Multi-modular patient-derived lung organoids to aid precision medicine

Supervisors:

• Professor Chris Brightling ceb17@le.ac.uk
• Dr Himanshu Kaul Himanshu.kaul@leicester.ac.uk

Project

Aim: To validate that patient-derived lung organoids can stratify patient-specific response to therapy.

Background: In the UK, 8 million people suffer from asthma. Precision medicine is a potentially powerful therapeutic strategy but they need to consider patients’ specific profiles. Organoids are 3D in-vitro cultures can help achieve this because they recapitulate human physiology and genomics more accurately vs animal models. My lab has developed a high-throughput protocol (unpublished) to create lung organoids using terminally differentiated cells that can undergo self-organisation and display aspects of lung physiology. Our hypothesis is that creating patient-derived organoids will capture patient-specific asthma mechanisms and response to therapy.

Methods:  

Objective 1: Optimise protocol to create patient-derived lung organoids
The candidate will extend the protocol to create organoids using patient-derived epithelial cells (currently, we use epithelial cell lines). A key challenge they will overcome is to culture human airway epithelial cells in media lacking steroid-based agents, by either altering organoid composition or adding supplements. Organoids will be assessed via imaging (markers: E-cadherin, α-smooth muscle actin, collagen II, collagen IV) and live-dead assay. Success indicator: asthmatics-derived organoids (vs healthy controls) yield hallmarks of asthma: e.g. higher collagen deposition, α-smooth muscle actin, etc.  

Objective 2: Capture patient-specific hallmarks of asthma pathophysiology
The candidate will create organoids and test in three groups: 1) no stimulation, 2) stimulation with Poly(I:C), and 3) control (H292 cells used with patient’s fibroblasts). The organoid eluent will be used to measure the presence of inflammatory molecules released in the short- (day 1) vs long-term (days 7–14). Success indicator: patients’ asthma severity is reflected in the intensity of organoid inflammatory signature. 

Objective 3: Capture patient-specific response to therapy
The candidate will expose organoids to media containing an anti-IL5 agent (e.g. mepolizumab – same as the original patient). Success indicator: patient-derived organoid response to anti-IL5 agent (reduction in the inflammatory pathways, collagen deposition, myofibroblast differentiation) consistent with known anti-IL5 pharmacodynamics.

Expected outcomes and impact: These personalised organoids will help study the progression of asthma mechanisms considering patient demographics and lifestyle behaviours in unprecedented detail and help tailor drugs to patients’ based on their individual characteristics/lifestyle.