Immune Cells to Discover Novel Principles in Cell- & Mechano-Biology. The immune system consists of an elaborate orchestration of cell types specialized for different molecular processes. Due to the spectrum of these specialisations, immune cells represent an illuminating cellular model to identify general principles in cell- and mechanical-biology. Particularly, we employ immune cells to unravel basic principles and mechanisms of cell motility, leukocyte trafficking, macropinocytosis, and host-pathogen interactions. Our lab interdisciplinary combines advanced live-cell microscopy, image analysis, genetic-engineering by CRISPR, custom-made micro-environments like microfluidics and 3D collagen matrices, and unbiased system-wide approaches like genome-wide CRISPR screening, transcriptomics, and proteomics. Together, we aim to identify fundamental principles in the cell- and mechano-biology of the immune system and their misregulation in disease.
Immune Cell Navigation. Immune cells are very fast moving cells. On their journey, these migratory cells have to find their way through crowded three-dimensional mazes (Kameritsch & Renkawitz, Trends in Cell Biology). Whereas some cell types such as mesenchymal cells proteolytically digest the environment on their path, immune cells typically migrate without digesting or remodelling their environment. Given the cumulative distance of all immune cells in the human body of more than 100,000 km per hour, they otherwise would perforate the body with more than two million kilometers of tunnels per day. To discover how these extremely fast cells navigate through the dense meshwork of interstitial tissue fibres without harming other cells, we built obstacle courses for immune cells in reconstituted environments (Kroll et al, Current Protocols). Thereby, we discovered that immune cells use their nucleus as a ruler to probe their surroundings for the largest pores — and thereby find the path of least resistance (Renkawitz et al, Nature). Further, we discovered that microtubules are functionally important to coordinate the exploratory fronts of dendritic cells that are simultaneously probing the 3D microenvironments (Kopf et al, Journal of Cell Biology; Renkawitz et al, Nature).
Kroll et al, EMBO J (2023)
Kroll et al, Current Protocols (2022)
Kameritsch & Renkawitz, Trends in Cell Biology (2020)
Renkawitz et al, Nature (2019)
Cytoskeletal Dynamics and Adaptability during Immune Cell Migration. Almost all moving cells employ the actin cytoskeleton as an intracellular force generator to move forward. We discovered that amoeboid cells (such as dendritic cells) adapt their actin cytoskeleton dynamics to the adhesiveness of the migratory substrate (Renkawitz et al, Nature Cell Biology). Thereby we could show that not tracks of adhesive substrates but gradients of chemoattractants dictate the path of amoeboid cells, endowing these cells with extraordinary flexibility and enabling them to traverse almost every type of tissue (Renkawitz & Sixt, EMBOreports).
Hons et al, Nature Immunology 2018
Renkawitz & Sixt, EMBOreports 2010
Lämmermann et al, Blood 2009
Renkawitz et al, Nature Cell Biology 2009
Three-dimensional Microenvironments. Whereas in vivo experiments are truly physiological, they do not allow for precise manipulation of environmental parameters and are elaborate for the discovery of detailed molecular mechanisms. In contrast, in vitro experiments on two-dimensional (2D) substrates (e.g. cell culture dishes) enable faster manipulations, but increasing knowledge points to substantial differences of cellular mechanisms in 2D and three-dimensional (3D) environments. To bridge this gap, we and others developed micro-engineered tissue-mimetic assays to combine the advantage of precise manipulations in 2D assays with the presence of complex 3D microenvironments. Specifically, we implemented the methodology of micro-engineered ”pillar-forests’ to study cell migration in vitro in 3D with precisely defined microenvironmental parameters (Renkawitz et al., Methods in Cell Biology). Shortly, these devices provide a 3D migration environment made of PDMS (polydimethylsiloxane), in which two closely adjacent surfaces are interconnected by micron-sized structures such as pillars (or micro-channels). Thereby, these devices represent a flattened approximation of complex 3D environments with the advantage of cellular confinement in one plane (XY) close to the imaging surface, enabling high-resolution single-cell imaging. We complement these assays with experimentation in collagen matrices, ex vivo tissue explants, and in vivo approaches.
Clausen et al, Eur J Immunology 2022
Holtkamp et al, Nature Immunology 2021
Renkawitz et al, Methods in Cell Biology 2018
Frick et al, PLoS One 2018
Genetic Engineering in Immune Cells. Current knowledge on the molecular mechanisms and cell biological principles of immune cell biology (such as cell migration) is often based on studies utilizing knockout mice, which however hampers screening of large numbers of candidate components due to its time and resource consuming nature. To circumvent this bottleneck, conditionally immortalised hematopoietic precursor cells (Hoxb8 cells) with myeloid and lymphoid potential have been established by the Häcker Lab (Redecke et al, Nature Methods 2013). We recently contributed in showing that Hoxb8 cells can be differentiated into migratory DCs functionally indistinguishable from their primary counterparts (bone-marrow derived DCs) (Leithner et al, Eur J Immunol). Importantly, Hoxb8-FL cells can be efficiently targeted via CRISPR mediated gene editing (Leithner et al, Eur J Immunol). Thus, we employ immortalised hematopoietic precursor cells (Hoxb8 cells) as an accessible cellular model for genetic engineering, including genetic knockouts and fluorescent tagging.
Leithner et al, Eur J Immunol 2018
Earlier Work on Homology Search during DNA Recombination. During his PhD research in the Lab of Prof. Stefan Jentsch, Jörg visualised homology search during DSB repair in vivo, using genome-wide analysis of chromatin immunoprecipitation of DSB repair factors in yeast. This showed that homology search is strongly influenced by the chromosomal architecture and nuclear organization (Renkawitz et al, Mol Cell). These findings led to a model, in which homology search during DSB repair proceeds by an accelerated random probing mechanism guided by genomic proximity (Renkawitz et al, Nat Rev Mol Cell Biol).
Lademann et al, Cell Reports 2017
Renkawitz, Lademann & Jentsch, Nature Reviews Molecular Cell Biology 2014
Renkawitz et al, Molecular Cell 2013