TY - JOUR
T1 - Serological autoantibody profiling of type 1 diabetes by protein arrays
AU - Miersch, Shane
AU - Bian, Xiaofang
AU - Wallstrom, Garrick
AU - Sibani, Sahar
AU - Logvinenko, Tanya
AU - Wasserfall, Clive H.
AU - Schatz, Desmond
AU - Atkinson, Mark
AU - Qiu, Ji
AU - LaBaer, Joshua
N1 - Funding Information:
Shane Miersch and Xiaofang Bian are the guarantors of this work and, as such, had full access to all the data in the study and take responsibility for the integrity of the data. Shane Miersch and Sahar Sibani performed the screening, Shane Miersch and Xiaofang Bian worked on the validation and wrote the manuscript. Garrick Wallstrom and Tanya Logvinenko are responsible for the accuracy of data analysis. Clive H. Wasserfall, Desmond Schatz, Mark Atkinson provide samples and valuable discussions. Ji Qiu and Joshua LaBaer contributed to experiment design and reviewed/edited manuscript. This study was supported by research grants from Juvenile Diabetes Research Foundation (JDRF) : 5-2005-1170 , 17-2007-1045 and 6-2012-513 . We thank Dr. John Hutton (Barbara Davis Center, Denver) for the provision of anti-ZnT8 antibodies and Dr. Catherine Cormier (Virginia G. Piper Center for Personalized Diagnostics) for critical reading of this manuscript.
PY - 2013/12/6
Y1 - 2013/12/6
N2 - The need for biomarkers that illuminate the pathophysiology of type 1 diabetes (T1D), enhance early diagnosis and provide additional avenues for therapeutic intervention is well recognized in the scientific community. We conducted a proteome-scale, two-stage serological AAb screening followed by an independent validation study. In the first stage, the immunoreactivity was compared between T1D cases and healthy controls against ~. 6000 human proteins using the nucleic acid programmable protein array (NAPPA). Genes identified with higher signal intensities in patients were challenged with a larger sample set during the second stage. Statistical analysis revealed 26 novel autoantigens and a known T1D-associated autoantigen. During validation, we verified the presence of AAbs to dual specificity tyrosine-phosphorylation-regulated kinase 2 (DYRK2) using the Luciferase ImmunoPrecipitation System (LIPS) assay (36% sensitivity, 98% specificity). The AUC for a combination of DYRK2A and the classical T1D AAb IA-2A was 0.90 compared to 0.72 for DYRK2A and 0.64 for IA-2A alone. This is the first systematic screening for seroreactivity against a large number of human proteins in T1D patients. We demonstrated the application of protein microarrays to identify novel autoantigens in T1D, expanded the current T1D "autoantigenome" and help fulfill the goal of searching for novel biomarker candidates for T1D. Biological significance: Protein microarrays provide a high-throughput platform that enables the profiling of serum antibodies to a large number of protein antigens. The value of AAb biomarkers in diagnosis, prognosis and treatment is well recognized in autoimmune diseases including T1D. We performed a systematic screening for new T1D-associated autoantigens by adapting the innovative protein array platform NAPPA. We believe that the discovery in this study will add information on candidate autoantigens that could potentially improve the diagnosis and help uncover the pathophysiology of T1D. The successful use of NAPPA for T1D AAb profiling will open the window for larger studies including more human antigen genes and other autoimmune diseases.
AB - The need for biomarkers that illuminate the pathophysiology of type 1 diabetes (T1D), enhance early diagnosis and provide additional avenues for therapeutic intervention is well recognized in the scientific community. We conducted a proteome-scale, two-stage serological AAb screening followed by an independent validation study. In the first stage, the immunoreactivity was compared between T1D cases and healthy controls against ~. 6000 human proteins using the nucleic acid programmable protein array (NAPPA). Genes identified with higher signal intensities in patients were challenged with a larger sample set during the second stage. Statistical analysis revealed 26 novel autoantigens and a known T1D-associated autoantigen. During validation, we verified the presence of AAbs to dual specificity tyrosine-phosphorylation-regulated kinase 2 (DYRK2) using the Luciferase ImmunoPrecipitation System (LIPS) assay (36% sensitivity, 98% specificity). The AUC for a combination of DYRK2A and the classical T1D AAb IA-2A was 0.90 compared to 0.72 for DYRK2A and 0.64 for IA-2A alone. This is the first systematic screening for seroreactivity against a large number of human proteins in T1D patients. We demonstrated the application of protein microarrays to identify novel autoantigens in T1D, expanded the current T1D "autoantigenome" and help fulfill the goal of searching for novel biomarker candidates for T1D. Biological significance: Protein microarrays provide a high-throughput platform that enables the profiling of serum antibodies to a large number of protein antigens. The value of AAb biomarkers in diagnosis, prognosis and treatment is well recognized in autoimmune diseases including T1D. We performed a systematic screening for new T1D-associated autoantigens by adapting the innovative protein array platform NAPPA. We believe that the discovery in this study will add information on candidate autoantigens that could potentially improve the diagnosis and help uncover the pathophysiology of T1D. The successful use of NAPPA for T1D AAb profiling will open the window for larger studies including more human antigen genes and other autoimmune diseases.
KW - Autoantibody
KW - NAPPA
KW - Protein array
KW - Type 1 diabetes
UR - http://www.scopus.com/inward/record.url?scp=84887524283&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84887524283&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2013.10.018
DO - 10.1016/j.jprot.2013.10.018
M3 - Article
C2 - 24148850
AN - SCOPUS:84887524283
SN - 1874-3919
VL - 94
SP - 486
EP - 496
JO - Journal of Proteomics
JF - Journal of Proteomics
ER -