Mevalonat Kinaz Eksikliğine Özgü Uyarılmış Pluripotent Kök Hücrelerden Türetilen Monositlerin Karakterizasyonu ve Hücre Göçünün Araştırılması

Abstract

Systemic autoinflammatory diseases are a group of disorders characterized by excessive activation of the innate immune system and recurring attacks of multisystemic inflammation. Familial Mediterranean fever (FMF) and Mevalonate Kinase Deficiency - Hyperimmunoglobulin D Syndrome (MKD-HIDS) are two systemic autoinflammatory diseases that result in the activation the pyrin inflammasome through different mechanisms. In previous works it has been demonstrated that pyrin and its protein partners colocalize with the actin cytoskeleton, potentially playing a role in actin rearrangement during cell migration. Increased cell migration, observed in cells from FMF patients where pyrin inflammasome is activated, supports these findings and points towards a link between pyrin activation and inflammatory cell migration. In contrast, primary cells from MKD-HIDS displayed defective cell migration. Due to the rarity of these diseases and difficulty accessing primary cells from patients, there is need for alternative models for autoinflammatory diseases. In light of this knowledge, in this thesis, primary fibroblast cultures were established from skin biopsies of 3 FMF and 3 MKD-HIDS patients and 1 healthy control. These primary fibroblasts were used to generate induced pluripotent stem cells (iPSC) for each individual. The iPSC lines were shown to express pluripotency markers and capacity to differentiate into three germ layers in vitro. The characterized iPSC lines were then differentiated into monocytes and characterized by gene expression and surface marker expression. Later, these iPSC-derived monocytes were utilized for cell migration and experiments which demonstrated a MKD-specific gene migration defect