleenan, A. et al. SUMOylation of the alpha-kleisin subunit of cohesin is required for DNA damage-induced cohesion. Curr. Biol. 22, 1564–1575 (2012).
pubmed: 22771042 doi: 10.1016/j.cub.2012.06.045
Alexandru, G., Uhlmann, F., Mechtler, K., Poupart, M. A. & Nasmyth, K. Phosphorylation of the cohesin subunit Scc1 by Polo/Cdc5 kinase regulates sister chromatid separation in yeast. Cell 105, 459–472 (2001).
pubmed: 11371343 doi: 10.1016/S0092-8674(01)00362-2
Kraus, W. L. & Lis, J. T. PARP goes transcription. Cell 113, 677–683 (2003).
pubmed: 12809599 doi: 10.1016/S0092-8674(03)00433-1
Ray Chaudhuri, A. & Nussenzweig, A. The multifaceted roles of PARP1 in DNA repair and chromatin remodelling. Nat. Rev. Mol. Cell Biol. 18, 610–621 (2017).
pubmed: 28676700 pmcid: 6591728 doi: 10.1038/nrm.2017.53
Mattiussi, S. et al. Inhibition of poly(ADP-ribose)polymerase impairs Epstein Barr virus lytic cycle progression. Infect. Agents Cancer 2, 18 (2007).
doi: 10.1186/1750-9378-2-18
Lang, F. et al. CTCF interacts with the lytic HSV-1 genome to promote viral transcription. Sci. Rep. 7, 39861 (2017).
pubmed: 28045091 pmcid: 5206630 doi: 10.1038/srep39861
Ertel, M. K., Cammarata, A. L., Hron, R. J. & Neumann, D. M. CTCF occupation of the herpes simplex virus 1 genome is disrupted at early times postreactivation in a transcription-dependent manner. J. Virol. 86, 12741–12759 (2012).
pubmed: 22973047 pmcid: 3497617 doi: 10.1128/JVI.01655-12
Washington, S. D., Musarrat, F., Ertel, M. K., Backes, G. L. & Neumann, D. M. CTCF binding sites in the herpes simplex virus 1 genome display site-specific ctcf occupation, protein recruitment, and insulator function. J. Virol. https://doi.org/10.1128/JVI.00156-18 (2018).
Watson, Z. L. et al. In vivo knockdown of the herpes simplex virus 1 latency-associated transcript reduces reactivation from latency. J Virol https://doi.org/10.1128/JVI.00812-18 (2018).
Washington, S. D. et al. Depletion of the insulator protein CTCF results in herpes simplex virus 1 reactivation in vivo. J. Virol. https://doi.org/10.1128/JVI.00173-18 (2018).
Stedman, W. et al. Cohesins localize with CTCF at the KSHV latency control region and at cellular c-myc and H19/Igf2 insulators. EMBO J. 27, 654–666 (2008).
pubmed: 18219272 pmcid: 2262040 doi: 10.1038/emboj.2008.1
Kang, H. & Lieberman, P. M. Cell cycle control of Kaposi’s sarcoma-associated herpesvirus latency transcription by CTCF-cohesin interactions. J. Virol. 83, 6199–6210 (2009).
pubmed: 19369356 pmcid: 2687369 doi: 10.1128/JVI.00052-09
Kang, H., Wiedmer, A., Yuan, Y., Robertson, E. & Lieberman, P. M. Coordination of KSHV latent and lytic gene control by CTCF-cohesin mediated chromosome conformation. PLoS Pathog. 7, e1002140, (2011).
pubmed: 21876668 pmcid: 3158054 doi: 10.1371/journal.ppat.1002140
Li, D., Mosbruger, T., Verma, D. & Swaminathan, S. Complex interactions between cohesin and CTCF in regulation of Kaposi’s Sarcoma-associated herpesvirus lytic transcription. J. Virol. https://doi.org/10.1128/JVI.01279-19 (2020).
Li, D. J., Verma, D., Mosbruger, T. & Swaminathan, S. CTCF and Rad21 act as host cell restriction factors for Kaposi’s sarcoma-associated herpesvirus (KSHV) lytic replication by modulating viral gene transcription. PLoS Pathog. 10, e1003880 (2014).
pubmed: 24415941 pmcid: 3887114 doi: 10.1371/journal.ppat.1003880
Martínez, F. P. et al. CTCF binding to the first intron of the major immediate early (MIE) gene of human cytomegalovirus (HCMV) negatively regulates MIE gene expression and HCMV replication. J. Virol. 88, 7389–7401 (2014).
pubmed: 24741094 pmcid: 4054410 doi: 10.1128/JVI.00845-14
Mehta, K., Gunasekharan, V., Satsuka, A. & Laimins, L. A. Human papillomaviruses activate and recruit SMC1 cohesin proteins for the differentiation-dependent life cycle through association with CTCF insulators. PLoS Pathog. 11, e1004763 (2015).
pubmed: 25875106 pmcid: 4395367 doi: 10.1371/journal.ppat.1004763
Paris, C. et al. CCCTC-binding factor recruitment to the early region of the human papillomavirus 18 genome regulates viral oncogene expression. J. Virol. 89, 4770–4785 (2015).
pubmed: 25694598 pmcid: 4403478 doi: 10.1128/JVI.00097-15
Pentland, I. et al. Disruption of CTCF-YY1-dependent looping of the human papillomavirus genome activates differentiation-induced viral oncogene transcription. PLoS Biol. 16, e2005752 (2018).
pubmed: 30359362 pmcid: 6219814 doi: 10.1371/journal.pbio.2005752
Martinez, M. P. et al. HTLV-1 CTCF-binding site is dispensable for in vitro immortalization and persistent infection in vivo. Retrovirology 16, 44 (2019).
pubmed: 31864373 pmcid: 6925871 doi: 10.1186/s12977-019-0507-9
Mandage, R. et al. Genetic factors affecting EBV copy number in lymphoblastoid cell lines derived from the 1000 Genome Project samples. PLoS ONE 12, e0179446 (2017).
pubmed: 28654678 pmcid: 5487016 doi: 10.1371/journal.pone.0179446
Houldcroft, C. J. et al. Host genetic variants and gene expression patterns associated with Epstein-Barr virus copy number in lymphoblastoid cell lines. PLoS ONE 9, e108384 (2014).
pubmed: 25290448 pmcid: 4188571 doi: 10.1371/journal.pone.0108384
Davies, M. L. et al. Cellular factors associated with latency and spontaneous Epstein-Barr virus reactivation in B-lymphoblastoid cell lines. Virology 400, 53–67 (2010).
pubmed: 20153012 doi: 10.1016/j.virol.2010.01.002
Zhang, Y. et al. Model-based analysis of ChIP-Seq (MACS). Genome Biol. 9, R137 (2008).
pubmed: 18798982 pmcid: 2592715 doi: 10.1186/gb-2008-9-9-r137
Feng, J., Liu, T., Qin, B., Zhang, Y. & Liu, X. S. Identifying ChIP-seq enrichment using MACS. Nat. Protoc. 7, 1728–1740 (2012).
pubmed: 22936215 doi: 10.1038/nprot.2012.101
Quinlan, A. R. & Hall, I. M. BEDTools: a flexible suite of utilities for comparing genomic features. Bioinformatics 26, 841–842 (2010).
pubmed: 20110278 pmcid: 2832824 doi: 10.1093/bioinformatics/btq033
Ramirez, F. et al. deepTools2: a next generation web server for deep-sequencing data analysis. Nucleic Acids Res. 44, W160–W165 (2016).
pubmed: 27079975 pmcid: 4987876 doi: 10.1093/nar/gkw257
Tanizawa, H., Kim, K. D., Iwasaki, O. & Noma, K. I. Architectural alterations of the fission yeast genome during the cell cycle. Nat. Struct. Mol. Biol. 24, 965–976 (2017).
pubmed: 28991264 pmcid: 5724045 doi: 10.1038/nsmb.3482
Imakaev, M. et al. Iterative correction of Hi-C data reveals hallmarks of chromosome organization. Nat. Methods 9, 999–1003 (2012).
pubmed: 22941365 pmcid: 3816492 doi: 10.1038/nmeth.2148
Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc. Ser. B (Methodol.) 57, 289–300 (1995).
Dobin, A. et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15–21 (2013).
pubmed: 23104886 doi: 10.1093/bioinformatics/bts635

Auteurs

Sarah M Morgan (SM)

The Wistar Institute, Philadelphia, PA, USA.
Fels Institute for Cancer Research and Molecular Biology, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, USA.

Hideki Tanizawa (H)

University of Oregon, Eugene, OR, USA.

Lisa Beatrice Caruso (LB)

The Wistar Institute, Philadelphia, PA, USA.

Michael Hulse (M)

Fels Institute for Cancer Research and Molecular Biology, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, USA.

Andrew Kossenkov (A)

The Wistar Institute, Philadelphia, PA, USA.

Jozef Madzo (J)

The Coriell Institute for Medical Research, Camden, NJ, USA.

Kelsey Keith (K)

The Coriell Institute for Medical Research, Camden, NJ, USA.

Yinfei Tan (Y)

Fox Chase Cancer Center, Philadelphia, PA, USA.

Sarah Boyle (S)

The Wistar Institute, Philadelphia, PA, USA.

Paul M Lieberman (PM)

The Wistar Institute, Philadelphia, PA, USA.

Italo Tempera (I)

The Wistar Institute, Philadelphia, PA, USA. itempera@wistar.org.

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