Hemotopoietic Stem Cell Transplantation Research Articles

IMUNNOPHENOTYPIC CHANGES IN JMML PATIENTS TREATED WITH 5-AZACITIDINE AND CORRELATION WITH CLINICAL RESPONSE

Background 5-azacitidine has been considered to be safe and efficient medication for use as a bridge to hemotopoietic stem cell transplantation in JMML patients. Patients under clinical treatment can be stratified as complete or partial response, stable disease or disease progression. Recently, the features found in JMML immunophenotyping (FCI) at diagnosis have been described and so, they could be useful for examining these characteristics for determining response of the patients to treatment. Aims to follow the immunophenotypic pattern of JMML patients during treatment with 5-azacitidine, with an already validated IFC protocol and also look for cytometric features that correlate with the type of clinical response. Materials and Methods An 8-color antibody panel described for pediatric population with JMML at diagnosis was used. Patients treated with cycles of 5-azacitidine 75 mg/m2 subcutaneous for consecutive 7 days, each 28 days were evaluated at diagnosis and after 3 and 6 cycles of medication. The findings were compared with the type of clinical response. Results 32 patients entered the study. Among them, only 28 patients could be analyzed by FCI after 3 cycles and 25 patients after 6 cycles. After treatment with 5-azacitidine, patients showed a reduction in CD34+ cells, with median CD34/CD117+ cells going from 3.35% to 2.8% after 3 cycles of medication. Values dropped to Conclusions FCI was a feasible tool in JMML patients after treatment with 5-azacitidine. Clinical response was associated with decrease of CD34+ myeloid progenitor and total monocytes and a rise in T and NK lymphocytes. But abnormal co-expressions remained, even in patients with complete clinical response and this speaks in favor of the persistence of the leukemic clone during the whole clinical treatment. The largest effect was seen after 3 cycles. CD34/117+ cell and T lymphocytes are variables that could be incorporated into the assessment of response to clinical treatment of JMML patients. A more detailed study of the several subsets of T cells should be included. The same MFC protocol could be used pre and post transplantation identifying patients with beneficial use of 5-aza after transplant, as it has recently been shown according to methylation status. Prospective studies are necessary to confirm the data with a larger number of patients, and including assessment of the pre and post transplantation status.

Chimeric Antigen Receptor (CAR) T Cell Therapy in Acute Myeloid Leukemia (AML).

Despite high response rates after initial chemotherapy in patients with acute myeloid leukemia (AML), relapses occur frequently, resulting in a five-year-survival by <30% of the patients. Hitherto, allogeneic hemotopoietic stem cell transplantation (allo-HSCT) is the best curative treatment option in intermediate and high risk AML. It is the proof-of-concept for T cell-based immunotherapies in AML based on the graft-versus-leukemia (GvL)-effect, but it also bears the risk of graft-versus-host disease. CD19-targeting therapies employing chimeric antigen receptor (CAR) T cells are a breakthrough in cancer therapy. A similar approach for myeloid malignancies is highly desirable. This article gives an overview on the state-of-the art of preclinical and clinical studies on suitable target antigens for CAR T cell therapy in AML patients.

Efficacy observation of hemocoagulase for the treatment of severe hemorrhagic cystitis following allogeneic hematopoietic stem cell transplantation

Objective To investigate the clinical efficacy of hemocoagulase for severe hemorrhagic cystitis (HC) following allogeneic hemotopoietic stem cell transplantation (HSCT). Methods Twenty patients undergoing allogeneic HSCT developed severe HC with an onset time of 14 to 70 days, all patients received the treatment of hemocoagulase (1 U ivgtt q12 h × 5 d). The urine speciments reserved before and after hemocoagulase were examined by naked eye and microscope to evaluate the efficacy. Results Twenty patients received the treatment of hemocoagulase. The HC was cured in 18 patients, improved in 1 patient and uncontrolled in 1 patient. For the patients with response, macroscopic hematuria disappeared at a median of 28 days (4-127 days) after the treatment. All procedures were tolerated well and no severe adverse effect was observed. Conclusion Hemocoagulase seems to be a safe and effective drug for severe HC following HSCT. Key words: Hemorrhagic cystitis; Hemotopoietic stem cell transplantation; Hemocoagulase

Donor KIR Genotype BX1 had a negative effect while centromeric B-Specific gene motifs had a positive effect of survival for standard risk AML Patients after unrelated donor hemotopoietic stem cell transplantation

Aim To investigate the impact of donor Killer Immunoglobulin-like Receptors (KIR) genotypes and centromeric motif B (Cen-B) on clinical outcomes after unrelated donor (URD) hematopoietic stem cell transplantation (HSCT) for Chinese population. Method A total of 210 Chinese patients underwent URD-HSCT were investigated in this 4-year retrospective study. KIR genotyping and HLA typing were performed for the population. Results The risk of transplant from Bx1 donors and the benefit of the presence of Cen-B (ignoring the number) were observed for standard risk AML/MDS patients Donor KIR genotype Bx was associated with significant improved OS (P = .026) and RFS (P = .021), and reduced NRM (P = .017) for AML/MDS patients. A much worse survival was observed for transplants from Bx1donors compared to Bx2, Bx3, and Bx4 donors for patients in CR1, ( n = 82; OS: P = .024; RFS: P = .021). Transplant form donors with Cen-B improved OS (HR = .256; 95% CI = .084 to .774; P = .016) and RFS (HR = .252; 95% CI = .084 to .758; P = .014) for AML/MDS patients in CR1. However, the effect did not increase with the number of Cen-B motifs (cB/B vs. cA/B; OS: P = .755; RFS: P = .768). No effect was observed for high risk AML/MDS and ALL/NHL, CML patients. Conclusions Avoiding the selection of HSCT donors with KIR genotype Bx1 is strongly advisable for AML/MDS patients of standard risk. The presence of cen-B motif rather than the number was more important in donor selection for Chinese population.

Donor KIR B Haplotype Is Associated with a Increased Rate of Leukemia Relapse and Worse Overall Survival after HLA Match Sibling Pediatric Stem Cell Transplantation

IntroductionGraft versus leukemia (GvL) effect after hemotopoietic stem cell transplantation (HSCT) is mediated by donor immune cells recovery. Natural Killer (NK) cell alloreactivity is controlled by the interaction of activatory receptors and inhibitory killer-immunoglobulin-like receptors (KIRs) with major histocompatibility locus class I antigens on the leukemia cells. Haploidentical setting is a plattform for NK cell alloreactivity however HLA identical setting remains unclear.Methods and PatientsWe performed KIR-genotyping of HLA-identical sibling donors in 35 pediatric CD34 selection peripheral blood stem cell transplantations to identify genetic factors affecting leukemia relapse and overall survival. Univariate analysis of leukemia relapse and KIR genotyping was performed in order to identify independent variables predictive of outocome for pediatric acute lymphoblastic leukemia (ALL), (n=20) and acute myeloblastic leukemia (AML), (n=15).ResultsDonor B haplotype was observed in 21 cases (60%). Statistical analysis shown that donor B haplotype was associated with significantly more relapse in leukemia pediatric patients (38% vs. 0%) and worse overall survival (40% vs. 7%). Further analysis revealed that 2DL5a, 2DS3 and 2DS5 were associated with an increased rate of leukemia relapse (47% vs. 6%, 54% vs. 10% and 58% vs. 13%, respectively) and 2DL5a, 2DS1 and 3DS1 were associated with a worse overall survival (48% vs. 6%, 64% vs. 10% and 58% vs. 13%, respectively). No difference was observed in patients KIR haplotype or donor-recipient KIR haplotype mismatch.ConclusionIn our study, which included only reduced intensity conditioning without antithymocyte globulin followed by related peripheral blood stem cell transplantation, we found a significant worse impact of donor KIR B haplotype in stem cell transplantation outcome. Our results suggest that donor KIR B haplotype (2DL5a, 2DS3 and 2DS5) increases leukemia relapse and also donor KIR B haplotype (2DL5a, 2DS1 and 3DS1) confer significant survival damage to HLA-identical sibling HSCT. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Incidence and Clinical Features of Extramedullary Multiple Myeloma in Patients Who Underwent Stem Cell Transplantation

Background: Extramedullary disease (EMD), strictly defined as an infiltrate of clonal plasma cells at an anatomic site distant from the bone marrow or adjacent soft tissue in a patient with underlying multiple myeloma, is an uncommon manifestation of multiple myeloma. Comparatively little is known about this disease entity, with no large case series published in the last decade. Patients and Methods: 663 consecutive patients with multiple myeloma who underwent autologous or allogeneic stem cell transplantation at a single, large, academic medical center in the United States from January 2005 to December 2011 were assessed for the presence or absence of EMD, as well as baseline demographic and biochemical characteristics, treatment regimens, and response to therapy. Results: A cohort of 55 patients with biopsy-proven EMD was identified, comprising 8.3% of the total study population. Among the patients with EMD, 13 (23.6%) were found to have EMD at the time of initial presentation, while the remainder developed EMD during the course of their illness. Patients with EMD received a median of 5 different treatment regimens during the course of their illness, most commonly with combinations of dexamethasone, thalidomide, lenalidomide, and bortezomib, as well as autologous hemotopoietic stem cell transplantation. Patients had received a median of 3 lines of therapy prior to experiencing an extramedullary relapse. Patients with EMD had markedly elevated maximum serum LDH levels (median 613.5 units/L) and low minimum hemoglobin levels (median 7.8 g/dL). Common cytogenetic abnormalities included deletion 13q, deletion 11q, t(11;14), and deletion 17p. Available immunohistochemical data suggest that EMD specimens had frequent expression of CXCR4, CD44, and CD56. The median overall survival data of these patients was 3.2 years (range, 0.9-9.5) and the median time from diagnosis of EMD to death was 0.5 years (range, 0.002-3.2). Conclusions: This report describes a large series of multiple myeloma patients with EMD who were treated in the era of stem cell transplant at a single academic medical center. Further studies to examine the molecular characteristics of extramedullary multiple myeloma are necessary to better define this entity and characterize therapeutic options that can prolong survival in this otherwise very vulnerable population. DisclosuresGhobrial:Millennium/Takeda: Membership on an entity’s Board of Directors or advisory committees; Onyx: Membership on an entity’s Board of Directors or advisory committees; BMS: Membership on an entity’s Board of Directors or advisory committees; Celgene: Membership on an entity’s Board of Directors or advisory committees. Laubach:Novartis: Research Funding; Onyx Pharmaceuticals: Research Funding. Schlossman:Millennium: Consultancy. Mitsiades:Millennium Pharmaceuticals: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Amgen: Research Funding; Johnson & Johnson: Research Funding; DFCI: patent submission on stromal co-culture technologies Patents & Royalties.

Retrospective Assessment of the Stanford Integrated Psychosocial Assessment for Transplantation (SIPAT) in Hemotopoietic Stem Cell Transplantation (HCT) Recipients

Retrospective Assessment of the Stanford Integrated Psychosocial Assessment for Transplantation (SIPAT) in Hemotopoietic Stem Cell Transplantation (HCT) Recipients

Clinical analysis of 104 patients with hematological malignancy after allogeneic hemotopoietic stem cell transplantation

To study the efficacy of allogeneic hemotopoietic stem cell transplantation (allo-HSCT) for hematological malignancy. A total of 104 patients with hematological malignancy, who underwent allo-HSCT in Xiangya Hospital from December 1999 to January 2010, were retrospectively analyzed. Of the patients, the transplantation related mortality (TRM), relapse rate (RR), 5-year overall survival (OS) and disease free survival (DFS) were estimated by Kaplan-Meier analysis. The unfavorable prognostic factors were also statistically examined. Hematopoietic reconstitution was achieved in 101 patients. At the last data of follow-up, the incidences of severe acute graft versus host disease (aGVHD) and extensive chronic GVHD were 15.38% and 25.53%, and the TRM and RR were 15.66% and 21.76%, respectively. The estimated 5-year OS and DFS for all patients were (73.49±4.59)% and (63.10±5.32)%, respectively. Those for acute myeloid leukemia (AML) patients were (63.00±9.51)% and (49.30±9.96)%, and those for chronic myeloid leukemia (CML) patients were (83.87±5.06)% and (74.55±6.79)%, respectively. The survival analysis suggested the poor prognostic factors for allo-HSCT recipients including female sex, severe aGVHD and refractory hematological malignancy. Further multivariate analyses revealed that severe aGVHD and refractory hematological malignancy were the independent risk factors of poor prognosis for the recipients (P<0.05). The 5-year DFS of severe aGVHD and refractory hematological malignancy patients was (48.22±12.69)% and (42.09±12.31)%, respectively. The TRM of severe aGVHD, HLA-mismatched graft and unrelated donor transplant was significantly higher than that of the corresponding control groups (57.14% vs. 4.81%, 33.33% vs. 10.41%, 26.09% vs. 9.28%; P<0.05). The RR of refractory hematological malignancy was significantly higher than that of the control group (41.09% vs. 15.63%, P<0.05). The treatment of allo-HSCT can improve the disease free survival of patients with hematological malignany and is an important therapeutic method for hematological malignancy. Severe aGVHD and refractory hematological malignancy are the independent risk factors of poor prognosis for the allo-HSCT recipients with hematological malignancy.

The Impact of Rhg-CSF on the LFA-1 Conformation of CD4+T Cells in Mobilized Hematopoietic Stem Cell Allografts

In hemotopoietic stem cell transplantation (HSCT), the primary effects of G-CSF on cells of the hemotopoietic system include stimulation of proliferation and differentiation of HSCs, acceleration of neutrophil reconstitution after HSCT and mobilization of bone marrow HSCs into the peripheral blood. In recent years, several investigators have unraveled that G-CSF-mediated immune regulation including the switching T cell cytokine secretion profile to Th2 response, the altering adhesion activity of CD4+T cells to ICAM-1 and so on. Most of these cellular interactions of T cells are dependent on the integrin leukocyte function associated antigen-1 (LFA-1, αLβ2, CD11a/CD18) binding to intercellular adhesion molecule-1 (ICAM-1). Integrin αLβ2 adopts a bent, compact conformation in their low affinity state and an extended conformation in the high affinity state. The epitope of MEM148 is only expressed on the free β2 integrin subunit but is masked on LFA-1. The monoclonal antibody MEM148 reacts with an epitope exposed on free human CD18 chain as well as on high affinity state of LFA-1 that represents a reporter for αLβ2 active conformation. Our previous studies have demonstrated that rhG-CSF mobilization decreased the adhesion activity of CD4+T cells to ICAM-1, but did not find that rhG-CSF mobilization had effect on the level of CD11a expression. rhG-CSF maybe has effect on conformational change of LFA-1 on CD4+T cells. In order to explore the impact of rhG-CSF mobilization on LFA-1 conformation of CD4+ T cells, human CD4+T cells were isolated from peripheral blood mononuclear cells by positive selection with Miltenyi MACS. Isolated CD4+T cells were activated by OKT3+ICAM-1 and PMA+Ion separately. The slides with CD4+T cells were incubated with FITC-CD11a and FITC-MEM148 mAb, then were examined by using fluorescence microscope. The results show that the level of CD25, CD69 expression and MEM148 epitope exposure on activated CD4+T cells were significantly higher compared with unactivated CD4+T cells. rhG-CSF mobilization had no effect on the expression of CD11a, but could decrease expression of CD25, CD69 and exposure of MEM148 epitope on activated CD4+T cells. The percentage of LFA-1 polarized CD4+T cells before and after rhG-CSF mobilization was 85.32%, 61.86% respectively (p <0.01). No MEM148 epitope exposure on unactivated CD4+T cells was detected, but MEM148 epitope exposure on activated CD4+T cells before and after rhGCSF mobilization was 63.63%, 32.79% respectively (p <0.01). Overall, these data suggest that rhG-CSF can affect on the activation of CD4+T cells in mobilized hematopoietic stem cell allografts by altering the conformation of LFA-1.

High Anti-Tumor Activity of Natural Killer Cells after Non-Myeloablative Conditioning and Allogeneic Stem Cell Transplantation.

Background: It is hypothesized that enhanced graft-versus-leukemia activity of natural killer (NK) cells contributes to the clinical efficacy of non-myeloablative allogeneic hemotopoietic stem cell transplantation (HSCT). We compared NK cell cytotoxic activity after transplantation with full and reduced dose regimens with normal values obtained from healthy volunteers.Methods&Material: Using a flow cytometric assay that detects CD107a expression after coincubation with leukemia cell lines as a marker for NK cell degranulation we prospectively quantified and characterized NK cells mediating anti-tumor activity in patients after HSCT. Mononuclear cells (MNC) were isolated from peripheral blood of 17 healthy individuals and 31 patients transplanted with G-CSF mobilized peripheral blood stem cells at day +30. MNC were incubated with leukemia cell lines HL60 and K562 (effector/target ratio 1:1) and expression of CD107a was measured after 3 hours. The absolute number of degranulating (CD107a+) NK cells was calculated.Results: Thirty one patients (five with ALL, four with NHL, one with Aplastic Anemia and 21 with AML) were enrolled, of whome 22 were transplanted from a matched related donor, seven from a matched unrelated donor, and two from a haploidentical donor. Nine patients had been transplanted after conditioning with 2 Gy TBI only, whereas 22 patients had received various conventional dose regimens. NK cell counts at day +30 after HSCT with conventional conditioning were comparable to normal values, but percentage and number of degranulating cells were significantly reduced (2.4% and 7/μl vs. 6.2% and 18/μl; p<0.0001 and p=0.0003). In contrast, in patients after 2 Gy TBI the percentage of degranulating NK cells was comparable to healthy donors (7.6%; p=0.9), and interestingly, absolute numbers of all and of degranulating NK cells (41/μl; p=0.004) were higher than normal values and consequently higher than in patients with conventional dose conditioning.Conclusions: Using this new method we were able to exactly quantify tumor-reactive NK cells after HSCT in a feasible way. We found a high cytotoxic activity of NK cells towards leukemia cell lines in patients undergoing non-myeloablative conditioning with 2Gy TBI. Further studies to determine the clinical impact of these findings are needed.