Multidisciplinary approaches in developing immune-evasive β-cell replacement therapies for type 1 diabetes

Type 1 Diabetes (T1D) is an autoimmune disorder characterized by the destruction of insulin producing pancreatic β-cells, leading to chronic hyperglycemia and associated severe complications. Current therapeutic strategies, including insulin therapy and islet transplantation, are limited by incomplete glycemic control, donor scarcity, and immune rejection. This thesis aims to overcome these challenges by integrating biomaterials, genetic engineering, and computational modeling to develop innovative β-cell replacement therapies. Firstly, Gelatin Methacryloyl (GelMA) hydrogels were optimized using visible-light photocrosslinking techniques. An Artificial Neural Network (ANN)- based computational model was developed to predict key hydrogel properties, significantly enhancing biomaterial design efficiency and efficacy for encapsulation and transplantation of β-cells. Secondly, genetic modifications were explored using CRISPR/Cas9 technology, initially targeting single genes (RNLS and HIVEP2) implicated in immune modulation and cellular protection. Despite successful gene editing, these single-gene knockout approaches failed to protect β-cells from immune rejection, highlighting the need for more comprehensive strategies. Consequently, a multi faceted genetic approach was developed, combining knockout of major histocompatibility complex genes (β2M and CIITA) with overexpression of immunomodulatory molecules (murine H2-Kᵇ, CD47, CD55) to comprehensively evade immune detection. Overall, this thesis demonstrates that integrating computational modeling, genetic engineering, and biomaterial science provides a powerful, synergistic platform for developing next-generation cellular therapies for T1D. The methodologies established have broad translational potential for addressing immune rejection and improving graft survival, ultimately paving the way toward clinically viable, personalized regenerative medicine approaches.