Periodic Reporting for period 3 - PAINTBOX (Pancreatic islet dysfunction and type 2 diabetes – beyond the genome) Reporting period: 2020-10-01 to 2022-03-31 Summary of the context and overall objectives of the project The prevalence of type 2 diabetes (T2D) is rapidly increasing worldwide and despite great efforts to prevent and cure the disease, the increase persist. There is an urgent need to develop innovative and effective prevention and treatment strategies for T2D. Recent genome-wide association studies were expected to uncover disease-causing mechanisms in common metabolic disorders. Surprisingly, the identified genetic variants only explain a small proportion of the estimated heritability of T2D, the world’s most rapidly increasing metabolic disorder. Novel studies, going beyond the genome, are therefore needed to identify key mechanisms causing T2D. These studies should focus on pancreatic islets, the tissue of primary pathogenetic importance for diabetes. Also, mechanisms that allow environmental factors to come into play are likely to be important. Epigenetics fulfill such criteria, and our previous work has pioneered epigenetic discoveries in T2D. PAINTBOX is a multidisciplinary program of research through which I will combine advanced sequencing technologies and novel epigenetic methods with mathematical modeling and clinical medicine to develop new prediction and treatment strategies for T2D. Innovative techniques and access to some of the world´s best patient cohorts permit us to address the role of epigenetics in the development of T2D in radically new ways. I expect this novel interdisciplinary project to generate groundbreaking information that opens up new horizons for prediction, prevention and treatment of T2D and it will thereby help many patient with the disease and reduce health-related costs for society.The overall objective is to find new mechanisms that contribute to the development of T2D as well as new biomarkers that predict the diease and potential new therapies. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far We have generated ATAC-seq data in human pancreatic islets from donors with T2D and non-diabetic controls. We have related ATAC-seq data and studied the interplay between genomic locations of open chromatin, DNA-binding proteins, RNA-sequencing, DNA methylation and histone modifications, which has been published (Bysani et al, doi: 10.1038/s41598-019-44076-8).We have also studied risk factors for T2D on human islets and found that both high glucose and high glucose plus palmitate exposure for 48 hours impair glucose-stimulated insulin secretion and alter gene expression and DNA methylation. These data have been published in two papers by Hall et al (doi: 10.2337/db18-0900 and doi: 10.1016/j.mce.2017.11.019).We have also been working on inhibitors of epigenetic enzymes in relation to islet function and T2D. This part of the project is also going well and we have published several paper related to this part (Backe et al and Daneshpajooh et al. doi: 10.1016/j.mce.2017.07.001 doi: 10.1155/2019/5451038 and doi: 10.1007/s00592-018-1201-4). Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) All studies described above are beyond state of the art and we expect to further build our model of identified defects in human T2D pancreatic islets until the end of the project. Example of ATAC-seq peaks in human islets.