Haploidentical hematopoietic stem cell transplantation (h-HSCT) represents an efficient curative approach for patients affected by hematologic malignancies in which the reduced intensity conditioning induces a state of immunologic tolerance between donor and recipient. However, opportunistic viral infections greatly affect h-HSCT clinical outcomes. NK cells are the first lymphocytes that recover after transplant and provide a prompt defense against human cytomegalovirus (HCMV) infection/reactivation. By undertaking a longitudinal single-cell computational profiling of multiparametric flow cytometry, we show that HCMV accelerates NK cell immune reconstitution together with the expansion of CD158b1b2j ^pos /NKG2A ^neg /NKG2C ^pos /NKp30 ^lo NK cells. The frequency of this subset correlates with HCMV viremia, further increases in recipients experiencing multiple episodes of viral reactivations, and persists for months after the infection. The transcriptional profile of FACS-sorted CD158b1b2j ^pos NK cells confirmed the ability of HCMV to deregulate NKG2C, NKG2A, and NKp30 gene expression, thus inducing the expansion of NK cells with adaptive traits. These NK cells are characterized by the downmodulation of several gene pathways associated with cell migration, the cell cycle, and effector-functions, as well as by a state of metabolic/cellular exhaustion. This profile reflects the functional impairments of adaptive NK cells to produce IFN-γ, a phenomenon also due to the viral-induced expression of lymphocyte-activation gene 3 (LAG-3) and programmed cell death protein 1 (PD-1) checkpoint inhibitors.

Risk factors for cytomegalovirus (CMV) reactivation and the impact of CMV reactivation on patient outcomes have been extensively investigated after matched related or unrelated donor transplantation, but little is known in the setting of haploidentical stem cell transplantation (Haplo-SCT) with post-transplantation cyclophosphamide (PT-Cy), in which recipients are considered more severely immunocompromised. We retrospectively analyzed a cohort of 554 consecutive patients undergoing Haplo-SCT with PT-Cy at 3 different centers. Early CMV reactivation (occurring within the first 120 days post-transplantation) occurred in 242 patients, for an estimated cumulative incidence of 44%. Among those patients, 74 (30%) had recurrent CMV and 20 (8%) had CMV disease. On multivariable analysis, positive recipient CMV serostatus (hazard ratio [HR] >2.5; P < .001), disease histology (lymphoid versus myeloid: HR, 0.66; P = .003) and increasing recipient age (HR, 1.01; P = .015) were independent predictors of CMV reactivation. At a 4-month landmark analysis, CMV reactivation was associated with higher 1-year and 5-year cumulative incidence of nonrelapse mortality (NRM) relative to patients without reactivation: 13% versus 5% and 22% versus 9%, respectively (P < .001). On multivariable analysis, CMV reactivation was an independent negative predictor of NRM (HR, 2.69; P < .001) and was close to statistically significant for overall survival (HR, 1.38; P = .062). Our results suggest that CMV reactivation plays an important role at determining NRM. Because patient CMV serostatus is the main predictor of CMV reactivation, it should be considered when evaluating strategies for preventing CMV reactivation. 2022 American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc.

Current guidelines recommend pre-therapeutic UGT1A1 genotyping to guide irinotecan dosing, but the usefulness of this approach remains to be clarified. In 247 patients with advanced gastrointestinal cancers undergoing irinotecan-based chemotherapy, we prospectively performed UGT1A1*28 genotyping and we analyzed the incidence of severe neutropenia according to genotype-guided dose reductions. Overall, 28 (11.3%) and 92 (37.2%) patients were homozygous or heterozygous UGT1A1*28 carriers, respectively. Grade ≥ 3 neutropenia was reported in 39% of homozygous patients receiving an upfront dose reduction of irinotecan (median 40%, range 22–58%), in 20% of heterozygous or wild-type patients receiving full dose (OR_{vs*28/*28 genotype} = 0.38; 95% CI: 0.14–1.03; p = 0.058), and in 15.3% of those receiving a reduced dose for clinical reasons (OR _{vs*28/*28 genotype} = 0.28, 95% IC: 0.12–0.67; p = 0.004). Occurrence of severe neutropenia was inversely associated with dose reduction in UGT1A1*28 homozygous carriers (OR_{x10 unit} = 0.62, 95% CI: 0.27–1.40, p = 0.249) and UGT1A1 heterozygous or wild-type patients (OR_{x10 unit} = 0.87, 95% CI: 0.59–1.28, p = 0.478). Incidence of severe neutropenia was related to irinotecan doses and UGT1A1 polymorphisms. Upfront irinotecan dose reductions do not reduce the burden of grade ≥ 3 neutropenia in UGT1A1*28 homozygous carriers.

Aims: The current diagnostic approach to inherited cardiac conditions (ICCs) is primarily focused on the analysis of single-nucleotide variants (SNVs) and small insertions/deletions (InDels). However, as recommended for other inherited diseases, the analysis of Copy Number Variants (CNVs) should be equally considered. In cardiology, the diagnostic contribution of CNVs remains insufficiently studied, with limited evidence and no standardized recommendations for analytical workflows. This study aims to assess the prevalence of pathogenic CNVs in an Italian multicentre cohort of ICC patients, providing recommendations for integrating CNV analysis into routine genetic testing. Methods and results: A total of 203 ICC probands with prior negative or inconclusive results for SNVs and InDels testing were included. The tertiary bioinformatic analysis was performed by eVai enGenome software, and putative CNVs were validated by Multiplex Ligation-dependent Probe Amplification (MLPA) assays. MLPA confirmed only one of the five initially suspected CNVs, identifying a deletion in MYBPC3 gene classified as pathogenic according to ACMG/AMP guidelines. This result confirmed a limited diagnostic yield of 0.50%. Considering the substantial costs, time constraints, and specialized expertise required, we propose a strategy to prioritize selected ICC patients for CNV analysis. Conclusion: Evidence from this real-world cohort suggests the incorporation of CNV analysis as a second-tier test in patients with ICC and specific clinical or molecular indications.

The monitoring of HHV-6 after allogeneic hematopoietic stem cell transplantation (HSTC) has proven to be useful in preventing life-threatening complications; however, the pathogenic role of HHV-6 after autologous HSCT is not well-characterized, although viral reactivation might be responsible for significant complications even after autologous HSCT. Here we report for the first time to our knowledge the case of a patient with genomic integration of HHV-6, presenting with high titers of HHV-6 after autologous HSCT, mimicking HHV-6 reactivation. The presence of viral DNA in the follicle bulb confirmed the genomic integration and allowed the discontinuation of the antiviral treatment to the patient. This case might be helpful for discriminate HHV-6 reactivation from viral integration, thus avoiding unnecessary and potentially toxic antiviral therapy once the genomic integration is confirmed.