The development of chimeric antigen receptor (CAR)-T cell therapy has revolutionized the treatment of hematological diseases. However, approximately 60% of patients relapse after CAR-T cell therapy, and no clear cause for this failure has been identified. The objective of the Bio-CAR-T BS study (ClinicalTrials.gov: NCT05366569) is to improve our understanding of the lymphocyte harvest to maximize the quality of the CAR-T cell product. Of the 14 patients enrolled, 11 were diagnosed with DLBCL, 2 with PMBCL, and 1 with ALL. Five of 11 DLBCL patients met the criteria for “pre-emptive” Lymphocytes-apheresis (being at high risk of second relapse), and 6 were included in the standard-of-care Lymphocytes-apheresis group. Previous autologous stem cell transplantation (ASCT) and age were significantly different between the two groups. At the time of Lymphocyte-apheresis, patients in the “pre-emptive” group had more “fit” lymphocytes (higher CD4+/CD8+ ratio; higher naïve T cells levels) compared with standard group, probably due to the impact of ASCT. At the same time, also being older than 60 years results in a more “exhausted” lymphocyte profile. Overall, “pre-emptive” Ly-apheresis in DLBCL patients at high risk of relapse appears to be feasible and may allow the timely collection of “fit” lymphocytes for CAR-T cell manufacturing.

Exosomes are extracellular vesicles playing a pivotal role in the intercellular communication. They shuttle different cargoes, including nucleic acids from their cell of origin. For this reason, they have been studied as carriers of tumor markers in different liquid biopsy approaches, in particular for solid tumors. Few data are available concerning exosomes as markers of myeloid neoplasia. To better understand their real potential and the best approach to investigate leukemic exosomes, we present the results of a pilot feasibility study evaluating the application of next-generation sequencing analysis of dsDNA derived from exosomes isolated in 14 adult patients affected by acute myeloid leukemias. In particular, leukemia-derived exosome fractions have been analyzed. The concentration of dsDNA co-extracted with exosomes and the number and types of mutations detected were considered and compared with ones identified in the Bone Marrow (BM) and Peripheral Blood (PB) cells. Exosomal DNA concentration, both considering the cargo and the DNA surrounding the lipid membrane resulted in a linear correlation with leukemic burden. Moreover, exosomal DNA mutation status presented 86.5% of homology with BM and 75% with PB. The results of this pilot study confirmed the feasibility of a leukemia-derived exosome enrichment approach followed by exosomal dsDNA NGS analysis for AML biomarker detection. These data point to the use of liquid biopsy in myeloid neoplasia for the detection of active leukemic cells resident in the BM via a painless procedure.

Background: Disease relapse after allogeneic stem cell transplantation (allo-SCT) is the main challenge for curing acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). We investigated the overall survival (OS) after allo-SCT relapse according to different therapeutic approaches. Methods: We analyzed 134 patients who relapsed after allo-SCT performed between 2015 and 2021 at Saint-Antoine University Hospital, Paris and Spedali Civili di Brescia, Brescia. Of these, 103 (77%) were treated, comprising 69/103 (67%) who received therapy in overt relapse and 34/103 (33%) who were treated in a pre-emptive manner when molecular/cytogenetics recurrence or mixed chimerism occurred. The treatment was donor lymphocyte infusion (DLI)-based for 40/103 (39%) patients. Results: The 1-, 2-, and 5-year OS of patients treated with DLI (n = 40) was 67%, 34%, and 34%, respectively, for those treated preventively (n = 20) and 43%, 20%, and 20%, respectively, for those treated in overt relapse (n = 20) (p < 0.01). The 1-, 2-, and 5-year OS of patients treated without DLI (n = 63) was 54%, 40%, and 26%, respectively, for those treated preventively (n = 14) and 17%, 5%, and 0%, respectively, for those treated in overt relapse (n = 49) (p < 0.01). Conclusions: Relapse treatment with a pre-emptive strategy was associated with improved outcomes, particularly when DLI was employed.

Background: The Comprehensive Geriatric Assessment (CGA) has proven to be a valuable tool for providing a more comprehensive health evaluation of allogeneic stem cell transplantation (allo-SCT) recipients. Methods: We prospectively developed and tested a new Simplified Geriatric Score-4 (SGS-4) on 135 consecutive patients aged ≥50 years who underwent allo-SCT between 2020 and 2023. Each CGA component was individually analyzed for its association with overall survival (OS), non-relapse mortality (NRM), and cumulative incidence of relapse (CIR). Then, we performed a two-factor analysis (FA) using oblimin rotation and Bartlett estimation on all CGA components and sex. Based on component weights, a simplified geriatric score-4 score (SGS-4) was created: [Gait Speed] + 2 × [Hand Grip] + Geriatric 8 + 1.5 × [Sex]. ROC analysis defined three fitness groups, frail (≤13), prefrail (>13–22.5), and fit (>22.5). Results: Reduced hand grip strength and impaired mini mental state examination (MMSE) were associated with worse OS and higher NRM. Vulnerable Elders Survey (VES-13) and Fondazione Italiana Linfomi (FIL) scores also indicated poorer OS, though with uneven group sizes. Other CGA domains and the Hematopoietic Cell Transplantation–Comorbidity Index (HCT-CI) showed no significant prognostic value. The SGS-4 effectively stratified patients into three fitness groups, with those in the frail category experiencing lower OS and an increased risk of relapse. Conclusions: The new Simplified Geriatric Score-4 (SGS-4) based on three CGA domains (gait speed, hand grip, Geriatric 8) and sex effectively predicts OS and CIR risk in patients aged ≥50 years undergoing allo-SCT. The study’s small sample size and disease heterogeneity warrant further validation in larger cohorts.