Modeling MyD88 Deficiency In Vitro Provides New Insights in Its Function
| Autor: | Craig-Mueller, Nils, Hammad, Ruba, Elling, Roland, Alzubi, Jamal, Timm, Barbara, Kolter, Julia, Knelangen, Nele, Bednarski, Christien, Gläser, Birgitta, Ammann, Sandra, Ivics, Zoltán, Fischer, Judith, Speckmann, Carsten, Schwarz, Klaus, Lachmann, Nico, Ehl, Stephan, Moritz, Thomas, Henneke, Philipp, Cathomen, Toni |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
IRAK4
Genetic therapy transposon Cell- and tissue-based therapy Primary Immunodeficiency Diseases Immunology Induced Pluripotent Stem Cells induced pluripotent stem cells (iPSC) Monocytes MyD88 Toll-like receptor cell therapy iMac gene therapy Induzierte pluripotente Stammzelle Cell Line Gentherapie Humans Myeloid Cells ddc:610 Original Research iPSC Macrophages Cell Differentiation Toll-like receptors Induced pluripotent stem cells HEK293 Cells Interleukin-1 Receptor-Associated Kinases Myeloid Differentiation Factor 88 Cytokines Toll-like-Rezeptoren Signal Transduction |
| Zdroj: | Frontiers in immunology, 11:608802 Frontiers in Immunology |
| DOI: | 10.3389/fimmu.2020.608802/full#h11 |
| Popis: | Inherited defects in MyD88 and IRAK4, two regulators in Toll-like receptor (TLR) signaling, are clinically highly relevant, but still incompletely understood. MyD88- and IRAK4-deficient patients are exceedingly susceptible to a narrow spectrum of pathogens, with ∼50% lethality in the first years of life. To better understand the underlying molecular and cellular characteristics that determine disease progression, we aimed at modeling the cellular response to pathogens in vitro. To this end, we determined the immunophenotype of monocytes and macrophages derived from MyD88- and IRAK4-deficient patients. We recognized that macrophages derived from both patients were particularly poorly activated by streptococci, indicating that both signaling intermediates are essential for the immune response to facultative pathogens. To characterize this defect in more detail, we generated induced pluripotent stem cells (iPSCs) of fibroblasts derived from an MyD88-deficient patient. The underlying genetic defect was corrected using Sleeping Beauty transposon vectors encoding either the long (L) or the short (S) MYD88 isoform, respectively. Macrophages derived from these iPSC lines (iMacs) expressed typical macrophage markers, stably produced either MyD88 isoform, and showed robust phagocytic activity. Notably, iMacs expressing MyD88-L, but not MyD88-S, exhibited similar responses to external stimuli, including cytokine release patterns, as compared to genetically normal iMacs. Thus, the two MyD88 isoforms assume distinct functions in signaling. In conclusion, iPSC technology, in combination with efficient myeloid differentiation protocols, provides a valuable and inexhaustible source of macrophages, which can be used for disease modeling. Moreover, iPSC-derived macrophages may eventually aid in stabilizing MyD88-deficient patients during pyogenic infections. |
| Databáze: | OpenAIRE |
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