werkbezoek University of Colorado, Denver

  • Aanvrager: Mevrouw C. Ciminieri

werkbezoek; Research at University of Colorado, Denver (UCD) Ms. Chiara Ciminiere; University of Groningen Department of Molecular Pharmacology

Research background
Chronic obstructive pulmonary disease (COPD) is characterized by a progressive loss of lung function with airflow obstruction that is not fully reversible. The key problem underlying COPD is defective airway and alveolar repair in response to inflammation and oxidative stress, causing bronchitis, small airways remodeling and emphysema. As current therapies do not modify the course of the disease in COPD, new therapies need to be developed.
Epithelial progenitor cells of the distal lung interact with structural cells such as fibroblasts to contribute to lung repair, which is driven by canonical WNT/?-catenin signaling. This mechanism is impaired in COPD, in part because chronic inflammation and oxidative stress provide a hostile local microenvironment in the COPD lung, not only causing damage, but also hampering repair responses. We now demonstrate that inflammation and oxidative stress directly and indirectly affect canonical WNT/?-catenin signaling (responsible for repair) and that reversal of this process using targeted pharmacological inhibition is possible. We hypothesize that the distortion of regenerative signaling is driven by a canonical to non-canonical WNT signaling switch, skewing signaling towards JNK, p38 and Rho-kinase, which lead to halted repair. These pathways provide rational drug targets for alveolar repair and we propose that it is possible to restore lung repair by targeting these pathways.

Research at University of Colorado, Denver (UCD)
This move to Denver is part of the PhD project as indicated in the grant application: “Two PhD students will be recruited for this project, with PhD student 1 based at the University of Groningen (RuG) and the University of Colorado, Denver (UCD) and PhD student 2 based at the Maastricht University Medical Center+ (MUMC+) and the University of Vermont (UVM). PhD student 1 will focus mostly on the experimental aims related to WNT signaling and perform at RuG the gene expression studies, mass spectrometry studies and ligand/receptor interaction studies from aim 1. In addition, PhD student 1 will use organoids and lung slices to study the impact of non-canonical signaling and its interactions with inflammation and oxidative stress. For those studies, PhD student 1 will move to UCD after 6 months. Their work on aim 1 and aim 2b will be continued by a technician at RuG, while PhD student 1 resides at UCD.”

In detail, my work at UCD will give me several opportunities to: 1) Enrich my expertise in techniques I have already learned, like the organoids assay, using different tools, and transfer the knowledge to other people in UCD’s lab. After learning the organoids technique in Groningen, I will keep performing this assay in a different context and this will help me to supplement the experiments that I have already performed at RuG on the role of WNT signaling in COPD. Here, in fact, I have access to novel pharmacological tools, such as one recently discovered compound (FzM1.8), an allosteric modulator of the Fzd4 receptor in WNT signaling [1], which could potentially restore lung regeneration in COPD and that is only accessible here at UCD since prof. Melanie Koenigshoff has a direct collaboration with the people who discovered the compound. In fact, in the past 3 months at UCD I got the chance to work alongside the researcher responsible for this discovery to perform preliminary studies, and now I will get to test FzM1.8 on different models to assess its effect on WNT/?-catenin activation.
In addition, other people at UCD have started working with organoids and they value my expertise on the subject. This scientific exchange will help establish this technique in the daily research at UCD and will also help me to expand my knowledge by having access for example to human samples that can provide material to create patient-derived organoids, which are a unique tool to study regeneration and test pharmacological compounds on patient material.
2) Learn new techniques and procedures through direct access, which I don’t have in Groningen.
The lab at UCD has direct access to human tissue from donor and COPD patients, which we receive frequently, and on which I am learning to perform several procedures useful for my experimental studies, including epithelial cells and fibroblasts isolation, which will be crucial to perform in vitro studies both in 2D culture and 3D culture with organoids to test FzM1.8. I also plan on learning how to use human and mouse tissue to generate 3D-PCLS (precision cut lung slices) which are a novel and promising tool for ex-vivo research; in fact, patient-derived 3D-LTCs enable validation of potential drugs (like FzM1.8) in tissue from COPD patients in an individualized fashion and thus might enhance translation into clinical practice. At UCD I also have direct access to state-of-the-art techniques such as single-cell RNA analysis and FACS (Fluorescence activated cell sorting) which give us unique information on the cell populations we are studying in both physiological and disease context. All the people working in the lab at UCD have direct expertise on the previously mentioned techniques which are performed on a daily basis, and they are therefore teaching me through hands-on experience. Direct access to these techniques is not possible in Groningen at the moment, since they are not part of the daily lab routine and are therefore only accessible through third parties. These techniques give an added value to my research and to my skill set.

3) Get in contact with and learn from people with unique expertise in lung research. At UCD I will have access to the unique expertise of Prof. Dr. Melanie Koenigshoff, who has performed extensive research on lung disease both in Europe and in the USA and will be my supervisor. In addition, I will get in touch with researchers working on different aspects of lung disease in the same department through collaborations, conferences, and weekly seminars.

In the end, this experience at UCD will help me not only to enrich my research path by learning new techniques and expertise that I can bring back to Groningen, but it will also enrich me as a person by performing research in a different and advanced environment, and this is the basis of scientific exchange that guarantees the growth of a researcher.