bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic Value of BCR-ABL Transcript Amount for Early Relapse Detection in Chronic Myeloid Leukemia Patients Treated with Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Solarska Iwona , Majewski Miroslaw , Nasilowska-Adamska Barbara , Zaucha Jan M. , Rzepecki Piotr , Calbecka Malgorzata , Sulek Kazimierz , Hellmann Andrzej , Marianska Bozena , Warzocha Krzysztof
Published: October 2019

The detection of BCR-ABL mRNA following allogeneic hematopoietic stem cell transplantation (allo-HSCT) is associated with relapse but not absolutely predictive. Early molecular relapse can be detected using real-time quantitative PCR (RQ-PCR) assay, which provides an accurate and reliable method of BCR-ABL expression evaluation. Using this method we analyzed 66 CML patients 3 months to 12 years (median 50 months) after allo-HSCT, who had BCR-ABL transcript measured in peripheral blood or bone marrow samples every 3, 6 and 12 months after allo-HSCT. The patients were analyzed at monthly intervals in the case of rising BCR-ABL transcript level. The median age at diagnosis and median age at time of allo-HSCT were 38 years (range, 16–55). Fifty three patients were in the first chronic phase, and 13 were in the second chronic phase. The median time from diagnosis to allo-HSCT was 8 months (range, 2.5–59). Eighty two percent of patients received allo-HSCT from HLA-identical siblings and 18% from unrelated donors. Transplant conditioning and GvHD prophylaxis were performed according to standard protocols. Quantification of BCR-ABL mRNA transcript was performed according to ‘Europe Against Cancer’ protocol. The median number of assays per patient was 6 (range, 3–16). According to the amount of BCR-ABL transcript after allo-HSCT, patients were allocated into 3 categories, including 35 patients with stable/low level of BCR-ABL transcript (below 0.005%), 11 patients with fluctuating-low level of BCR-ABL (0.005%–0.01%), and 18 patients with high level of BCR-ABL transcript (0.01%–0.1%). We found strong correlation between the amount of BCR-ABL transcript after allo-HSCT and probability of molecular relapse (6% vs. 9% vs. 28%, respectively) as well as relapse-free survival (median time to relapse: 134 months vs. 106 vs. 89, respectively). In the group of patients with high level of BCR-ABL transcript cytogenetic relapse was observed in 75%. In one case BCR-ABL level spontaneously fall down to a low-lewel expression within 6 months. One patient received donor lymphocyte infusion without response, and then was successfully treated with imatinib. RQ-PCR is valuable method to quantitate BCR-ABL expression in CML patients after allo-HSCT, allows monitoring the kinetics of BCR-ABL mRNA transcript amount and is useful in prediction of the hematologic relapse. RQ-PCR can successfully distinguish the group of patients with high risk of relapse from two groups without need for additional therapy (persistent disease/lack of disease).
Haematopoietic Stem Cell Transplantation - An Experience from Developing World.
Authors: Ullah Khalil , Ahmed Parvez , Raza Shahid , Mahmood Tariq , Khan Badshah
Published: October 2019

One hundred and fifty four patients received allogeneic stem cell transplant from HLA matched siblings for various haematological disorders at Armed Forces Bone Marrow Transplant Centre, Rawalpindi, Pakistan from July 2001 to Sep 2006. Indications for transplant included aplastic anaemia (n=66), b-thalassaemia major (n=40), CML (n=33), acute leukaemia (n=8) and misc disorders (n=7). One hundred and twenty patients were male and thirty four were female. Median age of patient cohort was 14 yrs (range 1 ¼ −54 yrs). Pre-transplant infection surveillance was carried out and strict prophylaxis against infection was observed. The mean mononuclear cell dose was 4.2 x 109/kg of recipient. Post transplant complications encountered in our patients were: acute GvHD (grade II-IV) 28.5%, chronic GvHD 15.5%, haemorrhagic cystitis 9.7%, VOD liver 5.1%, acute renal failure 3.2%, bacterial infections 51.2%, fungal infections 15.0%, CMV infection 4%, herpes zoster 4%, tuberculosis 2.6%, pneumocytitis jirovici infection 0.6%, malaria 0.6% patient, graft rejection 5.2% patients and relapse in 4%patients. Certain unexpected rare post transplant complications were also observed in our patients. These included hickman catheter embolization, GB syndrome, deep vein thrombosis, haemorrhagic pericarditis with clots leading to cardiac temponade, idiopathic polycythemia, dengue fever and status epilepticus. Mortality was observed in 27.2% patients. Major causes of mortality were GvHD, VOD, relapse, intracranial haemorrhage, acute renal failure, pseudomonas septicemia, tuberculosis, disseminated aspergillosis and CMV infection. At five years, the overall survival (OS) and disease free survival (DFS) was 72.5% & 70.7% respectively.
Efficacy Comparison of Oral Versus Intravenous Busulfan in a BuCy Conditioning Regimen for Allogeneic BMT.
Authors: Lee Je-Hwan , Choi Seong-Jun , Lee Jung-Hee , Seol Miee , Lee Young-Shin , Ryu Seong-Gil , Kang Young-Ah , Lee Kyoo-Hyung
Published: October 2019

Intravenous administration of busulfan (I-Bu) or targeted busulfan dosing has replaced oral administration of busulfan (O-Bu) in a conditioning therapy for BMT mainly due to erratic gastrointestinal absorption of oral busulfan. Several retrospective study results have demonstrated that use of I-Bu resulted in lower transplant-related morbidity and mortality compared to O-Bu, but the efficacy of I-Bu and O-Bu has not been compared. In this retrospective study, we compared the efficacy of I-Bu vs. O-Bu in a BuCy conditioning regimen for allogeneic BMT in a single institute. To avoid the bias of retrospective comparison, we restricted the patient eligibility to the followings:

patients with AML having intermediate or poor cytogenetic risk groups at the time of diagnosis, those in the first complete remission at the time of BMT, those who were treated with BuCy conditioning regimen, and those who received bone marrow as a stem cell source.

A total of 80 patients were eligible in this study among the patients who were transplanted at the Asan Medical Center between December 1993 and July 2007: 42 patients received O-Bu (4 mg/kg/d x 4d) and 38 patients received I-Bu (Busulfex, 3.2 mg/kg/d x 4d). The same dose of cyclophosphamide (60 mg/kg/d x 2d) was given to all the patients. We collected clinical and laboratory data from the BMT database of the Asan Medical Center. The efficacy was compared in terms of cumulative incidence of relapse (CIR), event-free survival (EFS), and overall survival (OS). Propensity score analysis was used to adjust the confounding factors between O-Bu and I-Bu groups. Several baseline clinico-laboratory factors at the time of BMT such as age, number of consolidation therapy prior to BMT, time from diagnosis to BMT, use of methotrexate for GVHD prophylaxis, and transfusion requirement before BMT were significantly different between 2 groups. Times to engraftment, transfusion requirements, infectious complications, hepatic VOD, and acute and chronic GVHD were similar between 2 groups. The cumulative incidence of non-relapse mortality was 14.3% in O-Bu group and 10.3% in I-Bu group (adjusted P-value, 0.990). The CIR at 4-year was 28.6% in O-Bu group and 16.9% in I-Bu group (adjusted P-value, 0.430). The 4-year probabilities of EFS and OS were 57.1% and 64.3% in O-Bu group and 72.6% and 69.3% in I-Bu group (adjusted P-values, 0.496 and 0.634, respectively). In summary, EFS and CIR were superior in I-Bu group compared to O-Bu group without statistical significance. Our results suggest that intravenous busulfan is as effective as oral busulfan in a conditioning therapy for BMT.
Outcome of Sibling Reduced-Intensity Conditioning Allogeneic Stem Cell Transplantations in Low-Grade B Cell Lymphoproliferative Disorders. A Single Centre Experience.
Authors: Desmond Ronan , David Marianna , Conneally Eibhlin , Browne Paul , Conghaile Mairead Ni , McCann Shaun , Vandenberghe Elizabeth
Published: October 2019

Reduced intensity conditioning allogeneic stem cell transplantations (allo-SCT) have been developed to exploit graft-versus-malignancy effect while reducing the toxicity of allo-SCT. We report the outcome of 21 patients with low-grade B cell lymphoproliferative disorders transplanted in a single center. The mean age of the patients was 54 years (range 32–66) with 11 males, 10 females. Follicular lymphoma was the diagnosis in 10 patients, CLL in 5, mantle cell lymphoma in 4, MALT lymphoma in 1 and lymphoplasmocytoid lymphoma in 1 patient. The median number of prior treatment lines was 2. All patients received peripheral blood stem cell transplants from sibling donors and no patients had undergone a prior autologous SCT. 12/21 patients (57%) received a ATG based conditioning regimen while the remainder, 9/21 (42%) received a Campath containing conditioning regimen. Full donor chimerism was confirmed in 17/19 (89%) patients. Acute graft versus host disease (GVHD) developed in 5/21 (23%) of the patients. Chronic GVHD occurred in 7/21 (33%) with 5/21 occuring post donor lymphocyte infusion (DLI). DLI has been administered in 11/21 patients (52%), 2 for progressive disease and 9 for re-emerging mixed chimerism. Non-relapse mortality (NRM) at 100 days and 1 year were 5% and 12.5% respectively. With a median follow up of 21 months (range 3–54) the overall survival rates (OS) at 1 year and 2 years were 72% and 58% respectively. Progression free survival (PFS) at 1 year and 2 years were 66% and 50%. We have shown that reduced intensity conditioning allogeneic stem cell transplantation is associated with a low TRM comparible to that of autologous transplantation and allows long term control of low grade B-cell lymphoproliferative disorders.
Nonsense-Mediated mRNA Decay Is Essential for the Hematopietic Compartement.
Authors: Weischenfeldt Joachim , Damgaard Inge , Bryder David , Nerlov Claus , Porse Bo
Published: October 2019

Nonsense-mediated mRNA decay (NMD) is a conserved cellular surveillance system that degrades mRNAs with premature termination codons (PTCs). PTC-containing transcripts can arise from faulty events such as erroneous mRNA processing events as well as mutations, and their translation may lead to the synthesis of deleterious proteins. In addition to serving as a genomic protection system, experiments in tissue culture cells have demonstrated that NMD regulates 5% of the normal mRNA pool suggesting that the NMD pathway may have a broader role in gene regulation. Finally, NMD has also been proposed to be important during lymphocyte development as a tool of riding the cells of transcripts resulting from unproductive re-arrangements events of T cell receptor and immunoglobulin genes. Although NMD has been studied extensively at the biochemical level, the actual role and importance of NMD in the mammalian organism has not been investigated. We therefore generated a conditional Upf2 knock-out mouse line (UPF2 being an essential NMD factor) which we crossed to different hematopoietic relevant Cre expressing lines. Full ablation of UPF2 (using the inducible Mx1-Cre deleter) led to complete loss of all nucleated cells in the bone marrow and death of the animals within 10 days. A similar phenotype was observed when Upf2fl/fl; Mx1Cre BM cells were transplanted into lethally irradiated WT recipients and induced with poly-IC, demonstrating the cell autonomous nature of the phenotype. Deletion of UPF2 in the myeloid lineage using the LysM-Cre deleter resulted in efficient ablation of UPF2 and the absence of NMD in reporter transfected bone marrow derived macrophages (BMDMs). However, the steady state levels of myeloid cells appeared unaltered. Finally, deletion of UPF2 in T cells using a Lck-Cre deleter led to a marked reduction of both CD4/CD8 double-positive and single-positive T cells and accumulation of PTC containing transcripts. Gene expression profiling experiments of BMDM and thymocytes from WT and UPF2-ablated animals identified a common core set of 27 up-regulated genes consistent with the role of NMD as a mRNA degrading system. The gene expression profiling data suggest that ablation of NMD leads to accumulation of unfolded proteins. In summary, these studies demonstrate the vital and cell-autonomous role of NMD in the hematopoietic system.
Cotransplantation of HLA-Identical Mesenchymal Stem Cells and Hematopoietic Stem Cells in Chinese Patients with Hematologic Diseases.
Authors: Zhang Xiaoyan , Li Jianyong , Cao Kejiang , Wu Hanxin , Lu Hua , Liu Peng , Qian Sixuan , Xu Wei , Shen Wenyi , Zhu Yu
Published: October 2019

Background: Mesenchymal stem cells(MSCs) can be isolated from bone-marrow and expanded ex-vivo, may support hematopoietic reconstruction and mitigate graft-versus-host disease (GVHD) in hematopoietic stem cells transplantation (HSCT). We hope to explore the feasibility and safety of cotransplantation culture-expanded MSCs and hematopoietic stem cells (HSCs) from the same human leucocyte antigen (HLA)-identical sibling donor in patients with hematologic diseases.

Methods: Bone marrow mononuclear cells from healthy donors were cultured and expanded ex-vivo. Immunophenotype, karyotye, immunosuppressive property of the harvested MSCs were characterized. Patients were cotransplanted with HSCs and MSCs from the same donor. Hematopoietic reconstruction, complications and clinical outcomes after transplantation in these patients were observed.

Results: (1.77±0.40)×106/kg (donor’s weight) MSCs were successfully expanded from 23.6±5.96ml bone marrow samples. They had normal karyotype and were CD73, CD90, CD105 positive and CD34, CD45, HLA-DR negative. They can inhibit mixed lymphocyte reactions (MLRs). Twelve patients were undergoing cotransplantation. No adverse response was observed during and after the infusion of allogenic MSCs. Hematopoietic reconstruction were rapid. Two patients developed grade II∼IV acute GVHD. Two patients developed systermatic chronic GVHD. Four patients suffered from cytomegalovirus (CMV) infection but were cured at last. Till now, seven patients have been alive for 29∼57 months and five patients died.

Conclusion: MSCs identified by immunophenotype analysis can be isolated from human bone marrow, expanded effectively by culture. Their quality and quantity are suitable for clinical use. It is safe and feasible to cotransplant patients with allogenic culture-expanded MSCs and HSCs.
Successful Engraftment of Cord Blood Transplantation Following Reduced-Intensity Conditioning Regimen Using Fludarabine and Busulfan for Advanced Hematologic Malignancies.
Authors: Satou Yuji , Nagasawa Toshiro , Azuma Takayuki , Miura Yuji , Komatsu Tsunehiko
Published: October 2019

Background: The potential role of reduced-intensity cord blood transplantation (RI-CBT) without total body irradiation (TBI) in adults remains unclear. We investigated the feasibility of RI-CBT using non- TBI regimen for the treatment of patients with advanced hematologic malignancies.

Methods: Twenty-three patients (median age, 61, range, 38–74) with advanced hematologic malignancies were enrolled in this study (18 patients in refractory or relapsed phase,5 patients in remission or chronic phase). Conditioning regimen comprised of fludarabine 30 mg/m2 on days −8 to −3, busulfan 4 mg/kg p.o. or 3.2 mg/kg i.v. on days −6 to −5. In one cases. we administered busulfan 3.2mg/kg i.v. on days −6 to −3, because of blastic crisis. Graft-versus-host disease (GVHD) prophylaxis was tacrolimus alone. Engraftment was defined as an absolute neutrophil count > 0.5 x 10E9/l. Primary graft failure was defined as the complete loss of donor-type hematopoiesis without engraftment. Secondary graft failure was defined as the loss of donor-type hematopoiesis after primary engraftment. Median follow-up of surviving patients was 1096 days (range, 53–1207). Primary endpoint was engraftment. All patients provided informed consent in accordance with the requirements of Institutional Review Board.

Results: All the patients tolerated the conditioning regimen. Median dose of infused nuclear cells was 2.7x10E7 /kg (range, 1.9–4.6). Twelve patients achieved engraftment at a median of day 20.5 (range, 10–36), but two of them developed secondary graft failure. Complete donor-type chimerism was documented within 30 days of transplant in six patients. Primary graft failure was diagnosed in remaining eleven patients. Six of them, underlying disease progressed despite conditioning regimens. As of August 2007, five patients survived (3 patients in complete remission, one patients relapsed after transplantation, the other one not reached remission). Estimated 1-year overall survival rate was 20.7% (95% confidence interval, 3.0–38.4%).

Conclusions: Though the pre-transplant condition of patients were not good, engraftment was obtained with approximately the half patients. This study demonstrated the feasibility of RI-CBT using non-TBI regimen for adult patients with advanced hematological diseases; however, high incidences of disease progression before engraftment was significant problem. RI-CBT may become the choice of treatment for patients with advanced hematologic malignancies that are incurable with conventional treatments.
Long-Term Survival after HLA-Haploidentical Stem Cell Transplantation from Non-Inherited Maternal Antigen-Mismatched Family Donors: Impact of Chronic Graft-Versus-Host Disease.
Authors: Ichinohe Tatsuo , Shimazaki Chihiro , Hamaguchi Motohiro , Watanabe Arata , Ishida Hiroyuki , Yoshihara Takao , Morimoto Akira , Uoshima Nobuhiko , Adachi Souichi , Inukai Takeshi , Sawada Akihisa , Oka Koji , Itoh Mitsuru , Hino Masayuki , Kanda Junya , Uchiyama Takashi , Kodera Yoshihisa
Published: October 2019

Background: We previously reported that T-cell-replete HLA-haploidentical hematopoietic stem cell transplantation (SCT) from a microchimeric noninherited maternal HLA antigen (NIMA)-mismatched related donor offers a feasible treatment option in patients with poor-risk hematologic malignancies who lack an immediately available stem cell source (Ichinohe T, et al. Blood2004;104:3821). However, long-term outcomes and late effects among patients undergoing such transplantation are largely unknown.

Methods: We studied 16 patients who had survived more than 3 years after T-cell-replete HLA-haploidentical NIMA-mismatched SCT to evaluate the impact of late complications on morbidity and mortality. The patients received bone marrow (n=5) or peripheral blood stem cell (n=11) between 01/2001 and 07/2004 at 11 centers with a median age of 19 years (range, 2 to 56) as treatment for acute myeloid leukemia (n=6), acute lymphoblastic leukemia (n=3), chronic myeloid leukemia (n=4), and other B-cell neoplasms (n=3). At the time of SCT, 6 patients had a chemosensitive disease and 10 had a chemorefractory disease. Type of donor was NIMA-mismatched sibling in 9, mother in 6, and daughter in 1; all patient-donor pairs had two or three serologic mismatches at HLA-A, −B, and −DR in the graft-versus-host direction. Organ-specific symptoms related to chronic graft-versus-host disease (cGVHD) and their severity were evaluated by clinical scoring system proposed by the National Institutes of Health consensus development project.

Results: At a median follow-up of 56 months (range, 38 to 74), 13 (81%) of 16 patients were alive and free of their primary disease. One patient was alive with relapsed disease; 2 died from bronchiolitis obliterans at 51 and 52 months in continuous remission. Karnofsky or Lansky performance score among 14 survivors was 100% in 6 (43%), 80–90% in 5 (36%), 70% in 2 (14%), and less than 70% in 1 (7%). Thirteen (81%) patients developed extensive cGVHD and eleven of them experienced organ symptoms corresponding to the consensus score greater than 1. The commonly involved organs (score>1) were lungs (n=5), skin (n=4), eyes (n=3), and liver (n=3). Seven patients were successfully withdrawn from immunosuppressive agents between 3 and 46 months (median, 19) after transplantation.

Conclusions: T-cell-replete HLA-haploidentical SCT from a microchimeric NIMA-mismatched donor confers long-term survival in selected patients without compromising their performance status, although a high incidence of moderate to severe cGVHD may limit its usefulness as compared with conventional SCT.
Inclusion of High-Dose Treosulfan Followed by Autologous Transplantation in the Tandem Transplantation for Patients with Multiple Myeloma <60 Years.
Authors: Reichle Albrecht , Burgmayer Nina , Holler Ernst , Berand Anna , Andreesen Reinhard
Published: October 2019

High-dose treosulfan followed by autologous transplantation is a tolerable and efficacious regimen in quite different tumor types. Aim of the present study was to proof safety and efficacy of high-dose treosulfan in an intraindividual comparison with melphalan. Between August 2003 and July 2006, 20 patients with multiple myeloma, age< 60 years were enrolled onto an unicentric phase II trial. The median age was 55 years. All patients received pretreatment with 3 to 4 cycles idarubicin and dexamethasone (90%) or CAD (10%). After induction the peripheral blood stem cells were mobilized with a combination of ifosfamide, epirubicin, etoposide (IEV) followed by G-CSF. Stem cell harvest was successful in all patients. Thereafter, the patients received tandem transplantation in an interval of 2 months, first after conditioning with high-dose treosulfan on day −4 to −2, 14g/m2 daily, then after high-dose melphalan 140 mg/m2. Maintenance therapy with interferon-alpha was administered in 30% of the cases. For all patients the completion rate of the first transplantation was 100%, for the second 85%. Three patients did not proceed to the second autologous transplantation due to a severe ischemic colitis (n= 1) or a high-risk profile (allogeneic transplantation in 2 cases). Two patients received an allogeneic transplantation during the further course of the disease. Overall the conditioning regimens were well tolerable. Hematotoxicity grade 4 was observed after each high-dose cycle. Grade 3/4 infections and stomatitis were present in 5%/5% after treosulfan and in 18%/6% after melphalan. An acute coronary heart syndrome occurred after Mel140. The duration of severe leukopenia (< 1.0 leukocytes) was significantly shorter after treosulfan (6.4 days vs. 7.9 days, p= 0.009), whereas time to leukocyte recovery did not significantly differ between the regimens. No treatment related deaths occurred. Best response after transplantation was CR 30%, PR 60%, NC 5%, and PD 5%. In comparison to the preceding chemotherapy a further >50% decline of the paraprotein was achieved by treosulfan in 3 cases, by melphalan in 2 cases. In the intraindividual comparison treosulfan and melphalan, respectively, have shown two times superior response. The median event-free survival was 28 months, and the overall survival at 4 years was 81%. In conclusion, high-dose treosulfan may be more favorable for patients at risk for infections than melphalan, and seems to be as efficacious as melphalan for the treatment of multiple myeloma. Treosulfan should be further investigated in multiple myeloma patients.
Darbepoietin in Autologous Peripheral Blood Stem Cell Transplantation (ASCT): A Pilot Study.
Authors: Chierichini Anna , Anaclerico Barbara , Borza Paola Anticoli , Fenu Susanna , Nobile Carolina , Cedrone Michele , Bongarzoni Velia , Bartolini Maurizio , Ronci Benedetto , Annino Luciana
Published: October 2019

In patients undergoing ASCT, RBC transfusions are depending on donor availability and correlated with early and late side effects such as immune reactions and infections. Three weeks long acting Darbepoietin is currently available, but its role in ASCT in terms of efficacy, safety and cost has not yet been established.

Aim: This pilot study was activated in ASCT patients to test

the impact of Darbepoietin on the haemoglobin level; RBC transfusion requirement; the total cost of Darbepoietin therapy during the first 30 days after ASCT.

Material and methods. From September 2006 to June 2007, 10 patients - 5 males, median age 57 years (range 20–71) - with Multiple Myeloma (MM n=7) in 1st response, Non-Hodgkin (NHL n=2) and Hodgkin (HL n=1) Lymphoma in 2nd complete remission entered the study. As conditioning regimens, the patients received Mel 200 (n=8) or BEAM (n=2). Baseline Hb and Epo median values were: 12,1g/L (range 10.3 – 16.2 g/L), and 28.7mU/mL (range, 4.1–220 mU/L), respectively. The median value of CD34+ cells infused was 4.99 x109/L (range 3.04 – 5.07) and the median time to engraftment was 10 days (range 8–11). To prevent anaemia, patients were given i.c. single dose 500 U Darbepoietin on day +1, associated with iron therapy and B12 vitamin /weekly, plus folinic acid and B6 vitamin/daily. RBC transfusion was mandatory for Hb level ≤ 8g/L.

Results: At day +30, in 9/10 patients (7 MM, 1 NHL and 1HL) the median Hb value was 13.2 g/L (range 10.2 – 15.1g/L), and none of them were transfused. Only 1 NHL received 1 RBC unit on day +7 because of Hb was 8g/L; in this case, the baseline dosage of Epo was high (220 mU/L). On the follow-up (median time 4 months, range 1–11), no thromboembolic events or other side effects occurred. Finally, in our Country the price of 1 vial Darbepoietin and 1 RBC transfusion (packed, filtered and irradiated) is 744.60 and 340 euro, respectively. Therefore,, the cost of the entire supportive therapy in our 10 patients has been 7.786 euro. Comments Despite the small number of patients, from this pilot study we can drawn the conclusion that Darbepoietin seems an effective and safe therapy and could represent an appropriate support in ASCT patients with baseline low/normal Epo level. Only a prospective, randomized study on a large number of patients will be able to answer the question on the Darbepoietin cost-efficacy in ASCT.
Human Herpesvirus 8 K1 Derived Peptides Reduced the Levels of Apoptosis Inhibitory Fas-K1 Complex: A Strategy for Stimulating Death Receptor Mediated Apoptosis.
Authors: Wang Shu , Berkova Zuzana , Wang Suizhao , Pharm Jaehoon Jung D. , Maeng Hoyoung , Samaniego Felipe
Published: October 2019

Background: Human herpesvirus 8 (HHV-8) infection is associated with the development of primary effusion lymphoma, Kaposi’s sarcoma and multicentric Castleman’s disease. The K1 gene of HHV-8 is expressed in some of these tumor cells as a transmembrane protein with an immunoglobulin like domain in its ectodomain and an immunoreceptor tyrosine-based activation motif (ITAM). We demonstrated that K1 protein activates nuclear factor of kappa B (NF-κB), and K1 expression in transgenic mice stimulated accumulation of lymphatic cells and development of lymphoma. How K1 blocks apoptosis, induces hyperplasia and lymphomas is not known. We hypothesized that K1 contributes to lymphoma development partly by suppressing apoptosis and this suppression combined with its NF-κB activation produces lymphoma.

Results: We mapped the region K1 uses to bind to Fas as an immunoglobulin (Ig) chain-like domain by expressing deletion mutants of K1. Overexpression of an Ig domain-contain protein CD79b competed with K1-Fas binding that was dose-dependent. Two 20 amino acid peptides (N251, N253) representing the Ig domain of K1 competed with K1-Fas binding in immunoprecipitation/immunoblotting analysis. The N251 and N253 peptides (100 μM) enhanced anti-Fas-(Ch-11, 50 ng/ml) induced apoptosis of BJAB lymphoma cells that expressed K1 but not of vector transfected BJAB cells. Ig deleted K1 (K1dIg) transfected mice were not protected (0/6) and K1 transfected mice were protected (7/10), P<0.01) against the lethal effects of agonistic anti-Fas (Jo2) antibody. K1dIg expressed in mice did not form complexes with Fas suggesting that the Ig is essential for K1-Fas binding and suppression of apoptosis.

Conclusion: Collectively, these results indicate that K1 potently blocks apoptosis and this effect is mediated through the Ig-like domain of K1. Because viral proteins mimic cellular proteins these results predict the presence of functional cellular homologues of K1 that have key roles in death receptor regulation.
Impact of the Addition of Rituximab to Initial CHOP Chemotherapy Compared with CHOP Alone in Patients with Relapsed Diffuse Large B-Cell Lymphoma Who Underwent Autologous Stem Cell Transplantation.
Authors: Chen Yi-Bin , Hochberg Ephraim P. , Feng Yang , Motyckova Gabriela , Neuberg Donna , Spitzer Thomas R. , Fisher David C. , LaCasce Ann S.
Published: October 2019

Standard therapy for chemosensitive relapsed diffuse large B-cell lymphoma (DLBCL) is high-dose chemotherapy followed by autologous stem cell transplantation (ASCT) based on the results of the Parma study (NEJM1995;333:1540–5). However, the addition of rituximab to initial chemotherapy has improved progression-free and overall survival compared with CHOP alone. Patients who relapse after R-CHOP may have disease which is less sensitive to second line therapy or, alternatively, the benefit of ASCT may be independent of the impact of rituxuimab, and, therefore, the true benefit of ASCT in current patients with relapsed DLBCL is unclear. We performed a retrospective cohort analysis of patients who received ASCT for DLBCL during the period of 1997–2006. Our goal was to evaluate the benefit of ASCT for patients with relapsed DLBCL who received R-CHOP as initial chemotherapy compared to those who were treated with CHOP alone. A total of 145 patients at first relapse with chemotherapy sensitive disease who underwent ASCT were identified. 111 received a CHOP-like regimen, and 34 received R-CHOP as their initial chemotherapy regimen. All patients received salvage chemotherapy prior to ASCT. 86.5% of CHOP patients and 88.2% of R-CHOP patients achieved a CR/CRu with their initial chemotherapy (p=0.99). At initial diagnosis, Ann Arbor stage and ECOG performance status (PS) between the two groups were also similar. The two groups of patients had significant differences in the following characteristics: median age at ASCT (51 for CHOP vs 57 for R-CHOP, p=0.002), Ann Arbor stage of disease at relapse (22.5% localized disease for CHOP vs 55.9% localized disease for R-CHOP, p=0.001), and percentage of patients who underwent nuclear imaging to evaluate response to salvage chemotherapy (37.8% in CHOP vs 79.4% in R-CHOP, p<0.0001). The majority of patients in both groups received either CBV or Bu/Cy conditioning chemotherapy at ASCT. 2-year relapse-free survival (RFS) was 57% in the CHOP group versus 35% in the R-CHOP/ASCT group (p=0.10). 2-year OS was 75% (CHOP) and 67% (R-CHOP/ASCT), respectively (p=0.09). Longer follow-up is essential and further analysis examining the potential impact of rituximab containing salvage therapy in rituximab naïve patients is needed. In conclusion, although this is a retrospective comparison, the trend toward inferior RFS and OS in the patients who initially received R-CHOP suggests that the benefit of ASCT in patients who subsequently relapse after R-CHOP may be inferior compared to patients who received CHOP alone.

Figure Figure
Low Predictive Value of Functional Imaging with 18FDG-PET or Gallium Scanning in Relapsed or Refractory Hodgkin Lymphoma Prior to Autologous Stem Cell Transplantation.
Authors: Peinert Stefan , Hoyt Rosemary , Szer Jeff , Roberts Andrew , Grigg Andrew , Prince H. Miles , Rigacci Luigi , Seymour John F. , Ritchie David S.
Published: October 2019

Introduction: 18FDG-PET early during first-line chemotherapy of Hodgkin Lymphoma (HL) has a high positive and negative predictive value. The predictive value of functional imaging (FI) in refractory or relapsed HL after salvage treatment and prior to high-dose (HD) chemotherapy and autologous stem cell transplantation (AUSCT) is less well established.

Methods: A retrospective analysis was undertaken of the clinical characteristics, FI results, event free survival (EFS) and over all survival (OS) data for 66 consecutive patients (pts) with refractory or relapsed HL treated with salvage chemotherapy and AUSCT between 1997 and 2007.

Results: The median follow-up was 52 months (range 0 to 125 months). Following salvage chemotherapy, and prior to AUSCT, 63 of the 66 evaluable pts (96%) had response assessment with a conventional CT scan, 53 pts also had FI with a gallium scan (n=18), 18FDG-PET scan (n=31) or both (n=4). Of the 53 pts with FI prior to AUSCT, 21 (40%) were in metabolic complete remission (FI-negative) and 32 pts (60%) had residual disease (FI-positive). The median EFS and OS for the whole cohort were 29 and 52 months, respectively. Median EFS was 31 and 13 months (p=0.096), and median OS was 60 and 39 months (p=0.35) for those with negative and positive FI, respectively. Significantly more pts with negative FI achieved CR with conventional imaging compared to those with positive FI (9/19 = 47% vs. 2/32 = 6%, p=0.001). Male pts were disproportionately represented in the cohort with positive FI (20/32) compared to the pts with negative FI (7/21, p=0.038). There was no association between FI-positivity and the clinical features of age, short duration (<12 months) of initial response, presence of B-symptoms or extranodal disease at the time of relapse, or the type of AUSCT conditioning used.

Conclusion: We did not demonstrate a strong predictive value of FI outcome prior to AUSCT for relapsed or refractory HL despite a trend for improved EFS for pts with negative FI. We did demonstrate that males had a higher rate of FI positivity suggesting this is also an adverse risk factor for failure of salvage therapy in relapsed/refractory HL.
Novel Efficient Primary Human Peripheral Blood Progenitor Cell-Targeted Recombinant Adeno-Associated Viral Vectors.
Authors: Veldwijk Marlon R. , Sellner Leopold , Stiefelhagen Marius , Kleinschmidt Jürgen A. , Laufs Stephanie , Topaly Julian , Wenz Frederik , Zeller W. Jens , Fruehauf Stefan
Published: October 2019

Introduction: G-CSF mobilized peripheral blood progenitor cells (PBPC) are an excellent target for the cure of acquired and inherited disorders of the hematopoietic system. AAV-based vectors have an advantageous safety profile, due to their low immunogenicity, the lack of any associated human disease and its episomal localization. Unfortunately, currently standard rAAV2-based vectors lack the required efficiency for gene transfer into human blood progenitor cells. To increase the gene transfer efficiency on primary hematopoietic cells we applied an AAV random peptide library on primary human CD34+ PBPC.

Materials and Methods: The AAV random peptide library was applied on primary human CD34+ PBPC samples (3 selection rounds). Several mutant blood progenitor-targeted rAAV2 capsid clones were obtained and sequenced. Five new rAAV vectors with the most efficient clones were produced. Titers were verified using real time PCR-based titration. To determine efficiency and specificity, a panel of leukemia (CML and AML) and non-leukemia cell lines, as well as primary human CD34+ PBPC were transduced. GFP expression was determined against uninfected control cells.

Results: On a panel of leukemia cell lines at least one capsid mutant was significantly more efficient (%GFP+; p<0.01) than standard rAAV2 vectors. On solid tumor cell lines rAAV2 was more efficient, suggesting the increased specificity of our capsid mutants for hematopoietic cells. Significantly higher gene transfer efficiency (up to 16% GFP+ cells) could be obtained with the newly generated vectors compared to standard rAAV2 (up to 9-fold higher; p<0.01). Using a different approach, up to 25% gene transfer into CD34+ PBPC was observed.

Conclusion: These novel vectors may finally enable efficient (>20%) gene transfer using rAAV-based vectors into primary human PBPC and may offer a safe alternative to other vector systems which is applicable in vivo.
Evaluation of a Powered Intraosseous Device for Bone Marrow Sampling.
Authors: Cohen Stephen C. , Soroka Jill M.
Published: October 2019

The importance of bone marrow examination in the evaluation of leukemia, multiple myeloma, anemia, pancytopenia, and other disorders is well established. The objective for this study was to evaluate the ability of a powered bone marrow aspiration device to penetrate the intraosseous medullary space of the iliac crest, and to aspirate bone marrow samples for the ultimate purpose of diagnosing disease and monitoring the course of disease and medical therapy. The device was used to obtain bone marrow samples in accordance with accepted practice guidelines and device’s directions for use. Among other data, insertion success, time to insertion, and complications were recorded. Patient pain levels were rated from 0 to 10 (10=extreme pain). Device operators rated the use of the device from 0 to 10 (10=outstanding). There were 55 patients in the study from three centers. Successful insertion and aspiration of bone marrow samples were achieved in 54 of the 55 patients (98.1%). Mean insertion time was 4.9±3.0 seconds; significantly faster than the 7.3 minutes reported by Kuball et al* (one-sample t-test, p<0.001). There were no complications. The mean insertion pain score was 2.5±2.2 and the mean aspiration pain score was 3.7±2.5. On a scale of 0 to 10, the six operators rated the ease of use of the device at a mean score of 8.3±1.7. Findings suggest that the powered aspiration device is safe and effective for bone marrow aspirations.
Nonhematological Tumor Metastasis in the Bone Marrow: An Analysis of 10112 Unselected Plastic-Embedded Biopsy Sections.
Authors: Li Xiao , Tao Ying , Liu Yizhi , Song Luxi , Wu Lingyun , Ying Shaoxu , Chang Chunkang , Pu Quan
Published: October 2019

We investigated the frequency of bone marrow (BM) involvement by nonhematological tumors in unselected BM biopsy samples from 10,112 patients. Plastic embedded BM biopsy sections were retrospectively analyzed, with special focus on

Frequency of BM involvement by nonhematological malignancies, the advantage of plastic-embedded BM biopsy specimens over paraffin-embedded samples, Complementarity of marrow biopsy to aspiration smears. the causes of false-negative results when using aspiration smears, and analysis of primary sites of metastatic solid tumors.

Overall 101 of the 10112 BM samples showed nonhematological tumor metastases, accounting for 1.0% of all analyzed specimens, which was higher than data reported on the basis of paraffin embedded samples. Of 74 aspiration smears obtained concurrently with the 101 biopsy sections, 55 were also positive for metastatic carcinoma. That is, the detection rate of metastatic tumors based on the aspiration smears was 74.3% relative to the rate observed on biopsy sections. All 101 sections with metastatic tumors showed various degrees of myelofibrosis (including 67cases with severe myelofibrosis [+++]) which was thought to be partly responsible for false negative results on aspiration samples. Primary tumors were documented ante-mortem in 50 of the 101 BM biopsy-positive patients (49.5%). The most frequent primary sites were lung, gastroinyestinal tract, and breast, in good agreement with data in the literature. In conclusion, nonhematological tumor metastases in unselected bone marrow are not exceptional findings. Plastic embedded BM biopsy examination is helpful in detecting unknown metastatic nonhematological malignancies and apparently more sensitive than paraffin embedded BM biopsies.

Figure Figure Figure Figure
The Association between Hematological and Inflammatory Factors and Stress Fractures among Female Military Recruits.
Authors: Merkel Drorit , Moran Daniel S. , Yanovich Ran , Evans Rachel K. , Finestone Aharon S. , Constantini Naama , Israeli Eran
Published: October 2019

Background: With growing number of females accepted for combat-related military duties in Israel, the special needs of these women should be addressed. Previous studies on women in combat training have found a high prevalence of iron deficiency at recruitment, as well as an increased rate of Stress Fractures (SF) and overuse injuries during training in comparison with men. The aim of this study was to assess the association between hematological and inflammatory variables and stress fracture occurrence among military recruits during basic training.

Methods: We followed 3 companies of females and males inducted to a gender integrated light infantry unit, prospectively. Females inducted for a medics and dental assistants’ courses served as controls. Hemoglobin, iron, transferrin, ferritin, CRP and IL-6 levels were measured for all participants, at recruitment, and after 2 and 4 months of training. SF’s were diagnosed radiographically or scintigraphically according to the IDF protocol.

Results: 227 female combatants, 83 male combatants and 128 female controls were recruited. At induction, 18% of the female combatants had anemia compared with 8% of the males and 19% of the female controls. Iron deficiency was noted in 40%, 6%, and 38% respectively. There were no clinically significant changes during training. Twelve percent of female combatants developed stress fractures, while there were no occurrences of SF among the male combatants or female controls. Subjects with SF had significantly lower levels of serum iron and iron saturation.

Conclusions: A relatively high incidence of anemia as well as iron deficiency was found in this young asymptomatic cohort, with no significant change during training. The lower stores of iron in the female combatants with stress fractures warrants further investigation.
Canonical Correlation Analysis (CCA) of Imatinib Treatment (ImRx) Nonadherence (NA) with Associated Patient Variables (APVs) in Chronic Myeloid Leukemia (CML) - Results from the ADAGIO Study.
Authors: van Lierde Marie-Anne , De Rop Larissa , Serra Filipo , Strobbe Eric , MacDonald Karen , Vancayzeele Stefaan , De Geest Sabina , Letvak Laurie , Albrecht Tara , Abraham Ivo
Published: October 2019

BACKGROUND. ImRx for CML is a long-term treatment potentially compromised by NA. Identifying APVs may assist in reducing NA and optimizing treatment outcomes.

OBJECTIVE. To model the relationships between two NA measures and selected APVs using CCA, a multivariate analog of multiple regression to accommodate multiple criterion variables.

DESIGN AND PATIENTS. Data subset from prospective, 90d observational, open-label, multicenter study. 169 evaluable pts on ImRx for minimum 30d at enrollment [1].

MEASUREMENTS. NA at 90d vector: Basel Assessment of Adherence Scale for pts (pBAAS; 0/1 with 1=NA); and % of ImRx taken per pill count (%ImRx, subtracted from 100 to reflect NA). APVs at BL vector: age; months since CML diagnosis (mCML); months since ImRx initiation (mImRx); knowledge of CML disease, treatment, and ImRx (KCMLRx); and general health (SF-8).

RESULTS. The criterion (dependent) vector of NA indicators included pBAAS and %ImRX. The predictor (independent) vector of APVs included: age, mCML, mImRx, KCMLRx, and SF-8. Two canonical correlations were generated: 0.389 (Bartlett Chi-squared=23.564, P=0.009) and 0.170 (Bartlett Chi-squared= 3.590, P=0.464); the second correlation was deleted due to nonsignificance from zero. The canonical loadings (or structure coefficients) for the retained model were: age 0.951, mCML 0.205, mImRx 0.145, SF-8 0.016, and KCMLRx -0.367. Redundancy analysis showed that 22.1% of variance in the predictor set was explained by variables within that set.

CONCLUSIONS. The patient NA vector (composed of a binary assessment of NA per pBAAS 0/1 with 1=NA) and continuous quantification of % of ImRx not taken was related to the APV vector as follows: NA increased as patients were older, had been diagnosed with CML for a longer period of time, had been on imatinib treatment for a longer period time, and were in slightly better health at enrollment. These may be considered warning signs for NA for clinicians to consider in practice, given the long-term nature of ImRx. Importantly, better patient knowledge of disease and treatment, a clinically modifiable APV, was associated with a decrease in NA. The initial insights in patient NA with ImRX provided by these findings, though some are counterintuitive to the NA literature at large, must be studied further to better understand the dynamics of NA in the CML population.
LMO2 Protein Expression Predicts Survival in Patients with Diffuse Large B-Cell Lymphoma in the Pre- and Post-Rituximab Treatment Eras.
Authors: Natkunam Yasodha , Farinha Pedro , Hsi Eric D. , Hans Christine P. , Tibshirani Robert , Sehn Laurie H. , Connors Joseph M. , Zhao Shuchun , Pohlman Brad , Spinelli John , Bast Martin , Nagler Arnon , Levy Ronald , Gascoyne Randy D. , Lossos Izidore S.
Published: October 2019

Finding new prognostic markers in diffuse large B-cell lymphoma (DLBCL) is of importance because the disease is heterogeneous and current therapies fail to cure many affected patients. Previously, we showed that the expression of LMO2 mRNA was the strongest single predictor of superior outcome in patients with DLBCL in a multivariate model based on the expression of six genes (Lossos et al, NEJM 2004). Subsequently, we generated an anti-LMO2 monoclonal antibody and showed that LMO2 protein is expressed in a subset of germinal center-derived B-cell lymphomas including DLBCL, and that its expression predicted a superior outcome in DLBCL patients who were treated with anthracycline-based regimens (Natkunam et al, Blood 2006 & 2007). We have now expanded this cohort to include DLBCL patients treated at five international medical centers and have also tested the prognostic impact of LMO2 in patients treated with the addition of rituximab to CHOP (R-CHOP), a regimen which was recently demonstrated to improve the outcome of DLBCL patients (Coiffier et al, NEJM 2002; Sehn et al, JCO 2005; Habermann et al, JCO 2006; Pfreundschuh et al, Lancet Oncol 2006). Immunohistochemistry for LMO2 was performed on tissue microarrays containing cores of diagnostic biopsy specimens from patients with de novo DLBCL treated with anthracycline-based chemotherapy (280 patients) or with R-CHOP (80 patients) who had been followed for clinical outcome. The results show that in the pre-rituximab group staining for LMO2 was strongly correlated with overall survival (OS) and progression-free survival (PFS) (p=0.0018 and p=0.00071, respectively). Similarly, in the post-rituximab group LMO2 protein expression was associated with a significantly longer OS (p=0.0048) and PFS (p=0.0087). In multivariate Cox regression analyses, the expression of the LMO2 protein was found to be independent of the clinical IPI and most strikingly, was more robust than the conventional IPI in predicting OS and PFS, particularly in the post-rituximab DLBCL group of patients (OS p = 0.03, PFS p = 0.01). The capacity of LMO2 protein to identify DLBCL patients with improved outcome in an IPI-independent manner indicates its potential role in risk stratification of this disease. We conclude that the prognostic value of LMO2 protein expression remains significant in the era of R-CHOP treatment and recommend its assessment in all newly diagnosed DLBCL patients to confirm these results and eventually to optimize patient management.