Research Stijn Verleden

Abstract

Hierarchical Phase-Contract Tomography (HiP-CT) has been developed as an innovative way to study an intact human organ in 3D resolution on a near-microscopic scale. Within this project we want to leverage HiP-CT to investigate human lungs from patients with COPD and lung fibrosis. We aim to identify and quantify disease specific features and unravel the role of the vasculature in the pathophysiology of chronic lung diseases. HiP-CT will provide us with the advantage of comparing disease specific features across the entire organ (and thereby allow us to compare none vs mild vs severely affected tissue with the same lung) and quantify this in 3 dimensions which is not possible with any of the existing technologies. Using this highly detailed information we will segment and quantify the entire bronchial and vascular system and compare this in health and disease. We next want to leverage this knowledge to also obtain deeper biologic information by performing spatial analysis in the same organs using multiplex immunofluorescence and spatial transcriptomics. Lastly, we want to validate our findings in a larger collection of organs, as well as organs or biopsies with early disease from our own biorepository to validate the importance of our findings. We are convinced that by combining state-of-the-art imaging technology with next generation biological tools, we will get one step closer in our understanding of chronic lung diseases which is desperately needed given their poor outcome.

Researcher(s)

• Promoter: Verleden Stijn

Research team(s)

• Antwerp Surgical Training, Anatomy and Research Centre (ASTARC)

Project type(s)

• Research Project

Abstract

Visualisation of intact organs with microscopic resolution can learn us a lot about the morphological and molecular changes in health and disease. Specifically for the lung, we do not have sufficient insights about the changes that are associated with chronic lung disease. Therefore within this project, we want to leverage unique methodology (hierarchic phase contrast tomography) to image the complete lung at a microscopic resolution. We aim to better understand the changes associated with disease. Unique with this technology is that the vasculature can also be directly visualised. Therefore we want to utilize this tool to better understand what happens with the airways, the vasculature, the alveoli, the pleura and most interestingly how these interact. We also aim to associate our findings with the in vivo pre-operative omaging, but also to the pathological diagnosis. These insights will be crucial to better understand the mechanisms of chronic lung diseases.

Researcher(s)

• Promoter: Verleden Stijn

Research team(s)

• Antwerp Surgical Training, Anatomy and Research Centre (ASTARC)

Project type(s)

• Research Project

Abstract

Pulmonary diseases are a major cause of morbidity and mortality. A diversity of different lung diseases exists each associated with their own presentation, prognosis, and pathophysiologic mechanisms. Leveraging a macro to micro approach using state-of-the-art technologies such as HiP-CT, microCT and spatial transcriptome analysis on wellcharacterized human lungs and innovative animal models such as primary bronchial epithelial culture and a murine lung transplant model, the aim of this application is to better diagnose and better understand the pathophysiological mechanisms of lung diseases. Several topics are detailed where the overall aim is to improve the diagnosis of lung disease. Firstly, we seek to consolidate the tumor classification of surgically resected lung lobes more adequately by investigating micro-lesions in the lung. Secondly, we want to assess suitability of donor lungs for lung transplantation via ex vivo imaging. Thirdly, we aim to find better markers of small airway disease in patients with smoking induced lung changes. Fourthly, we seek to directly correlate in vivo CT data to ex vivo histological and functional data to construct a virtual biopsy. Next to better diagnosis, it is also important to improve the knowledge on the pathophysiological mechanisms of lung diseases. Therefore, the morphological changes and pathophysiological mechanisms of several lung diseases will be further explored in comparison to physiological lung aging. HiP-CT analysis will be used on aging and end-stage lung specimens to provide an unprecedented view on the bronchial and vasculature changes at extreme high resolution on an entire specimen. As this approach is logistically difficult in early disease specimens, microCT will be used to compare bronchial and vasculature changes in early to late-stage disease and aging. To further investigate the mechanisms of this lung remodeling, primary bronchial epithelial cells will be harvested and used to investigate the immunoreactivity to infectious and non-infectious triggers. The lung tissue itself will also be subjected to in-depth investigation by staining for different inflammatory markers, by scanning electron microscopy to investigate collateral ventilation and by spatial transcriptome/single nuclear sequencing analysis. Lastly, the role of microvascular alterations will be further investigated using a murine orthotopic lung transplant model where the sequence of vascular changes will be investigated via bulk transcriptome analysis and in/ex vivo imaging techniques. Lastly, pharmacological inhibition will be attempted to confirm the involvement of the microvasculature. Ultimately, this ambitious and innovative project will lead to tremendous new insights which could help in the diagnosis of (early stage) lung disease and will definitely be important to better understand the pathophysiology of respiratory diseases. This will assist in assessing disease prognosis and eventually therapy, which is desperately needed given the high- morbidity and mortality of lung diseases.

Researcher(s)

• Promoter: Verleden Stijn
• Fellow: Verleden Stijn

Research team(s)

• Antwerp Surgical Training, Anatomy and Research Centre (ASTARC)

Project type(s)

• Research Project