Network Terapia Avanzata (LHS-TA) Nell’ambito del Piano Nazionale di Ripresa e Resilienza (PNRR), Investimento E.3 “Ecosistema innovativo della salute”

Task 1: Validation and sharing of standardized and homogeneously employed protocols for monitoring the persistence and expansion of CAR T-cells and their detection in different biological fluids (leader INT, co-leaders Federico II, S. Orsola).  

Given our previous experience in treating and monitoring patients receiving commercial CAR T cells (Monfrini et al. CCR 2022),  our unit will coordinate the efforts of the different spokes involved in CAR T cell administration,  and work at the optimization, standardization, and implementation of protocols for monitoring the persistence, expansion and features of CART cells. In particular we will develop standardized procedures for the: 

• Evaluation of the cellular kinetics and persistence of CART cell products using both flow-cytometry and ddPCR.  
• Evaluation of the infusion product phenotypes (flow-cytometry and CyTOF) 
• Evaluation of the phenotype of expanding CART cells (flow-cytometry and CyTOF) 
• Evaluation of the dynamics of different immune cell subsets in peripheral blood  upon CART cell infusion (flow-cytometry and CyTOF) 
• Evaluation of the impact of leukapheresis characteristics (flow-cytometry and Nanostring) 
• Evaluation of CART cell trafficking and biodistribution in different body fluids (flow-cytometry and ddPCR). 

Task 2: Use of liquid biopsy for monitoring of residual tumor cells and for validation of predictive biomarkers of response, including cell-free DNA and extracellular vesicles (leader INT, co-leaders S. Orsola, Pisa).  

Our Unit will work on longitudinal circulating tumor DNA (ctDNA) evaluation and correlation with imaging for response assessment and will identify a panel of genes and the relevant methodology to be used for these analyses.  

Task 3: Development of an integrated platform for managing Clinical/Regulatory aspects, including submission of IMPD and interaction with regulatory agencies (leader INT, co-leaders OPBG, OSR).  

Our study coordinators and data managers have extensive experience in clinical studies with CART cells and they will work for the development of guidelines for the Clinical/Regulatory aspects related to ATMPs clinical use.