Cell migration is a complicated multistep process that is central to the pathogenesis of diverse disease processes. Although neutrophils are an essential part of the innate immune response, their inappropriate recruitment is central to chronic inflammatory disorders including asthma, arthritis and inflammatory bowel disease;and also contributes to the pathogenesis of other diseases such as cardiovascular disease, tumor progression and Alzheimer's disease. Chemotaxis, the movement of cells within a chemical gradient, is the fundamental process underlying neutrophil recruitment. Despite its importance, current tools limit progress towards understanding the molecular mechanisms that regulate cell migration. In particular, current methods are largely limited to 2D environments, require relatively large blood draws and use stimuli of questionable physiological relevance. Our goal is to use and further develop microscale methods and microscale in vitro models that overcome these challenges, enhancing our ability to identify signaling pathways that regulate cell polarization and directed motility in the context of disorders involving the innate immune system - specifically the role of Hax1 signaling in neutrophil chemotaxis in the context of severe congential neutropenia (SCN). Currently, chemotaxis studies are very time and labor intensive limiting the number of experimental conditions that can be explored. Our approach fundamentally changes the way a researcher can approach chemotaxis studies. Many more experimental conditions can be examined with the same time/effort. A key strength of this application lies in the use of novel (but simple) microfluidic devices that generate defined and stable chemical gradients that do not require laminar flow for gradient formation or maintenance and can be adapted to high throughput screening.
In Aim 1, we will develop improved 3D assays that provide a more physiologically relevant context for cell migration and develop methods that allow the use of small volume finger stick samples.
In Aim 2, we will develop microfluidic-based wound assays to analyze neutrophil recruitment through cell sourced gradients to determine factors that regulate cell polarization and directed cell migration in multicelluar environments In Aim 3, we propose to study how Hax1/HS1/G113 signaling modulates neutrophil motility and recruitment in more complex 3D and cell sourced gradients to mimic in vivo conditions. Using these systems, we will dissect how Hax1 regulates gradient sensing and directed cell migration with Hax1-deficient neutrophil-like PLB987 cells. Future applications will include analysis of neutrophil motility from patients presenting with neutropenia using 2D and 3D/cell sourced microfluidic systems to understand disease pathogenesis and identify novel therapeutic targets.

Public Health Relevance

We anticipate that the research proposed in this grant will not only help to elucidate the basic mechanisms that regulate cell motility, but will also provide insight into the development of novel therapeutic approaches to treat inflammation in general. These studies will therefore have implications for treating many different disorders where cell motility is central to disease pathogenesis, including arthritis, cardiovascular disease and cancer.

National Institute of Health (NIH)
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Research Project (R01)
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Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
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Hunziker, Rosemarie
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University of Wisconsin Madison
Biomedical Engineering
Schools of Engineering
United States
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Li, Chao; Yu, Jiaquan; Paine, Paxton et al. (2018) Double-exclusive liquid repellency (double-ELR): an enabling technology for rare phenotype analysis. Lab Chip 18:2710-2719
Powell, Davalyn; Tauzin, Sebastien; Hind, Laurel E et al. (2017) Chemokine Signaling and the Regulation of Bidirectional Leukocyte Migration in Interstitial Tissues. Cell Rep 19:1572-1585
Barkal, Layla J; Procknow, Clare L; Álvarez-García, Yasmín R et al. (2017) Microbial volatile communication in human organotypic lung models. Nat Commun 8:1770
Moussavi-Harami, S F; Mladinich, K M; Sackmann, E K et al. (2016) Microfluidic device for simultaneous analysis of neutrophil extracellular traps and production of reactive oxygen species. Integr Biol (Camb) 8:243-52
Zanotelli, Matthew R; Ardalani, Hamisha; Zhang, Jue et al. (2016) Stable engineered vascular networks from human induced pluripotent stem cell-derived endothelial cells cultured in synthetic hydrogels. Acta Biomater 35:32-41
Jiménez-Torres, José A; Peery, Stephen L; Sung, Kyung E et al. (2016) LumeNEXT: A Practical Method to Pattern Luminal Structures in ECM Gels. Adv Healthc Mater 5:198-204
Jiménez-Torres, José A; Beebe, David J; Sung, Kyung E (2016) A Microfluidic Method to Mimic Luminal Structures in the Tumor Microenvironment. Methods Mol Biol 1458:59-69
Guckenberger, David J; Berthier, Erwin; Beebe, David J (2015) High-density self-contained microfluidic KOALA kits for use by everyone. J Lab Autom 20:146-53
Bischel, Lauren L; Beebe, David J; Sung, Kyung E (2015) Microfluidic model of ductal carcinoma in situ with 3D, organotypic structure. BMC Cancer 15:12
Theberge, Ashleigh B; Yu, Jiaquan; Young, Edmond W K et al. (2015) Microfluidic multiculture assay to analyze biomolecular signaling in angiogenesis. Anal Chem 87:3239-46

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