Use of human intestinal organoids for theratyping in cystic fibrosis
Universita degli Studi di Verona
Department of Medicine, General Pathology Division Strada Le Grazie 8, 37134 Verona, Italy
Claudio Sorio, Virginia Lotti, Karina Kleinfelder, Jessica Conti
Claudio Sorio, Virginia Lotti, Karina Kleinfelder, Jessica Conti
English; Italian
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No Yes Yes No No No No No
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Drugs targeting specific CFTR mutations entered the clinic, however the amount of rescued CFTR function is still limited and variable responses to these therapies are recorded in Cystic Fibrosis (CF) patients due to still undefined mechanisms of action. Therefore, prediction of individual drug responses in CF is challenging. The challenge is mainly based on very limited availability of primary cell models for screening of compounds able to restore the function of defective CFTR. Organoids are a new research tool with remarkable basic and translational potential: they could be useful for assessing the efficacy of new therapies and provide personalized therapies for (CF) subjects
What is the aim of the project?
We aim at developing a personalized medicine approach in CF patients by collecting, expanding, storing and analyzing human intestinal organoids. A functional assay will be applied in order to measure the response to CFTR modulators available in clinic (Ivacaftor, Lumacafotr, Tezacaftor) as well as experimental ones. Our experiment will be made up of three main tasks: Task 1: To measure CFTR function in organoids of individual patients undergoing treatments with 1) Ivacaftor (Kalydeco), 2) Orkambi by FIS assay. Task 2: To study CFTR expression and localization and CFTR channel in organoids. CFTR expression and subcellular localization will be studied by confocal microscopy analysis. Task 3: Evaluate all clinically relevant data recorded in the clinical database in relation to the outcome of FIS assay.
What techniques and methods are used?
Crypts will be isolated and cultured from human intestinal biopsies of Cystic Fibrosis patients carrying various mutations. We will measure CFTR (Cystic fibrosis transmembrane conductance regulator) activity by forskolin induced swelling (FIS) assay. Organoids of patients from both groups undergoing treatment with Ivacaftor or Orkambi will be analysed. CFTR expression and subcellular localization will be studied by confocal microscopy analysis. All clinical relevant data recorded in the clinical database will be evaluated in relation to the outcome of FIS assay.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Understanding the value of translational research for future clinical applications of personalized medicine, by performing cell culture activity and analysis of gene and protein expression by PCR, western blotting and immunofluorescence. According to the basic scientific knowledge and the capability demonstrated by the student, he/she will be allowed to personally perform some of these tasks under the supervison of experienced personnel.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Relevant literature will be provided to the student
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Interest in laboratory work. Subjects passed: pathology courses, knowledge of basic and clinical aspects of cystic fibrosis and genetic diseases
Are there any legal limitations in the student’s involvement
Insurance is required for access, non disclosure agreement of sensitive data eventually acquired by student is requested.
Type of students accepted
This project accepts:
- Medical students
- Cyst Fibros. 2019 Nov 28. pii: S1569-1993(19)30964-6. doi: 10.1016/j.jcf.2019.11.002. Intestinal organoids for Cystic Fibrosis research. de Poel E; Lefferts JW; Beekman JM.
- M. de Winter – de Groot; G. Berkers and R.E.P. Marck – van der Wilt et al.; Forskolin-induced swelling of intestinal organoids correlates with disease severity in adults with cystic fibrosis and homozygous F508del mutations; Journal of Cystic Fibrosis;