Understanding the Role of Large Extracellular Vesicles in Lymphomas and Lymphoproliferative Disorders: the "Off the Beaten Track" Liquid Biopsy

It is a 3-year monocentric in vitro study on human tissue. 50 patients with aggressive and indolent Non-Hodgkin Lymphomas (Diffuse Large B-Cell Lymphoma (DLBCL; Group A; n=10), Follicular Lymphoma (FL; Group B; n=10), Marginal Zone Lymphoma (MZL; Group C; n=10), Peripheral T-Cell Lymphoma (PTCL; Group D; n=10), Mycosis Fungoides (MF; Group E; n=10)), and patients with Chronic Lymphocytic Leukaemia (CLL; N=10; Group F; n=10), Hairy Cell Leukemia (HCL; Group G; n=10), and Multiple Myeloma (MM; N=10; Group H; n=10) will be enrolled over a period of 24 months. Follow-up: 12 months. Patients will be enrolled at the Complex Operative Unit of Haematology-IRCCS Azienda Ospedaliero-Universitaria di Bologna. The peripheral blood samples of patients (40 ml) will be collected only at diagnosis within normal clinical practice, outside of clinical study protocols, and following the treatment guidelines in use at the Centre. The analysis of the samples will be performed at the Unità Operativa Complessa di Ematologia-IRCCS Azienda Ospedaliero-Universitaria di Bologna in collaboration with the Istituto Romagnolo per lo Studio dei Tumori 'Dino Amadori' - IRST IRCCS and with the University of Perugia.

Primary outcomes

• Extracellular vesicle characterization [2 years]: Comparison of concentration (particles/ml), phenotype, and cargo of extracellular vesicles from patients and healthy donors.
• Validation of extracellular vesicles as biomarker [2 years]: Correlation of extracellular vesicles' characterization results with biomarkers of disease aggressiveness and short-term response to standard therapy.

Secondary outcomes

• Functional effects of extracellular vesicles [2 years]: Comparison of functional effects of extracellular vesicles from patients and healthy donors on selected immune cells. The isolated vesicles will be used to treat the immune cells (T cells, B cells, monocytes) to evaluate viability, phenotype, migration ability, and inflammatory cytokine production of the target cells.

Other outcomes

• Extracellular vesicles' cell of origin [1 years]: Correlation of extracellular vesicles' phenotype with the corresponding cells of origin.

Sex: All, ages: 18 Years to any, accepts healthy volunteers

Inclusion criteria for patients:

* Age ≥ 18 years.
* Patients with WHO 2017-defined diagnosis of indolent and aggressive Non-Hodgkin Lymphoma (specifically Diffuse Large B-Cell Lymphoma, Follicular Lymphoma, Marginal Zone Lymphoma, Peripheral T-Cell Lymphoma, Mycosis Fungoides) or Chronic Lymphocytic Leukemia or Hairy Cell Leukemia or Multiple Myeloma.
* Signed informed consent.

Exclusion criteria for patients:

\- Concomitant secondary neoplasia.

Inclusion criteria for healthy donors:

* Age ≥ 18 years.
* Signed informed consent.

Exclusion criteria for healthy donors:

\- Healthy donors with neoplasia.

• Lucia Catani, PhD, Principal investigator, IRCCS Azienda Ospedaliero-Universitaria di Bologna