PMCC PMCC

Search tips
Search criteria

Advanced
Results 1-25 (25)
 

Clipboard (0)
None

Select a Filter Below

Journals
more »
Year of Publication
Document Types
1.  Global analysis of the nuclear processing of transcripts with unspliced U12-type introns by the exosome 
Nucleic Acids Research  2014;42(11):7358-7369.
U12-type introns are a rare class of introns in the genomes of diverse eukaryotes. In the human genome, they number over 700. A subset of these introns has been shown to be spliced at a slower rate compared to the major U2-type introns. This suggests a rate-limiting regulatory function for the minor spliceosome in the processing of transcripts containing U12-type introns. However, both the generality of slower splicing and the subsequent fate of partially processed pre-mRNAs remained unknown. Here, we present a global analysis of the nuclear retention of transcripts containing U12-type introns and provide evidence for the nuclear decay of such transcripts in human cells. Using SOLiD RNA sequencing technology, we find that, in normal cells, U12-type introns are on average 2-fold more retained than the surrounding U2-type introns. Furthermore, we find that knockdown of RRP41 and DIS3 subunits of the exosome stabilizes an overlapping set of U12-type introns. RRP41 knockdown leads to slower decay kinetics of U12-type introns and globally upregulates the retention of U12-type, but not U2-type, introns. Our results indicate that U12-type introns are spliced less efficiently and are targeted by the exosome. These characteristics support their role in the regulation of cellular mRNA levels.
doi:10.1093/nar/gku391
PMCID: PMC4066798  PMID: 24848017
2.  Genome-wide association studies identify four ER negative–specific breast cancer risk loci 
Garcia-Closas, Montserrat | Couch, Fergus J | Lindstrom, Sara | Michailidou, Kyriaki | Schmidt, Marjanka K | Brook, Mark N | orr, Nick | Rhie, Suhn Kyong | Riboli, Elio | Feigelson, Heather s | Le Marchand, Loic | Buring, Julie E | Eccles, Diana | Miron, Penelope | Fasching, Peter A | Brauch, Hiltrud | Chang-Claude, Jenny | Carpenter, Jane | Godwin, Andrew K | Nevanlinna, Heli | Giles, Graham G | Cox, Angela | Hopper, John L | Bolla, Manjeet K | Wang, Qin | Dennis, Joe | Dicks, Ed | Howat, Will J | Schoof, Nils | Bojesen, Stig E | Lambrechts, Diether | Broeks, Annegien | Andrulis, Irene L | Guénel, Pascal | Burwinkel, Barbara | Sawyer, Elinor J | Hollestelle, Antoinette | Fletcher, Olivia | Winqvist, Robert | Brenner, Hermann | Mannermaa, Arto | Hamann, Ute | Meindl, Alfons | Lindblom, Annika | Zheng, Wei | Devillee, Peter | Goldberg, Mark S | Lubinski, Jan | Kristensen, Vessela | Swerdlow, Anthony | Anton-Culver, Hoda | Dörk, Thilo | Muir, Kenneth | Matsuo, Keitaro | Wu, Anna H | Radice, Paolo | Teo, Soo Hwang | Shu, Xiao-Ou | Blot, William | Kang, Daehee | Hartman, Mikael | Sangrajrang, Suleeporn | Shen, Chen-Yang | Southey, Melissa C | Park, Daniel J | Hammet, Fleur | Stone, Jennifer | Veer, Laura J Van’t | Rutgers, Emiel J | Lophatananon, Artitaya | Stewart-Brown, Sarah | Siriwanarangsan, Pornthep | Peto, Julian | Schrauder, Michael G | Ekici, Arif B | Beckmann, Matthias W | Silva, Isabel dos Santos | Johnson, Nichola | Warren, Helen | Tomlinson, Ian | Kerin, Michael J | Miller, Nicola | Marme, Federick | Schneeweiss, Andreas | Sohn, Christof | Truong, Therese | Laurent-Puig, Pierre | Kerbrat, Pierre | Nordestgaard, Børge G | Nielsen, Sune F | Flyger, Henrik | Milne, Roger L | Perez, Jose Ignacio Arias | Menéndez, Primitiva | Müller, Heiko | Arndt, Volker | Stegmaier, Christa | Lichtner, Peter | Lochmann, Magdalena | Justenhoven, Christina | Ko, Yon-Dschun | Muranen, Taru A | Aittomäki, Kristiina | Blomqvist, Carl | Greco, Dario | Heikkinen, Tuomas | Ito, Hidemi | Iwata, Hiroji | Yatabe, Yasushi | Antonenkova, Natalia N | Margolin, Sara | Kataja, Vesa | Kosma, Veli-Matti | Hartikainen, Jaana M | Balleine, Rosemary | Tseng, Chiu-Chen | Van Den Berg, David | Stram, Daniel O | Neven, Patrick | Dieudonné, Anne-Sophie | Leunen, Karin | Rudolph, Anja | Nickels, Stefan | Flesch-Janys, Dieter | Peterlongo, Paolo | Peissel, Bernard | Bernard, Loris | Olson, Janet E | Wang, Xianshu | Stevens, Kristen | Severi, Gianluca | Baglietto, Laura | Mclean, Catriona | Coetzee, Gerhard A | Feng, Ye | Henderson, Brian E | Schumacher, Fredrick | Bogdanova, Natalia V | Labrèche, France | Dumont, Martine | Yip, Cheng Har | Taib, Nur Aishah Mohd | Cheng, Ching-Yu | Shrubsole, Martha | Long, Jirong | Pylkäs, Katri | Jukkola-Vuorinen, Arja | Kauppila, Saila | knight, Julia A | Glendon, Gord | Mulligan, Anna Marie | Tollenaar, Robertus A E M | Seynaeve, Caroline M | Kriege, Mieke | Hooning, Maartje J | Van den Ouweland, Ans M W | Van Deurzen, Carolien H M | Lu, Wei | Gao, Yu-Tang | Cai, Hui | Balasubramanian, Sabapathy P | Cross, Simon S | Reed, Malcolm W R | Signorello, Lisa | Cai, Qiuyin | Shah, Mitul | Miao, Hui | Chan, Ching Wan | Chia, Kee Seng | Jakubowska, Anna | Jaworska, Katarzyna | Durda, Katarzyna | Hsiung, Chia-Ni | Wu, Pei-Ei | Yu, Jyh-Cherng | Ashworth, Alan | Jones, Michael | Tessier, Daniel C | González-Neira, Anna | Pita, Guillermo | Alonso, M Rosario | Vincent, Daniel | Bacot, Francois | Ambrosone, Christine B | Bandera, Elisa V | John, Esther M | Chen, Gary K | Hu, Jennifer J | Rodriguez-gil, Jorge L | Bernstein, Leslie | Press, Michael F | Ziegler, Regina G | Millikan, Robert M | Deming-Halverson, Sandra L | Nyante, Sarah | Ingles, Sue A | Waisfisz, Quinten | Tsimiklis, Helen | Makalic, Enes | Schmidt, Daniel | Bui, Minh | Gibson, Lorna | Müller-Myhsok, Bertram | Schmutzler, Rita K | Hein, Rebecca | Dahmen, Norbert | Beckmann, Lars | Aaltonen, Kirsimari | Czene, Kamila | Irwanto, Astrid | Liu, Jianjun | Turnbull, Clare | Rahman, Nazneen | Meijers-Heijboer, Hanne | Uitterlinden, Andre G | Rivadeneira, Fernando | Olswold, Curtis | Slager, Susan | Pilarski, Robert | Ademuyiwa, Foluso | Konstantopoulou, Irene | Martin, Nicholas G | Montgomery, Grant W | Slamon, Dennis J | Rauh, Claudia | Lux, Michael P | Jud, Sebastian M | Bruning, Thomas | Weaver, Joellen | Sharma, Priyanka | Pathak, Harsh | Tapper, Will | Gerty, Sue | Durcan, Lorraine | Trichopoulos, Dimitrios | Tumino, Rosario | Peeters, Petra H | Kaaks, Rudolf | Campa, Daniele | Canzian, Federico | Weiderpass, Elisabete | Johansson, Mattias | Khaw, Kay-Tee | Travis, Ruth | Clavel-Chapelon, Françoise | Kolonel, Laurence N | Chen, Constance | Beck, Andy | Hankinson, Susan E | Berg, Christine D | Hoover, Robert N | Lissowska, Jolanta | Figueroa, Jonine D | Chasman, Daniel I | Gaudet, Mia M | Diver, W Ryan | Willett, Walter C | Hunter, David J | Simard, Jacques | Benitez, Javier | Dunning, Alison M | Sherman, Mark E | Chenevix-Trench, Georgia | Chanock, Stephen J | Hall, Per | Pharoah, Paul D P | Vachon, Celine | Easton, Douglas F | Haiman, Christopher A | Kraft, Peter
Nature genetics  2013;45(4):392-398e2.
Estrogen receptor (ER)-negative tumors represent 20–30% of all breast cancers, with a higher proportion occurring in younger women and women of African ancestry1. The etiology2 and clinical behavior3 of ER-negative tumors are different from those of tumors expressing ER (ER positive), including differences in genetic predisposition4. To identify susceptibility loci specific to ER-negative disease, we combined in a meta-analysis 3 genome-wide association studies of 4,193 ER-negative breast cancer cases and 35,194 controls with a series of 40 follow-up studies (6,514 cases and 41,455 controls), genotyped using a custom Illumina array, iCOGS, developed by the Collaborative Oncological Gene-environment Study (COGS). SNPs at four loci, 1q32.1 (MDM4, P = 2.1 × 10−12 and LGR6, P = 1.4 × 10−8), 2p24.1 (P = 4.6 × 10−8) and 16q12.2 (FTO, P = 4.0 × 10−8), were associated with ER-negative but not ER-positive breast cancer (P > 0.05). These findings provide further evidence for distinct etiological pathways associated with invasive ER-positive and ER-negative breast cancers.
doi:10.1038/ng.2561
PMCID: PMC3771695  PMID: 23535733
3.  A meta-analysis of genome-wide association studies of breast cancer identifies two novel susceptibility loci at 6q14 and 20q11 
Siddiq, Afshan | Couch, Fergus J. | Chen, Gary K. | Lindström, Sara | Eccles, Diana | Millikan, Robert C. | Michailidou, Kyriaki | Stram, Daniel O. | Beckmann, Lars | Rhie, Suhn Kyong | Ambrosone, Christine B. | Aittomäki, Kristiina | Amiano, Pilar | Apicella, Carmel | Baglietto, Laura | Bandera, Elisa V. | Beckmann, Matthias W. | Berg, Christine D. | Bernstein, Leslie | Blomqvist, Carl | Brauch, Hiltrud | Brinton, Louise | Bui, Quang M. | Buring, Julie E. | Buys, Saundra S. | Campa, Daniele | Carpenter, Jane E. | Chasman, Daniel I. | Chang-Claude, Jenny | Chen, Constance | Clavel-Chapelon, Françoise | Cox, Angela | Cross, Simon S. | Czene, Kamila | Deming, Sandra L. | Diasio, Robert B. | Diver, W. Ryan | Dunning, Alison M. | Durcan, Lorraine | Ekici, Arif B. | Fasching, Peter A. | Feigelson, Heather Spencer | Fejerman, Laura | Figueroa, Jonine D. | Fletcher, Olivia | Flesch-Janys, Dieter | Gaudet, Mia M. | Gerty, Susan M. | Rodriguez-Gil, Jorge L. | Giles, Graham G. | van Gils, Carla H. | Godwin, Andrew K. | Graham, Nikki | Greco, Dario | Hall, Per | Hankinson, Susan E. | Hartmann, Arndt | Hein, Rebecca | Heinz, Judith | Hoover, Robert N. | Hopper, John L. | Hu, Jennifer J. | Huntsman, Scott | Ingles, Sue A. | Irwanto, Astrid | Isaacs, Claudine | Jacobs, Kevin B. | John, Esther M. | Justenhoven, Christina | Kaaks, Rudolf | Kolonel, Laurence N. | Coetzee, Gerhard A. | Lathrop, Mark | Le Marchand, Loic | Lee, Adam M. | Lee, I-Min | Lesnick, Timothy | Lichtner, Peter | Liu, Jianjun | Lund, Eiliv | Makalic, Enes | Martin, Nicholas G. | McLean, Catriona A. | Meijers-Heijboer, Hanne | Meindl, Alfons | Miron, Penelope | Monroe, Kristine R. | Montgomery, Grant W. | Müller-Myhsok, Bertram | Nickels, Stefan | Nyante, Sarah J. | Olswold, Curtis | Overvad, Kim | Palli, Domenico | Park, Daniel J. | Palmer, Julie R. | Pathak, Harsh | Peto, Julian | Pharoah, Paul | Rahman, Nazneen | Rivadeneira, Fernando | Schmidt, Daniel F. | Schmutzler, Rita K. | Slager, Susan | Southey, Melissa C. | Stevens, Kristen N. | Sinn, Hans-Peter | Press, Michael F. | Ross, Eric | Riboli, Elio | Ridker, Paul M. | Schumacher, Fredrick R. | Severi, Gianluca | dos Santos Silva, Isabel | Stone, Jennifer | Sund, Malin | Tapper, William J. | Thun, Michael J. | Travis, Ruth C. | Turnbull, Clare | Uitterlinden, Andre G. | Waisfisz, Quinten | Wang, Xianshu | Wang, Zhaoming | Weaver, JoEllen | Schulz-Wendtland, Rüdiger | Wilkens, Lynne R. | Van Den Berg, David | Zheng, Wei | Ziegler, Regina G. | Ziv, Elad | Nevanlinna, Heli | Easton, Douglas F. | Hunter, David J. | Henderson, Brian E. | Chanock, Stephen J. | Garcia-Closas, Montserrat | Kraft, Peter | Haiman, Christopher A. | Vachon, Celine M.
Human Molecular Genetics  2012;21(24):5373-5384.
Genome-wide association studies (GWAS) of breast cancer defined by hormone receptor status have revealed loci contributing to susceptibility of estrogen receptor (ER)-negative subtypes. To identify additional genetic variants for ER-negative breast cancer, we conducted the largest meta-analysis of ER-negative disease to date, comprising 4754 ER-negative cases and 31 663 controls from three GWAS: NCI Breast and Prostate Cancer Cohort Consortium (BPC3) (2188 ER-negative cases; 25 519 controls of European ancestry), Triple Negative Breast Cancer Consortium (TNBCC) (1562 triple negative cases; 3399 controls of European ancestry) and African American Breast Cancer Consortium (AABC) (1004 ER-negative cases; 2745 controls). We performed in silico replication of 86 SNPs at P ≤ 1 × 10-5 in an additional 11 209 breast cancer cases (946 with ER-negative disease) and 16 057 controls of Japanese, Latino and European ancestry. We identified two novel loci for breast cancer at 20q11 and 6q14. SNP rs2284378 at 20q11 was associated with ER-negative breast cancer (combined two-stage OR = 1.16; P = 1.1 × 10−8) but showed a weaker association with overall breast cancer (OR = 1.08, P = 1.3 × 10–6) based on 17 869 cases and 43 745 controls and no association with ER-positive disease (OR = 1.01, P = 0.67) based on 9965 cases and 22 902 controls. Similarly, rs17530068 at 6q14 was associated with breast cancer (OR = 1.12; P = 1.1 × 10−9), and with both ER-positive (OR = 1.09; P = 1.5 × 10−5) and ER-negative (OR = 1.16, P = 2.5 × 10−7) disease. We also confirmed three known loci associated with ER-negative (19p13) and both ER-negative and ER-positive breast cancer (6q25 and 12p11). Our results highlight the value of large-scale collaborative studies to identify novel breast cancer risk loci.
doi:10.1093/hmg/dds381
PMCID: PMC3510753  PMID: 22976474
4.  The role of genetic breast cancer susceptibility variants as prognostic factors 
Fasching, Peter A. | Pharoah, Paul D.P. | Cox, Angela | Nevanlinna, Heli | Bojesen, Stig E. | Karn, Thomas | Broeks, Annegien | van Leeuwen, Flora E. | van 't Veer, Laura J. | Udo, Renate | Dunning, Alison M. | Greco, Dario | Aittomäki, Kristiina | Blomqvist, Carl | Shah, Mitul | Nordestgaard, Børge G. | Flyger, Henrik | Hopper, John L. | Southey, Melissa C. | Apicella, Carmel | Garcia-Closas, Montserrat | Sherman, Mark | Lissowska, Jolanta | Seynaeve, Caroline | Huijts, Petra E.A. | Tollenaar, Rob A.E.M. | Ziogas, Argyrios | Ekici, Arif B. | Rauh, Claudia | Mannermaa, Arto | Kataja, Vesa | Kosma, Veli-Matti | Hartikainen, Jaana M. | Andrulis, Irene L. | Ozcelik, Hilmi | Mulligan, Anna-Marie | Glendon, Gord | Hall, Per | Czene, Kamila | Liu, Jianjun | Chang-Claude, Jenny | Wang-Gohrke, Shan | Eilber, Ursula | Nickels, Stefan | Dörk, Thilo | Schiekel, Maria | Bremer, Michael | Park-Simon, Tjoung-Won | Giles, Graham G. | Severi, Gianluca | Baglietto, Laura | Hooning, Maartje J. | Martens, John W.M. | Jager, Agnes | Kriege, Mieke | Lindblom, Annika | Margolin, Sara | Couch, Fergus J. | Stevens, Kristen N. | Olson, Janet E. | Kosel, Matthew | Cross, Simon S. | Balasubramanian, Sabapathy P. | Reed, Malcolm W.R. | Miron, Alexander | John, Esther M. | Winqvist, Robert | Pylkäs, Katri | Jukkola-Vuorinen, Arja | Kauppila, Saila | Burwinkel, Barbara | Marme, Frederik | Schneeweiss, Andreas | Sohn, Christof | Chenevix-Trench, Georgia | Lambrechts, Diether | Dieudonne, Anne-Sophie | Hatse, Sigrid | van Limbergen, Erik | Benitez, Javier | Milne, Roger L. | Zamora, M. Pilar | Pérez, José Ignacio Arias | Bonanni, Bernardo | Peissel, Bernard | Loris, Bernard | Peterlongo, Paolo | Rajaraman, Preetha | Schonfeld, Sara J. | Anton-Culver, Hoda | Devilee, Peter | Beckmann, Matthias W. | Slamon, Dennis J. | Phillips, Kelly-Anne | Figueroa, Jonine D. | Humphreys, Manjeet K. | Easton, Douglas F. | Schmidt, Marjanka K.
Human Molecular Genetics  2012;21(17):3926-3939.
Recent genome-wide association studies identified 11 single nucleotide polymorphisms (SNPs) associated with breast cancer (BC) risk. We investigated these and 62 other SNPs for their prognostic relevance. Confirmed BC risk SNPs rs17468277 (CASP8), rs1982073 (TGFB1), rs2981582 (FGFR2), rs13281615 (8q24), rs3817198 (LSP1), rs889312 (MAP3K1), rs3803662 (TOX3), rs13387042 (2q35), rs4973768 (SLC4A7), rs6504950 (COX11) and rs10941679 (5p12) were genotyped for 25 853 BC patients with the available follow-up; 62 other SNPs, which have been suggested as BC risk SNPs by a GWAS or as candidate SNPs from individual studies, were genotyped for replication purposes in subsets of these patients. Cox proportional hazard models were used to test the association of these SNPs with overall survival (OS) and BC-specific survival (BCS). For the confirmed loci, we performed an accessory analysis of publicly available gene expression data and the prognosis in a different patient group. One of the 11 SNPs, rs3803662 (TOX3) and none of the 62 candidate/GWAS SNPs were associated with OS and/or BCS at P<0.01. The genotypic-specific survival for rs3803662 suggested a recessive mode of action [hazard ratio (HR) of rare homozygous carriers=1.21; 95% CI: 1.09–1.35, P=0.0002 and HR=1.29; 95% CI: 1.12–1.47, P=0.0003 for OS and BCS, respectively]. This association was seen similarly in all analyzed tumor subgroups defined by nodal status, tumor size, grade and estrogen receptor. Breast tumor expression of these genes was not associated with prognosis. With the exception of rs3803662 (TOX3), there was no evidence that any of the SNPs associated with BC susceptibility were associated with the BC survival. Survival may be influenced by a distinct set of germline variants from those influencing susceptibility.
doi:10.1093/hmg/dds159
PMCID: PMC3412377  PMID: 22532573
5.  Gene Expression Patterns Underlying the Reinstatement of Plasticity in the Adult Visual System 
Neural Plasticity  2013;2013:605079.
The nervous system is highly sensitive to experience during early postnatal life, but this phase of heightened plasticity decreases with age. Recent studies have demonstrated that developmental-like plasticity can be reactivated in the visual cortex of adult animals through environmental or pharmacological manipulations. These findings provide a unique opportunity to study the cellular and molecular mechanisms of adult plasticity. Here we used the monocular deprivation paradigm to investigate large-scale gene expression patterns underlying the reinstatement of plasticity produced by fluoxetine in the adult rat visual cortex. We found changes, confirmed with RT-PCRs, in gene expression in different biological themes, such as chromatin structure remodelling, transcription factors, molecules involved in synaptic plasticity, extracellular matrix, and excitatory and inhibitory neurotransmission. Our findings reveal a key role for several molecules such as the metalloproteases Mmp2 and Mmp9 or the glycoprotein Reelin and open up new insights into the mechanisms underlying the reopening of the critical periods in the adult brain.
doi:10.1155/2013/605079
PMCID: PMC3710606  PMID: 23936678
6.  Drug repositioning: a machine-learning approach through data integration 
Existing computational methods for drug repositioning either rely only on the gene expression response of cell lines after treatment, or on drug-to-disease relationships, merging several information levels. However, the noisy nature of the gene expression and the scarcity of genomic data for many diseases are important limitations to such approaches. Here we focused on a drug-centered approach by predicting the therapeutic class of FDA-approved compounds, not considering data concerning the diseases. We propose a novel computational approach to predict drug repositioning based on state-of-the-art machine-learning algorithms. We have integrated multiple layers of information: i) on the distances of the drugs based on how similar are their chemical structures, ii) on how close are their targets within the protein-protein interaction network, and iii) on how correlated are the gene expression patterns after treatment. Our classifier reaches high accuracy levels (78%), allowing us to re-interpret the top misclassifications as re-classifications, after rigorous statistical evaluation. Efficient drug repurposing has the potential to significantly impact the whole field of drug development. The results presented here can significantly accelerate the translation into the clinics of known compounds for novel therapeutic uses.
doi:10.1186/1758-2946-5-30
PMCID: PMC3704944  PMID: 23800010
Drug repositioning; Connectivity map; CMap; ATC code; Mode of action; Machine learning; SVM; Integrative genomics; SMILES; Anthelmintics; Antineoplastic; Oxamniquine; Niclosamide
7.  Gene Expression Profiling of Gliadin Effects on Intestinal Epithelial Cells Suggests Novel Non-Enzymatic Functions of Pepsin and Trypsin 
PLoS ONE  2013;8(6):e66307.
Gliadin triggers T-cell mediated immunity in celiac disease, and has cytotoxic effects on enterocytes mediated through obscure mechanisms. In addition, gliadin transport mechanisms, potential cell surface receptors and gliadin-activated downstream signaling pathways are not completely understood. In order to screen for novel downstream gliadin target genes we performed a systematic whole genome expression study on intestinal epithelial cells. Undifferentiated Caco-2 cells were exposed to pepsin- and trypsin- digested gliadin (PT-G), a blank pepsin-trypsin control (PT) and to a synthetic peptide corresponding to gliadin p31-43 peptide for six hours. RNA from four different experiments was used for hybridization on Agilent one color human whole genome DNA microarray chips. The microarray data were analyzed using the Bioconductor package LIMMA. Genes with nominal p<0.01 were considered statistically significant. Compared to the untreated cells 1705, 1755 and 211 probes were affected by PT-G, PT and p31-43 respectively. 46 probes were significantly different between PT and PT-G treated cells. Among the p31-43 peptide affected probes, 10 and 21 probes were affected by PT-G and PT respectively. Only PT-G affected genes could be validated by quantitative real-time polymerase chain reaction. All the genes were, nonetheless, also affected to a comparable level by PT treated negative controls. In conclusion, we could not replicate previously reported direct effects of gliadin peptides on enterocytes. The results rather suggest that certain epitopes derived from pepsin and trypsin may also affect epithelial cell gene transcription. Our study suggests novel non-enzymatic effects of pepsin and trypsin on cells and calls for proper controls in pepsin and trypsin digested gliadin experiments. It is conceivable that gliadin effects on enterocytes are secondary mediated through oxidative stress, NFkB activation and IL-15 up-regulation.
doi:10.1371/journal.pone.0066307
PMCID: PMC3688875  PMID: 23824913
8.  A common variant at the TERT-CLPTM1L locus is associated with estrogen receptor–negative breast cancer 
Haiman, Christopher A | Chen, Gary K | Vachon, Celine M | Canzian, Federico | Dunning, Alison | Millikan, Robert C | Wang, Xianshu | Ademuyiwa, Foluso | Ahmed, Shahana | Ambrosone, Christine B | Baglietto, Laura | Balleine, Rosemary | Bandera, Elisa V | Beckmann, Matthias W | Berg, Christine D | Bernstein, Leslie | Blomqvist, Carl | Blot, William J | Brauch, Hiltrud | Buring, Julie E | Carey, Lisa A | Carpenter, Jane E | Chang-Claude, Jenny | Chanock, Stephen J | Chasman, Daniel I | Clarke, Christine L | Cox, Angela | Cross, Simon S | Deming, Sandra L | Diasio, Robert B | Dimopoulos, Athanasios M | Driver, W Ryan | Dünnebier, Thomas | Durcan, Lorraine | Eccles, Diana | Edlund, Christopher K | Ekici, Arif B | Fasching, Peter A | Feigelson, Heather S | Flesch-Janys, Dieter | Fostira, Florentia | Försti, Asta | Fountzilas, George | Gerty, Susan M | Giles, Graham G | Godwin, Andrew K | Goodfellow, Paul | Graham, Nikki | Greco, Dario | Hamann, Ute | Hankinson, Susan E | Hartmann, Arndt | Hein, Rebecca | Heinz, Judith | Holbrook, Andrea | Hoover, Robert N | Hu, Jennifer J | Hunter, David J | Ingles, Sue A | Irwanto, Astrid | Ivanovich, Jennifer | John, Esther M | Johnson, Nicola | Jukkola-Vuorinen, Arja | Kaaks, Rudolf | Ko, Yon-Dschun | Kolonel, Laurence N | Konstantopoulou, Irene | Kosma, Veli-Matti | Kulkarni, Swati | Lambrechts, Diether | Lee, Adam M | Le Marchand, Loïc | Lesnick, Timothy | Liu, Jianjun | Lindstrom, Sara | Mannermaa, Arto | Margolin, Sara | Martin, Nicholas G | Miron, Penelope | Montgomery, Grant W | Nevanlinna, Heli | Nickels, Stephan | Nyante, Sarah | Olswold, Curtis | Palmer, Julie | Pathak, Harsh | Pectasides, Dimitrios | Perou, Charles M | Peto, Julian | Pharoah, Paul D P | Pooler, Loreall C | Press, Michael F | Pylkäs, Katri | Rebbeck, Timothy R | Rodriguez-Gil, Jorge L | Rosenberg, Lynn | Ross, Eric | Rüdiger, Thomas | Silva, Isabel dos Santos | Sawyer, Elinor | Schmidt, Marjanka K | Schulz-Wendtland, Rüdiger | Schumacher, Fredrick | Severi, Gianluca | Sheng, Xin | Signorello, Lisa B | Sinn, Hans-Peter | Stevens, Kristen N | Southey, Melissa C | Tapper, William J | Tomlinson, Ian | Hogervorst, Frans B L | Wauters, Els | Weaver, JoEllen | Wildiers, Hans | Winqvist, Robert | Van Den Berg, David | Wan, Peggy | Xia, Lucy Y | Yannoukakos, Drakoulis | Zheng, Wei | Ziegler, Regina G | Siddiq, Afshan | Slager, Susan L | Stram, Daniel O | Easton, Douglas | Kraft, Peter | Henderson, Brian E | Couch, Fergus J
Nature Genetics  2011;43(12):1210-1214.
Estrogen receptor (ER)-negative breast cancer shows a higher incidence in women of African ancestry compared to women of European ancestry. In search of common risk alleles for ER-negative breast cancer, we combined genome-wide association study (GWAS) data from women of African ancestry (1,004 ER-negative cases and 2,745 controls) and European ancestry (1,718 ER-negative cases and 3,670 controls), with replication testing conducted in an additional 2,292 ER-negative cases and 16,901 controls of European ancestry. We identified a common risk variant for ER-negative breast cancer at the TERT-CLPTM1L locus on chromosome 5p15 (rs10069690: per-allele odds ratio (OR) = 1.18 per allele, P = 1.0 × 10−10). The variant was also significantly associated with triple-negative (ER-negative, progesterone receptor (PR)-negative and human epidermal growth factor-2 (HER2)-negative) breast cancer (OR = 1.25, P = 1.1 × 10−9), particularly in younger women (<50 years of age) (OR = 1.48, P = 1.9 × 10−9). Our results identify a genetic locus associated with estrogen receptor negative breast cancer subtypes in multiple populations.
doi:10.1038/ng.985
PMCID: PMC3279120  PMID: 22037553
9.  19p13.1 is a triple negative-specific breast cancer susceptibility locus 
Stevens, Kristen N. | Fredericksen, Zachary | Vachon, Celine M. | Wang, Xianshu | Margolin, Sara | Lindblom, Annika | Nevanlinna, Heli | Greco, Dario | Aittomäki, Kristiina | Blomqvist, Carl | Chang-Claude, Jenny | Vrieling, Alina | Flesch-Janys, Dieter | Sinn, Hans-Peter | Wang-Gohrke, Shan | Nickels, Stefan | Brauch, Hiltrud | Ko, Yon-Dschun | Fischer, Hans-Peter | Schmutzler, Rita K. | Meindl, Alfons | Bartram, Claus R. | Schott, Sarah | Engel, Christof | Godwin, Andrew K. | Weaver, JoEllen | Pathak, Harsh B. | Sharma, Priyanka | Brenner, Hermann | Müller, Heiko | Arndt, Volker | Stegmaier, Christa | Miron, Penelope | Yannoukakos, Drakoulis | Stavropoulou, Alexandra | Fountzilas, George | Gogas, Helen J. | Swann, Ruth | Dwek, Miriam | Perkins, Annie | Milne, Roger L. | Benítez, Javier | Zamora, M Pilar | Pérez, José Ignacio Arias | Bojesen, Stig E. | Nielsen, Sune F. | Nordestgaard, Børge G | Flyger, Henrik | Guénel, Pascal | Truong, Thérèse | Menegaux, Florence | Cordina-Duverger, Emilie | Burwinkel, Barbara | Marmé, Frederick | Schneeweiss, Andreas | Sohn, Christof | Sawyer, Elinor | Tomlinson, Ian | Kerin, Michael J. | Peto, Julian | Johnson, Nichola | Fletcher, Olivia | Silva, Isabel dos Santos | Fasching, Peter A. | Beckmann, Matthias W. | Hartmann, Arndt | Ekici, Arif B. | Lophatananon, Artitaya | Muir, Kenneth | Puttawibul, Puttisak | Wiangnon, Surapon | Schmidt, Marjanka K | Broeks, Annegien | Braaf, Linde M | Rosenberg, Efraim H | Hopper, John L. | Apicella, Carmel | Park, Daniel J. | Southey, Melissa C. | Swerdlow, Anthony J. | Ashworth, Alan | Orr, Nicholas | Schoemaker, Minouk J. | Anton-Culver, Hoda | Ziogas, Argyrios | Bernstein, Leslie | Dur, Christina Clarke | Shen, Chen-Yang | Yu, Jyh-Cherng | Hsu, Huan-Ming | Hsiung, Chia-Ni | Hamann, Ute | Dünnebier, Thomas | Rüdiger, Thomas | Ulmer, Hans Ulrich | Pharoah, Paul P. | Dunning, Alison M | Humphreys, Manjeet K. | Wang, Qin | Cox, Angela | Cross, Simon S. | Reed, Malcom W. | Hall, Per | Czene, Kamila | Ambrosone, Christine B. | Ademuyiwa, Foluso | Hwang, Helena | Eccles, Diana M. | Garcia-Closas, Montserrat | Figueroa, Jonine D. | Sherman, Mark E. | Lissowska, Jolanta | Devilee, Peter | Seynaeve, Caroline | Tollenaar, R.A.E.M. | Hooning, Maartje J. | Andrulis, Irene L. | Knight, Julia A. | Glendon, Gord | Mulligan, Anna Marie | Winqvist, Robert | Pylkäs, Katri | Jukkola-Vuorinen, Arja | Grip, Mervi | John, Esther M. | Miron, Alexander | Alnæs, Grethe Grenaker | Kristensen, Vessela | Børresen-Dale, Anne-Lise | Giles, Graham G. | Baglietto, Laura | McLean, Catriona A | Severi, Gianluca | Kosel, Matthew L. | Pankratz, V.S. | Slager, Susan | Olson, Janet E. | Radice, Paolo | Peterlongo, Paolo | Manoukian, Siranoush | Barile, Monica | Lambrechts, Diether | Hatse, Sigrid | Dieudonne, Anne-Sophie | Christiaens, Marie-Rose | Chenevix-Trench, Georgia | Beesley, Jonathan | Chen, Xiaoqing | Mannermaa, Arto | Kosma, Veli-Matti | Hartikainen, Jaana M. | Soini, Ylermi | Easton, Douglas F. | Couch, Fergus J.
Cancer Research  2012;72(7):1795-1803.
The 19p13.1 breast cancer susceptibility locus is a modifier of breast cancer risk in BRCA1 mutation carriers and is also associated with risk of ovarian cancer. Here we investigated 19p13.1 variation and risk of breast cancer subtypes, defined by estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) status, using 48,869 breast cancer cases and 49,787 controls from the Breast Cancer Association Consortium (BCAC). Variants from 19p13.1 were not associated with breast cancer overall or with ER-positive breast cancer but were significantly associated with ER-negative breast cancer risk [rs8170 Odds Ratio (OR)=1.10, 95% Confidence Interval (CI) 1.05 – 1.15, p=3.49 × 10-5] and triple negative (TN) (ER, PR and HER2 negative) breast cancer [rs8170 OR=1.22, 95% CI 1.13 – 1.31, p=2.22 × 10-7]. However, rs8170 was no longer associated with ER-negative breast cancer risk when TN cases were excluded [OR=0.98, 95% CI 0.89 – 1.07, p=0.62]. In addition, a combined analysis of TN cases from BCAC and the Triple Negative Breast Cancer Consortium (TNBCC) (n=3,566) identified a genome-wide significant association between rs8170 and TN breast cancer risk [OR=1.25, 95% CI 1.18 – 1.33, p=3.31 × 10-13]. Thus, 19p13.1 is the first triple negative-specific breast cancer risk locus and the first locus specific to a histological subtype defined by ER, PR, and HER2 to be identified. These findings provide convincing evidence that genetic susceptibility to breast cancer varies by tumor subtype and that triple negative tumors and other subtypes likely arise through distinct etiologic pathways.
doi:10.1158/0008-5472.CAN-11-3364
PMCID: PMC3319792  PMID: 22331459
genetic susceptibility; association study; subtype; neoplasms; common variant
10.  Differential DNA Methylation in Purified Human Blood Cells: Implications for Cell Lineage and Studies on Disease Susceptibility 
PLoS ONE  2012;7(7):e41361.
Methylation of cytosines at CpG sites is a common epigenetic DNA modification that can be measured by a large number of methods, now even in a genome-wide manner for hundreds of thousands of sites. The application of DNA methylation analysis is becoming widely popular in complex disorders, for example, to understand part of the “missing heritability”. The DNA samples most readily available for methylation studies are derived from whole blood. However, blood consists of many functionally and developmentally distinct cell populations in varying proportions. We studied whether such variation might affect the interpretation of methylation studies based on whole blood DNA. We found in healthy male blood donors there is important variation in the methylation profiles of whole blood, mononuclear cells, granulocytes, and cells from seven selected purified lineages. CpG methylation between mononuclear cells and granulocytes differed for 22% of the 8252 probes covering the selected 343 genes implicated in immune-related disorders by genome-wide association studies, and at least one probe was differentially methylated for 85% of the genes, indicating that whole blood methylation results might be unintelligible. For individual genes, even if the overall methylation patterns might appear similar, a few CpG sites in the regulatory regions may have opposite methylation patterns (i.e., hypo/hyper) in the main blood cell types. We conclude that interpretation of whole blood methylation profiles should be performed with great caution and for any differences implicated in a disorder, the differences resulting from varying proportions of white blood cell types should be considered.
doi:10.1371/journal.pone.0041361
PMCID: PMC3405143  PMID: 22848472
11.  Comparison of Dorsocervical With Abdominal Subcutaneous Adipose Tissue in Patients With and Without Antiretroviral Therapy–Associated Lipodystrophy 
Diabetes  2011;60(7):1894-1900.
OBJECTIVE
Combination antiretroviral therapy (cART) is associated with lipodystrophy, i.e., loss of subcutaneous adipose tissue in the abdomen, limbs, and face and its accumulation intra-abdominally. No fat is lost dorsocervically and it can even accumulate in this region (buffalo hump). It is unknown how preserved dorsocervical fat differs from abdominal subcutaneous fat in HIV-1–infected cART-treated patients with (cART+LD+) and without (cART+LD−) lipodystrophy.
RESEARCH DESIGN AND METHODS
We used histology, microarray, PCR, and magnetic resonance imaging to compare dorsocervical and abdominal subcutaneous adipose tissue in cART+LD+ (n = 21) and cART+LD− (n = 11).
RESULTS
Albeit dorsocervical adipose tissue in cART+LD+ seems spared from lipoatrophy, its mitochondrial DNA (mtDNA; copies/cell) content was significantly lower (by 62%) than that of the corresponding tissue in cART+LD−. Expression of CD68 mRNA, a marker of macrophages, and numerous inflammatory genes in microarray were significantly lower in dorsocervical versus abdominal subcutaneous adipose tissue. Genes with the greatest difference in expression between the two depots were those involved in regulation of transcription and regionalization (homeobox genes), irrespective of lipodystrophy status. There was negligible mRNA expression of uncoupling protein 1, a gene characteristic of brown adipose tissue, in either depot.
CONCLUSIONS
Because mtDNA is depleted even in the nonatrophic dorsocervical adipose tissue, it is unlikely that the cause of lipoatrophy is loss of mtDNA. Dorsocervical adipose tissue is less inflamed than lipoatrophic adipose tissue. It does not resemble brown adipose tissue. The greatest difference in gene expression between dorsocervical and abdominal subcutaneous adipose tissue is in expression of homeobox genes.
doi:10.2337/db11-0075
PMCID: PMC3121420  PMID: 21602514
12.  Common breast cancer susceptibility loci are associated with triple negative breast cancer 
Stevens, Kristen N. | Vachon, Celine M. | Lee, Adam M. | Slager, Susan | Lesnick, Timothy | Olswold, Curtis | Fasching, Peter A. | Miron, Penelope | Eccles, Diana | Carpenter, Jane E. | Godwin, Andrew K. | Ambrosone, Christine | Winqvist, Robert | Schmidt, Marjanka K. | Cox, Angela | Cross, Simon S. | Sawyer, Elinor | Hartmann, Arndt | Beckmann, Matthias W. | Schulz-Wendtland, Rüdiger | Ekici, Arif B. | Tapper, William J | Gerty, Susan M | Durcan, Lorraine | Graham, Nikki | Hein, Rebecca | Nickels, Stephan | Flesch-Janys, Dieter | Heinz, Judith | Sinn, Hans-Peter | Konstantopoulou, Irene | Fostira, Florentia | Pectasides, Dimitrios | Dimopoulos, Athanasios M. | Fountzilas, George | Clarke, Christine L. | Balleine, Rosemary | Olson, Janet E. | Fredericksen, Zachary | Diasio, Robert B. | Pathak, Harsh | Ross, Eric | Weaver, JoEllen | Rüdiger, Thomas | Försti, Asta | Dünnebier, Thomas | Ademuyiwa, Foluso | Kulkarni, Swati | Pylkäs, Katri | Jukkola-Vuorinen, Arja | Ko, Yon-Dschun | Van Limbergen, Erik | Janssen, Hilde | Peto, Julian | Fletcher, Olivia | Giles, Graham G. | Baglietto, Laura | Verhoef, Senno | Tomlinson, Ian | Kosma, Veli-Matti | Beesley, Jonathan | Greco, Dario | Blomqvist, Carl | Irwanto, Astrid | Liu, Jianjun | Blows, Fiona M. | Dawson, Sarah-Jane | Margolin, Sara | Mannermaa, Arto | Martin, Nicholas G. | Montgomery, Grant W | Lambrechts, Diether | dos Santos Silva, Isabel | Severi, Gianluca | Hamann, Ute | Pharoah, Paul | Easton, Douglas F. | Chang-Claude, Jenny | Yannoukakos, Drakoulis | Nevanlinna, Heli | Wang, Xianshu | Couch, Fergus J.
Cancer Research  2011;71(19):6240-6249.
Triple negative breast cancers are an aggressive subtype of breast cancer with poor survival, but there remains little known about the etiological factors which promote its initiation and development. Commonly inherited breast cancer risk factors identified through genome wide association studies (GWAS) display heterogeneity of effect among breast cancer subtypes as defined by estrogen receptor (ER) and progesterone receptor (PR) status. In the Triple Negative Breast Cancer Consortium (TNBCC), 22 common breast cancer susceptibility variants were investigated in 2,980 Caucasian women with triple negative breast cancer and 4,978 healthy controls. We identified six single nucleotide polymorphisms (SNPs) significantly associated with risk of triple negative breast cancer, including rs2046210 (ESR1), rs12662670 (ESR1), rs3803662 (TOX3), rs999737 (RAD51L1), rs8170 (19p13.11) and rs8100241 (19p13.11). Together, our results provide convincing evidence of genetic susceptibility for triple negative breast cancer.
doi:10.1158/0008-5472.CAN-11-1266
PMCID: PMC3327299  PMID: 21844186
genetic susceptibility; neoplasms; association study; subtypes; common variant
13.  Gene expression analysis of Drosophilaa Manf mutants reveals perturbations in membrane traffic and major metabolic changes 
BMC Genomics  2012;13:134.
Background
MANF and CDNF are evolutionarily conserved neurotrophic factors that specifically support dopaminergic neurons. To date, the receptors and signalling pathways of this novel MANF/CDNF family have remained unknown. Independent studies have showed upregulation of MANF by unfolded protein response (UPR). To enlighten the role of MANF in multicellular organism development we carried out a microarray-based analysis of the transcriptional changes induced by the loss and overexpression of Drosophila Manf.
Results
The most dramatic change of expression was observed with genes coding membrane transport proteins and genes related to metabolism. When evaluating in parallel the ultrastructural data and transcriptome changes of maternal/zygotic and only zygotic Manf mutants, the endoplasmic reticulum (ER) stress and membrane traffic alterations were evident. In Drosophila Manf mutants the expression of several genes involved in Parkinson's disease (PD) was altered as well.
Conclusions
We conclude that besides a neurotrophic factor, Manf is an important cellular survival factor needed to overcome the UPR especially in tissues with high secretory function. In the absence of Manf, the expression of genes involved in membrane transport, particularly exocytosis and endosomal recycling pathway was altered. In neurodegenerative diseases, such as PD, correct protein folding and proteasome function as well as neurotransmitter synthesis and uptake are crucial for the survival of neurons. The degeneration of dopaminergic neurons is the hallmark for PD and our work provides a clue on the mechanisms by which the novel neurotrophic factor MANF protects these neurons.
doi:10.1186/1471-2164-13-134
PMCID: PMC3364883  PMID: 22494833
14.  Variants on the promoter region of PTEN affect breast cancer progression and patient survival 
Breast Cancer Research : BCR  2011;13(6):R130.
Introduction
The PTEN gene, a regulator of the phosphatidylinositol-3-kinase (PI3K)/Akt oncogenic pathway, is mutated in various cancers and its expression has been associated with tumor progression in a dose-dependent fashion. We investigated the effect of germline variation in the promoter region of the PTEN gene on clinical characteristics and survival in breast cancer.
Methods
We screened the promoter region of the PTEN gene for germline variation in 330 familial breast cancer cases and further determined the genotypes of three detected PTEN promoter polymorphisms -903GA, -975GC, and -1026CA in a total of 2,412 breast cancer patients to evaluate the effects of the variants on tumor characteristics and disease outcome. We compared the gene expression profiles in breast cancers of 10 variant carriers and 10 matched non-carriers and performed further survival analyses based on the differentially expressed genes.
Results
All three promoter variants associated with worse prognosis. The Cox's regression hazard ratio for 10-year breast cancer specific survival in multivariate analysis was 2.01 (95% CI 1.17 to 3.46) P = 0.0119, and for 5-year breast cancer death or distant metastasis free survival 1.79 (95% CI 1.03 to 3.11) P = 0.0381 for the variant carriers, indicating PTEN promoter variants as an independent prognostic factor. The breast tumors from the promoter variant carriers exhibited a similar gene expression signature of 160 differentially expressed genes compared to matched non-carrier tumors. The signature further stratified patients into two groups with different recurrence free survival in independent breast cancer gene expression data sets.
Conclusions
Inherited variation in the PTEN promoter region affects the tumor progression and gene expression profile in breast cancer. Further studies are warranted to establish PTEN promoter variants as clinical markers for prognosis in breast cancer.
doi:10.1186/bcr3076
PMCID: PMC3326572  PMID: 22171747
15.  MiR-34a Expression Has an Effect for Lower Risk of Metastasis and Associates with Expression Patterns Predicting Clinical Outcome in Breast Cancer 
PLoS ONE  2011;6(11):e26122.
MiR-34a acts as a candidate tumour suppressor gene, and its expression is reduced in several cancer types. We aimed to study miR-34a expression in breast cancer and its correlation with tumour characteristics and clinical outcome, and regulatory links with other genes. We analysed miR-34a expression in 1,172 breast tumours on TMAs. 25% of the tumours showed high, 43% medium and 32% low expression of miR-34a. High miR-34a expression associated with poor prognostic factors for breast cancer: positive nodal status (p = 0.006), high tumour grade (p<0.0001), ER-negativity (p = 0.0002), HER2-positivity (p = 0.0002), high proliferation rate (p<0.0001), p53-positivity (p<0.0001), high cyclin E (p<0.0001) and γH2AX (p<0.0001). However, multivariate analysis adjusting for conventional prognostic factors indicated that high miR-34a expression in fact associated with a lower risk of recurrence or death from breast cancer (HR = 0.63, 95% CI = 0.41–0.96, p = 0.031). Gene expression analysis by differential miR-34a expression revealed an expression signature with an effect on both the 5-year and 10-year survival of the patients (p<0.001). Functional genomic analysis highlighted a novel regulatory role of the transcription factor MAZ, apart from the known control by p53, on the expression of miR-34a and a number of miR-34a targets. Our findings suggest that while miR-34a expression activation is a marker of aggressive breast tumour phenotype it exerts an independent effect for a lower risk of recurrence or death from breast cancer. We also present an expression signature of 190 genes associated with miR-34a expression. Our analysis for regulatory loops suggest that MAZ and p53 transcription factors co-operate in modulating miR-34a, as well as miR-34a targets involved in several cellular pathways. Taken together, these results suggest that the network of genes co-regulated with and targeted by miR-34a form a group of down-stream effectors that maybe of use in predicting clinical outcome, and that highlight novel regulatory mechanisms in breast cancer.
doi:10.1371/journal.pone.0026122
PMCID: PMC3213093  PMID: 22102859
16.  Breast tumors from CHEK2 1100delC-mutation carriers: genomic landscape and clinical implications 
Introduction
Checkpoint kinase 2 (CHEK2) is a moderate penetrance breast cancer risk gene, whose truncating mutation 1100delC increases the risk about twofold. We investigated gene copy-number aberrations and gene-expression profiles that are typical for breast tumors of CHEK2 1100delC-mutation carriers.
Methods
In total, 126 breast tumor tissue specimens including 32 samples from patients carrying CHEK2 1100delC were studied in array-comparative genomic hybridization (aCGH) and gene-expression (GEX) experiments. After dimensionality reduction with CGHregions R package, CHEK2 1100delC-associated regions in the aCGH data were detected by the Wilcoxon rank-sum test. The linear model was fitted to GEX data with R package limma. Genes whose expression levels were associated with CHEK2 1100delC mutation were detected by the bayesian method.
Results
We discovered four lost and three gained CHEK2 1100delC-related loci. These include losses of 1p13.3-31.3, 8p21.1-2, 8p23.1-2, and 17p12-13.1 as well as gains of 12q13.11-3, 16p13.3, and 19p13.3. Twenty-eight genes located on these regions showed differential expression between CHEK2 1100delC and other tumors, nominating them as candidates for CHEK2 1100delC-associated tumor-progression drivers. These included CLCA1 on 1p22 as well as CALCOCO1, SBEM, and LRP1 on 12q13. Altogether, 188 genes were differentially expressed between CHEK2 1100delC and other tumors. Of these, 144 had elevated and 44, reduced expression levels.
Our results suggest the WNT pathway as a driver of tumorigenesis in breast tumors of CHEK2 1100delC-mutation carriers and a role for the olfactory receptor protein family in cancer progression. Differences in the expression of the 188 CHEK2 1100delC-associated genes divided breast tumor samples from three independent datasets into two groups that differed in their relapse-free survival time.
Conclusions
We have shown that copy-number aberrations of certain genomic regions are associated with CHEK2 mutation 1100delC. On these regions, we identified potential drivers of CHEK2 1100delC-associated tumorigenesis, whose role in cancer progression is worth investigating. Furthermore, poorer survival related to the CHEK2 1100delC gene-expression signature highlights pathways that are likely to have a role in the development of metastatic disease in carriers of the CHEK2 1100delC mutation.
doi:10.1186/bcr3015
PMCID: PMC3262202  PMID: 21542898
17.  Human Papillomavirus 16 E5 Modulates the Expression of Host MicroRNAs 
PLoS ONE  2011;6(7):e21646.
Human papillomavirus (HPV) infection is a prerequisite of developing cervical cancer, approximately half of which are associated with HPV type 16. HPV 16 encodes three oncogenes, E5, E6, and E7, of which E5 is the least studied so far. Its roles in regulating replication and pathogenesis of HPV are not fully understood. Here we utilize high-throughput screening to coordinately investigate the effect of E5 on the expression of host protein-coding and microRNA genes. MicroRNAs form a class of 22nt long noncoding RNAs with regulatory activity. Among the altered cellular microRNAs we focus on the alteration in the expression of miR-146a, miR-203 and miR-324-5p and their target genes in a time interval of 96 hours of E5 induction. Our results indicate that HPV infection and subsequent transformation take place through complex regulatory patterns of gene expression in the host cells, part of which are regulated by the E5 protein.
doi:10.1371/journal.pone.0021646
PMCID: PMC3128596  PMID: 21747943
18.  MicroRNA Expression Profiling Reveals MiRNA Families Regulating Specific Biological Pathways in Mouse Frontal Cortex and Hippocampus 
PLoS ONE  2011;6(6):e21495.
MicroRNAs (miRNAs) are small regulatory molecules that cause post-transcriptional gene silencing. Although some miRNAs are known to have region-specific expression patterns in the adult brain, the functional consequences of the region-specificity to the gene regulatory networks of the brain nuclei are not clear. Therefore, we studied miRNA expression patterns by miRNA-Seq and microarrays in two brain regions, frontal cortex (FCx) and hippocampus (HP), which have separate biological functions. We identified 354 miRNAs from FCx and 408 from HP using miRNA-Seq, and 245 from FCx and 238 from HP with microarrays. Several miRNA families and clusters were differentially expressed between FCx and HP, including the miR-8 family, miR-182|miR-96|miR-183 cluster, and miR-212|miR-312 cluster overexpressed in FCx and miR-34 family overexpressed in HP. To visualize the clusters, we developed support for viewing genomic alignments of miRNA-Seq reads in the Chipster genome browser. We carried out pathway analysis of the predicted target genes of differentially expressed miRNA families and clusters to assess their putative biological functions. Interestingly, several miRNAs from the same family/cluster were predicted to regulate specific biological pathways. We have developed a miRNA-Seq approach with a bioinformatic analysis workflow that is suitable for studying miRNA expression patterns from specific brain nuclei. FCx and HP were shown to have distinct miRNA expression patterns which were reflected in the predicted gene regulatory pathways. This methodology can be applied for the identification of brain region-specific and phenotype-specific miRNA-mRNA-regulatory networks from the adult and developing rodent brain.
doi:10.1371/journal.pone.0021495
PMCID: PMC3120887  PMID: 21731767
19.  Gene Expression Profiling of U12-Type Spliceosome Mutant Drosophila Reveals Widespread Changes in Metabolic Pathways 
PLoS ONE  2010;5(10):e13215.
Background
The U12-type spliceosome is responsible for the removal of a subset of introns from eukaryotic mRNAs. U12-type introns are spliced less efficiently than normal U2-type introns, which suggests a rate-limiting role in gene expression. The Drosophila genome contains about 20 U12-type introns, many of them in essential genes, and the U12-type spliceosome has previously been shown to be essential in the fly.
Methodology/Principal Findings
We have used a Drosophila line with a P-element insertion in U6atac snRNA, an essential component of the U12-type spliceosome, to investigate the impact of U12-type introns on gene expression at the organismal level during fly development. This line exhibits progressive accumulation of unspliced U12-type introns during larval development and the death of larvae at the third instar stage. Surprisingly, microarray and RT-PCR analyses revealed that most genes containing U12-type introns showed only mild perturbations in the splicing of U12-type introns. In contrast, we detected widespread downstream effects on genes that do not contain U12-type introns, with genes related to various metabolic pathways constituting the largest group.
Conclusions/Significance
U12-type intron-containing genes exhibited variable gene-specific responses to the splicing defect, with some genes showing up- or downregulation, while most did not change significantly. The observed residual U12-type splicing activity could be explained with the mutant U6atac allele having a low level of catalytic activity. Detailed analysis of all genes suggested that a defect in the splicing of the U12-type intron of the mitochondrial prohibitin gene may be the primary cause of the various downstream effects detected in the microarray analysis.
doi:10.1371/journal.pone.0013215
PMCID: PMC2952598  PMID: 20949011
20.  Bayesian integrated modeling of expression data: a case study on RhoG 
BMC Bioinformatics  2010;11:295.
Background
DNA microarrays provide an efficient method for measuring activity of genes in parallel and even covering all the known transcripts of an organism on a single array. This has to be balanced against that analyzing data emerging from microarrays involves several consecutive steps, and each of them is a potential source of errors. Errors tend to accumulate when moving from the lower level towards the higher level analyses because of the sequential nature. Eliminating such errors does not seem feasible without completely changing the technologies, but one should nevertheless try to meet the goal of being able to realistically assess degree of the uncertainties that are involved when drawing the final conclusions from such analyses.
Results
We present a Bayesian hierarchical model for finding differentially expressed genes between two experimental conditions, proposing an integrated statistical approach where correcting signal saturation, systematic array effects, dye effects, and finding differentially expressed genes, are all modeled jointly. The integration allows all these components, and also the associated errors, to be considered simultaneously. The inference is based on full posterior distribution of gene expression indices and on quantities derived from them rather than on point estimates. The model was applied and tested on two different datasets.
Conclusions
The method presents a way of integrating various steps of microarray analysis into a single joint analysis, and thereby enables extracting information on differential expression in a manner, which properly accounts for various sources of potential error in the process.
doi:10.1186/1471-2105-11-295
PMCID: PMC2894040  PMID: 20515463
21.  Gene expression analysis in SV-40 immortalized human corneal epithelial cells cultured with an air-liquid interface 
Molecular Vision  2010;16:2109-2120.
Purpose
To compare the global gene expression profile of stratified epithelia generated in vitro using simian virus 40 (SV40) immortalized human corneal epithelial cells with the previously reported gene expression of normal human corneal epithelia.
Methods
Immortalized cells expanded in submerged culture were grown in an air-liquid interface of liquid permeable collagen-coated filters to foster stratification and differentiation. Stratified epithelia displaying resistances exceeding 300 Ω · cm2 were dissolved in an RNA purification lysis buffer. Purified RNA was used to globally determine gene expression levels using high-density single-channel oligonucleotide microarrays. Raw hybridization readings were converted into relative gene expression levels using Robust Multi-array Average (RMA) algorithm. Expression levels for selected genes were validated by real-time RT-qPCR. The biologic significance of the gene expression profiles was interpreted with the help of several microarray software analysis tools and ad hoc thematical analysis.
Results
The stratified cell culture to native epithelial comparison identified over- and under-expression in 22% and 14% of the probed genes, respectively. The larger expression decreases occurred in genes intimately associated with both the stratified epithelial lineage at large such as keratin 14 and the corneal phenotype, such as keratin 12, connexin 43, aldehyde dehydrogenases (ALDHs), and paired box gene 6 (PAX6) and its whole downstream transcriptome. Overexpression related to genes associated with cell cycling stimulation.
Conclusions
The results indicate that the stratified corneal epithelial cell model generated using SV40 immortalized cells may be useful only in certain research applications. Extrapolations of studies with these cells to actual tissue cells should be done with a great deal of caution.
PMCID: PMC2994346  PMID: 21139686
22.  Correction: Comparison of Gene Expression Profile in Embryonic Mesencephalon and Neuronal Primary Cultures 
PLoS ONE  2009;4(5):10.1371/annotation/bb336408-6ec5-480e-ae37-85e9cefe0454.
doi:10.1371/annotation/bb336408-6ec5-480e-ae37-85e9cefe0454
PMCID: PMC2685749
23.  Comparison of Gene Expression Profile in Embryonic Mesencephalon and Neuronal Primary Cultures 
PLoS ONE  2009;4(3):e4977.
In the mammalian central nervous system (CNS) an important contingent of dopaminergic neurons are localized in the substantia nigra and in the ventral tegmental area of the ventral midbrain. They constitute an anatomically and functionally heterogeneous group of cells involved in a variety of regulatory mechanisms, from locomotion to emotional/motivational behavior. Midbrain dopaminergic neuron (mDA) primary cultures represent a useful tool to study molecular mechanisms involved in their development and maintenance. Considerable information has been gathered on the mDA neurons development and maturation in vivo, as well as on the molecular features of mDA primary cultures. Here we investigated in detail the gene expression differences between the tissue of origin and ventral midbrain primary cultures enriched in mDA neurons, using microarray technique. We integrated the results based on different re-annotations of the microarray probes. By using knowledge-based gene network techniques and promoter sequence analysis, we also uncovered mechanisms that might regulate the expression of CNS genes involved in the definition of the identity of specific cell types in the ventral midbrain. We integrate bioinformatics and functional genomics, together with developmental neurobiology. Moreover, we propose guidelines for the computational analysis of microarray gene expression data. Our findings help to clarify some molecular aspects of the development and differentiation of DA neurons within the midbrain.
doi:10.1371/journal.pone.0004977
PMCID: PMC2654915  PMID: 19305503
24.  Physiology, Pathology and Relatedness of Human Tissues from Gene Expression Meta-Analysis 
PLoS ONE  2008;3(4):e1880.
Background
Development and maintenance of the identity of tissues is of central importance for multicellular organisms. Based on gene expression profiles, it is possible to divide genes in housekeeping genes and those whose expression is preferential in one or a few tissues and which provide specialized functions that have a strong effect on the physiology of the whole organism.
Results
We have surveyed the gene expression in 78 normal human tissues integrating publicly available microarray gene expression data. A total amount of 1601 genes were identified as selectively expressed in one or more tissues. The tissue-selective genes covered a wide range of cellular and molecular functions, and could be linked to 361 human diseases with Mendelian inheritance. Based on the gene expression profiles, we were able to form a network of tissues reflecting their functional relatedness and, to certain extent, their development. Using co-citation driven gene network technique and promoter analysis, we predicted a transcriptional module where the co-operation of the transcription factors E2F and NF-kappaB can possibly regulate a number of genes involved in the neurogenesis that takes place in the adult hippocampus.
Conclusions
Here we propose that integration of gene expression data from Affymetrix GeneChip experiments is possible through re-annotation and commonly used pre-processing methods. We suggest that some functional aspects of the tissues can be explained by the co-operation of multiple transcription factors that regulate the expression of selected groups of genes.
doi:10.1371/journal.pone.0001880
PMCID: PMC2268968  PMID: 18382664
25.  Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy 
BMC Genomics  2007;8:268.
Background
The Down syndrome phenotype has been attributed to overexpression of chromosome 21 (Hsa21) genes. However, the expression profile of Hsa21 genes in trisomic human subjects as well as their effects on genes located on different chromosomes are largely unknown. Using oligonucleotide microarrays we compared the gene expression profiles of hearts of human fetuses with and without Hsa21 trisomy.
Results
Approximately half of the 15,000 genes examined (87 of the 168 genes on Hsa21) were expressed in the heart at 18–22 weeks of gestation. Hsa21 gene expression was globally upregulated 1.5 fold in trisomic samples. However, not all genes were equally dysregulated and 25 genes were not upregulated at all. Genes located on other chromosomes were also significantly dysregulated. Functional class scoring and gene set enrichment analyses of 473 genes, differentially expressed between trisomic and non-trisomic hearts, revealed downregulation of genes encoding mitochondrial enzymes and upregulation of genes encoding extracellular matrix proteins. There were no significant differences between trisomic fetuses with and without heart defects.
Conclusion
We conclude that dosage-dependent upregulation of Hsa21 genes causes dysregulation of the genes responsible for mitochondrial function and for the extracellular matrix organization in the fetal heart of trisomic subjects. These alterations might be harbingers of the heart defects associated with Hsa21 trisomy, which could be based on elusive mechanisms involving genetic variability, environmental factors and/or stochastic events.
doi:10.1186/1471-2164-8-268
PMCID: PMC1964766  PMID: 17683628

Results 1-25 (25)