UOSD CLINICAL PATHOLOGY AND CANCER BIOBANK
Head: Laura Conti, MD, PhD
Diagnostic harmonization initiative on Multiple Myeloma for Gruppo Laziale Mieloma Multiplo (GLMM)
The project aims to reach a consensus among regional laboratories specialized on onco-haematology diagnosis regarding the flow cytometry antibodies panel, data analysis and clinical report for Multiple Myeloma (MM) diagnosis and monitoring.
The network is also focusing on the positive selection of the plasma cell population by immune-magnetic beads separation, for a better assessment of the cytogenetic profile in plasma cell disorders.
New forms have been designed, approved and introduced on RedCap, the clinical and laboratory data base shared among all the GLMM Centers. The new collection forms focus on cytogenetic and immunophenotype data of MM patients at diagnosis and on follow up.
A National project is ongoing to create an “Italian MM MRD network” that will share the same method for MRD analysis in MM patients across hub-Haematology centers in Italy. The most common employed methods for MRD evaluation (both flow cytometry and molecular biology-based) will be harmonized among the involved “start-up” centers.
Role of Che-1 in transgenic mouse model of Multiple Myeloma
In collaboration with the SAFU laboratory, we investigated the role of Che-1, a RNA binding protein witch is involved in the control of transcription and cellular proliferation by regulating the state of the chromatin and by increasing its accessibility in Multiple Myeloma (MM). In particular we performed Serum Protein Electrophoresis (SPEP) to detect the levels of monoclonal immunoglobulins in serum of the Vk*Myc transgenic mouse model, which, through activating c-Myc oncogene in maturing B cells, recapitulates the pathogenesis and clinical manifestations of human MM, including progression from MGUS to plasma cell expansions (Chesi et al.; Cancer Cell 2008). At this purpose CD138+ neoplastic cells were isolated from the bone marrow (BM) of these mice and manipulated for knockdown of Che1 by siRna and transplanted into 5 recipient wild-type mice for each group. The delay in disease progression in Che-1 depleted MM cells, it was been recognize by analyzing the levels of monoclonal immunoglobulins in murine serum, as a distinct band (M-spike).
Urine monoclonal Free Light Chains
In collaboration with the Protein Study Group of Italian Society of Clinical Biochemistry and Molecular Biology (SIBioC) in the last year we participated in the revision and update of the “Consent document for the research and quantification of the BENCE JONES protein”. The document is now being published. . With same national Group we started in 2020 a collaboration on new project for the harmonization of serum Electrophoresys and Immunofixation reporting aimed at drafting a national consensus document
Upper extremity venous thrombosis in cancer patients with peripherally inserted central inserted catheters
Symptomatic PICC related deep venous Thrombosis (DVT) are frequent in cancer patients receiving chemotherapy. In collaboration with the Vascular Access Management Team we conducted a retrospective cohort study in cancer patients who underwent PICC placement for the administration of chemotherapy to evaluate the incidence of upper extremity venous thrombosis (UEVT) and establish the most predictive risk factors for the development of PICC-related thrombosis in cancer patients during chemotherapic treatment, for the future design of an integrated care pathway (ICT) that could be used to prevent thrombotic events. All patients were followed for a minimum of 6 months after PICC insertion, unless they died during this period. Factors previously associated with catheter-related thrombosis, including side of catheter placement, tip location, tumor type, inherited and acquired thrombophilia and environmental factors have been evaluated. The data collection study allowed to highlight a decrease in the incidence of the number of DVT events from 6.8% to 2.9%.
Accurate dosimetry and biomarkers improve survival in HCC patients treated with resin 90Y-μspheres: a randomized trial
Hepatocellular Carcinoma (HCC) is the most frequent liver primary tumor worldwide. An increasing amount of evidence supports the effectiveness of Yttrium-90 (90Y) labeled microspheres to treat intermediate and advanced disease in these patients. The trial (in collaboration with Medical Physics Laboratory, Nuclear Medicine and Radiology Departments) aims at demonstrating that a robust patient-specific dosimetry associated with PIVKA-II analysis improves the patients’ survival compared with standard BSA (Body Surface Area) method.
Radio-induced modifications of lymphoid subpopulations involved in resistance and escape mechanisms to the treatment of localized prostate cancer
Aim of the study is to evaluate the effect of radiotherapy (RT) on immuno-regulatory B and T lymphocyte subpopulations (Breg and Treg) and plasma cells and possible correlations with the clinical course of the disease and acute and late toxicity.
The flow cytometry characterization developed in our laboratory allowed the identification of Treg and Breg peripheral blood subpopulations through the acquisition of a high number (> 5000) of regulatory cells using innovative acquisition and analysis strategies. Twenty-one patients were analyzed before treatment (T0), 3 hours after the first RT session (T1), after an average dose of 24 Gy (T2), after the last RT session (T3), at + 6 months (T4) and at + 12 months (T5) after RT. Six patients entered the study and were analyzed also before hormono-therapy (T-1) for a total of 135 characterization. Analysis of the regulatory sub-populations modulation and modifications are ongoing.
Cerebrospinal fluid (CSF) flow cytometry in the diagnosis of leptomeningeal disease in onco-haematology
Cerebrospinal fluid (CSF) flow cytometry has a crucial role in the diagnosis of leptomeningeal disease in onco-haematology. We are evaluating the cytometry characterization of 138 CSF samples from patients affected by non-Hodgkin lymphoma, negative for disease infiltration. The aim is to focus on the CSF non-neoplastic population, to compare the cellular composition of the CSF with paired peripheral blood samples and to document the feasibility of flow cytometry in hypocellular samples.
Preliminary results have documented that T lymphocytes aree the most abundant subset in CSF with a predominance of CD4-positive over CD8-positive T cells (CD4/CD8 ratio = 2) together with a minority of monocytes. No B cells are present. The differences between CSF and paired peripheral blood lymphoid phenotype is under evaluation to investigate the existence of an active mechanism of lymphoid migration through the meninges.
Coagulation/complement activation and cerebral hypoperfusion in relapsing-remitting multiple sclerosis
Multiple sclerosis (MS) is a demyelinating disease of the central nervous system with an underlying immune-mediated and inflammatory pathogenesis. Innate immunity, in addition to the adaptive immune system, plays a relevant role in MS pathogenesis. It represents the immediate non-specific defense against infections through the intrinsic effector mechanism “immunothrombosis” linking inflammation and coagulation. Moreover, decreased cerebral blood volume (CBV), cerebral blood flow (CBF), and prolonged mean transit time (MTT) have been widely demonstrated by MRI in MS patients. We hypothesized that coagulation/complement and platelet activation during MS relapse, likely during viral infections, could be related to CBF decrease. Our specific aims are to evaluate whether there are differences in serum/plasma levels of coagulation/complement factors between relapsing-remitting (RR) MS patients (RRMS) in relapse and those in remission and healthy controls as well as to assess whether brain hemodynamic changes detected by MRI occur in relapse compared with remission. This will allow us to correlate coagulation status with perfusion and demographic/clinical features in MS patients. This is a multi-center, prospective, controlled study. RRMS patients (1° group: 30 patients in relapse; 2° group: 30 patients in remission) and age/sex-matched controls (3° group: 30 subjects) will be enrolled in the study. Patients and controls will be tested for either coagulation/complement (C3, C4, C4a, C9, PT, aPTT, fibrinogen, factor II, VIII, and X, D-dimer, antithrombin, protein C, protein S, von-Willebrand factor), soluble markers of endothelial damage (thrombomodulin, Endothelial Protein C Receptor), antiphospholipid antibodies, lupus anticoagulant, complete blood count, viral serological assays, or microRNA microarray. Patients will undergo dynamic susceptibility contrast-enhanced MRI using a 3.0-T scanner to evaluate CBF, CBV, MTT, lesion number, and volume. Our work aims to identify a link between activation of the coagulation / complement system and cerebral hypoperfusion that could improve the search for new biomarkers and molecular and / or imaging targets, leading to the development of new effective therapeutic strategies in MS.
Coagulation/complement activation and cerebral hypoperfusion in relapsing-remitting multiple sclerosis
Multiple sclerosis (MS) is a chronic immune-mediated inflammatory demyelinating and degenerative disease of the central nervous system. It has been demonstrated that not only adaptive immunity but also innate immune system plays a relevant role in MS pathogenesis. There are several studies supporting coagulation/complement and platelet involvement in the innate immune response in MS by linking inflammation and coagulation. Beyond, decreased MRI cerebral blood volume and flow (CBV, CBF), and its prolonged mean transit time (MTT) have been demonstrated in all forms of MS.
This is a multicenter, prospective, controlled study. Informed and consenting MS patients [1st group: 30 relapsing patients; 2nd group: 30 patients in remission] and 30 age and gender controls (3rd group) will be enrolled in the study.
Our study is based on hypothesis that coagulation/complement activation due to inflammatory-thrombotic processes in the course of MS relapse could determine cerebral blood flow deceleration.
The specific aims of the study are to evaluate both in patients and controls:
- The serum/plasma concentrations of coagulation/complement factors
- Absolute cerebral blood flow (CBF), blood volume (CBV) and mean transit time (MTT), by dynamic susceptibility contrast-enhanced 3.0-T MRI
- The correlation between the serum/plasma levels of coagulation/complement factors with both MRI perfusion data and demographic/clinical (age, gender, disease duration, disability) features of MS patients
The relationships between the laboratory and clinical data and the MRI perfusion findings could lead to the development of new effective therapeutic strategies in MS.
Use of antibody tests for screening and prevention of the transmission of the infection from Pilot Observational Study
The aim of this study, according to the WHO guidelines, is the implementation ofa validation study of serological tests for the search for antibodies to SARS-Cov-2 and the possible relationship with an asymptomatic carrier status, ongoing infection or complete recovery with protective immunity.
The first phase for validation will be the evaluation of the prevalence of the positive examination in some populations of subjects belonging to the IFO and the comparison of the results between the different types of test.
The data collected will allow to evaluate the effectiveness of the immunological diagnostics of the infection and contribute to validate the immunological profiles correlated with the status of asymptomatic infection or with pictures of complete recovery with acquisition of protective immunity and they will also be able to provide key elements for defining the procedures necessary to ensure COVID-FREE nosocomial realities.
At first 300 informed and consenting health workers will be enrolled and only in a later time we will examine 150 serum of cancer patients stored in our Biobank and enrolled previously.