Search

Your search keyword '"Wyatt, Rebecca C."' showing total 12 results
Start Over Author "Wyatt, Rebecca C."
12 results on '"Wyatt, Rebecca C."'

2. Non-coding variants disrupting a tissue-specific regulatory element in HK1 cause congenital hyperinsulinism

Author

Wakeling, Matthew N., Owens, Nick D. L., Hopkinson, Jessica R., Johnson, Matthew B., Houghton, Jayne A. L., Dastamani, Antonia, Flaxman, Christine S., Wyatt, Rebecca C., Hewat, Thomas I., Hopkins, Jasmin J., Laver, Thomas W., van Heugten, Rachel, Weedon, Michael N., De Franco, Elisa, Patel, Kashyap A., Ellard, Sian, Morgan, Noel G., Cheesman, Edmund, Banerjee, Indraneel, Hattersley, Andrew T., Dunne, Mark J., Richardson, Sarah J., and Flanagan, Sarah E.

Published
2022
Full Text
View/download PDF

5. Improved Specificity of Glutamate Decarboxylase 65 Autoantibody Measurement Using Luciferase-Based Immunoprecipitation System Assays.

Author

Wyatt, Rebecca C., Grace, Sian L., Brigatti, Cristina, Marzinotto, Ilaria, Gillard, Ben T., Shoemark, Deborah K., Chandler, Kyla, Achenbach, Peter, Piemonti, Lorenzo, Long, Anna E., Gillespie, Kathleen M., Lampasona, Vito, and Williams, Alistair J. K.

Subjects
*GLUTAMATE decarboxylase, *AUTOANTIBODIES, *TYPE 1 diabetes, *IMMUNOPRECIPITATION, *ANTIGENS
Abstract

Autoantibodies to glutamate decarboxylase (GADA) are widely used in the prediction and classification of type 1 diabetes. GADA radiobinding assays (RBAs) using N-terminally truncated antigens offer improved specificity, but radioisotopes limit the high-throughput potential for population screening. Luciferase-based immunoprecipitation system (LIPS) assays are sensitive and specific alternatives to RBAs with the potential to improve risk stratification. The performance of assays using the Nanoluc luciferase (Nluc)-conjugated GAD65 constructs, Nluc-GAD65(96-585) and full length Nluc-GAD65(1-585), were evaluated in 434 well-characterized serum samples from patients with recent-onset type 1 diabetes and first-degree relatives. Nonradioactive, high-throughput LIPS assays are quicker and require less serum than RBAs. Of 171 relatives previously tested single autoantibody positive for autoantibodies to full-length GAD65 by RBA but had not progressed to diabetes, fewer retested positive by LIPS using either truncated (n = 72) or full-length (n = 111) antigen. The Nluc-GAD65(96-585) truncation demonstrated the highest specificity in LIPS assays overall, but in contrast to RBA, N-terminus truncations did not result in a significant increase in disease-specificity compared with the full-length antigen. This suggests that binding of nonspecific antibodies is affected by the conformational changes resulting fromaddition of the Nluc antigen. Nluc-GAD65(96-585) LIPS assays offer low-blood-volume, high-specificity GADA tests for screening and diagnostics. [ABSTRACT FROM AUTHOR]

Published
2024
Full Text
View/download PDF

7. A novel LIPS assay for insulin autoantibodies

Author

Liberati, Daniela, Wyatt, Rebecca C., Brigatti, Cristina, Marzinotto, Ilaria, Ferrari, Maurizio, Bazzigaluppi, Elena, Bosi, Emanuele, Gillard, Ben T., Gillespie, Kathleen M., Gorus, Frans, Weets, Ilse, Balti, Eric, Piemonti, Lorenzo, Achenbach, Peter, Williams, Alistair J. K., and Lampasona, Vito

Published
2018
Full Text
View/download PDF

9. Circulating C-Peptide Levels in Living Children and Young People and Pancreatic β-Cell Loss in Pancreas Donors Across Type 1 Diabetes Disease Duration

Author

Carr, Alice L.J., primary, Inshaw, Jamie R.J., additional, Flaxman, Christine S., additional, Leete, Pia, additional, Wyatt, Rebecca C., additional, Russell, Lydia A., additional, Palmer, Matthew, additional, Prasolov, Dmytro, additional, Worthington, Thomas, additional, Hull, Bethany, additional, Wicker, Linda S., additional, Dunger, David B., additional, Oram, Richard A., additional, Morgan, Noel G., additional, Todd, John A., additional, Richardson, Sarah J., additional, and Besser, Rachel E.J., additional

Published
2022
Full Text
View/download PDF

10. Circulating C-Peptide Levels in Living Children and Young People and Pancreatic Beta Cell Loss in Pancreas Donors Across Type 1 Diabetes Disease Duration

Author

Carr, Alice L.J., primary, Inshaw, Jamie R.J., primary, Flaxman, Christine S., primary, Leete, Pia, primary, Wyatt, Rebecca C., primary, Russell, Lydia A., primary, Palmer, Matthew, primary, Prasolov, Dmytro, primary, Worthington, Thomas, primary, Hull, Bethany, primary, Wicker, Linda S., primary, Dunger, David B., primary, Oram, Richard A., primary, Morgan, Noel G., primary, Todd, John A., primary, Richardson, Sarah J., primary, and Besser, Rachel E.J., primary

Published
2022
Full Text
View/download PDF

11. Non-coding variants disrupting a tissue-specific regulatory element in HK1cause congenital hyperinsulinism

Author

Wakeling, Matthew N., Owens, Nick D. L., Hopkinson, Jessica R., Johnson, Matthew B., Houghton, Jayne A. L., Dastamani, Antonia, Flaxman, Christine S., Wyatt, Rebecca C., Hewat, Thomas I., Hopkins, Jasmin J., Laver, Thomas W., van Heugten, Rachel, Weedon, Michael N., De Franco, Elisa, Patel, Kashyap A., Ellard, Sian, Morgan, Noel G., Cheesman, Edmund, Banerjee, Indraneel, Hattersley, Andrew T., Dunne, Mark J., Richardson, Sarah J., and Flanagan, Sarah E.

Abstract

Gene expression is tightly regulated, with many genes exhibiting cell-specific silencing when their protein product would disrupt normal cellular function1. This silencing is largely controlled by non-coding elements, and their disruption might cause human disease2. We performed gene-agnostic screening of the non-coding regions to discover new molecular causes of congenital hyperinsulinism. This identified 14 non-coding de novo variants affecting a 42-bp conserved region encompassed by a regulatory element in intron 2 of the hexokinase 1 gene (HK1). HK1 is widely expressed across all tissues except in the liver and pancreatic beta cells and is thus termed a ‘disallowed gene’ in these specific tissues. We demonstrated that the variants result in a loss of repression of HK1in pancreatic beta cells, thereby causing insulin secretion and congenital hyperinsulinism. Using epigenomic data accessed from public repositories, we demonstrated that these variants reside within a regulatory region that we determine to be critical for cell-specific silencing. Importantly, this has revealed a disease mechanism for non-coding variants that cause inappropriate expression of a disallowed gene.

Published
2022
Full Text
View/download PDF

12. A novel LIPS assay for insulin autoantibodies

Author

Frans Gorus, Elena Bazzigaluppi, Cristina Brigatti, Vito Lampasona, Peter Achenbach, Ilaria Marzinotto, Daniela Liberati, Ilse Weets, Emanuele Bosi, Eric V. Balti, Rebecca Wyatt, Alistair J K Williams, Ben Gillard, Maurizio Ferrari, Kathleen M Gillespie, Lorenzo Piemonti, Liberati, Daniela, Wyatt, Rebecca C., Brigatti, Cristina, Marzinotto, Ilaria, Ferrari, Maurizio, Bazzigaluppi, Elena, Bosi, Emanuele, Gillard, Ben T., Gillespie, Kathleen M., Gorus, Fran, Weets, Ilse, Balti, Eric, Piemonti, Lorenzo, Achenbach, Peter, Williams, Alistair J. K., Lampasona, Vito, Pathology/molecular and cellular medicine, Diabetes Pathology & Therapy, Vriendenkring VUB, Clinical Biology, Faculty of Medicine and Pharmacy, and Internal Medicine

Subjects
0301 basic medicine, medicine.medical_specialty, Insulin Antibodies, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, 030209 endocrinology & metabolism, medicine.disease_cause, Sensitivity and Specificity, Autoimmunity, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Antigen, Predictive Value of Tests, Internal medicine, Diabetes mellitus, Internal Medicine, medicine, Humans, Immunoprecipitation, Insulin, Child, Luciferases, Fluorescent Dyes, Autoantibodies, Proinsulin, Immunoassay, Type 1 diabetes, medicine.diagnostic_test, business.industry, Autoantibody, General Medicine, Prognosis, medicine.disease, Autoantibodie, Diabetes Mellitus, Type 1, HEK293 Cells, 030104 developmental biology, Case-Control Studies, T1D, LIPS, Prediction, business, Biomarkers
Abstract

Aims: Insulin autoantibodies (IAA) are often the first marker of autoimmunity detected in children in the preclinical phase of type 1 diabetes (T1D). Currently, the vast majority of laboratories adopt the radiobinding micro-assay (RBA) for measuring IAA. Our aim was to replace RBA with a novel non-radioactive IAA Luciferase Immuno Precipitation System (LIPS) assay with improved performance. Methods: We developed (pro)insulin antigens with alternative placements of a NanoLuc™ luciferase reporter (NLuc). Performance in LIPS was evaluated by testing sera from new onset T1D (n = 80), blood donors (n = 123), schoolchildren (n = 186), first-degree relatives (FDRs) from the Bart’s Oxford family study (n = 53) and from the Belgian Diabetes Registry (n = 136), coded sera from the Islet Autoantibody Standardization Program (IASP) (T1D n = 50, blood donors n = 90). Results: IAA LIPS based on B chain-NLuc proinsulin or B chain-NLuc insulin, in which NLuc was fused at the C-terminus of the insulin B chain, required only 2 μL of serum and a short incubation time, showed high concordance with RBA (Spearman r = 0.866 and 0.833, respectively), high assay performance (B chain-NLuc proinsulin ROC-AUC = 0.894 and B chain-NLuc insulin ROC-AUC = 0.916), and an adjusted sensitivity at 95% specificity ranking on par with the best assays submitted to the two most recent IASP workshops. In FDRs, the IAA LIPS showed improved discrimination of progressors to T1D compared to RBA. Conclusions: We established a novel high-performance non-radioactive IAA LIPS that might replace the current gold standard RBA and find wide application in the study of the IAA response in T1D.

Published
2018

Catalog

Books, media, physical & digital resources
See catalog results