An antioxidant signature as a newer prognostic model in chronic lymphocytic leukemia
Universita degli Studi di Verona
Dept. of Neurciences, Biomedicine and Movement, Strada Le Grazie,8 – 37134 Verona, ITALY Research Center LURM, Ple LA Scuro,10 - 37134 Verona, ITALY
Maria Teresa Scupoli Chiara Cavallini Marilisa Galasso
Maria Teresa Scupoli Chiara Cavallini Marilisa Galasso
English; Italian
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No No Yes No No No No No
Type of Research Project
- Basic science
What is the background of the project?
Chronic lymphocytic leukemia (CLL) is characterized by an extremely variable clinical course. While many prognostic markers have been associated with CLL clinical behavior, few have been demonstrated to be useful in the daily management of patients. We have recently shown that higher levels of catalase and glutathione peroxidase (GPX) expression in CLL cells are linked to unfavorable prognostic parameters (UM-IGHV) and faster progression of the disease. Based on these results, the goal of our proposal is to develop newer models that correlate antioxidant signature with time to first treatment (TTFT) to identify high-risk patients for progressive disease and shortened TTFT, particularly for patients with early stage disease. Our central hypothesis is that altered antioxidant enzymes could mediate antitumor signals in leukemic cells, i.e. cell death or susceptibility to apoptosis, thus influencing disease progression and evolution.
What is the aim of the project?
This mechanistic role of antioxidant enzymes provides the rationale for developing a prognostic model based on an antioxidant signature in CLL. We will test this hypothesis by pursuing the following specific objectives: 1. Investigate the expression of catalase and GPX in normal and CLL B cells. 2. Study the biological effects and mechanisms of catalase and GPX on leukemic cells. 3. Validate the prognostic value of catalase and GPX expression in CLL.
What techniques and methods are used?
1. Investigate the expression of catalase and GPX in normal and CLL B cells. Real-time quantification (qRT-PCR) for CAT (catalase) and GPX1 (glutathione peroxidase1) on B cells isolated from CLL and healthy-donor PBMCs by negative selection. 2. Study the biological effects and mechanisms of catalase and GPX on leukemic cells. In leukemic cells transfected with catalase, GPX siRNA or scramble controls, we will quantify siRNA-mediated down-regulation of enzymes by qRT-PCR and flow cytometry. Moreover, in antioxidant enzyme-silenced cells we will measure ROS production using flow cytometry and analyze spontaneous apoptosis measuring cell membrane phosphatidylserine (PS) exposure and propidium iodide (PI) staining with flow cytometry. 3. Validate the prognostic value of catalase and GPX expression in CLL. We will associate flow-cytometry data of enzyme expression with TTFT and generate time-to-events models that include flow cytometry data and currently used standard prognostic factors, using Cox proportional hazard regression.
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 of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Cell culture experiments; Signal transduction analysis using phospho-specific flow cytometry; Immuno-phenotype analysis using flow cytometry analysis; Apoptosis measuring; ROS quantification; Real-time PCR; Cell transfection experiments; Data analysis.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will read specific scientific literature and attend to seminar and lab meetings during the internship period.
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?
Knowledge of cell biology
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Graduated students (less than 6 months)
- Pre-Medical students from the American-British system
- Cavallini C; Chignola R; Dando I; et al. Low catalase expression confers redox hypersensitivity and identifies an indolent clinical behavior in CLL. Blood 2018;131:1942-54 Cesano A; Perbellini O; Evensen E; et al. Association between B-cell receptor responsiveness and disease progression in B-cell chronic lymphocytic leukemia: results from single cell network profiling studies. Haematologica. 2013;98(4):626–634