Search

Your search keyword '"Parnell, Euan"' showing total 35 results
Start Over Author "Parnell, Euan"
35 results on '"Parnell, Euan"'

1. Excitatory Dysfunction Drives Network and Calcium Handling Deficits in 16p11.2 Duplication Schizophrenia Induced Pluripotent Stem Cell–Derived Neurons

Author

Parnell, Euan, Culotta, Lorenza, Forrest, Marc P., Jalloul, Hiba A., Eckman, Blair L., Loizzo, Daniel D., Horan, Katherine K.E., Dos Santos, Marc, Piguel, Nicolas H., Tai, Derek J.C., Zhang, Hanwen, Gertler, Tracy S., Simkin, Dina, Sanders, Alan R., Talkowski, Michael E., Gejman, Pablo V., Kiskinis, Evangelos, Duan, Jubao, and Penzes, Peter

Published
2023
Full Text
View/download PDF

2. Partial Loss of USP9X Function Leads to a Male Neurodevelopmental and Behavioral Disorder Converging on Transforming Growth Factor β Signaling.

Author

Johnson, Brett V, Kumar, Raman, Oishi, Sabrina, Alexander, Suzy, Kasherman, Maria, Vega, Michelle Sanchez, Ivancevic, Atma, Gardner, Alison, Domingo, Deepti, Corbett, Mark, Parnell, Euan, Yoon, Sehyoun, Oh, Tracey, Lines, Matthew, Lefroy, Henrietta, Kini, Usha, Van Allen, Margot, Grønborg, Sabine, Mercier, Sandra, Küry, Sébastien, Bézieau, Stéphane, Pasquier, Laurent, Raynaud, Martine, Afenjar, Alexandra, Billette de Villemeur, Thierry, Keren, Boris, Désir, Julie, Van Maldergem, Lionel, Marangoni, Martina, Dikow, Nicola, Koolen, David A, VanHasselt, Peter M, Weiss, Marjan, Zwijnenburg, Petra, Sa, Joaquim, Reis, Claudia Falcao, López-Otín, Carlos, Santiago-Fernández, Olaya, Fernández-Jaén, Alberto, Rauch, Anita, Steindl, Katharina, Joset, Pascal, Goldstein, Amy, Madan-Khetarpal, Suneeta, Infante, Elena, Zackai, Elaine, Mcdougall, Carey, Narayanan, Vinodh, Ramsey, Keri, Mercimek-Andrews, Saadet, Pena, Loren, Shashi, Vandana, Undiagnosed Diseases Network, Schoch, Kelly, Sullivan, Jennifer A, Pinto E Vairo, Filippo, Pichurin, Pavel N, Ewing, Sarah A, Barnett, Sarah S, Klee, Eric W, Perry, M Scott, Koenig, Mary Kay, Keegan, Catherine E, Schuette, Jane L, Asher, Stephanie, Perilla-Young, Yezmin, Smith, Laurie D, Rosenfeld, Jill A, Bhoj, Elizabeth, Kaplan, Paige, Li, Dong, Oegema, Renske, van Binsbergen, Ellen, van der Zwaag, Bert, Smeland, Marie Falkenberg, Cutcutache, Ioana, Page, Matthew, Armstrong, Martin, Lin, Angela E, Steeves, Marcie A, Hollander, Nicolette den, Hoffer, Mariëtte JV, Reijnders, Margot RF, Demirdas, Serwet, Koboldt, Daniel C, Bartholomew, Dennis, Mosher, Theresa Mihalic, Hickey, Scott E, Shieh, Christine, Sanchez-Lara, Pedro A, Graham, John M, Tezcan, Kamer, Schaefer, GB, Danylchuk, Noelle R, Asamoah, Alexander, Jackson, Kelly E, Yachelevich, Naomi, Au, Margaret, Pérez-Jurado, Luis A, and Kleefstra, Tjitske

Subjects
Undiagnosed Diseases Network, Animals, Humans, Mice, Ubiquitin Thiolesterase, Transforming Growth Factor beta, Developmental Disabilities, Signal Transduction, Phenotype, Female, Male, Haploinsufficiency, Intellectual Disability, Brain malformation, Deubiquitylating enzyme, Hippocampus, Neurodevelopmental disorder, TGFβ, USP9X, Congenital Structural Anomalies, Genetics, Neurosciences, Pediatric, Mental Health, Behavioral and Social Science, Brain Disorders, Clinical Research, 2.1 Biological and endogenous factors, Aetiology, Neurological, TGF beta, Biological Sciences, Medical and Health Sciences, Psychology and Cognitive Sciences, Psychiatry
Abstract

BackgroundThe X-chromosome gene USP9X encodes a deubiquitylating enzyme that has been associated with neurodevelopmental disorders primarily in female subjects. USP9X escapes X inactivation, and in female subjects de novo heterozygous copy number loss or truncating mutations cause haploinsufficiency culminating in a recognizable syndrome with intellectual disability and signature brain and congenital abnormalities. In contrast, the involvement of USP9X in male neurodevelopmental disorders remains tentative.MethodsWe used clinically recommended guidelines to collect and interrogate the pathogenicity of 44 USP9X variants associated with neurodevelopmental disorders in males. Functional studies in patient-derived cell lines and mice were used to determine mechanisms of pathology.ResultsTwelve missense variants showed strong evidence of pathogenicity. We define a characteristic phenotype of the central nervous system (white matter disturbances, thin corpus callosum, and widened ventricles); global delay with significant alteration of speech, language, and behavior; hypotonia; joint hypermobility; visual system defects; and other common congenital and dysmorphic features. Comparison of in silico and phenotypical features align additional variants of unknown significance with likely pathogenicity. In support of partial loss-of-function mechanisms, using patient-derived cell lines, we show loss of only specific USP9X substrates that regulate neurodevelopmental signaling pathways and a united defect in transforming growth factor β signaling. In addition, we find correlates of the male phenotype in Usp9x brain-specific knockout mice, and further resolve loss of hippocampal-dependent learning and memory.ConclusionsOur data demonstrate the involvement of USP9X variants in a distinctive neurodevelopmental and behavioral syndrome in male subjects and identify plausible mechanisms of pathogenesis centered on disrupted transforming growth factor β signaling and hippocampal function.

Published
2020

3. Partial Loss of USP9X Function Leads to a Male Neurodevelopmental and Behavioral Disorder Converging on Transforming Growth Factor β Signaling

Author

Pena, Loren, Shashi, Vandana, Schoch, Kelly, Sullivan, Jennifer A., Acosta, Maria T., Adams, David R., Aday, Aaron, Alejandro, Mercedes E., Allard, Patrick, Ashley, Euan A., Azamian, Mahshid S., Bacino, Carlos A., Bademci, Guney, Baker, Eva, Balasubramanyam, Ashok, Baldridge, Dustin, Barbouth, Deborah, Batzli, Gabriel F., Beggs, Alan H., Bellen, Hugo J., Bernstein, Jonathan A., Berry, Gerard T., Bican, Anna, Bick, David P., Birch, Camille L., Bivona, Stephanie, Bonnenmann, Carsten, Bonner, Devon, Boone, Braden E., Bostwick, Bret L., Briere, Lauren C., Brokamp, Elly, Brown, Donna M., Brush, Matthew, Burke, Elizabeth A., Burrage, Lindsay C., Butte, Manish J., Carrasquillo, Olveen, Peter Chang, Ta Chen, Chao, Hsiao-Tuan, Clark, Gary D., Coakley, Terra R., Cobban, Laurel A., Cogan, Joy D., Cole, F. Sessions, Colley, Heather A., Cooper, Cynthia M., Cope, Heidi, Craigen, William J., D'Souza, Precilla, Dasari, Surendra, Davids, Mariska, Davidson, Jean M., Dayal, Jyoti G., Dell'Angelica, Esteban C., Dhar, Shweta U., Dorrani, Naghmeh, Dorset, Daniel C., Douine, Emilie D., Draper, David D., Dries, Annika M., Duncan, Laura, Eckstein, David J., Emrick, Lisa T., Eng, Christine M., Enns, Gregory M., Esteves, Cecilia, Estwick, Tyra, Fernandez, Liliana, Ferreira, Carlos, Fieg, Elizabeth L., Fisher, Paul G., Fogel, Brent L., Forghani, Irman, Friedman, Noah D., Gahl, William A., Godfrey, Rena A., Goldman, Alica M., Goldstein, David B., Gourdine, Jean-Philippe F., Grajewski, Alana, Groden, Catherine A., Gropman, Andrea L., Haendel, Melissa, Hamid, Rizwan, Hanchard, Neil A., High, Frances, Holm, Ingrid A., Hom, Jason, Huang, Alden, Huang, Yong, Isasi, Rosario, Jamal, Fariha, Jiang, Yong-hui, Johnston, Jean M., Jones, Angela L., Karaviti, Lefkothea, Kelley, Emily G., Koeller, David M., Kohane, Isaac S., Kohler, Jennefer N., Krakow, Deborah, Krasnewich, Donna M., Korrick, Susan, Koziura, Mary, Krier, Joel B., Kyle, Jennifer E., Lalani, Seema R., Lam, Byron, Lanpher, Brendan C., Lanza, Ian R., Lau, C. Christopher, Lazar, Jozef, LeBlanc, Kimberly, Lee, Brendan H., Lee, Hane, Levitt, Roy, Levy, Shawn E., Lewis, Richard A., Lincoln, Sharyn A., Liu, Pengfei, Liu, Xue Zhong, Loo, Sandra K., Loscalzo, Joseph, Maas, Richard L., Macnamara, Ellen F., MacRae, Calum A., Maduro, Valerie V., Majcherska, Marta M., Malicdan, May Christine V., Mamounas, Laura A., Manolio, Teri A., Markello, Thomas C., Marom, Ronit, Martin, Martin G., Martínez-Agosto, Julian A., Marwaha, Shruti, May, Thomas, McCauley, Jacob, McConkie-Rosell, Allyn, McCormack, Colleen E., McCray, Alexa T., Merker, Jason D., Metz, Thomas O., Might, Matthew, Morava-Kozicz, Eva, Moretti, Paolo M., Morimoto, Marie, Mulvihill, John J., Murdock, David R., Nath, Avi, Nelson, Stan F., Newberry, J. Scott, Newman, John H., Nicholas, Sarah K., Novacic, Donna, Oglesbee, Devin, Orengo, James P., Pak, Stephen, Pallais, J. Carl, Palmer, Christina GS., Papp, Jeanette C., Parker, Neil H., Phillips, John A., III, Posey, Jennifer E., Postlethwait, John H., Potocki, Lorraine, Pusey, Barbara N., Renteri, Genecee, Reuter, Chloe M., Rives, Lynette, Robertson, Amy K., Rodan, Lance H., Rosenfeld, Jill A., Rowley, Robb K., Sacco, Ralph, Sampson, Jacinda B., Samson, Susan L., Saporta, Mario, Schaechter, Judy, Schedl, Timothy, Scott, Daryl A., Shakachite, Lisa, Sharma, Prashant, Shields, Kathleen, Shin, Jimann, Signer, Rebecca, Sillari, Catherine H., Silverman, Edwin K., Sinsheimer, Janet S., Smith, Kevin S., Solnica-Krezel, Lilianna, Spillmann, Rebecca C., Stoler, Joan M., Stong, Nicholas, Sweetser, David A., Tamburro, Cecelia P., Tan, Queenie K.-G., Tekin, Mustafa, Telischi, Fred, Thorson, Willa, Tifft, Cynthia J., Toro, Camilo, Tran, Alyssa A., Urv, Tiina K., Vogel, Tiphanie P., Waggott, Daryl M., Wahl, Colleen E., Walley, Nicole M., Walsh, Chris A., Walker, Melissa, Wambach, Jennifer, Wan, Jijun, Wang, Lee-kai, Wangler, Michael F., Ward, Patricia A., Waters, Katrina M., Webb-Robertson, Bobbie-Jo M., Wegner, Daniel, Westerfield, Monte, Wheeler, Matthew T., Wise, Anastasia L., Wolfe, Lynne A., Woods, Jeremy D., Worthey, Elizabeth A., Yamamoto, Shinya, Yang, John, Yoon, Amanda J., Yu, Guoyun, Zastrow, Diane B., Zhao, Chunli, Zuchner, Stephan, Gahl, William, Johnson, Brett V., Kumar, Raman, Oishi, Sabrina, Alexander, Suzy, Kasherman, Maria, Vega, Michelle Sanchez, Ivancevic, Atma, Gardner, Alison, Domingo, Deepti, Corbett, Mark, Parnell, Euan, Yoon, Sehyoun, Oh, Tracey, Lines, Matthew, Lefroy, Henrietta, Kini, Usha, Van Allen, Margot, Grønborg, Sabine, Mercier, Sandra, Küry, Sébastien, Bézieau, Stéphane, Pasquier, Laurent, Raynaud, Martine, Afenjar, Alexandra, Billette de Villemeur, Thierry, Keren, Boris, Désir, Julie, Van Maldergem, Lionel, Marangoni, Martina, Dikow, Nicola, Koolen, David A., VanHasselt, Peter M., Weiss, Marjan, Zwijnenburg, Petra, Sa, Joaquim, Reis, Claudia Falcao, López-Otín, Carlos, Santiago-Fernández, Olaya, Fernández-Jaén, Alberto, Rauch, Anita, Steindl, Katharina, Joset, Pascal, Goldstein, Amy, Madan-Khetarpal, Suneeta, Infante, Elena, Zackai, Elaine, Mcdougall, Carey, Narayanan, Vinodh, Ramsey, Keri, Mercimek-Andrews, Saadet, Pinto e Vairo, Filippo, Pichurin, Pavel N., Ewing, Sarah A., Barnett, Sarah S., Klee, Eric W., Perry, M. Scott, Koenig, Mary Kay, Keegan, Catherine E., Schuette, Jane L., Asher, Stephanie, Perilla-Young, Yezmin, Smith, Laurie D., Bhoj, Elizabeth, Kaplan, Paige, Li, Dong, Oegema, Renske, van Binsbergen, Ellen, van der Zwaag, Bert, Smeland, Marie Falkenberg, Cutcutache, Ioana, Page, Matthew, Armstrong, Martin, Lin, Angela E., Steeves, Marcie A., Hollander, Nicolette den, Hoffer, Mariëtte J.V., Reijnders, Margot R.F., Demirdas, Serwet, Koboldt, Daniel C., Bartholomew, Dennis, Mosher, Theresa Mihalic, Hickey, Scott E., Shieh, Christine, Sanchez-Lara, Pedro A., Graham, John M., Jr., Tezcan, Kamer, Schaefer, G.B., Danylchuk, Noelle R., Asamoah, Alexander, Jackson, Kelly E., Yachelevich, Naomi, Au, Margaret, Pérez-Jurado, Luis A., Kleefstra, Tjitske, Penzes, Peter, Wood, Stephen A., Burne, Thomas, Pierson, Tyler Mark, Piper, Michael, Gécz, Jozef, and Jolly, Lachlan A.

Published
2020
Full Text
View/download PDF

5. EPAC isoform specificity : drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan

Subjects
571.6, QH301 Biology, QH345 Biochemistry, RM Therapeutics. Pharmacology
Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shape are unknown. Therefore, we have attempted to identify pathways involved in EPAC1-mediated morphological change by assessing the effects of various inhibitors on cell spreading. Interestingly, we found that EPAC1 and PKA synergise to produce maximal cell spreading in HEK293T cells. Recent reports suggest that the cortical actin-membrane linker protein ezrin is required for the cell spreading effects of EPAC1. Here, we demonstrate that ezrin responds to elevations in intracellular cAMP in HEK293T cells in a PKA-dependent manner. Indeed, PKA activation promotes the post translational modification of ezrin and alters the response of EPAC1-expressing cells to cAMP. These results suggests that the PKA pathway is able to regulate ezrin by post translational modification and that this is required for PKA and EPAC1 to synergise and produce maximal cell spreading. In addition to agents which directly activate the catalytic activity of EPAC1, there is a body of evidence that supports the idea that compartmentalisation of cAMP effectors is an important mechanism for the determination of downstream signalling events leading to cellular responses, such as cell spreading. As such, we have attempted to identify the regions within EPAC proteins that determine their subcellular distribution. This was done through a combination of subcellular fractionation and the immunofluorescent detection of the localisation of EPAC isoforms. In particular, mutational analysis of EPAC1 revealed a carboxy terminal (C-terminal) nuclear localisation domain that is required for the perinuclear distribution of EPAC1 alongside the nuclear pore protein, RANBP2. Structural analyses suggest that this domain appears to be conserved within EPAC2 despite EPAC2 adopting a distinct cytoplasmic distribution. One explanation for this observation is steric interference within EPAC2 which blocks access to the conserved nuclear localisation domain. We have observed that the additional amino-terminal (N-terminal) CNB of EPAC2 appears to disrupt nuclear localisation and promote a cytoplasmic distribution within the cell. Indeed, the absence of the CNB1 promotes nuclear accumulation of EPAC2, with a pattern similar to that of EPAC1. The presence of this domain within EPAC2, absent in EPAC1, may represent a mechanism which regulates the subcellular distribution, and therefore function, of EPACs within the cellular environment. In summary, we have developed a screening cascade to identify small molecules which may form the basis of therapeutic agents able to selectively target EPAC1 to promote the beneficial effects of EPAC1. In addition, a secondary screen involving EPAC1 induced morphological change was developed and characterised as an effective assay in which to test the agonist properties of compounds identified by primary HTS screening. We have confirmed that HEK293T cell spreading in response to cAMP elevation requires the expression of EPAC1, but that a secondary pathway involving interactions between PKA and ezrin is able to supplement the primary cell spreading effects of EPAC1. Finally, we have identified a potential mechanism for the different subcellular localisation of EPAC1 and EPAC2: EPAC1 is targeted to the perinuclear compartment via a previously undiscovered C-terminal nuclear localisation domain.

Published
2015

6. Missense variant contribution to USP9X-female syndrome

Author

Jolly, Lachlan A., Parnell, Euan, Gardner, Alison E., Corbett, Mark A., Pérez-Jurado, Luis A., Shaw, Marie, Lesca, Gaetan, Keegan, Catherine, Schneider, Michael C., Griffin, Emily, Maier, Felicitas, Kiss, Courtney, Guerin, Andrea, Crosby, Kathleen, Rosenbaum, Kenneth, Tanpaiboon, Pranoot, Whalen, Sandra, Keren, Boris, McCarrier, Julie, Basel, Donald, Sadedin, Simon, White, Susan M., Delatycki, Martin B., Kleefstra, Tjitske, Küry, Sébastien, Brusco, Alfredo, Sukarova-Angelovska, Elena, Trajkova, Slavica, Yoon, Sehoun, Wood, Stephen A., Piper, Michael, Penzes, Peter, and Gecz, Jozef

Published
2020
Full Text
View/download PDF

7. 19. Excitatory Dysfunction Drives Synchrony Deficits in a Schizophrenia 16p11.2 Duplication iPSC Derived Neuron Model

Author

Parnell, Euan, primary, Culotta, Lorenza, additional, Forrest, Marc, additional, Jalloul, Hiba, additional, Eckman, Blair, additional, Loizzo, Daniel, additional, Santos, Marc Dos, additional, Piguel, Nicolas, additional, Tai, Derek, additional, Zhang, Hanwen, additional, Gertler, Tracy, additional, Simkin, Dina, additional, Sanders, Alan, additional, Talkowski, Michael, additional, Gejman, Pablo, additional, Kiskinis, Evangelos, additional, Duan, Jubao, additional, and Penzes, Peter, additional

Published
2023
Full Text
View/download PDF

9. Excitatory Dysfunction Drives Network and Calcium Handling Deficits in 16p11.2 Duplication Schizophrenia Induced Pluripotent Stem Cell–Derived Neurons

Author

Parnell, Euan, primary, Culotta, Lorenza, additional, Forrest, Marc P., additional, Jalloul, Hiba A., additional, Eckman, Blair L., additional, Loizzo, Daniel D., additional, Horan, Katherine K.E., additional, Dos Santos, Marc, additional, Piguel, Nicolas H., additional, Tai, Derek J.C., additional, Zhang, Hanwen, additional, Gertler, Tracy S., additional, Simkin, Dina, additional, Sanders, Alan R., additional, Talkowski, Michael E., additional, Gejman, Pablo V., additional, Kiskinis, Evangelos, additional, Duan, Jubao, additional, and Penzes, Peter, additional

Published
2022
Full Text
View/download PDF

10. Shed CNTNAP2 ectodomain is detectable in CSF and regulates Ca2+ homeostasis and network synchrony via PMCA2/ATP2B2

Author

Martín-de-Saavedra, M. Dolores, primary, Dos Santos, Marc, additional, Culotta, Lorenza, additional, Varea, Olga, additional, Spielman, Benjamin P., additional, Parnell, Euan, additional, Forrest, Marc P., additional, Gao, Ruoqi, additional, Yoon, Sehyoun, additional, McCoig, Emmarose, additional, Jalloul, Hiba A., additional, Myczek, Kristoffer, additional, Khalatyan, Natalia, additional, Hall, Elizabeth A., additional, Turk, Liam S., additional, Sanz-Clemente, Antonio, additional, Comoletti, Davide, additional, Lichtenthaler, Stefan F., additional, Burgdorf, Jeffrey S., additional, Barbolina, Maria V., additional, Savas, Jeffrey N., additional, and Penzes, Peter, additional

Published
2022
Full Text
View/download PDF

15. Usp9X Controls Ankyrin-Repeat Domain Protein Homeostasis during Dendritic Spine Development

Author

Yoon, Sehyoun, primary, Parnell, Euan, additional, Kasherman, Maria, additional, Forrest, Marc P., additional, Myczek, Kristoffer, additional, Premarathne, Susitha, additional, Sanchez Vega, Michelle C., additional, Piper, Michael, additional, Burne, Thomas H.J., additional, Jolly, Lachlan A., additional, Wood, Stephen A., additional, and Penzes, Peter, additional

Published
2020
Full Text
View/download PDF

17. Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1

Author

Beck, Elizabeth M., primary, Parnell, Euan, additional, Cowley, Angela, additional, Porter, Alison, additional, Gillespie, Jonathan, additional, Robinson, John, additional, Robinson, Lindsay, additional, Pannifer, Andrew D., additional, Hamon, Veronique, additional, Jones, Philip, additional, Morrison, Angus, additional, McElroy, Stuart, additional, Timmerman, Martin, additional, Rutjes, Helma, additional, Mahajan, Pravin, additional, Wiejak, Jolanta, additional, Luchowska-Stańska, Urszula, additional, Morgan, David, additional, Barker, Graeme, additional, Rehmann, Holger, additional, and Yarwood, Stephen J., additional

Published
2019
Full Text
View/download PDF

22. The future of EPAC-targeted therapeutics: agonism versus antagonism

Author

Parnell, Euan, Palmer, Timothy M., and Yarwood, Stephen J.

Abstract

Pharmaceutical manipulation of cyclic adenosine monophosphate (cAMP) levels exerts beneficial effects through the regulation of the exchange protein activated by cAMP (EPAC) and protein kinase A (PKA) signalling routes. Recent attention has turned to the specific regulation of EPAC isoforms (EPAC1 and EPAC2) as a more targeted approach to cAMP-based therapeutics. For example, EPAC2-selective agonists could promote insulin secretion from pancreatic beta-cells, whereas EPAC1-selective agonists may be useful in the treatment of vascular inflammation. In contrast, EPAC1 and EPAC2 antagonists could both be useful in the treatment of heart failure. Here we discuss whether the best way forward is to design EPAC-selective agonists or antagonists and the current strategies being used to develop isoform-selective, small molecule regulators of EPAC1 and EPAC2 activity.

Published
2015

26. Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1.

Author

M. Beck, Elizabeth, Parnell, Euan, Cowley, Angela, Porter, Alison, Gillespie, Jonathan, Robinson, John, Robinson, Lindsay, Pannifer, Andrew D., Hamon, Veronique, Jones, Philip, Morrison, Angus, McElroy, Stuart, Timmerman, Martin, Rutjes, Helma, Mahajan, Pravin, Wiejak, Jolanta, Luchowska-Stańska, Urszula, Morgan, David, Barker, Graeme, and Rehmann, Holger

Subjects
*BENZOFURAN, *CYCLIC adenylic acid, *GUANINE nucleotide exchange factors, *BENZOFURANS, *LIGANDS (Biochemistry), *BINDING site assay
Abstract

Cyclic AMP promotes EPAC1 and EPAC2 activation through direct binding to a specific cyclic nucleotide-binding domain (CNBD) within each protein, leading to activation of Rap GTPases, which control multiple cell responses, including cell proliferation, adhesion, morphology, exocytosis, and gene expression. As a result, it has become apparent that directed activation of EPAC1 and EPAC2 with synthetic agonists may also be useful for the future treatment of diabetes and cardiovascular diseases. To identify new EPAC agonists we have developed a fluorescent-based, ultra-high-throughput screening (uHTS) assay that measures the displacement of binding of the fluorescent cAMP analogue, 8-NBD-cAMP to the EPAC1 CNBD. Triage of the output of an approximately 350,000 compound screens using this assay identified a benzofuran oxaloacetic acid EPAC1 binder (SY000) that displayed moderate potency using orthogonal assays (competition binding and microscale thermophoresis). We next generated a limited library of 91 analogues of SY000 and identified SY009, with modifications to the benzofuran ring associated with a 10-fold increase in potency towards EPAC1 over SY000 in binding assays. In vitro EPAC1 activity assays confirmed the agonist potential of these molecules in comparison with the known EPAC1 non-cyclic nucleotide (NCN) partial agonist, I942. Rap1 GTPase activation assays further demonstrated that SY009 selectively activates EPAC1 over EPAC2 in cells. SY009 therefore represents a novel class of NCN EPAC1 activators that selectively activate EPAC1 in cellulae. [ABSTRACT FROM AUTHOR]

Published
2019
Full Text
View/download PDF

27. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

28. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

29. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

30. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

31. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

32. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

33. EPAC isoform specificity: drug development, subcellular targeting and relevance to cell morphology

Author

Parnell, Euan and Parnell, Euan

Abstract

Cyclic adenosine monophosphate (cAMP) is a second messenger signalling molecule that has been reported to exert beneficial effects within the vasculature and other physiological systems. cAMP produces its effects within the cell through two key downstream effector molecules: exchange protein activated by cAMP (EPAC) and protein kinase A (PKA). Many of the effects of cAMP have been attributed to PKA, however there is a growing appreciation of the potential of EPAC, particularly isoform 1 (EPAC1), based therapies for the regulation of inflammatory responses within the vasculature, thereby promoting cardiovascular health. Furthermore, side effects associated with global cAMP elevating agents may be avoided by isoform selective EPAC regulation. To date no small molecule agonists have been discovered to effectively or selectively promote EPAC1 activity. In order to address this, we have developed a fluorescence based competition assay able to identify compounds which interact with the cyclic nucleotide binding domains (CNBs) of both EPAC1 and EPAC2. Rigorous testing of the assay has confirmed that it is able to reliably and reproducibly identify EPAC interacting compounds within high throughput screening (HTS) of small molecule libraries. Furthermore, dual screening of EPAC1 and EPAC2 has allowed isoform selective compounds to be identified from a small compound library, confirming the suitability of this assay for HTS. This HTS assay is likely to facilitate the discovery of EPAC1-selective interacting molecules with the potential to be effective, small molecule regulators of EPAC1. In order to classify small molecules isolated by HTS as either agonists or antagonists of EPAC1, we developed a secondary screen that is able to detect EPAC1 activation in vivo. This assay is based on the ability of EPAC1 to produce a rapid, cell spreading response in HEK293T cells stably transfected with EPAC1. However, the precise signalling pathways which produce these changes in cell shap

34. Shed CNTNAP2 ectodomain is detectable in CSF and regulates Ca 2+ homeostasis and network synchrony via PMCA2/ATP2B2.

Author

Martín-de-Saavedra MD, Dos Santos M, Culotta L, Varea O, Spielman BP, Parnell E, Forrest MP, Gao R, Yoon S, McCoig E, Jalloul HA, Myczek K, Khalatyan N, Hall EA, Turk LS, Sanz-Clemente A, Comoletti D, Lichtenthaler SF, Burgdorf JS, Barbolina MV, Savas JN, and Penzes P

Subjects
Cell Membrane metabolism, Homeostasis, Humans, Neurons metabolism, Signal Transduction, Autism Spectrum Disorder cerebrospinal fluid, Autism Spectrum Disorder genetics, Autism Spectrum Disorder metabolism, Membrane Proteins metabolism, Nerve Tissue Proteins metabolism, Plasma Membrane Calcium-Transporting ATPases cerebrospinal fluid, Plasma Membrane Calcium-Transporting ATPases genetics, Plasma Membrane Calcium-Transporting ATPases metabolism
Abstract

Although many neuronal membrane proteins undergo proteolytic cleavage, little is known about the biological significance of neuronal ectodomain shedding (ES). Here, we show that the neuronal sheddome is detectable in human cerebrospinal fluid (hCSF) and is enriched in neurodevelopmental disorder (NDD) risk factors. Among shed synaptic proteins is the ectodomain of CNTNAP2 (CNTNAP2-ecto), a prominent NDD risk factor. CNTNAP2 undergoes activity-dependent ES via MMP9 (matrix metalloprotease 9), and CNTNAP2-ecto levels are reduced in the hCSF of individuals with autism spectrum disorder. Using mass spectrometry, we identified the plasma membrane Ca 2+ ATPase (PMCA) extrusion pumps as novel CNTNAP2-ecto binding partners. CNTNAP2-ecto enhances the activity of PMCA2 and regulates neuronal network dynamics in a PMCA2-dependent manner. Our data underscore the promise of sheddome analysis in discovering neurobiological mechanisms, provide insight into the biology of ES and its relationship with the CSF, and reveal a mechanism of regulation of Ca 2+ homeostasis and neuronal network synchrony by a shed ectodomain., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Published
2022
Full Text
View/download PDF

35. The future of EPAC-targeted therapies: agonism versus antagonism.

Author

Parnell E, Palmer TM, and Yarwood SJ

Subjects
Animals, Cyclic AMP agonists, Cyclic AMP antagonists & inhibitors, Cyclic AMP metabolism, Forecasting, Guanine Nucleotide Exchange Factors metabolism, Humans, Protein Structure, Secondary, Protein Structure, Tertiary, Drug Delivery Systems trends, Guanine Nucleotide Exchange Factors agonists, Guanine Nucleotide Exchange Factors antagonists & inhibitors
Abstract

Pharmaceutical manipulation of cAMP levels exerts beneficial effects through the regulation of the exchange protein activated by cAMP (EPAC) and protein kinase A (PKA) signalling routes. Recent attention has turned to the specific regulation of EPAC isoforms (EPAC1 and EPAC2) as a more targeted approach to cAMP-based therapies. For example, EPAC2-selective agonists could promote insulin secretion from pancreatic β cells, whereas EPAC1-selective agonists may be useful in the treatment of vascular inflammation. By contrast, EPAC1 and EPAC2 antagonists could both be useful in the treatment of heart failure. Here we discuss whether the best way forward is to design EPAC-selective agonists or antagonists and the current strategies being used to develop isoform-selective, small-molecule regulators of EPAC1 and EPAC2 activity., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Published
2015
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results