Genetic manipulation of neutrophil granulocytes

For a long time, it was assumed that neutrophil granulocytes, in their function as cells of the unspecific immune system, are only involved in the defence against infections and guarantee this solely by phagocytosis and chemical toxicity. Studies of the last 10 years, however, paint a more differentiated picture of this cell type. Thus, we now know that neutrophils have very complex functional patterns based on their heterogeneity and plasticity. Neutrophils are not only associated with infectious diseases or their defence, but are also involved in the pathogenesis of e.g. pancreatic tumours (Reid et al., 2011), cystic fibrosis and COPD (Kruger et al., 2015). It is surprising, however, that a massive infiltration of neutrophil granulocytes into lung or tumor tissue in connection with these diseases has exclusively negative effects, leading to a worse prognosis for the affected patient. The molecular mechanisms that determine why neutrophil granulocytes cause more damage in some patients or certain diseases instead of protecting the body are still largely unknown.

HoxB8 cells represent a suitable system for the analysis of precisely these molecular mechanisms (Wang et al., 2006). In their morphology, inflammatory behaviour and, as far as known, their expression pattern, they largely correspond to primary, naive neutrophils. However, these cells can be genetically manipulated in vitro using the CRISPR/Cas9 system (Jinek et al., 2012). Genes suspected of being involved in important regulatory processes of granulocytes can be knocked out by these genetic scissors. The aim of the present project is to investigate their significance for neutrophil function by specifically switching off individual genes in order to draw conclusions about the molecular causes of diseases and, in the long term, to understand the plasticity and regulation of a neutrophil response in its entirety.

Researcher

• Prof. Dr. Matthias Gunzer
• Andreas Kraus