Search

Your search keyword '"Hawkins, R David"' showing total 19 results
Start Over Author "Hawkins, R David" Publication Type Electronic Resources
19 results on '"Hawkins, R David"'

1. An Activating STAT3 Mutation Causes Neonatal Diabetes through Premature Induction of Pancreatic Differentiation

Author

University of Helsinki, Research Programs Unit, University of Helsinki, Research Program of Molecular Neurology, University of Helsinki, Institute of Biotechnology, University of Helsinki, Clinicum, Saarimaki-Vire, Jonna, Balboa, Diego, Russell, Mark A., Saarikettu, Juha, Kinnunen, Matias, Keskitalo, Salla, Malhi, Amrinder, Valensisi, Cristina, Andrus, Colin, Eurola, Solja, Grym, Heli, Ustinov, Jarkko, Wartiovaara, Kirmo, Hawkins, R. David, Silvennoinen, Olli, Varjosalo, Markku, Morgan, Noel G., Otonkoski, Timo, University of Helsinki, Research Programs Unit, University of Helsinki, Research Program of Molecular Neurology, University of Helsinki, Institute of Biotechnology, University of Helsinki, Clinicum, Saarimaki-Vire, Jonna, Balboa, Diego, Russell, Mark A., Saarikettu, Juha, Kinnunen, Matias, Keskitalo, Salla, Malhi, Amrinder, Valensisi, Cristina, Andrus, Colin, Eurola, Solja, Grym, Heli, Ustinov, Jarkko, Wartiovaara, Kirmo, Hawkins, R. David, Silvennoinen, Olli, Varjosalo, Markku, Morgan, Noel G., and Otonkoski, Timo

Abstract

Activating germline mutations in STAT3 were recently identified as a cause of neonatal diabetes mellitus associated with beta-cell autoimmunity. We have investigated the effect of an activating mutation, STAT3(K392R,) on pancreatic development using induced pluripotent stem cells (iPSCs) derived from a patient with neonatal diabetes and pancreatic hypoplasia. Early pancreatic endoderm differentiated similarly from STAT3(K392R) and healthy-control cells, but in later stages, NEUROG3 expressionwas upregulated prematurely in STAT3(K392R) cells together with insulin (INS) and glucagon (GCG). RNA sequencing (RNA-seq) showed robust NEUROG3 downstream targets upregulation. STAT3 mutation correction with CRISPR/Cas9 reversed completely the disease phenotype. STAT3(K392R) -activating properties were not explained fully by altered DNA-binding affinity or increased phosphorylation. Instead, reporter assays demonstrated NEUROG3 promoter activation by STAT3 in pancreatic cells. Furthermore, proteomic and immunocytochemical analyses revealed increased nuclear translocation of STAT3(K392R). Collectively, our results demonstrate that the STAT3(K392R) mutation causes premature endocrine differentiation through direct induction of NEUROG3 expression.

Published
2017

2. The metabolome regulates the epigenetic landscape during naive-to-primed human embryonic stem cell transition.

Author

Sperber, Henrik, Sperber, Henrik, Mathieu, Julie, Wang, Yuliang, Ferreccio, Amy, Hesson, Jennifer, Xu, Zhuojin, Fischer, Karin A, Devi, Arikketh, Detraux, Damien, Gu, Haiwei, Battle, Stephanie L, Showalter, Megan, Valensisi, Cristina, Bielas, Jason H, Ericson, Nolan G, Margaretha, Lilyana, Robitaille, Aaron M, Margineantu, Daciana, Fiehn, Oliver, Hockenbery, David, Blau, C Anthony, Raftery, Daniel, Margolin, Adam A, Hawkins, R David, Moon, Randall T, Ware, Carol B, Ruohola-Baker, Hannele, Sperber, Henrik, Sperber, Henrik, Mathieu, Julie, Wang, Yuliang, Ferreccio, Amy, Hesson, Jennifer, Xu, Zhuojin, Fischer, Karin A, Devi, Arikketh, Detraux, Damien, Gu, Haiwei, Battle, Stephanie L, Showalter, Megan, Valensisi, Cristina, Bielas, Jason H, Ericson, Nolan G, Margaretha, Lilyana, Robitaille, Aaron M, Margineantu, Daciana, Fiehn, Oliver, Hockenbery, David, Blau, C Anthony, Raftery, Daniel, Margolin, Adam A, Hawkins, R David, Moon, Randall T, Ware, Carol B, and Ruohola-Baker, Hannele

Abstract

For nearly a century developmental biologists have recognized that cells from embryos can differ in their potential to differentiate into distinct cell types. Recently, it has been recognized that embryonic stem cells derived from both mice and humans exhibit two stable yet epigenetically distinct states of pluripotency: naive and primed. We now show that nicotinamide N-methyltransferase (NNMT) and the metabolic state regulate pluripotency in human embryonic stem cells (hESCs). Specifically, in naive hESCs, NNMT and its enzymatic product 1-methylnicotinamide are highly upregulated, and NNMT is required for low S-adenosyl methionine (SAM) levels and the H3K27me3 repressive state. NNMT consumes SAM in naive cells, making it unavailable for histone methylation that represses Wnt and activates the HIF pathway in primed hESCs. These data support the hypothesis that the metabolome regulates the epigenetic landscape of the earliest steps in human development.

Published
2015

3. Identification of global regulators of T-helper cell lineage specification.

Author

Kanduri, Kartiek, Kanduri, Kartiek, Tripathi, Subhash, Larjo, Antti, Mannerström, Henrik, Ullah, Ubaid, Lund, Riikka, Hawkins, R David, Ren, Bing, Lähdesmäki, Harri, Lahesmaa, Riitta, Kanduri, Kartiek, Kanduri, Kartiek, Tripathi, Subhash, Larjo, Antti, Mannerström, Henrik, Ullah, Ubaid, Lund, Riikka, Hawkins, R David, Ren, Bing, Lähdesmäki, Harri, and Lahesmaa, Riitta

Abstract

BackgroundActivation and differentiation of T-helper (Th) cells into Th1 and Th2 types is a complex process orchestrated by distinct gene activation programs engaging a number of genes. This process is crucial for a robust immune response and an imbalance might lead to disease states such as autoimmune diseases or allergy. Therefore, identification of genes involved in this process is paramount to further understand the pathogenesis of, and design interventions for, immune-mediated diseases.MethodsWe aimed at identifying protein-coding genes and long non-coding RNAs (lncRNAs) involved in early differentiation of T-helper cells by transcriptome analysis of cord blood-derived naïve precursor, primary and polarized cells.ResultsHere, we identified lineage-specific genes involved in early differentiation of Th1 and Th2 subsets by integrating transcriptional profiling data from multiple platforms. We have obtained a high confidence list of genes as well as a list of novel genes by employing more than one profiling platform. We show that the density of lineage-specific epigenetic marks is higher around lineage-specific genes than anywhere else in the genome. Based on next-generation sequencing data we identified lineage-specific lncRNAs involved in early Th1 and Th2 differentiation and predicted their expected functions through Gene Ontology analysis. We show that there is a positive trend in the expression of the closest lineage-specific lncRNA and gene pairs. We also found out that there is an enrichment of disease SNPs around a number of lncRNAs identified, suggesting that these lncRNAs might play a role in the etiology of autoimmune diseases.ConclusionThe results presented here show the involvement of several new actors in the early differentiation of T-helper cells and will be a valuable resource for better understanding of autoimmune processes.

Published
2015

4. Integrative analysis of 111 reference human epigenomes

Author

Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, Kellis, Manolis, Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, and Kellis, Manolis

Abstract

The reference human genome sequence set the stage for studies of genetic variation and its association with human disease, but epigenomic studies lack a similar reference. To address this need, the NIH Roadmap Epigenomics Consortium generated the largest collection so far of human epigenomes for primary cells and tissues. Here we describe the integrative analysis of 111 reference human epigenomes generated as part of the programme, profiled for histone modification patterns, DNA accessibility, DNA methylation and RNA expression. We establish global maps of regulatory elements, define regulatory modules of coordinated activity, and their likely activators and repressors. We show that disease-and trait-associated genetic variants are enriched in tissue-specific epigenomic marks, revealing biologically relevant cell types for diverse human traits, and providing a resource for interpreting the molecular basis of human disease. Our results demonstrate the central role of epigenomic information for understanding gene regulation, cellular differentiation and human disease. © 2015 Macmillan Publishers Limited.

Published
2015

5. Integrative analysis of 111 reference human epigenomes

Author

Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, Kellis, Manolis, Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, and Kellis, Manolis

Abstract

The reference human genome sequence set the stage for studies of genetic variation and its association with human disease, but epigenomic studies lack a similar reference. To address this need, the NIH Roadmap Epigenomics Consortium generated the largest collection so far of human epigenomes for primary cells and tissues. Here we describe the integrative analysis of 111 reference human epigenomes generated as part of the programme, profiled for histone modification patterns, DNA accessibility, DNA methylation and RNA expression. We establish global maps of regulatory elements, define regulatory modules of coordinated activity, and their likely activators and repressors. We show that disease-and trait-associated genetic variants are enriched in tissue-specific epigenomic marks, revealing biologically relevant cell types for diverse human traits, and providing a resource for interpreting the molecular basis of human disease. Our results demonstrate the central role of epigenomic information for understanding gene regulation, cellular differentiation and human disease. © 2015 Macmillan Publishers Limited.

Published
2015

6. The metabolome regulates the epigenetic landscape during naive-to-primed human embryonic stem cell transition.

Author

Sperber, Henrik, Sperber, Henrik, Mathieu, Julie, Wang, Yuliang, Ferreccio, Amy, Hesson, Jennifer, Xu, Zhuojin, Fischer, Karin A, Devi, Arikketh, Detraux, Damien, Gu, Haiwei, Battle, Stephanie L, Showalter, Megan, Valensisi, Cristina, Bielas, Jason H, Ericson, Nolan G, Margaretha, Lilyana, Robitaille, Aaron M, Margineantu, Daciana, Fiehn, Oliver, Hockenbery, David, Blau, C Anthony, Raftery, Daniel, Margolin, Adam A, Hawkins, R David, Moon, Randall T, Ware, Carol B, Ruohola-Baker, Hannele, Sperber, Henrik, Sperber, Henrik, Mathieu, Julie, Wang, Yuliang, Ferreccio, Amy, Hesson, Jennifer, Xu, Zhuojin, Fischer, Karin A, Devi, Arikketh, Detraux, Damien, Gu, Haiwei, Battle, Stephanie L, Showalter, Megan, Valensisi, Cristina, Bielas, Jason H, Ericson, Nolan G, Margaretha, Lilyana, Robitaille, Aaron M, Margineantu, Daciana, Fiehn, Oliver, Hockenbery, David, Blau, C Anthony, Raftery, Daniel, Margolin, Adam A, Hawkins, R David, Moon, Randall T, Ware, Carol B, and Ruohola-Baker, Hannele

Abstract

For nearly a century developmental biologists have recognized that cells from embryos can differ in their potential to differentiate into distinct cell types. Recently, it has been recognized that embryonic stem cells derived from both mice and humans exhibit two stable yet epigenetically distinct states of pluripotency: naive and primed. We now show that nicotinamide N-methyltransferase (NNMT) and the metabolic state regulate pluripotency in human embryonic stem cells (hESCs).  Specifically, in naive hESCs, NNMT and its enzymatic product 1-methylnicotinamide are highly upregulated, and NNMT is required for low S-adenosyl methionine (SAM) levels and the H3K27me3 repressive state. NNMT consumes SAM in naive cells, making it unavailable for histone methylation that represses Wnt and activates the HIF pathway in primed hESCs. These data support the hypothesis that the metabolome regulates the epigenetic landscape of the earliest steps in human development.

Published
2015

7. Identification of global regulators of T-helper cell lineage specification.

Author

Kanduri, Kartiek, Kanduri, Kartiek, Tripathi, Subhash, Larjo, Antti, Mannerström, Henrik, Ullah, Ubaid, Lund, Riikka, Hawkins, R David, Ren, Bing, Lähdesmäki, Harri, Lahesmaa, Riitta, Kanduri, Kartiek, Kanduri, Kartiek, Tripathi, Subhash, Larjo, Antti, Mannerström, Henrik, Ullah, Ubaid, Lund, Riikka, Hawkins, R David, Ren, Bing, Lähdesmäki, Harri, and Lahesmaa, Riitta

Abstract

BackgroundActivation and differentiation of T-helper (Th) cells into Th1 and Th2 types is a complex process orchestrated by distinct gene activation programs engaging a number of genes. This process is crucial for a robust immune response and an imbalance might lead to disease states such as autoimmune diseases or allergy. Therefore, identification of genes involved in this process is paramount to further understand the pathogenesis of, and design interventions for, immune-mediated diseases.MethodsWe aimed at identifying protein-coding genes and long non-coding RNAs (lncRNAs) involved in early differentiation of T-helper cells by transcriptome analysis of cord blood-derived naïve precursor, primary and polarized cells.ResultsHere, we identified lineage-specific genes involved in early differentiation of Th1 and Th2 subsets by integrating transcriptional profiling data from multiple platforms. We have obtained a high confidence list of genes as well as a list of novel genes by employing more than one profiling platform. We show that the density of lineage-specific epigenetic marks is higher around lineage-specific genes than anywhere else in the genome. Based on next-generation sequencing data we identified lineage-specific lncRNAs involved in early Th1 and Th2 differentiation and predicted their expected functions through Gene Ontology analysis. We show that there is a positive trend in the expression of the closest lineage-specific lncRNA and gene pairs. We also found out that there is an enrichment of disease SNPs around a number of lncRNAs identified, suggesting that these lncRNAs might play a role in the etiology of autoimmune diseases.ConclusionThe results presented here show the involvement of several new actors in the early differentiation of T-helper cells and will be a valuable resource for better understanding of autoimmune processes.

Published
2015

8. Integrative analysis of 111 reference human epigenomes

Author

Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, Kellis, Manolis, Roadmap Epigenomics Consortium, Kundaje, Anshul, Meuleman, Wouter, Ernst, Jason, Bilenky, Misha, Yen, Angela, Heravi-Moussavi, Alireza, Kheradpour, Pouya, Zhang, Zhizhuo, Wang, Jianrong, Ziller, Michael J., Amin, Viren, Whitaker, John W., Schultz, Matthew D., Ward, Lucas D., Sarkar, Abhishek, Quon, Gerald, Sandstrom, Richard S., Eaton, Matthew L., Wu, Yi-Chieh, Pfenning, Andreas R., Wang, Xinchen, Claussnitzer, Melina, Liu, Yaping, Coarfa, Cristian, Harris, R. Alan, Shoresh, Noam, Epstein, Charles B., Gjoneska, Elizabeta, Leung, Danny, Xie, Wei, Hawkins, R. David, Lister, Ryan, Hong, Chibo, Gascard, Philippe, Mungall, Andrew J., Moore, Richard, Chuah, Eric, Tam, Angela, Canfield, Theresa K., Hansen, R. Scott, Kaul, Rajinder, Sabo, Peter J., Bansal, Mukul S., Carles, Annaick, Dixon, Jesse R., Farh, Kai-How, Feizi, Soheil, Karlic, Rosa, Kim, Ah-Ram, Kulkarni, Ashwinikumar, Li, Daofeng, Lowdon, Rebecca, Elliott, GiNell, Mercer, Tim R., Neph, Shane J., Onuchic, Vitor, Polak, Paz, Rajagopal, Nisha, Ray, Pradipta, Sallari, Richard C., Siebenthall, Kyle T., Sinnott-Armstrong, Nicholas A., Stevens, Michael, Thurman, Robert E., Wu, Jie, Zhang, Bo, Zhou, Xin, Beaudet, Arthur E., Boyer, Laurie A., De Jager, Philip L., Farnham, Peggy J., Fisher, Susan J., Haussler, David, Jones, Steven J.M., Li, Wei, Marra, Marco A., McManus, Michael T., Sunyaev, Shamil, Thomson, James A., Tlsty, Thea D., Tsai, Li-Huei, Wang, Wei, Waterland, Robert A., Zhang, Michael Q., Chadwick, Lisa H., Bernstein, Bradley E., Costello, Joseph F., Ecker, Joseph R., Hirst, Martin, Meissner, Alexander, Milosavljevic, Aleksandar, Ren, Bing, Stamatoyannopoulos, John A., Wang, Ting, and Kellis, Manolis

Abstract

The reference human genome sequence set the stage for studies of genetic variation and its association with human disease, but epigenomic studies lack a similar reference. To address this need, the NIH Roadmap Epigenomics Consortium generated the largest collection so far of human epigenomes for primary cells and tissues. Here we describe the integrative analysis of 111 reference human epigenomes generated as part of the programme, profiled for histone modification patterns, DNA accessibility, DNA methylation and RNA expression. We establish global maps of regulatory elements, define regulatory modules of coordinated activity, and their likely activators and repressors. We show that disease-and trait-associated genetic variants are enriched in tissue-specific epigenomic marks, revealing biologically relevant cell types for diverse human traits, and providing a resource for interpreting the molecular basis of human disease. Our results demonstrate the central role of epigenomic information for understanding gene regulation, cellular differentiation and human disease. © 2015 Macmillan Publishers Limited.

Published
2015

9. Epigenomic analysis of multilineage differentiation of human embryonic stem cells

Author

Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., Ren, Bing, Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., and Ren, Bing

Abstract

Epigenetic mechanisms have been proposed to play crucial roles in mammalian development, but their precise functions are only partially understood. To investigate epigenetic regulation of embryonic development, we differentiated human embryonic stem cells into mesendoderm, neural progenitor cells, trophoblast-like cells, and mesenchymal stem cells and systematically characterized DNA methylation, chromatin modifications, and the transcriptome in each lineage. We found that promoters that are active in early developmental stages tend to be CG rich and mainly engage H3K27me3 upon silencing in nonexpressing lineages. By contrast, promoters for genes expressed preferentially at later stages are often CG poor and primarily employ DNA methylation upon repression. Interestingly, the early developmental regulatory genes are often located in large genomic domains that are generally devoid of DNA methylation in most lineages, which we termed DNA methylation valleys (DMVs). Our results suggest that distinct epigenetic mechanisms regulate early and late stages of ES cell differentiation. © 2013 Elsevier Inc.

Published
2013

10. Epigenomic analysis of multilineage differentiation of human embryonic stem cells

Author

Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., Ren, Bing, Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., and Ren, Bing

Abstract

Epigenetic mechanisms have been proposed to play crucial roles in mammalian development, but their precise functions are only partially understood. To investigate epigenetic regulation of embryonic development, we differentiated human embryonic stem cells into mesendoderm, neural progenitor cells, trophoblast-like cells, and mesenchymal stem cells and systematically characterized DNA methylation, chromatin modifications, and the transcriptome in each lineage. We found that promoters that are active in early developmental stages tend to be CG rich and mainly engage H3K27me3 upon silencing in nonexpressing lineages. By contrast, promoters for genes expressed preferentially at later stages are often CG poor and primarily employ DNA methylation upon repression. Interestingly, the early developmental regulatory genes are often located in large genomic domains that are generally devoid of DNA methylation in most lineages, which we termed DNA methylation valleys (DMVs). Our results suggest that distinct epigenetic mechanisms regulate early and late stages of ES cell differentiation. © 2013 Elsevier Inc.

Published
2013

11. Epigenomic analysis of multilineage differentiation of human embryonic stem cells

Author

Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., Ren, Bing, Xie, Wei, Schultz, Matthew D., Lister, Ryan, Hou, Zhonggang, Rajagopal, Nisha, Ray, Pradipta, Whitaker, John W., Tian, Shulan, Hawkins, R. David, Leung, Danny, Yang, Hongbo, Wang, Tao, Lee, Ah Young, Swanson, Scott A., Zhang, Jiuchun, Zhu, Yun, Kim, Audrey, Nery, Joseph R., Urich, Mark A., Kuan, Samantha, Yen, Chia-an, Klugman, Sarit, Yu, Pengzhi, Suknuntha, Kran, Propson, Nicholas E., Chen, Huaming, Edsall, Lee E., Wagner, Ulrich, Li, Yan, Ye, Zhen, Kulkarni, Ashwinikumar, Xuan, Zhenyu, Chung, Wen-Yu, Chi, Neil C., Antosiewicz-Bourget, Jessica E., Slukvin, Igor, Stewart, Ron, Zhang, Michael Q., Wang, Wei, Thomson, James A., Ecker, Joseph R., and Ren, Bing

Abstract

Epigenetic mechanisms have been proposed to play crucial roles in mammalian development, but their precise functions are only partially understood. To investigate epigenetic regulation of embryonic development, we differentiated human embryonic stem cells into mesendoderm, neural progenitor cells, trophoblast-like cells, and mesenchymal stem cells and systematically characterized DNA methylation, chromatin modifications, and the transcriptome in each lineage. We found that promoters that are active in early developmental stages tend to be CG rich and mainly engage H3K27me3 upon silencing in nonexpressing lineages. By contrast, promoters for genes expressed preferentially at later stages are often CG poor and primarily employ DNA methylation upon repression. Interestingly, the early developmental regulatory genes are often located in large genomic domains that are generally devoid of DNA methylation in most lineages, which we termed DNA methylation valleys (DMVs). Our results suggest that distinct epigenetic mechanisms regulate early and late stages of ES cell differentiation. © 2013 Elsevier Inc.

Published
2013

12. Histone Modifications at Human Enhancers Reflect Global Cell-Type-Specific Gene Expression

Author

Massachusetts Institute of Technology. Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Kellis, Manolis, Kheradpour, Pouya, Stark, Alexander, Heintzman, Nathaniel D., Hon, Gary C., Hawkins, R. David, Harp, Lindsey F., Ye, Zhen, Lee, Leonard K., Stuart, Rhona K., Ching, Christina W., Ching, Keith A., Antosiewicz-Bourget, Jessica E., Liu, Hui, Zhang, Xinmin, Green, Roland D., Lobanenkov, Victor V., Stewart, Ron, Thomson, James A., Crawford, Gregory E., Ren, Bing, Massachusetts Institute of Technology. Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Kellis, Manolis, Kheradpour, Pouya, Stark, Alexander, Heintzman, Nathaniel D., Hon, Gary C., Hawkins, R. David, Harp, Lindsey F., Ye, Zhen, Lee, Leonard K., Stuart, Rhona K., Ching, Christina W., Ching, Keith A., Antosiewicz-Bourget, Jessica E., Liu, Hui, Zhang, Xinmin, Green, Roland D., Lobanenkov, Victor V., Stewart, Ron, Thomson, James A., Crawford, Gregory E., and Ren, Bing

Abstract

The human body is composed of diverse cell types with distinct functions. Although it is known that lineage specification depends on cell-specific gene expression, which in turn is driven by promoters, enhancers, insulators and other cis-regulatory DNA sequences for each gene1, 2, 3, the relative roles of these regulatory elements in this process are not clear. We have previously developed a chromatin-immunoprecipitation-based microarray method (ChIP-chip) to locate promoters, enhancers and insulators in the human genome4, 5, 6. Here we use the same approach to identify these elements in multiple cell types and investigate their roles in cell-type-specific gene expression. We observed that the chromatin state at promoters and CTCF-binding at insulators is largely invariant across diverse cell types. In contrast, enhancers are marked with highly cell-type-specific histone modification patterns, strongly correlate to cell-type-specific gene expression programs on a global scale, and are functionally active in a cell-type-specific manner. Our results define over 55,000 potential transcriptional enhancers in the human genome, significantly expanding the current catalogue of human enhancers and highlighting the role of these elements in cell-type-specific gene expression.

Published
2012

13. An assessment of histone-modification antibody quality

Author

Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah C R, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, Lieb, Jason D, Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah C R, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, and Lieb, Jason D

Abstract

We have tested the specificity and utility of more than 200 antibodies raised against 57 different histone modifications in Drosophila melanogaster, Caenorhabditis elegans and human cells. Although most antibodies performed well, more than 25% failed specificity tests by dot blot or western blot. Among specific antibodies, more than 20% failed in chromatin immunoprecipitation experiments. We advise rigorous testing of histone-modification antibodies before use, and we provide a website for posting new test results (http://compbio.med.harvard.edu/antibodies/).

Published
2011
Full Text
View/download PDF

14. An assessment of histone-modification antibody quality.

Author

Egelhofer, Thea A, Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah CR, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, Lieb, Jason D, Egelhofer, Thea A, Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah CR, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, and Lieb, Jason D

Abstract

We have tested the specificity and utility of more than 200 antibodies raised against 57 different histone modifications in Drosophila melanogaster, Caenorhabditis elegans and human cells. Although most antibodies performed well, more than 25% failed specificity tests by dot blot or western blot. Among specific antibodies, more than 20% failed in chromatin immunoprecipitation experiments. We advise rigorous testing of histone-modification antibodies before use, and we provide a website for posting new test results (http://compbio.med.harvard.edu/antibodies/).

Published
2011

15. An assessment of histone-modification antibody quality.

Author

Egelhofer, Thea A, Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah CR, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, Lieb, Jason D, Egelhofer, Thea A, Egelhofer, Thea A, Minoda, Aki, Klugman, Sarit, Lee, Kyungjoon, Kolasinska-Zwierz, Paulina, Alekseyenko, Artyom A, Cheung, Ming-Sin, Day, Daniel S, Gadel, Sarah, Gorchakov, Andrey A, Gu, Tingting, Kharchenko, Peter V, Kuan, Samantha, Latorre, Isabel, Linder-Basso, Daniela, Luu, Ying, Ngo, Queminh, Perry, Marc, Rechtsteiner, Andreas, Riddle, Nicole C, Schwartz, Yuri B, Shanower, Gregory A, Vielle, Anne, Ahringer, Julie, Elgin, Sarah CR, Kuroda, Mitzi I, Pirrotta, Vincenzo, Ren, Bing, Strome, Susan, Park, Peter J, Karpen, Gary H, Hawkins, R David, and Lieb, Jason D

Abstract

We have tested the specificity and utility of more than 200 antibodies raised against 57 different histone modifications in Drosophila melanogaster, Caenorhabditis elegans and human cells. Although most antibodies performed well, more than 25% failed specificity tests by dot blot or western blot. Among specific antibodies, more than 20% failed in chromatin immunoprecipitation experiments. We advise rigorous testing of histone-modification antibodies before use, and we provide a website for posting new test results (http://compbio.med.harvard.edu/antibodies/).

Published
2011

16. Comparison of sequencing-based methods to profile DNA methylation and identification of monoallelic epigenetic modifications.

Author

Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, Costello, Joseph F, Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, and Costello, Joseph F

Abstract

Analysis of DNA methylation patterns relies increasingly on sequencing-based profiling methods. The four most frequently used sequencing-based technologies are the bisulfite-based methods MethylC-seq and reduced representation bisulfite sequencing (RRBS), and the enrichment-based techniques methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylated DNA binding domain sequencing (MBD-seq). We applied all four methods to biological replicates of human embryonic stem cells to assess their genome-wide CpG coverage, resolution, cost, concordance and the influence of CpG density and genomic context. The methylation levels assessed by the two bisulfite methods were concordant (their difference did not exceed a given threshold) for 82% for CpGs and 99% of the non-CpG cytosines. Using binary methylation calls, the two enrichment methods were 99% concordant and regions assessed by all four methods were 97% concordant. We combined MeDIP-seq with methylation-sensitive restriction enzyme (MRE-seq) sequencing for comprehensive methylome coverage at lower cost. This, along with RNA-seq and ChIP-seq of the ES cells enabled us to detect regions with allele-specific epigenetic states, identifying most known imprinted regions and new loci with monoallelic epigenetic marks and monoallelic expression.

Published
2010

17. Comparison of sequencing-based methods to profile DNA methylation and identification of monoallelic epigenetic modifications.

Author

Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, Costello, Joseph F, Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, and Costello, Joseph F

Abstract

Analysis of DNA methylation patterns relies increasingly on sequencing-based profiling methods. The four most frequently used sequencing-based technologies are the bisulfite-based methods MethylC-seq and reduced representation bisulfite sequencing (RRBS), and the enrichment-based techniques methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylated DNA binding domain sequencing (MBD-seq). We applied all four methods to biological replicates of human embryonic stem cells to assess their genome-wide CpG coverage, resolution, cost, concordance and the influence of CpG density and genomic context. The methylation levels assessed by the two bisulfite methods were concordant (their difference did not exceed a given threshold) for 82% for CpGs and 99% of the non-CpG cytosines. Using binary methylation calls, the two enrichment methods were 99% concordant and regions assessed by all four methods were 97% concordant. We combined MeDIP-seq with methylation-sensitive restriction enzyme (MRE-seq) sequencing for comprehensive methylome coverage at lower cost. This, along with RNA-seq and ChIP-seq of the ES cells enabled us to detect regions with allele-specific epigenetic states, identifying most known imprinted regions and new loci with monoallelic epigenetic marks and monoallelic expression.

Published
2010

18. Comparison of sequencing-based methods to profile DNA methylation and identification of monoallelic epigenetic modifications.

Author

Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, Costello, Joseph F, Harris, R Alan, Harris, R Alan, Wang, Ting, Coarfa, Cristian, Nagarajan, Raman P, Hong, Chibo, Downey, Sara L, Johnson, Brett E, Fouse, Shaun D, Delaney, Allen, Zhao, Yongjun, Olshen, Adam, Ballinger, Tracy, Zhou, Xin, Forsberg, Kevin J, Gu, Junchen, Echipare, Lorigail, O'Geen, Henriette, Lister, Ryan, Pelizzola, Mattia, Xi, Yuanxin, Epstein, Charles B, Bernstein, Bradley E, Hawkins, R David, Ren, Bing, Chung, Wen-Yu, Gu, Hongcang, Bock, Christoph, Gnirke, Andreas, Zhang, Michael Q, Haussler, David, Ecker, Joseph R, Li, Wei, Farnham, Peggy J, Waterland, Robert A, Meissner, Alexander, Marra, Marco A, Hirst, Martin, Milosavljevic, Aleksandar, and Costello, Joseph F

Abstract

Analysis of DNA methylation patterns relies increasingly on sequencing-based profiling methods. The four most frequently used sequencing-based technologies are the bisulfite-based methods MethylC-seq and reduced representation bisulfite sequencing (RRBS), and the enrichment-based techniques methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylated DNA binding domain sequencing (MBD-seq). We applied all four methods to biological replicates of human embryonic stem cells to assess their genome-wide CpG coverage, resolution, cost, concordance and the influence of CpG density and genomic context. The methylation levels assessed by the two bisulfite methods were concordant (their difference did not exceed a given threshold) for 82% for CpGs and 99% of the non-CpG cytosines. Using binary methylation calls, the two enrichment methods were 99% concordant and regions assessed by all four methods were 97% concordant. We combined MeDIP-seq with methylation-sensitive restriction enzyme (MRE-seq) sequencing for comprehensive methylome coverage at lower cost. This, along with RNA-seq and ChIP-seq of the ES cells enabled us to detect regions with allele-specific epigenetic states, identifying most known imprinted regions and new loci with monoallelic epigenetic marks and monoallelic expression.

Published
2010

19. Genome-wide profiling of interleukin-4 and STAT6 transcription factor regulation of human Th2 cell programming

Author

Elo, Laura L., Järvenpää, Henna, Tuomela, Soile, Raghav, Sunil, Ahlfors, Helena, Laurila, Kirsti, Gupta, Bhawna, Lund, Riikka J., Tahvanainen, Johanna, Hawkins, R. David, Oresic, Matej, Lähdesmäki, Harri, Rasool, Omid, Rao, Kanury V., Aittokallio, Tero, Lahesmaa, Riitta, Elo, Laura L., Järvenpää, Henna, Tuomela, Soile, Raghav, Sunil, Ahlfors, Helena, Laurila, Kirsti, Gupta, Bhawna, Lund, Riikka J., Tahvanainen, Johanna, Hawkins, R. David, Oresic, Matej, Lähdesmäki, Harri, Rasool, Omid, Rao, Kanury V., Aittokallio, Tero, and Lahesmaa, Riitta

Abstract

Dissecting the molecular mechanisms by which T helper (Th) cells differentiate to effector Th2 cells is important for understanding the pathogenesis of immune-mediated diseases, such as asthma and allergy. Because the STAT6 transcription factor is an upstream mediator required for interleukin-4 (IL-4)-induced Th2 cell differentiation, its targets include genes important for this process. Using primary human CD4(+) T cells, and by blocking STAT6 with RNAi, we identified a number of direct and indirect targets of STAT6 with ChIP sequencing. The integration of these data sets with detailed kinetics of IL-4-driven transcriptional changes showed that STAT6 was predominantly needed for the activation of transcription leading to the Th2 cell phenotype. This integrated genome-wide data on IL-4- and STAT6-mediated transcription provide a unique resource for studies on Th cell differentiation and, in particular, for designing interventions of human Th2 cell responses., Funding agencies:European Commission EC-FP7-SYBILLA-201106 EC-FP7-NANOMMUNE-214281 EC-FP7-DIABIMMUNE-202063 CIMO/SitraSigrid Juselius FoundationDepartment of Biotechnology, Government of IndiaTurku University HospitalNote: The Turku Centre for Biotechnology is a joint department of the Turku University and the Åbo Akademi University

Published
2010
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results