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  • 1
    In: Blood, American Society of Hematology, Vol. 140, No. Supplement 1 ( 2022-11-15), p. 7721-7722
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2022
    detail.hit.zdb_id: 1468538-3
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  • 2
    In: Blood, American Society of Hematology, Vol. 103, No. 6 ( 2004-03-15), p. 2284-2290
    Abstract: Imatinib is a tyrosine-kinase inhibitor that binds to ABL proteins and induces cytogenetic remissions in patients with chronic myeloid leukemia (CML). In these patients measuring response by molecular techniques is clearly required. We determined the cytogenetic and molecular response (CgR, MR) to imatinib in 191 patients with late chronic-phase Philadelphia-positive (Ph+) CML, previously treated with interferon α. MR was assessed with real-time quantitative (TaqMan) reverse transcription–polymerase chain reaction and was expressed as the ratio between BCR/ABL and β2-microglobulin × 100, the lowest level of detectability of the method being 0.00001. A complete CgR (CCgR) was achieved in 85 (44%) of 191 patients and was maintained for 2 years in 67 (79%) of 85 patients. A reduction of the transcript level of more than 2 logs was achieved in all but 9 patients with CCgR versus none of 23 with partial CgR. In the CCgRs the median value of the MR was 0.0008 after 12 months and 0.0001 after 24 months, with the transcript level undetectable in 22 cases. We conclude that in CCgRs the degree of MR may vary from 2 to more than 4 logs, and that there is a progressive decrease of transcript level by time. Only 1 of 22 negative cases has had a relapse as yet.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2004
    detail.hit.zdb_id: 1468538-3
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  • 3
    In: Blood Advances, American Society of Hematology, Vol. 7, No. 12 ( 2023-06-27), p. 2855-2871
    Abstract: Acute myeloid leukemia (AML) still represents an unmet clinical need for adult and pediatric patients. Adoptive cell therapy by chimeric antigen receptor (CAR)-engineered T cells demonstrated a high therapeutic potential, but further development is required to ensure a safe and durable disease remission in AML, especially in elderly patients. To date, translation of CAR T-cell therapy in AML is limited by the absence of an ideal tumor-specific antigen. CD123 and CD33 are the 2 most widely overexpressed leukemic stem cell biomarkers but their shared expression with endothelial and hematopoietic stem and progenitor cells increases the risk of undesired vascular and hematologic toxicities. To counteract this issue, we established a balanced dual-CAR strategy aimed at reducing off-target toxicities while retaining full functionality against AML. Cytokine-induced killer (CIK) cells, coexpressing a first-generation low affinity anti-CD123 interleukin-3–zetakine (IL-3z) and an anti-CD33 as costimulatory receptor without activation signaling domains (CD33.CCR), demonstrated a powerful antitumor efficacy against AML targets without any relevant toxicity on hematopoietic stem and progenitor cells and endothelial cells. The proposed optimized dual-CAR cytokine-induced killer cell strategy could offer the opportunity to unleash the potential of specifically targeting CD123+/CD33+ leukemic cells while minimizing toxicity against healthy cells.
    Type of Medium: Online Resource
    ISSN: 2473-9529 , 2473-9537
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2023
    detail.hit.zdb_id: 2876449-3
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  • 4
    In: Blood, American Society of Hematology, Vol. 106, No. 11 ( 2005-11-16), p. 225-225
    Abstract: Normal cytogenetics in AML is a major drawback to detect minimal residual disease. We have identified heterozygous NPM1 mutations as the most frequent genetic lesion in AML with normal karyotype (NEJM2005:352,254). Mutations A, B and D, all characterized by tetranucleotide insertions, account for more than 90% of all mutations. NPM1 mutations are new important prognostic markers in AML with normal cytogenetics, being predictive of successful response to induction treatment (NEJM2005:352,254) and longer overall survival in cases without FLT3 mutations (Dohner K et al, and Schnittger S et al, Blood 2005, on line). Their clinical impact prompted us to develop a Real Time Quantitative PCR assay for the identification and monitoring of NPM1 mutants. Tests were set up using either cDNA (13 adult cases with mutation A and 1 with mutation B) or genomic DNA (gDNA) (7 pediatric and 8 adult patients). cDNA RQ-PCR . Forward primer was designed in exon 11, probe in exon 11/exon 12 junction (5′-FAM-3′-MGB) and reverse primers in exon 12. Samples were analyzed in ABI PRISM 7700 Sequence Detection System (Applied Biosystems). ABL1 gene was used as control. For the absolute quantitative assessment of NPM1 mutation A copies a standard curve with serial dilutions of a plasmid containing the target sequences was used. The highest reproducible sensitivity of this RQ-PCR assay was 10 molecules. The highest reproducible sensitivity of the relative quantification RQ-PCR assay for mutation A and B was 10−4. Thirteen AML NPM mutation A patients were monitored at diagnosis and over follow up. The number of mutated copies was high in all cases at diagnosis and significantly decreased after induction treatment in all cases with complete hematological remission. Only a slight decrease was observed in the case who did not reach remission. Patients with a complete hematological remission but MRD decrease 〈 3log showed a higher risk of relapse. gDNA RQ-PCR . specific forward primers for six different mutations were designed using Primer Express software to anneal to the mutated region of NPM exon 12. Reverse primers were designed on NPM1 exon 12. The TaqMan core reagent kit (Applied Biosystems) was used for RQ-PCR. The albumin gene was used as control. A reproducible sensitivity of 10−4 was reached in all but one case. The mean Ct of undiluted DNA samples was 23.3 (range 22.1–24.8). The mean slope of the dilution curves was 3.6 (2.9–4.0). High correlation coefficients (0.99 in all but one) were obtained. Using both cDNA and gDNA we set up sensitive and reproducible systems to detect minimal residual disease in 60% of AML with normal karyotype and NPM gene mutations. While gDNA RQ-PCR has the advantage to be directly related to the number of residual leukemic cells, cDNA RQ-PCR can be easily applied in samples collected in the routine diagnostic testing for common translocations. Large prospective studies are necessary to clarify the clinical impact of NPM1-based MRD monitoring of AML with normal karyotype.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2005
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 5
    In: Blood, American Society of Hematology, Vol. 134, No. Supplement_1 ( 2019-11-13), p. 2042-2042
    Abstract: Introduction: Total skin electron beam (TSEB) therapy represents a valid treatment option in the management of patients with cutaneous T-Cell lymphoma (CTCL) with diffuse skin involvement. While the efficacy of TSEB for palliative treatment is well established, its inclusion as a debulking strategy before reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation (allo-HSCT) has been only recently reported. Due to the lack of hematopoietic toxicity, TSEB may also be effectively administered in patients relapsing after allo-HSCT. With this study we aimed to retrospectively investigate the use of TSEB among patients who underwent to allo-HSCT in our Center for advanced CTCL. Patients & Methods: As of July 2019, 45 CTCL patients (26 M and 19 F, median age 54 years, range 19-66) all having stage IIB to IV refractory Mycosis Fungoides (MF, n=33) or Sézary Syndrome (SS, n=12) underwent allo-HSCT from HLA-identical sibling (n=18), unrelated donors (n=23) or haploidentical related donors (n=4). Median time from diagnosis to HSCT was 46 months and median number of previous treatment lines was 6. Source of stem cells was peripheral blood in 40 patients, bone marrow in 4 patients and cord blood in 1 patient. Conditioning regimens included FC/TBI200, pentostatin/TBI200, fludarabine/melphalan and TT/CTX/Fluda/TBI200. Outcomes at 5-years were: OS 51% (33%-66%), DFS 40% (20%-50%), NRM 15% (4%-27%) and relapse incidence 48% (32%-65%). Clinical charts of the whole series of patients were extensively reviewed to collect information on the use of TSEB from diagnosis to the last follow up date. Results: Overall 21 patients of the series received TSEB with dose fractionation of 2 Gy in 2 days administered with the Stanford Technique as part of their treatment strategy. In six cases it was included among the lines of treatment administered over the disease course preceding transplant referral, with a total dose of 36 Gy in all of them. In 13 patients TSEB was part of the debulking strategy for patients with chemorefractory disease just prior to allo-HSCT with a median total dose of 24 Gy (range 10.8- 36). In 4 patients TSEB was administered after transplantation: as a salvage treatment for skin relapse during early post tranplant intense immunosuppression phase in 3 cases (12, 36, 21.6 Gy respectively) and as palliative treatment for disease progression in one case (36 Gy). Among patients who received TSEB as a bridge to transplant strategy, CR was achieved in 5 cases: at the last follow up visit (median time from transplantation 23 months, range 6-80) 4 were alive with persistent CR, whereas 1 patient who experienced graft failure died from disease progression 31 months after transplantation. Partial remission (PR) was documented in 7 (very good in 2): at the last follow up visit (median time from transplantation 14 months, range 6-52) 2 patients were alive in CR and 5 died: 3 from disease progression, 1 from GVHD and 1 from myocardial infarction 42 months after transplantion. In 1 patient, who only showed minimal response to TSEB therapy, death from disease progression occurred 9 months after an autologous-allogeneic tandem transplant strategy. All the 3 patients receiving TSEB after allo-HSCT for early relapse achieved a new and durable CR maintained at the last follow up date (42, 21 and 10 months after transplant respectively), suggesting the occurrence of a durable graft-versus-lymphoma effect following TSEB. As far as safety is concerned, only grade 1 skin toxicity (erythematous reactions) was observed in a minority of patients, while no case of haematological toxicity was documented. Conclusions: Our experience confirmed TSEB as a particularly safe and potentially effective treatment strategy in CTCL patients, both to induce remission prior to allo-HSCT and to rescue early post transplant relapse occurring before immunosuppression withdrawal. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2019
    detail.hit.zdb_id: 1468538-3
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  • 6
    In: Blood, American Society of Hematology, Vol. 104, No. 11 ( 2004-11-16), p. 4672-4672
    Abstract: Imatinib mesylate (STI571), a specific Bcr-Abl inhibitor, has shown a potent antileukemic activity in clinical studies of chronic myeloid leukemia (CML) patients. Early prediction of response to imatinib cannot be anticipated. We used a standardized quantitative reverse-transcriptase polymerase chain reaction (QRT-PCR) for bcr-abl transcripts on 191 out of 200 late-chronic phase CML patients enrolled in a phase II clinical trial with imatinib 400 mg/day. Bone marrow samples were collected before treatment, after 3, 6 and 12 months or at the end of study treatment (12 months) while peripheral blood samples were obtained after 2, 3, 6, 10, 14, 20 and 52 weeks of therapy. The amount of Bcr-Abl transcript was expressed as the ratio of Bcr-Abl to β2-microglobulin (β2M). We show that, following initiation of imatinib, the early Bcr-Abl level trends in both bone marrow and peripheral blood samples made it possible to predict the subsequent cytogenetic outcome after 6 and 12 months of treatment, and that these early trends were also predictive of progression-free survival.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2004
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 7
    In: Blood, American Society of Hematology, Vol. 98, No. 9 ( 2001-11-01), p. 2657-2663
    Abstract: Chronic hepatitis C virus (HCV) infection has been associated with development of mixed cryoglobulinemia type 2 (MC2), a lymphoproliferative disorder characterized by B cell monoclonal expansion and immunoglobulin M/k cryoprecipitable immunoglobulin production. A short sequence (codons 384-410) of the HCV E2 protein, which has the potential to promote B cell proliferation, was investigated in 21 patients with HCV-related MC2 and in a control group of 20 HCV carriers without MC2. In 6 of the 21 (29%) patients with MC2, all the clones isolated from plasma, peripheral blood mononuclear cells, and liver showed sequence length variation compared with the hypervariable region 1 (HVR1) consensus sequence; 5 patients had an insertion at codon 385, and 1 patient had a deletion at codon 384. Inserted residues at position 385 were different within and between patients. No such mutations were observed in any of the HVR1 clones from control patients without MC2, and the difference between the 2 groups was statistically significant (P = .02). Analysis of 1345 HVR1 sequences obtained from GenBank strongly supported the conclusion that the observed insertions and deletion represent a rare event in HCV-infected patients, suggesting that they are significantly associated with MC2. The physical and chemical profiles of the 385 inserted residues detected in the MC2 patients were consistent with the possibility that these mutations, which occurred in a region containing immunodominant epitopes for neutralizing antibodies and binding sites for B lymphocytes, may be selected by functional constraints for interaction with host cells.
    Type of Medium: Online Resource
    ISSN: 1528-0020 , 0006-4971
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2001
    detail.hit.zdb_id: 1468538-3
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  • 8
    In: Blood, American Society of Hematology, Vol. 132, No. Supplement 1 ( 2018-11-29), p. 4655-4655
    Abstract: Introduction: Although a few novel drugs have recently shown promising activity in mycosis fungoides (MF) and Sézary syndrome (SS), prognosis of patients with advanced stages or refractory disease remain poor, with median survival ranging from 1.4 to 3.4 years (Agar NS et al. JCO 2010). In selected patients, allogeneic hematopoietic stem cell transplantation (allo-HSCT) represents a potential curative strategy. By decreasing transplant-related mortality, reduced intensity (RIC) and nonmyeloablative conditioning (NMA) regimens lead to better outcomes in comparison to myeloablative ones. Here we report long-term outcome of our RIC allo-HSCT experimental program, initiated in 2001. Patients & Methods: As of July 2018, in our Center 40 patients underwent RIC allo-HSCT from HLA-identical sibling (n=16), unrelated donors (UD, n=22 - fully-matched in 10, 1 or 2-mismatch in 12) or haploidentical related donors (n=2). Median age was 52 years (range 19-66). All patients (24 M and 16 F) had stage IIB/IV refractory MF (n=27) or SS (n=13). Median number of previous treatment lines was 6 (range 2-12) while median time from diagnosis to transplantation was 46 months (range 11-264). The source of stem cells has been peripheral blood in 35 patients (87.5%), bone marrow in 4 (10%) and cord blood in 1 (2.5%). Conditioning regimens included FC/TBI200, pentostatin+TBI200, and fludarabine/melphalan in transplants from HLA-identical sibling donor or UD, whereas the TT/Flu/CTX/TBI200 regimen was used in the haploidentical setting. GvHD prophylaxis included CsA/MMF in all patients, with the addition of ATG in cases with UD and post-transplant CTX (50 mg/kg giorni +3 e +4) in haploidentical setting. Results: Full donor chimerism was obtained in 32 out of 37 evaluable patients, in a median time of 2 months (range 1-12). Acute GvHD occurred in 18 patients out of the 32 evaluable (56%), being of grade III-IV in 9 (28%). Chronic GvHD was observed in 10 patients (31%), being extensive in 4 (12%). Of note, the latter were all patients transplanted from HLA-identical sibling (i.e. without ATG). Following transplantation, a complete remission (CR) was achieved in 26 out of the 37 evaluable patients (70%), of whom 4 experienced relapse at +2 (2 pts), +25 and +35 months, respectively. At the last follow-up, 19 patients were alive and 17 (89%) maintained CR after a median follow-up of 80 months (range 4-210). Out of the 11 patients who did not achieve CR, 9 died from progressive disease (median follow-up of 12 months, range 3-31), 1 from a secondary malignancy, while 1 is still alive with disease 62 months after transplant. Transplant-related death occurred in 7 patients (17%), of whom 5 were in CR. In the whole population, the 5-year OS was 52% (95% CI 34-70) [Fig.1] and the 5-year DFS was 43% (95% CI 27-62). However, when MF and SS were analysed separately, 5-yrs DFS were 30% (95% CI 12-51) and 72% (95% CI 38-99), respectively (Fig.2). Apart from diagnosis, outcome appeared to be primarily associated with chemosensitivity and status of disease at transplantation. Conclusions: After a median follow-up longer than 6.5 years, we confirm the efficacy of RIC allo-HSCT as a powerful therapeutic strategy in inducing and maintaining remission in selected patients with chemosensitive advanced-stage CTCL, with results particularly encouraging in SS. Disclosures Cortelezzi: roche: Consultancy; abbvie: Consultancy; novartis: Consultancy; janssen: Consultancy.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2018
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    detail.hit.zdb_id: 80069-7
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  • 9
    In: Blood, American Society of Hematology, Vol. 140, No. Supplement 1 ( 2022-11-15), p. 10237-10238
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2022
    detail.hit.zdb_id: 1468538-3
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  • 10
    In: Blood, American Society of Hematology, Vol. 134, No. Supplement_1 ( 2019-11-13), p. 5744-5744
    Abstract: Introduction: In patients with advanced stage mycosis fungoides (MF) and Sézary syndrome (SS), allogeneic hematopoietic stem cell transplantation (allo-HSCT) continues to represent the only potentially curative treatment strategy. We have previously reported long-term outcome of our reduced intensity conditioning-based allo-HSCT program in MF and SS, including 38 patients who consecutively underwent transplantation from matched sibling or unrelated donor in the 2001-2018 time interval, with a 5-yr overall and disease-free survivals of 52% and 43%, respectively, and a 1-yr non relapse mortality of 17% (ASH annual meeting, 2018). In the most recent years, haploidentical donors are considered an increasing valid alternative for patients with haematological malignancies lacking a suitable matched donor. Here we report the results of our first series of haploidentical SCT (haplo-HSCT) in patients witn MF and SS. Patients & Methods: From May 2016 to June 2019, 4 patients (2 males and 2 females) underwent haplo-HSCT from family donors (3 siblings and 1 son) in our center. Median age was 53 years (range 19-62). Two patients had stage IV refractory MF - involving nodes and lungs in one case and blood in the other one, while two patients had SS. Median number of previous treatment lines was 4 in SS and 4.5 in MF (range 2-6) while median time from diagnosis to transplantation was 21 months (range 17-101). Two patients (1 SS and 1 MF) received 24Gy total skin electron beam (TSEB) therapy as a bridge to transplant, associated to brentuximab vedotin (5 cycles) in one case showing also lung CD30+ involvement. At the time of transplant two patients were in CR and 2 were in very good partial remission, with limited nodal involvement in one SS and limited skin disease in one MF patient, respectively. The source of stem cells was bone marrow in SS and peripheral blood in MF patients. A reduced-intensity conditioning regimen including Thiotepa 10 mg/kg, Cy 30 mg/kg, Fludarabine 120 mg/m2 (over 4 days) and low dose TBI (200 cGy) was used in all patients. GvHD prophylaxis included CsA/MMF and post-transplant CTX (50 mg/kg on days +3 e +4), with the addition of ATG 2.5 mg/kg in the most recently transplanted patient for whom donor peripheral blood was the selected source of stem cells. Results: Hematologic engraftment occurred in all patients, with a median time to ANC 〉 0.5 x 109/L of 16.5 days (range 14-18) and to PLT 〉 20 x 109/L of 15 days (range 13-45). At +100 days after transplantation, donor chimerism was 100% in 3 patients, and 90% in one. Acute GvHD occurred in 3 patients, always of grade II (involving skin in all, gastrointestinal in 2 and liver in 1 patient), with overlap characteristics in one case. Major early infectious complications included two cases of fungal pneumonia and 1 case of bacteremia from P. aeruginosa. Chronic GvHD was observed in 2 out of the 3 evaluable patients - i.e. with a follow-up longer than 100 days - being mild in one case (with joints involvement) and severe in the other case (skin). With all patients and their donors being CMV positive at baseline, CMV reactivations occurred in 3 cases, successfully treated with preemptive valganciclovir. Following transplantation, a complete remission (CR) was achieved in all the four patients. One patient with SS who experienced a skin biopsy-proven relapse 9 months after transplant, achieved a new and durable CR following the occurrence of a severe skin chronic GvHD triggered by an inadvertent sunburn, which required steroids + ECP treatment. At the last visit, all patients were alive in CR with a follow-up of 38, 36, 6 and 3 months, respectively. Conclusions: Even though with a limited follow-up time, our preliminary experience of haplo-HSCT appears particularly safe and highly encouraging in inducing and maintaining remission in patients with advanced MF/SS eligible to allo-HSCT. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2019
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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