bluuebunny/crossref_metadata_2025 huggingface dataset:
Primary Oral Cavity Lymphomas. Features of 29 Patients Assisted at Oncology Hospital Marie Curie, Buenos Aires Argentina.
Authors: Dragosky Marta O. , Frascino Nelly , Alcaraz Susana , Annetta Isabel , Devoto Laura , Luchetta Patricia , Marquez Marisa , Giaco Carmen , Gallo Karina , Acosta Gabriela , Alcoba Elsa , Stagnaro Jorgelina , Guerrero Laura
Published: October 2019

Primary oral cavity lymphomas (POCV) are uncommon and must be distinguish from widespread involvement of systemic lymphomas.

Countries with high incidence of oral malignancies report cancer predominance. HIV+ population have higher prevalence than immunocompetent patientes. Alcohol and tobacco have been mentioned like risk factors. The International Prognostic Index has been taken in consideration. Patients attended Hospital on referal from medical or odontology centers and others on their own. A total of 29 patientes were studied. Aged 21 to 70, media 47,3 years. Male predominance: 69%. Histopathology: NHL high grade B: 19 cases, NHL low grade B: 4, NHL high grade T: 3 and plasmoblastic 3. Sites: superior maxilla: 12, inferior maxilla 11, tongue 3, palate 3. No predominance for right or left locations. HIV+ 9 patients (31%) The prevalence among inmmunocompetent was markedly lower: in 662 diagnosed with NHL 20 cases=3,02%, in 32 NHL HIV+ 9 cases= 28,12%. More frequent complaints were: painless swell of the maxilla, movable teeth, dysphagia, facial edema. The stages were I and II, 28% with locoregional extension: adenopathies or bone infiltration. The IPI was 0 in17, 1 in 10 and 2 in 2 patients. All patients received chemotherapy (CVP, CHOP), with initial good response and early relapse in HIV+.

Conclusions: In correspondence with others reports, we found predominance in male and middle age, B cell lineage and connection with HIV infection. POCL may lead to the diagnosis of an up to then unknown HIV infection. The little specificity of the initial symptons tends to delay the diagnosis and the referal to a specialized center. IPI didn’t prove useful as a prognostic assessment.
Effects of Integrin α5β1-Mediated Signal TransductionPathways on the Proliferation and Apoptosis of K562 and CML Cells.
Authors: Pan Ling , Niu Zhiyun , Zhang Xuejun , Wang Fuxu , Suo Xiaohui , Dong Zuoren
Published: October 2019

Objective and methods: More than 90% of chronic myeloid leukemia (CML) patients have the Philadelphia (Ph) chromosome which produces a specific oncoprotein known as BCR/ABL (P210 protein). Having strong tyrosine kinase activity, the BCR/ABL oncoprotein can trigger several signal transduction pathways and inhibit the normal signal transduction pathways of CML progenitor cells. Presenting on cell surface, Integrins belong to the adhesive molecule family. They mediate adhesion of cell to cell or cell to extra cellular matrix (ECM) and take part in many physiological and pathological processes such as signaling transduction, cell differentiation, proliferation and apoptosis. Many studies showed that abnormal circulating of CML Ph+ cells in peripheral blood and unregulated proliferation of CML Ph+ cells in bone marrow were related closely to the abnormal adhesion function of integrins. It is also well known that Interferon (IFN)-α is one of the effective therapeutic agents for CML. Therefore the present study was designed, by cell culture, FCM, RT-PCR and Western Blot, to observe the effects of IFNα-2b on the proliferation of k562 cells and fresh leukemia cells from patients with CML-in blast crisis (CML cells), before and after integrin α5β1 having been neutralized by Anti-integrin α5β1 monoclonal antibody (Anti-α5β1), and to investigate the changes of mRNA and protein of several key molecules in integrin-mediated pathways, such as FAK, ERK, AKT, PI3K and Cychin D1, p21 and survivin before and after the treatment of Anti-α5β1 with or without IFNα-2b.

Results: The expression level of integrin α5β1 on K562 cells and CML cells was much higher than that on health volunteers’ BMMNC. And the adhesion capability of K562 cells and CML cells to fibronectin (FN) decreased significantly than that of health volunteers’ BMMNC. IFNα-2b improved the adhesion capability of K562 cells and CML cells to FN. Anti-α5β1 inhibited proliferation but induced apoptosis and G0/G1 arresting of K562 cells and CML cells. Anti-α5β1 downregulated the expression level of p-FAK-, p-ERK-, PI3K- and p-AKT-mRNA and protein. Anti-α5β1 also downregulated the expression level of Cyclin D1 mRNA, but upregulated the expression level of P21 mRNA. Moreover, the effect of Anti-α5β1 is stronger than that of IFNα-2b.

Conclusions: Anti-α5β1 inhibited integrin α5β1-mediated signal transduction pathways: FAK-ras-ERK pathway and PI3K-AKT-survivin pathway. This might be the mechanisms of Anti-α5β1 inhibiting proliferation and inducing apoptosis of K562 cellsand CML cells. This indicated that the dysfunction of integrin α5β1 pathway might have important meanings in the leukemogenesis of CML and might become a new target for the treatment of CM.
Blockage of IL-6 Mediated JAK2/STAT3 Cell Signaling Pathway by sIL-6R Effectively Inhibited the Growth of Multiple Myeloma Cell.
Authors: Park Ju-won , Ahn Kwang-Sung , Kim Young-Ju , Bae Eunkyung , Lee Yong-Yeol , Yoon Sung-Soo , Kim Inho , Park Seonyang , Kim Byoung-Kook
Published: October 2019

Interleukin-6 (IL-6) is a key growth factor and thus plays a pivotal role in the pathogenesis of multiple myeloma. IL-6 is absolutely necessary for the cellular signalling cascade via JAK/STAT and RAS/MAPK pathways involved in proliferation and viability. We found interleukin (IL)-6 induced JAK-2 phosphorylation and induced proliferation of multiple myeloma cell line. Also phosphorylation of Stat1 and Stat3 was increased by IL-6 stimulation. Silencing JAK2 expression by small interfering RNA (siRNA) abrogated IL-6 induced phosphorylation of STAT3. In addition, soluble interleukin-6 receptor (sIL-6R) antagonized the function of hIL-6 and efficiently induced the growth arrest and apoptosis of U266 cells in a dose-dependent manner. Increasing levels of sIL-6R suppressed the phosphorylation of JAK-2 and Stat3. This suggests a comprehensive knowledge of the signaling and survival pathways could help find additional molecular targets and lead to the development of novel and more effective treatment strategies in multiple myeloma.
Treatment of Diuretic Refractory Pleural Effusions with Bevacizumab in Four Patients with Primary Systemic Amyloidosis (AL).
Authors: Hoyer Robert J. , Leung Nelson , Witzig Thomas E. , Lacy Martha Q.
Published: October 2019

Introduction

Refractory pleural effusions present a challenging management problem and are associated with a poor prognosis in patients with primary systemic amyloidosis (AL). Vascular endothelial growth factor (VEGF) has been implicated to play a role in both malignant and non-malignant pleural effusions through its ability to regulate vascular permeability. VEGF can induce morphologic changes in vascular endothelial cells, regulate gene expression, stimulate cell proliferation and migration, and inhibit apoptosis. We report a series of four patients with AL who presented with bilateral pleural effusions that were refractory to diuretic therapy. After treatment with bevacizumab, an anti-VEGF antibody, three of the four patients had improvement in their pleural effusions, peripheral edema, and functional status.

Methods

A retrospective chart review was conducted four patients with AL and refractory pleural effusions who received bevacizumab. Three of the patients had documented renal amyloid by biopsy and two had suspected cardiac involvement. Serum creatinine ranged from 1.3–3.8 mg/dL (mean=2.15 mg/dL), 24 hour urine protein ranged from 190 mg – 12.5 grams per 24 hours (mean=6.1 grams). Bevacizumab 5 mg/kg was administered for 1–4 cycles (mean=2).

Results

Three of the four patients had demonstrable improvement in their pleural effusions, peripheral edema and functional status after a single dose of bevacizumab. One patient had no objective response to treatment. The three responders experienced a 2–7 kg (mean= 4.6 kg) decrease in weight within two weeks after receiving bevacizumab. This effect was observed in patients who were re-treated. All three patients received concurrent diuretic therapy, however, they had not previously responded to diuretic therapy. Two weeks after the first dose of bevacizumab, the 24 hour urine protein of the patient who received four cycles of bevacizumab increased to 18.2 grams per 24 hours, however it returned to baseline (12.5 grams per 24 hours) one week later, and increased slightly to 13.3 grams per 24 hours after the fourth dose. A second patient also experienced an increase in 24 hour protein excretion (6.9 grams per 24 hours to 9.2 grams per 24 hours) after the first dose of bevacizumab. One patient experienced a deep vein thrombosis after the third cycle of bevacizumab. No hemorrhagic complications were observed.

Conclusion

Bevacizumab may represent a novel non-invasive management strategy for patients with systemic amyloidosis and diuretic-refractory pleural effusions, however, additional studies are needed to further define the role of bevacizumab in the management of this group of patients.
Myeloablative Conditioning Followed by T-Cell Depletion Is Associated with Low Treatment Related Mortality.
Authors: Novitzky Nicolas , Thomas Valda , Toit Cecile du , McDonald Andrew
Published: October 2019

Introduction: Classically the conditioning for transplantation in haematological malignancies has been with myeloablative doses of radiation or chemotherapy, and was followed by variable immunosuppression for the prevention of GvHD. This strategy has been associated with substantial transplant related mortality from regimen toxicity and GvHD, which were most pronounced in older patients. Reduced intensity conditioning programs that are mainly immunosuppressive allow engraftment and predisposed to donor chimerism and appear to have lower TRM, but seem associated to higher rates of GvHD and disease recurrence. To better apportion conditioning strategies for patients undergoing allogeneic stem cell transplantation, we studied the outcome of 81 patients who received similar GvHD prophylaxis with T-cell depletion and immunosuppression, after myeloablative conditioning.

Patients & methods: Patients with haematological malignancies, in remission or still responsive to chemotherapy who had an HLA identical sibling were offered T-cell depleted stem cell grafts. Conditioning was with ablative doses of either chemotherapy or total body radiotherapy. Stem cells were mobilised into the blood (PBPC) with G-CSF (5–10ug/kg × 5) and grafts were harvested by large volume (30 litres) apheresis. GvHD prophylaxis was by ex vivo depletion of lymphocytes from the graft with CAMPATH-1 H antibodies followed by therapeutic doses of cyclosporin until day 90. End points were TRM, disease recurrence rate and overall survival time.

Results: 90 consecutive patients with median age of 45 (17–62) years were studied. The diagnosis included acute leukaemia (ALL in 7) in CR in 38, myeloproliferative disorders in 16, lymphoproliferative diseases in 26 and multiple myeloma in 10. Post transplantation, patients with CML received, in addition, escalating doses of lymphocytes at 6 months (maximum 1 × 107/kg CD3) or imatinib for 12 months. Median CD34+ cell number was 2.7 (1–12.3) and the median dose of campath-1H was 10 (range 7.5–45) mg. Median time to engraftment was 11 days. Overall, 20 (22%) individuals died while treatment related mortality occurred in 17% (n= 15; VOD 3, infections in 8, pneumonitis in 1, EBV lymphoma in 1 and GvHD in 2). GvHD (> grade1) occurred in 7 patients, was controlled with further immunosuppression but lead to death from infections in 6. Disease recurrence was seen in 12, but 7 with CML or myeloma responded to DLI. At a median follow up of 688 days, 77 survive; 75% in unsustained remission.

Conclusions: Myeloablative conditioning is well tolerated in patients receiving T-cell depleted grafts, and treatment related mortality of <20% can be expected consistently with this strategy. This information is useful to more precisely select patients who would benefit most from reduced intensity conditioning schedules.
Experiences with Caspofungin Plus Posaconazole in Immunocompromised Adults with Invasive Aspergillosis (IA) Refractory to Prior Standard Therapy.
Authors: Dahlke Joachim , Zabelina Tatjana , Lellek Heinrich , Muth Alexander , Waschke Olga , Wolschke Christine , Sobottka Ingo , Kroeger Nicolaus , Zander Axel R.
Published: October 2019

Background: Aspergillosis is a lethal infection of immunocompromised patients that respond poorly to antifungal therapy. Caspofungin (CAS) or Posaconazole (POS) as salvage monotherapy for IA have response rates of 45% and 42% respectively. Preliminary results of in vitro studies, animal models and clinical studies suggest that combination therapy with azoles and echinocandins may have additive activity against Aspergillus species.

Methods: Seven adult patients after unrelated haematopoietic stem cell transplantation, with proven, probable or possible IA (per EORTC/MSG criteria) and refractory to standard antifungal therapy received Caspofungin plus Posaconazole (standard monotherapy dosage). All patients had > 3 days of combination antifungal therapy. Underlying diseases were AML (n=5), Biphenotypic leukaemia (n=1), Mantle cell Lymphoma (n=1). Three patients were neutropenic (ANC<500), all patients received immunosuppressive medication. IA sites were lung (n=6) and liver (n=1). All patients were evaluable for toxicity. Efficacy was assessed by signs, symptoms and CT-scan/ultrasound. Favourable responses (complete or partial) required significant clinical and radiographic/ultrasound improvement.

Results: Success at end of combination therapy (EOCT) was seen in six patients (86%). One critically ill patient died (due to bacterial sepsis and severe acute GvHD) four days after first administration of combination therapy. One patient with complete radiographic response died 20 days after EOCT due to bacterial sepsis in relapse of AML. Combination therapy at 4 to 59 (mean 35.7) days was well tolerated. No patient discontinued combination therapy due to toxicity. Follow up from start of combination therapy was 4 to 204 (mean 79.8) days.

Conclusion: Randomized trials are warranted to determine whether this combination should be used as primary therapy for aspergillosis.
Favorable Impact of Short Term Methotrexate on Early Immune Reaction and Non-Relapse Mortality in Umbilical Cord Blood Transplantation for Adult Patients.
Authors: Watanabe Masato , Narimatsu Hiroto , Terakura Seitaro , Matsuo Keitaro , Oba Taku , Kasai Masanobu , Iida Hiroatsu , Hamaguchi Motohiro , Kohno Akio , Murata Makoto , Sawa Masashi , Miyamura Koichi , Morishita Yoshihisa ,   Nagoya BMT Group
Published: October 2019

Cord blood transplantation (CBT) represents an attractive alternative for patients with advanced hematological malignancy who lack matched related or unrelated donors. Adult patients receiving myeloablative or reduced-intensity CBT display a 90% chance of engraftment, but also experience a 50% rate of transplant-related mortality. Post-transplant immune disorders including early immune reactions and acute GVHD are problematic in CBT for adult patients. These reactions and/or additional immunosuppressive therapy might increase the risk of infection and organ dysfunction, leading to high rates of transplantation-related mortality. However, optimal prophylaxis and management for immune reactions have not been established in CBT. We investigated retrospectively whether intensive graft-versus-host disease (GVHD) prophylaxis has a prognostic impact on CBT. Post-CBT immune reactions were classified according to time course as pre-engraftment immune reaction (PIR), engraftment syndrome (ES) or acute GVHD. Between March 2001 and September 2003, a total of 77 patients underwent CBT at 8 centers of the Nagoya Blood and Marrow Transplantation Group (NBMTG). Median age of patients was 48 years (range, 18–69 years). Preparative regimens were myeloablative (n=31) or reduced-intensity (n=46). Acute GVHD prophylaxis included cyclosporine alone (n=23), tacrolimus alone (n=12), cyclosporine plus short-term methotrexate (n=17), tacrolimus plus short-term MTX (n=23) or cyclosporine plus methylprednisolone (n=2). Cumulative incidences of PIR, ES and grade I-IV GVHD were 38%, 12% and 29%, respectively. Short-term MTX exerted significant favorable effects on PIR in univariate analysis (hazard ratio, 0.41; 95% confidence interval, 0.19–0.89) and on non-relapse mortality (NRM) in multivariate analysis (hazard ratio, 0.36; 95% confidence interval, 0.16–0.81). On the other hand, PIR occurred in 35% of patients with cyclosporine and 37% with tacrolimus. Short-term MTX appears to offer optimal GVHD prophylaxis to reduce PIR and improve NRM.

Figure Figure
Oral Tipifarnib (R115777) Has Single Agent Anti-Tumor Activity in Patients with Relapsed Aggressive Non-Hodgkin Lymphoma (NHL): Results of a Phase II Trial in the University of Iowa/Mayo Clinic Lymphoma SPORE (CA97274).
Authors: Witzig Thomas E. , Maurer Matthew J. , Johnston Patrick B. , Colgan Joseph P. , Kaufmann Scott H. , Inwards David J. , Micallef Ivana N. , Ansell Stephen M. , Zent Clive S. , Allmer Cristine , Weiner George J. , Wooldridge James E. , Link Brian K. , Habermann Thomas M.
Published: October 2019

Background: Patients (pts) with aggressive NHL who relapse after conventional chemotherapy or stem cell transplant have a poor prognosis. Novel agents are needed for this important subset. Tipifarnib is an orally available methylquinolone derivative that is an inhibitor of the farnesyl transferase enzyme that transfers the farnesyl group from farnesyl pyrophosphate to specific polypeptides including small GTPases of the Ras, Rho-B, and Rheb families, as well as nuclear lamins and kinetochore proteins. Tipifarnib has documented activity in acute leukemia and produces stable disease in relapsed multiple myeloma. The goal of this study was to evaluate the activity of single-agent tipifarnib in patients with relapsed aggressive NHL in the context of a phase II trial.

Methods: Pts with relapsed diffuse large, follicular grade III, or mantle cell lymphoma were eligible for this study. Pts were treated with oral tipifarnib 300 mg twice daily d1–21 q 28 days until progression. Pts were restaged after 2 cycles, 6 cycles, and every 3 cycles thereafter. Tumor was sampled prior to tipifarnib and at day 8 in selected patients with accessible tumors. These paired samples were assayed for accumulation of unprocessed HDJ-2 and prelamin A, two polypeptides that require farnesylation for processing.

Results: Forty-two pts were accrued between March 2004 and June 2006. Thirty-seven pts had diffuse large cell, 1 follicular grade III, and 4 had mantle cell. Of the 38 patients evaluable for response, 7 (18%) had a partial response and 8 (21%) had stable disease. The 7 responders all had diffuse large cell histology. Median progression-free survival (PFS) for all pts was 2 months (range, 0–25+). The PFS of the 7 responders was noteworthy at 4.2, 4.9+, 6.7, 9.6, 14.9+, 21.6+, and 25.1+ months, respectively. The primary toxicity was myelosuppression and 33% of pts required dose reductions. The median dose received in all pts was 300 mg twice daily (mean 275, range, 100–400). The median dose received by the 7 responders was 200 mg twice daily (range, 100–400). The dose of 300 mg twice daily resulted in accumulation of unfarnesylated forms of one or both markers in all pts studied to date.

Conclusion: Oral tipifarnib has antitumor activity as a single agent with an excellent toxicity profile in pts with relapsed/refractory aggressive lymphoma. When given in this dose and schedule, tipifarnib is well-tolerated and can be taken for months without cumulative toxicity. This drug warrants further investigation in combination with conventional chemotherapy and other novel agents for the treatment of this important subset of patients.
Iron Overload in Acute Myelogenous Leukemia after Bone Marrow Transplantation.
Authors: Pakbaz Zahra , Fischer Roland , Noonan Nancy , Shiota Sherrie , Harmatz Paul , Vichinsky Elliott , Walters Mark C.
Published: October 2019

Children with acute leukemias typically receive RBC transfusions during the course of their treatment. However, the severity and significance of transfusional iron overload is not known in this patient population. Earlier, we reported elevated serum ferritin (SF) in 5 patients with AML who received HLA-identical sibling bone marrow transplantation (BMT). However, SF has a wide predictive interval for liver iron concentration (LIC) in thalassemia and sickle cell disease and the current recommendation is to measure LIC to estimate total body iron burden. Further exploration of the SF-to-LIC ratio (SF/LIC) to investigate the relationship between SF and LIC has shown ratio differences by specific disease (SCD, thalassemia, genetic hemochromatosis), transfusion status and use of chelation. The reasons for these differences are not presently known. In this study LIC was measured within 2 weeks of serum ferritin (SF), in 8 AML patients after transplantation, to explore the significance of the elevated SF and to determine the range and character of the SF/LIC ratio after BMT for AML. LIC was measured (1–4 year after BMT) by a low temperature SQUID biosusceptometer system (Ferritometer®) under the standardized Hamburg-Torino-Oakland protocol. The range for LIC in healthy individuals measured by SQUID is 90–340 mg/g wet weight. The median serum ferritin was 1227 (582–1723) μg/l and the median LIC was 1284 (751–1612) mg/g wet weight or approximately 4 times greater than the upper limit of normal. ALT was measured in 4 patients of which 2 were mildly elevated. Neither LIC nor SF changed over the interval of follow-up extending to 3 years in 2 patients (aged 11.5y and 14.5 y) who returned annually for LIC measurements. The ratio of SF/LIC ranged from 0.5 to 1.4 (median: 0.9) in the patients with AML. This compares to ratios of 1.2 (0.6–2.6) in regularly transfused sickle cell disease patients (n=45), 0.87 (0.23–2.7) in transfusion dependent thalassemia patients and 0.32 (0.05–0.57) in transfusion independent thalassemia patients. These preliminary observations suggest that children with acute leukemias who undergo bone marrow transplantation develop significant transfusion related iron accumulation. Additional investigation should be undertaken to determine if AML patients would benefit from iron reduction therapy by phlebotomy after BMT.
Transplantation for 37cases of Refractory Acute Lymphocytic Leukemia.
Authors: Ma Xiao , Wu De Pei , Su Ai - Ning
Published: October 2019

Objective: To explore the efficiency and toxicity of allo-geneic hematopoietic stem cell transplantation (Allo-HSCT) for refractory acute lymphocytic leukemia.

Patients and Methods: Thirty-seven patients of refractory acute lymphocytic leukemia received allo-geneic hematopoietic stem cell transplantation containing 18/37 cases from HLA-identical siblings donors, 8/37 cases from HLA-identical unrelated donors and 11/37 cases from haplo-identical donors of the mother. The tolerance, transplant related complications, survival rate were observed and analyzed. A myeloablative conditioning regimen included TBI and Cyclophosphamide. Cyclosporine A combined MTX were used to prevent graft versus host disease (GVHD) and G-CSF was administered after Allo-HSCT.

Result: All 37 patients tolerated the therapy well and Slight or moderate mucositis was happened. Nineteen patients had light abnormal liver function and two had renal disfunction lightly, one patient had epilepsy during the therapy. Fatal infection complications were occurred in 6 patients (including idiopathic pneumonia in 2 patients). Severe acute GVHD was presented in 5 patients. Chronic GVHD was occurred in 16 patients. Hemorrhagic cystitis was happened in 1 patient, and 9 patients relapsed again after transplantation. Venoocclusive liver disease, host versus graft disease, substantia alba encephalopathy, fatal hemorrhage, secondary tumor and dysphrenia were not occured. Hematopoietic reconstitution obtained on days 12 to 25(day 17 on average). In the follow-up duration of 9 to 55 months (34 months of mid position), most of these patients had a complete chimerical state. Twenty of 37 patients are alive (54.1%). Disease free survival is 48.6%.

Conclusion: Allo-HSCT is an effective therapy in the treatment of refractory acute lymphocytic leukemia. Relapse again after transplantation, fatal infection, and severe acute GVHD are the main reasons of failure.
A Low Intensity Protocol of Fludarabine, Busulfan and Cyclophosphamide Reduces Toxicity without Compromising Engraftment in Children Undergoing Allogeneic Bone Marrow Transplantation for Beta Thalassaemia Major.
Authors: George Biju , Mathews Vikram , Viswabandhya Auro , Chandy Mammen , Srivastava Alok
Published: October 2019

Busulfan (Bu) and Cyclophosphamide (Cy) based conditioning protocols are associated with significant morbidity and mortality in Class III (Pesaro classification) patients with beta thalassaemia major undergoing allogeneic stem cell transplantation. After IRB clearance and informed consent, we have developed a new low intensity protocol using fludarabine 30mg/m2/day for 5 days (Day - 15 to Day -11), busulfan 14 mg/kg over 4 days (Day -10 to -7) and cyclophosphamide 160 mg/kg over 4 days (Days -5 to -2). Graft versus host disease (GVHD) prophylaxis consisted of cyclosporine (5mg/kg/day) in 2 divided doses starting Day -4 and methotrexate 10 mg/m2 on Day +1 followed by 7 mg/m2 on Day +3, +6 and +11. Between January and July 2006, 14 children with Class II (n = 5) or Class III (n = 9) beta thalassaemia major were treated with this protocol. The median age was 10 years (range: 3 – 13). All patients had HLA matched related donors [sibling (13) or family (1)] with a median age of 4 years (range: 2 – 35). Ten (71%) of these transplants were sex mismatched. One patient expired on Day +10 due to an intracranial bleed while 13 (92.8%) engrafted. The median time to ANC > 500/mm3 was 17 days (range: 14–20) and platelet count > 20,000/mm3 was 25 days (range: 11 – 49). One patient had graft rejection. Two patients (14%) had grade 2 hemorrhagic cystitis and veno-occlusive disease each that resolved with conservative management. None of the patients had grade IV mucositis while 1 patient has a grade II gastro-intestinal bleed lasting 2 days that resolved with transfusion support. Acute GVHD (grade I – II) was seen in 5 patients (41.6%) and resolved in all with corticosteroids. Febrile neutropenia occurred in all but only 2 patients had a documented infection. The day 30 chimerism was complete in 10 patients (77%), > 90% donor chimerism in 2 (15.3%) with rejection in 1 patient (7.7%). At a median follow up of 4 months (range: 1–6), the overall survival (OS) is 92.8% and the disease free survival is 85.7%. We compared engraftment and toxicity with 167 patients with Class II and III Thalasssaemia treated with Bu16/Cy200 (n=116) or Bu600 mg/m2/ Cy200 (n=51) and found that the engraftment was not delayed but there was significant reduction in VOD (14% vs 53%; p=0.005) and gastrointestinal bleeding (7% vs 39%; p=0.021) with this new protocol.

Conclusion: A low intensity protocol incorporating reduced doses of busulfan and cyclophosphamide in combination with fludarabine is associated with reduced toxicity and improved survival in children undergoing allogeneic BMT for Thalassaemia. Long term follow up is required to assess late toxicity and graft function.
Comparative Analysis of Matched Related Donor (MRD) and Matched Unrelated Donor (MUD) Hematopoietic Stem Cell Transplantation in Treatment of Patients with Chronic Myeloid Leukemia.
Authors: Styczynski Jan , Gil Lidia , Zaucha Jan Maciej , Czyz Anna , Bieniaszewska Maria , Wysocki Mariusz , Hellmann Andrzej , Komarnicki Mieczyslaw
Published: October 2019

Allogeneic HSCT offers a long term remission for CML patients. We compared results of MRD and MUD HSCT in 82 pts with CML, treated between 1999 and 2005. In MRD group, 61 pts (median age 39, range 8–54 yrs) were conditioned with BuCy2 regimen. 50 pts were in chronic, 7 in acceleration and 4 in 2nd chronic phase. 38 pts received peripheral blood (PB) and 23 bone marrow (BM) cells. Prophylaxis of graft versus host disease (GVHD) consisted of cyclosporin (CsA) and methotrexate (MTX). 21 pts (median age 31, range 11–51 yrs) transplanted from MUD were conditioned with BuCy2 (n=13) or TBI-based regimen (n=8). All MUD patients received thymoglobulin. This group comprised of 14 pts in chronic and 3 with accelerated, 1 in blastic phase and 3 in 2nd chronic phase. 10 pts received PB and 11 BM cells. CsA+MTX was given as GVHD prophylaxis. MUD group differed from MRD group by following factors: younger age (p=0.020), longer time to HSCT (p<0.001), use of imatinib before HSCT (p=0.039), and higher pre-transplant (Gratwohl) risk score (p<0.001). Results: Hematopoietic recovery occurred in all pts in MRD group, and in 95% in MUD group. Probability of overall survival for all pts was OS=0.72±0.05. 42 pts developed acute GVHD >2 grade (26 in MFD and 16 in MUD) and 27 pts extensive chronic GVHD (23 in MFD and 4 in MUD). Mean survival, estimated 5-yrs OS, probabilities of GVHD incidence, relapse and non-relapse mortality (Kaplan-Meier method) with respect to transplantation method, are shown in Table.

TABLE: Results of transplantation in MRD and MUD groups Probability MRD-HSCT (n=61) MUD-HSCT (n=21) P Mean survival (years) 5.6 (95%CI=4.9–6.3) 3.6 (95%CI=2.4–4.7) 0.0020 Overall survival 0.75±0.05 0.60±0.11 0.0759 Relapse incidence 0.12±0.04 0.25±0.12 0.1950 Non-relapse mortality 0.22±0.05 0.41±0.11 0.0311 Acute GVHD≥2 0.45±0.06 0.79±0.10 0.0175 Chronic extensive GVHD 0.47±0.07 (n=57) 0.35±0.14 (n=15) 0.3103

Factors predicting negative outcome by multivariate analysis were: acute GVHD≥2 (p=0.0097, HR=1.81, 95%CI=1.12−2.94), and pre-transplant risk score >2 (p=0.0280, HR=1.61, 95%CI=1.06−2.00). MUD-HSCT or use of imatinib before HSCT did not predict for OS. Conclusion: In the era of tyrosine kinase inhibitors, results of MRD− and MUD-HSCT show that HSCT still remains an important therapeutic option for CML patients.
The Long Term Survival of the Patients with Refractory Leukemia Treated by Related HLA Haploidentical Stem Cells Transplantation.
Authors: Wu Bingyi , Kunyuan Guo , Hu Haiyan , Yang Yuliang , Deng Lan
Published: October 2019

To assess the Long Term Survival of patients with refractory leukemia treated by HLA haploidentical stem cells transplantation. We analysed the outcomes of 35 patients with refractory leukemia who underwent HLA haploidentical peripheral blood stem cells transplantation from August, 1998 to December, 2005. Thirty five patients with refractory leukemia including 16 cases of acute non-lymphocytes leukemia, 11 cases of acute lymphocytes leukemia, 7 cases of chronic myeloid leukemia and 1 case of IV phase of non-Hodgkin lymphoma underwent HLA haploidentical peripheral blood stem cells transplantation. The median age was 25 years old (3– 52 years old). Fourteen patients received stem cells from parent donors, four from daughter and one from son donor, the others from sibling donors. Three HLA locus mismatched in thirteen cases, Two HLA locus mismatched donors in fifteen cases, One HLA locus mismatched in six cases. The conditioning regime consisted of fludara (25mg/m2 ×5d), busufan (4mg/kg ×4d) and cyclophsphamide(60mg/kg ×2d). Rabbit anti-human lymphocyte globulin (5mg/kg ×5d) was added in some patients in conditioning regime. A mean of 5.0 × 108/kg (2.97×108/kg-8.0×108/kg) mono-nucleated cells was grafted. The mean CD34+ cells number was 5.5 ×106/Kg (3–6.5 ×106/kg). Ten patients received 400ml donor derived bone marrow beside donor peripheral stem cells. Thirty-two patients were successfully grafted and one failed to graft and one died with severe fung infection at day 2, and one died from severe veno-occlusive disease at day 28. The mean time of white cell count more than 1×109/L was 14(11–18) days and platelet count more than 20×109/L was 15 (11–18) days. All patients got complete remission in 32 successful grafted patients. Severe acute graft versus host disease occurred in seven patients, and four died. Eight patients suffered from chronic graft versus host disease. Eight patients relapsed and died. The median relapse time was 10 (3–24) months. Fourteen patients are still disease free survival. The three years disease free survival rate was 40%. We concluded from our observation that HLA haploidentical peripheral blood stem cells transplantation may be an effective therapy for refractory leukemia. Some patients with refractory leukemia treated by HLA haploidentical stem cells transplantation could survive with free diseases; Though graft versus leukemia effect may be strong in HLA haploidentical blood stem cells transplantation, leukemia will probably be relapse when the patient without complete remission at transplant.
Hematopoietic Stem Cell Transplantation (HSCT) for Malignancies Using Plasma Depleted Umbilical Cord Blood (UCB) That Were Not Red Blood Cell Depleted.
Authors: Petz Lawrence , Nademanee Auayporn , Rosenthal Joseph , Graham Michael , Eames Gretchen , Karanes Chatchada , Chan Lee Lee , Tan Ah Moy , Tan Poh-Lin , Wang Brian , Tseng Peggy , Chow Tiffany , Wu Tina , Jaing Tang-Her , Gjertson David , Tan Patrick , Chow Robert
Published: October 2019

Cell dosage is a limiting factor for UCB HSCT, especially for adult patients. Most UCB banks practice red cell depletion (RCD) techniques to save storage space, which incur significant nucleated cell loss after processing. One method to minimize cell loss and still reduce volume after processing is to deplete plasma (PD), but not the red blood cells. Not washing UCB (NW) after thawing also minimizes cell loss. A large racially diverse PD UCB inventory of over 25,000 units is now available on stem cell registries. There were 70 ALL, 54 AML, 19 CML, and 45 others transplanted for malignant indications. A retrospective audited analysis was performed on 170 patients with engraftment and/or survival information. Of the ALL/AML/CML cases with available information, there were 32 1CR/CP, 33 2CR, and 13 3CR/CP. The median age of patients was 13 years old (range 05–59); median weight 50 kg (range 5–137); male 45%. Transplant characteristics indicated a median # HLA ABDR matches of 4.0; median pre-freeze TNC dose 4.1 × 107/kg; median post-thaw TNC dose as reported by TC 3.7 × 107/kg; median pre-freeze CD34 dose 1.4 × 105/kg; transplants outside of U.S. 34%; double unit transplant 30%; non-myeloablative 16%. Sixty-one percent of the transplanted UCB were washed post-thaw (W), 39% were infused without post-thaw wash (NW), with 20% of the units without available post-thaw data. Kaplan-Meier estimates of 3-month ANC500 and 6-month platelet 20K and 50K engraftment are 89±3%, 81±5%, and 73±5% respectively. Median time to engraftment for ANC 500, platelet 20K, and 50K were 22 days (range 7–69 days), 52.5 days (range 12–181 days), and 64 days (range 25–374 days) respectively. Median time to engraftment for post-thaw washed units (W) versus PD CBU not washed and infused directly after thawing (NW) were 24 vs. 22 days for ANC500, and 57 vs. 48 days for platelet 20K respectively. The incidence of reported grade II-IV and III-IV acute GVHD were 32% and 14% respectively. Twelve percent developed limited chronic GVHD and 14% developed extensive chronic GVHD. With a median follow-up of 170 days (range 4–1402 days), the Kaplan-Meier estimates of 1-year TRM, OS and relapse-free survival were 31±4%, 51±4% and 47±4% respectively. These results demonstrate that HSCT using unrelated PD UCB can be performed safely and effectively in patients with malignancies, and post-thaw wash may not be necessary.
Results of High Dose Chemotherapy and Autologous Stem Cell Transplantation in Patients with Acute Myeloid Leukemia.
Authors: Ault Kenneth A. , Case Delvyn Caedren , Boyd Marjorie A. , Ervin Thomas J. , Aronson Frederick , Ebrahim Kurt S. , Hedlund Jacquelyn A.
Published: October 2019

43 patients with acute myeloid leukemia have undergone transplantation at our institution over the past 14 years. Patient selection criteria included age less than 70 years, creatinine less than 2mg/ml, no active infection, cardiac ejection fraction >40%, DLCO > 50% of predicted and no other co-morbid conditions that would jeopardize survival. 39 patients were in first remission, 4 were in second or higher remission. 3 patients had favorable cytogenetics, 40 had intermediate or unfavorable cytogenetics. After achieving remission for at least 30 days, patients were consolidated with Etoposide and AraC, followed by G-CSF. Hematopoietic stem cells were collected when the WBC rebounded to at least 10,000/μl. The target dose of CD34 positive cells was 5×106/kg. The minimum dose given was 2.3 × 106/kg). High dose therapy consisted of Busulfan 1mg/kg and Etoposide 60mg/kg. The average age at transplantation was 42 years (range 20 to 61). Days of neutropenia (AGC<500/μl) ranged from 2 to 10 (average 5.2). The median length of follow up is 4.0 years. Kaplan-Meier progression-free survival is 38% at 5 years and 35% at 10 years. Currently 26 patients are alive, and 23 are free from progression. Overall survival is 60%.

Maine Medical Center Autologous HPC Transplant Program Acute Myelogenous Leukemia Maine Medical Center Autologous HPC Transplant Program Acute Myelogenous Leukemia
Pretreatment of the Esophagus with Manganese Superoxide Dismutase Plasmid/Liposome Complex (MnSOD-PL) before Irradiation Results in Increased Migration and Proliferation of Marrow-Derived Stem Cell Progenitors in the Esophageal Squamous Epithelium.
Authors: Epperly Michael W. , Niu Yunyun , Shen Hongmei , Greenberger Joel S.
Published: October 2019

Irradiation of the mouse esophagus increases production of superoxide, nitric oxide, peroxynitrite, and other reactive oxygen species (ROS) for weeks after exposure. ROS production in the esophageal microenvironment may be toxic to cells migrating in to repair irradiation damage. MnSOD-PL gene therapy decreases ROS production. We tested whether such treatment increased the engraftment of circulating marrow origin cells involved in repair of esophageal irradiation damage. C57BL/6NHsd female mice were treated with intraesophageal MnSOD-PL and irradiated 24 hr later along with control mice to 29 Gy to the thoracic cavity. Subgroups of mice were sacrificed at days 1, 2, 3, 4, or 5 days after irradiation, esophagus removed, frozen in OCT and sectioned. Sections were stained and apoptosis quantitated. Low level apoptosis was detected for the first 3 days with significant apoptosis on day 5. Mice pretreated with MnSOD-PL had reduced apoptosis (3 ± 1%) compared to control irradiated mice (20 ± 7%) (p= 0.010). Female mice in each 29 Gy esophagus irradiated group were injected I.V. 5 days later with 1 × 105 bone marrow cells from male ROSA mice. Subgroups received MnSOD-PL (100 ug plasmid DNA in 100 ul) 24 hr before irradiation. ROSA donor cells were differentiated from recipient C57BL/6NHsd cells by Y probe and LacZ positive staining and resistance to G418 follows explants in vitro. At sacrifice 14 days after irradiation, esophagus was removed and sorted into side population (SP) and non-side population cells (NSP). Mice that received MnSOD-PL before irradiation, had 42.3 ± 5.3% of NSP cells LacZ+ compared to 10.9 ± 1% in the irradiation only group (p < 0.0001). SP cells from MnSOD-PL treated mice demonstrated 26.5 ± 4.8% LacZ+ cells compared to 9.3 ± 2.0% in the irradiation control group (p = 0.0021). Serial transfer of donor ROSA marrow origin esophageal “stem cell candidates” was carried out by I.V. transfer of 1st generation SP and NSP harvests to 29 Gy thoracic irradiated female recipients. Second generation recipients pretreated with intraesophageal MnSOD-PL who received ROSA male SP cells isolated from first generation mice (also pretreated with MnSOD-PL) demonstrated the highest number of LacZ+ cells (4.7 ± 1.6%). In contrast, no LacZ+, ROSA cells were detected in second generation irradiation control recipients. Therefore, intraesophageal delivery of MnSOD-PL increases stable migration of bone marrow derived progenitors of esophageal squamous epithelium and improves the stability of serially transferred marrow origin, esophageal stem cell candidates.
Development of an Effective Safety Switch for Selective Elimination of Human T Cells In Vivo after Adoptive Transfer.
Authors: van Meerten Tom , Rozemuller Henk , Mackus Wendy J.M. , Parren Paul W.H.I. , van de Winkel Jan G.J. , Claessen Marie-Jose , van Rijn Rozemarijn S. , Hagenbeek Anton , Martens Anton C.M. , Ebeling Saskia B.
Published: October 2019

Adoptive transfer of T cells is frequently associated with unwanted side effects. In order to tackle these effects one could introduce a safety switch into the cells that permits their selective in vivo elimination. The human CD20 gene in combination with CD20 antibodies was recently proposed as a novel safety switch. In such a system, T cells may be genetically modified with a CD20-encoding vector prior to adoptive transfer. If necessary, CD20-transgenic cells can be eliminated in vivo through administration of CD20 antibodies, such as the chimeric antibody rituximab (RTX) that is currently used to treat CD20+ lymphoma. RTX activates the complement system and recruits immune effector cells, resulting in rapid death of CD20+ cells. Recently, a novel human CD20 antibody, Humab 7D8, was shown to have superior activity over RTX.

In this study a set of CD20-encoding retroviral vectors was generated, which either lacked or contained one or both of two regulatory elements:

the woodchuck posttranscriptional regulatory element (WPRE) to increase CD20 expression, and the chicken hypersensitivity site 4 insulator element (INS) to achieve a position independent expression of CD20 and to increase the safety profile of the vector by preventing activation of cellular (onco)genes by the retroviral enhancer.

We found that the level of CD20 expression obtained with vectors containing INS was 2-fold lower than with vectors lacking INS. Additional inclusion of WPRE restored the level to that of the vector without INS. In addition, INS greatly enhanced the homogeneity of CD20 expression in T cells. Moreover, after 3 months in culture, all cells generated with CD20-INS had retained CD20 expression, while 60% of cells transduced with the control CD20 vector had lost CD20 expression.

Complement dependent cell kill (CDC) of both RTX and HuMab 7D8 was dependent on the level of CD20 expression (p<0.01). However, while very low CD20-expressing cells were completely resistant against RTX they could be effectively killed by HuMab 7D8. For maximal kill of CD20-high cells, a 100-fold lower dose of HuMab 7D8 was required, compared to RTX. In vivo efficacy was studied through bioluminescent imaging of luciferase+ CD20-transgenic T cells. After transfer of CD20+ cells in immune deficient RAG2−/−gamma c−/− mice, both CD20 antibodies were capable of eliminating >99% of CD20+ cells, prolonging survival of mice from 20 till 42 days.

In conclusion, we developed a safe vector that leads to homogeneous and stable expression of CD20 on human T cells. These cells can be killed effectively in vivo with HuMab 7D8, a recently developed CD20 antibody. This system will be applicable to other approaches that require inclusion of a safety switch in ex vivo modified cells.
Use of Erythropoietin (EPO) in Children with Malignancies.
Authors: Kovacs Gabor T. , Muller Judit , Csoka Monika , Vonnak Eszter , Erlaky Hajna , Farkas Andrea , Somlai Agnes , Fekete Gyorgy
Published: October 2019

Recombinant erythropoietin is widely used for the treatment of anemia in malignant diseases in adults. There are only limited data of its use in pediatric population.

In this study we analysed the effectiveness and tolerability of recombinant human erythropoietin (NeoRecormon) in children with malignant diseases.

80 children with malignant diseases were analysed. 40 patients (15 girls, 25 boys) received EPO in a mean dosage of 144.5±14.1 IU/kg three times a week. The mean age of the EPO-treated patients was 8.8 (2.5–16) years. 26 children had acute lymphoblastic leukemia and 14 patients had solid tumor. Match-paired, retrospective control patients (n=40) with similar diagnosis were used for the data analysis as control group (C).

The mean duration of EPO treatment was 5.8 months (3–8 mo). In 6 patients the therapy was ceased due to elevated serum hemoglobin (Hb) (>130 g/L), in 6 patients the dose was increased up to 200 IU/kg three times a week, and 5 patients discontinued the therapy (2 died, 3 unsuccessful treatment). The mean amount of erythrocyte transfusion in the first 3 months of chemotherapy (CT) was 4.1±3.1 U/patient in the EPO group, and 8.0±4.2 in C, and during 6 months of CT 4.5±3.4 with EPO, and 11.6±7.1 in C (p<0.05).

Soluble transferrine receptor (STFR) levels in serum increased in the EPO group after 2 weeks of therapy from 3.2±2.0 up to 4.8±2.9 (p<0.05). In general in 26/40 patients a significant elevation of the Hb levels and decrease of the need of erythrocyte transfusions could be detected. In 22 patients the STFR levels increased more than 50 % after 2 weeks of therapy. In this subgroup 18/22 children responded to EPO therapy. All patients tolerated the therapy well, no severe side effects were detected. In summary, EPO treatment is effective in about 2/3 of pediatric oncology patients. The therapy is well-tolerated. Increase in the STFR serum levels might be a useful marker for the effectiveness of EPO in children.