Search

Altres ajuts: Torsten Haferlach Leukemia Diagnostics Foundation; US National Institute of Health (NIH) grants R35 HL135795, R01HL123904, R01 HL118281, R01 HL128425, R01 HL132071, R01 CA217992, R01 LM013067, R21 CA248138; Edward P. Evans Foundation; Blood Cancer UK, grants 13042 and 19004; Fundació Internacional Josep Carreras and "la Caixa" Foundation. Background: Haploinsufficiency (HI) resulting from deletion of the long arm of chromosome 5 [del(5q)] and the accompanied loss of heterozygosity are likely key pathogenic factors in del(5q) myeloid neoplasia (MN) although the consequences of del(5q) have not been yet clarified. Methods: Here, we explored mutations, gene expression and clinical phenotypes of 388 del(5q) vs. 841 diploid cases with MN [82% myelodysplastic syndromes (MDS)]. Findings: Del(5q) resulted as founder (better prognosis) or secondary hit (preceded by TP53 mutations). Using Bayesian prediction analyses on 57 HI marker genes we established the minimal del(5q) gene signature that distinguishes del(5q) from diploid cases. Clusters of diploid cases mimicking the del(5q) signature support the overall importance of del(5q) genes in the pathogenesis of MDS in general. Sub-clusters within del(5q) patients pointed towards the inherent intrapatient heterogeneity of HI genes. Interpretation: The underlying clonal expansion drive results from a balance between the "HI-driver" genes (e.g., CSNK1A1, CTNNA1, TCERG1) and the proapoptotic "HI-anti-drivers" (e.g., RPS14, PURA, SIL1). The residual essential clonal expansion drive allows for selection of accelerator mutations such as TP53 (denominating poor) and CSNK1A1 mutations (with a better prognosis) which overcome pro-apoptotic genes (e.g., p21, BAD, BAX), resulting in a clonal expansion. In summary, we describe the complete picture of del(5q) MN identifying the crucial genes, gene clusters and clonal hierarchy dictating the clinical course of del(5q) patients.

Context: Chronic lymphoproliferative disorder of natural killer cells (CLPD-NK) is a subset of large granular lymphocyte (LGL) leukemia included as a provisional entity in the 2016 WHO classification of lymphoid neoplasms, which is characterized by clonal expansion of NK cells with no standardized therapy. STAT3 and TET2 mutations were previously each reported in 15%–30% of cases; however, the genomic basis of the remaining cases is still unknown. Objective: We aimed to perform a comprehensive genomic and transcriptomic analysis of CLPD-NK to elucidate the genomic landscape, investigate mechanisms of pathogenesis, and improve diagnostic classification and treatment strategies. Methods: CLPD-NK samples (n=121) were examined by whole-genome, targeted, and/or whole-transcriptome sequencing. Structural modeling, G-protein signaling, β-arrestin signaling, G-protein-coupled receptor (GPCR) for CCL22, CCR4 internalization, and chemotaxis were examined. Cell line-derived xenografts were used to examine cell proliferation and phenotype. Results: Somatic mutations in the chemokine CCL22 were found in 21.5% of CLPD-NK patients but not in 2837 cases of other hematological malignancies, including T-LGL and other NK leukemias. Clinically, there was a trend to an association with cutaneous complications and/or neurological symptoms. CCL22 mutations were enriched at the highly conserved residues Leu45, Pro46, and Pro79 and were mutually exclusive of STAT3 mutations. CCL22 mutations were associated with dysregulated gene expression, similar to normal CD56bright NK cells, with enrichment for activation of cytokine signaling pathways, representing a distinct subgroup. Most CCL22 mutations were substitutions to Arg, a basic residue, and Leu45Arg was predicted to promote electrostatic interactions with CCR4-Tyr22/Glu23 acidic residues. The mutations resulted in ligand-biased CCR4 signaling, in which activation of G-protein signaling was unchanged, but recruitment of β-arrestin, which mediates receptor internalization, was impaired, which, in turn, resulted in enhanced cell chemotaxis. CCL22 mutations drove enhanced cell proliferation in vivo in hIL-15-transgenic mice and induced an expression profile resembling that of primary samples, indicating the importance of microenvironmental crosstalk in pathogenesis. Conclusions: Somatic CCL22 mutations defined a new subgroup of CLPD-NK similar to CD56bright normal NK cells and represented a novel mechanism of leukemogenesis in which gain-of-function chemokine mutations drive tumorigenesis by biased GPCR signaling and dysregulation of microenvironmental crosstalk.

Background: Clonal hematopoiesis of indeterminate potential (CHIP) is associated with increased risk of cancers and inflammation-related diseases. The phenomenon becomes very common in oldest-old individuals, in whom the implications of CHIP are not well defined. Here we investigate the relationships between CHIP and associated pathologies in people aged >80 years. Methods: We performed a mutational screening in 1794 oldest-old individuals enrolled in two population-based studies (“Health_&_Anemia” and “Monzino_80+”). Findings: Clonal mutations were observed in one third of oldest-old individuals and were associated with reduced survival and increased risk of cancers. Mutations in JAK2 and splicing genes (SF3B1, SRSF2, U2AF1, ZRSR2), multiple mutations (DNMT3A, TET2, ASXL1) combined with additional genetic lesions) and variant allele frequency ≥0.14 had positive predictive value for myeloid neoplasms. Combining mutation profiles with abnormalities in red blood cell indices improved the ability of myeloid neoplasm prediction. On this basis, we defined a predictive model that identifies 3 risk groups with different probabilities of developing myeloid neoplasms. Mutations in DNMT3A, TET2, ASXL1 or JAK2 (most occurring as single lesion) were associated with coronary heart disease and rheumatoid arthritis. Cytopenia (most including anemia) was a common finding in oldest-old population (>20%), the underlying cause remaining unexplained in 30% of cases. Among individuals with unexplained cytopenia, the presence of highly-specific mutation patterns was associated with a myeloid neoplasms-like phenotype and a probability of survival comparable to that of myeloid neoplasms. Accordingly, 7.5% of oldest-old subjects with cytopenia had presumptive evidence of myeloid neoplasm. Interpretation: Specific mutational patterns define different risk of developing cancers vs. inflammatory-associated diseases in oldest-old population. In individuals with unexplained cytopenia, mutational status may identify those subjects with presumptive evidence of myeloid neoplasms. Trial Registration: (ClinicalTrials.gov number:NCT03907553) Funding: Supported by European Union; Cariplo Foundation, Italy; Italian Association for Cancer Research; Italian Ministry of Health and Research. Declaration of Interests: The Authors declare no competing interests. Ethics Approval Statement: Study procedures were in accordance with the Declaration of Helsinki. Ethics Committees of Humanitas Research Hospital and Mario Negri Pharmacological Institute, Milan Italy approved the study. Written informed consent was obtained prior to blood sampling.

Background No standardized and widely accepted cytogenetic classification with prognostic impact for adult T-ALL has been proposed to date. Methods Patients with abnormal karyotypes (65/139, 47%) were classified according to the number of chromosomal alterations (Chun K. et al., 2009). Cohort 216 adults T-ALL patients/ NCT00853008 - NCT01540812 /PETHEMA cooperative group. Prognostic impact of karyotype on event-free survival (EFS), overall survival (OS), and cumulative incidence of relapse (CIR) were assessed. Additionally, next-generation sequencing (NGS) experiments were done. Results Greater than three cytogenetic abnormalities were associated with lower rates of both complete remission (CR, 77% vs. 94%; p=0.032) and minimal residual disease (MRD) level Conclusions Compared to BCP-ALL, a lower cut-off to define complex karyotypes based on the presence of ≥3 cytogenetic alterations allows the identification of T-ALL patients with poor prognosis. Interestingly, molecular analyses of patients carrying ≥3 cytogenetic alterations revealed a unique molecular profile that could contribute to understanding the underlying molecular mechanisms of resistance and to evaluate novel targeted therapies (e.g. IL7R-directed) that might improve the response to treatment and outcome of adult T-ALL patients. Funding ISCIII (PI19/01828), co-funded by ERDF/ESF, “A way to make Europe”/”Investing in your future”.

Context The clonal architecture of KRAS/NRAS mutations (RASMT) in Core-Binding Factor AML (CBF-AML). Objective To dissect the impact of RASMT on CBF-AML. Design/Setting Clinical and molecular data and survival outcomes (OS) of the patients were collected at The Cleveland Clinic Foundation and Munich Leukemia Laboratory. Patients A cohort of 368 CBF-AML patients was identified, in whom inv(16) and t(8;21) represented 64% (n=237) and 36% (n=131) of the cases. Thirty-eight percent (141/368) carried RASMT (KRAS=22; NRAS=105) with 14 patients harboring both KRAS/NRAS. Main outcome measures Clinical and molecular characteristics and OS of RASMT CBF-AML. Results Leukopenia and anemia were significantly higher in RASMT and RASWT. RASMT had more +8 (14 vs. 7%) but less -Y (7 vs. 15%; P=0.02) and -X (2 vs. 7%; P=0.03) abnormalities. RASMT were less associated with TET2MT (6 vs. 12%; P=0.02) and KITMT (9 vs. 31%; P Conclusions RAS MT account for one-third of CBF-AML and their prognostic relevance is still a matter of debate. Similar to previous studies, our analysis demonstrated no impact on OS for RASMT CBF-AML. However, the dissection of clonal succession and mutational ranking concluded that the acquisition of multiple hits in epigenetic regulators (e.g., IDH2, RUNX1) determined poorer OS in the RASMT CBF-AML.

Context: Myeloid neoplasia (MN) phenotype and outcome associates with the clonal architecture of SF3B1 mutations (SF3B1MT). Objective: To explore the implications of the clonal architecture of SF3B1MT in MN. Design/Setting: Clinical characteristics, mutational profile, and the clonal hierarchy of MN patients were reviewed at The Cleveland Clinic. Genomic studies were conducted by whole-exome and targeted deep sequencing. Patients: A cohort of 5102 patients was studied for detailed clinical and molecular annotation. Main outcome measures: To describe whether the clonal nature of SF3B1MT might alter the cellular trajectories in patients with MN. Results: The clonal hierarchy was resolved using our in-house designed VAF-based bioanalytic method and confirmed by the PyClone analysis. We first assigned clonal hierarchy to SF3B1MT by using VAFs (adjusted for copy number and zygosity) and classifying the mutations into dominant (if a cutoff of at least 5% difference between VAFs existed), secondary (any subsequent sub-clonal hit) and co-dominant hits (if the difference of VAFs between two mutations was Conclusions: Our study of the clonal architecture of SF3B1MT highlights that clonal progression of cases with MN harboring SF3B1MT might be inferred by the rank of additional genetic lesions cooperating with SF3B1.

Purpose: In this study we determined the frequency of U2AF1 mutations (mut) in different MDS entities and evaluated the correlation of U2AF1mut with mutations in other genes coding for splicing components, cytogenetics and clinical features. Materials and Methods: The total cohort of 406 patients comprised of RAEB-1/RAEB-2: n=55, RARS/RARS-T: n=22, RCMD/RCMDRS: n=53, and CMML-1/CMML-2: n=276. U2AF1mut at Ser34 and Gln157 were analyzed by melting curve analyses and subsequent sequencing of positive cases. Mutational analyses of SF3B1 (n=404) and SRSF2 (n=398) were performed by Sanger sequencing. Cytogenetics was available in 398/406 cases. Results: Overall, U2AF1 was mutated in 38/406 cases (9.4%). Missense mutation in Ser34 occurred more frequently (76%) than missense mutation in Gln157 (24%). MDS patients were more frequently affected by U2AF1mut than patients of theMDS/MPN overlap category CMML (18.5% inMDS vs. 5.1% in CMML; p