bluuebunny/crossref_metadata_2025 huggingface dataset:
C3435T Polymorphism of MDR1 Gene Is Not Associated with p-Glycoprotein Function of Leukemic Blasts and Clinical Outcomes in Patients with Acute Myeloid Leukemia.
Authors: Lee Michael Jinpyo , Hur Eun-Hye , Lee Je-Hwan , Choi Seong-Jun , Lee Jung-Hee , Seol Miee , Kim Keun-Hee , Lee Hee-Jung , Kang Ip-Sol , Shim Kyung-Soo , Ryu Seong-Gil , Kang Young-Ah , Lee Young-Shin , Park Chan-Jeoung , Chi Hyung-Sook , Lee Kyoo-Hyung
Published: April 2020

P-glycoprotein (P-gp) function of leukemic blasts is associated with chemotherapy resistance in acute myeloid leukemia (AML). The C3435T polymorphism in the human MDR1 gene has been studied in AML and contradictory results were reported regarding clinical implications of the polymorphism. We investigated the association of C3435T polymorphism of MDR1 gene with P-gp function of leukemic blasts and clinical outcomes in patients with AML excluding M3 subtype. A total of 200 patients, 127 males and 73 females, were included in this study. Median age was 44 years (range, 14–75). All patients were newly diagnosed and were treated with standard ‘7+3’ induction chemotherapy regimens at the Asan Medical Center, Seoul, Korea between May 1999 and November 2006. The C3435T polymorphism was analyzed with PCR/RFLP method. P-gp function of leukemic blasts was measured at diagnosis by the rhodamine-123 efflux assay. The clinico-laboratory data at diagnosis and data on clinical outcomes were obtained from the AMC Leukemia Registry. Genotype frequency of C3435T polymorphism was 71 (35.5%) for CC, 93 (46.5%) for CT, and 36 (18.0%) for TT. There were no significant differences between different genotypes of C3435T polymorphism regarding age, sex, FAB subtypes, initial leukocyte counts, percentages of blasts in bone marrow or blood at diagnosis, and cytogenetic risk groups. P-gp function of leukemic blasts was not significantly different according to the genotype of C3435T polymorphism: 32.2 ± 24.0% for CC, 35.8 ± 21.4% for CT, and 29.1 ± 24.0% for TT (P=0.370). Complete remission was induced in 79.2% for CC, 79.8% for CT, and 77.8% for TT (P=0.969). Overall, relapse-free and event-free survival probabilities at 5 years were 47.6%, 55.4% and 51.0% for CC, 51.3%, 51.0% and 42.5% for CT, and 53.1%, 60.0% and 55.0% for TT (P= 0.594, 0.932 and 0.764, respectively). In conclusion, C3435T polymorphism does not have significant clinical implications in AML regarding P-gp function of leukemic blasts and clinical outcomes.
Breast Nodules: First Manifestation of LLA.
Authors: Avila Estela Est. , Macias Mirian Mir , De Salvo cardullo Luigi Lui , Nunez Alonso Alo , Romero Esmeira Esm. , Weir Jesus Jes , Hassanieh Angelina Ang
Published: April 2020

INTRODUCTION: Malignant hematological process that affects breast are infrequent in lymphoma and leukemia, only 0.25%, being most common in non Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), and very rare in acute lymphoblastic leukemia (ALL), with very few cases reported. The infiltration of breast due to leukemia cells may occur initially, during relapse, after bone marrow transplantation (ABMT) or after radiotherapy in cancer. Affect young people presenting single or multiple mass, with diffuse increment of breast, unilateral or bilateral. The following report belongs to one case of ALL with initial presentation of nodules in breast.

DESCRIPTION: 35 years old female patient who presented increased volume in breasts, intermittent fever and generalized weakness. The echogram showed solid nodules in both breasts. The needle biopsy reported diffuse infiltration of lymphoid cells, being confirmed by biopsy and inmunohistochemical study, which results were: bcl-2, cd99, cd10, cd34, cd43 and cd45ro positives in neoplastic lymphocytes. The diagnosis of ALL/lymphoblastic lymphoma was made. The Oncologist referred the case to the Hematology Service and blood peripheral studies were made: Hemoglobin: 9.4grs/dl, hematocrit 29%, WBC; 86000 dl., 80% of blasts. Bone marrow was infiltrated by immature lymphoid cells. It was confirmed by morphology and flow cytometry studies. The diagnosis was ALL L1 CALLA positive. Cytogenetic studies revealed t(2:7)(q11:q11), t(9:22)(q34:q11) [20] Philadelphia chromosome positive. The patient was treated with chemotherapy without remission, allogenic bone marrow transplant with recidive in breasts, including the scalp and bone marrow by lymphoid blasts and confirmed by inmunophenotype. The patient received Imatinib Mesylato and Hyper CVAD as reinduction. She, eventually, died.

CONCLUSION: The importance of this case is the atypical and unusual presentation of ALL Ph (+) and the association of Philadelphia chromosome with balanced translocation t(2:7)(q11:q11), which has been rarely reported in hematological malignances. The biopsy of nodules allowed the early diagnosis. This is the first case reported in Venezuela.
Post-Transplant Lymphoproliferative Diseases after Renal Transplantation. Long Term Experience over 35 Years in Peru.
Authors: Beltran Brady , Morales Domingo , Malaga Jose , Carrasco-Yalan Antonio A.
Published: April 2020

BACKGROUND: Post-transplant lymphoproliferative diseases (PTLD) are a combination of diseases characterized by the proliferation of lymphoid cells that occur in the context of post-transplant immunosuppression. Primary infection by Epstein Barr Virus (EBV) is the main risk factor for the development of PTLD. A total of 11% of all the cancers recorded in renal transplant recipients are PTLD.

OBJECTIVE: To determine prevalence of PTLD in renal transplant recipients.

PATIENTS AND METHODS: We retrospectively reviewed the records of 1038 renal transplant recipients who received allograft transplants between 1972 and 2007 and recorded the incidence and types of de novo malignancies that developed in these patients. The study was carried out from two centers in Peru.

RESULTS: Forty-seven neoplasms were diagnosed in 43 recipients. 26 were men and 16 women, mean age was 46.0, range (17–67 years). Malignancy was diagnosed 2 to 240 months after transplantation. The tumors included Kaposi’s sarcoma (KS) in ten patients, squamous cell carcinoma (SCC) in seven, basal cell carcinoma (BCC) in six, cervix cancer in four, colon and breast cancer in three and hepatocarcinoma in two .Also were reported one case of small intestine cancer, osteosarcoma, pancreatic cancer, lung cancer, bladder cancer, schwanoma, melanoma, tongue cancer, pelvis renal cancer and carcinomatosis from unknown primary. PTLD were 4.2% (two cases) from all neoplasm found in renal transplant patients. First case corresponds a female 53 years old with four year from renal transplant. ATG, ALG OKT3 were not used in the patient during transplant induction. She was receiving cyclosporin and azatrioprine as inmunosupresors. She had a bulky diffuse large B cell lymphoma primary from retroperitoneal node, stage IVB, IPI high risk and bone marrow involved. She received CHOP-21 regimen for four cycles with progressive disease. She died from meningeal involvement, six months after from diagnosis. Second case was a male, 74 years old with ten years from renal transplant. He has a diffuse large B cell lymphoma from neck skin, stage IB. ATG, ALG OKT3 were not used in the patient during transplant induction. He was receiving cyclosporine and azathioprine as inmunosupresors, currently the patient continues CHOP-21 regimen with good response.

Conclusions: A low incidence of PTLD after renal transplantation occurred in Peru.
RASSF8 Overexpression Expands KLS Cells in Vitro and Blocks Murine Hematopoietic Stem Cells in a Primitive Phenotype.
Authors: Schoemans Helene M. , Snoeckx Rikkert , Buckley Shannon , Mimiola Elda , Schouteden Sarah , Verfaillie Catherine M.
Published: April 2020

A recent transcriptome analysis of human umbilical cord blood and bone marrow (BM) CD34+CD33−CD38−ckit+Rholo (Rholo) vs. CD34+CD33−CD38−ckit+Rhohi (Rhohi) cells identified the gene RASSF8, to be more highly expressed in the hematopoietic stem cell (HSC) rich compartment (Rholo) versus committed progenitor cells (Rhohi). In a subsequent screen for the function of differentially expressed genes in hematopoiesis, the knockdown of RASSF8 using morpholinos in zebrafish embryos resulted in decreased blood formation and reduced expression of early (scl and gata1) and late (hbae1 and lcp1) hematopoietic markers. To test the function of RASSF8 in mammalian hematopoiesis, we overexpressed RASSF8 cDNA in lineage depleted murine bone marrow cells (Lin−) by retroviral transduction using the MSCV-RASSF8-IRES-GFP vector (rMIG-RASSF8). Sorted transduced GFP+ cells were cultured in serum-free medium (supplemented with SCF, TPO, Flt3L and Il-3) for three to five days. No significant differences were seen in overall cell expansion, cell death (7-AAD staining) or cell proliferation (thymidine incorporation assay and propidium iodide staining) between rMIG-RASSF8 transduced cells and cells transduced with the control vector (rMIG). However, we noted a significant accumulation of the stem cell enriched cKit+Lin−Sca1+ (KLS) population in rMIG-RASSF8 transduced cells (43+/−9%) compared to control cells transduced with rMIG (13+/−8%, p=0.003; n=4) after five days. The total number of colony forming cells (CFCs) produced by 750 rMIG-RASSF8 transduced Lin− cells (24+/−7) was significantly lower than in rMIG transduced cells (54+/−8, p=0.008; n=3). CFCs from rMIG-RASSF8 transduced cells were morphologically smaller and more compact, consistent with the morphology of primitive blast-forming colonies. Competitive repopulation assays using 25 000 rMIG-RASSF8 or rMIG transduced C57/Bl6 CD45.1 Lin- cells versus 100 000 C57/Bl6 CD45.2 mononuclear BM cells were performed to evaluate HSC engraftment potential. Four weeks after transplantation only 1+/−0.96% CD45.1 cells were found in the peripheral blood (PB) of the animals receiving rMIG-RASSF8 transduced cells vs. 23+/−12% CD45.1 cells in animals that had received rMIG transduced cells (p< 0.05; n=3). Likewise, PB analysis at 3 months demonstrated nearly no reconstitution of rMIG-RASSF8 transduced CD45.1 cells (1+/−1.7%) vs 30+/−23% rMIG transduced CD45.1 cells (p=0.02). In conclusion, constitutive overexpression of RASSF8 increases the retention/expansion of KLS cells in vitro, blocks differentiation of progenitors giving rise to CFC and inhibits engraftment of HSC. These studies thus suggest that short term overexpression of RASSF8 might ultimately lead to new methods of HSC expansion.
A Phase II Trial of Darinaparsin in Advanced Lymphomas: Report on Safety and Activity.
Authors: Craig Michael , Shah Sandip , Tallman Martin , Patil Poonam , Easow Jose , Buck Jill , Shah Sachin , Lewis Jonathan , Lossos Izidore S.
Published: April 2020

Darinaparsin (ZIO-101) is a novel organic arsenic molecule that has a mechanism of action mediated by targeted disruption of mitochondrial function, modified signal transduction, and antiangiogenesis. It is active against diverse cancers in vitro. This Phase II multi-center trial is being conducted in patients diagnosed with advanced lymphomas that have received at least 1 prior therapy and require additional treatment. Patients receive 300 mg/m2 of darinaparsin intravenously for 5 consecutive days every 28 days (1 cycle) and are then evaluated for efficacy and safety by standard criteria. Treatment continues until toxicity or progression. To date the study has accrued 22 patients (15 non-Hodgkin’s [NHL], 7 Hodgkin’s); 12 are male and 10 are female. Median age at baseline was 60.5 years (range: 28–80), ECOG performance status was ≤2, and median number of prior therapies was 4 (range 2–6). Thirteen subjects have received at least 2 cycles of darinaparsin and are evaluable for efficacy. Of these, 1 (diagnosed with PTCL) has achieved a complete response (CR) and 3 (diagnosed with marginal zone transformed to diffuse large B-cell, marginal zone, and Hodgkin’s nodular sclerosis, respectively) have achieved partial responses (PRs). These patients had been heavily pretreated (PTCL: CHOP, ICE, and EPOCH; marginal zone transformed to diffuse B-cell: RCHOP, RICE, RT and autologous bone marrow transplantation; marginal zone: rituximab, RCVP, and gemcitabine; and Hodgkin’s: ICE, CBV, gemcitabine+ MDX-060, and stem cell transplant). In the patient with transformed diffuse large B-cell lymphoma who achieved PR, no evidence for macroscopic disease was present, but microscopic low-grade marginal zone lymphoma was detectable in random biopsies from normal-appearing gastric mucosa. In addition, 2 patients (diagnosed with PTCL and Hodgkin’s, respectively) have achieved stable disease (SD). The only Grade 3 adverse event (AE) considered drug-related was wheezing. A total of 12 subjects have reported 37 serious adverse events (SAEs) while on study. Of these, only 2 had SAEs that were considered drug-related (neutropenic fever, fall). In conclusion, darinaparsin has been very well tolerated and has demonstrated promising activity in heavily pretreated patients diagnosed with advanced lymphoma. Initial responses (1 CR, 3 PRs, 2 SDs) have been observed among 13 evaluable patients. Accrual continues; additional safety and efficacy data will be reported.
Disruption of T Cell Suppression in Chronic Lymphocytic Leukemia by CD200 Blockade.
Authors: Pallasch Christian P , Ulbrich Susanne , Brinker Reinhild , Uger Robert A , Hallek Michael , Wendtner Clemens- Martin
Published: April 2020

Suppression of patients’ T-cells is a key event in CLL pathogenesis and was demonstrated to be mediated by direct cell-cell contact of malignant CLL cells with T-cells. CD200 plays a critical role in regulating the immune system and has been shown to be up-regulated on the surface of different tumors including CLL. In this study we addressed the effects of CD200 over-expression on CLL cells on autologous T cells in a mixed lymphocyte reaction system. We used native and CD40 ligand (CD40L)- stimulated CLL cells as antigen-presenting cells (APCs) to expand autologous T cells of 14 patients. T-cell proliferation was analyzed over 3 weeks of in vitro culture. A functional anti- CD200 antibody (1B9) was added to reveal CD200-mediated immunosuppression in the autologous system. Expansion of patient T-cells was assessed by flow cytometry including intracellular staining of FOXP3. Specificity towards CLL-specific antigens was monitored applying fibromodulin derived peptides for detection of specific T-cells by ELISPOT analysis. T-cell proliferation over 3 weeks of in vitro culture was significantly enhanced compared to control cells when using CD40L-stimulated APCs and an anti-CD200 antibody (p=0.0004). CD200 blockade was further shown to stimulate antigen-specific T-cell responses towards the F2 and F4 peptides of the CLL-associated antigen fibromodulin (p=0.04). Finally, the number of CD4+/CD25high/FOXP3+ T cells (Treg) was significantly decreased in CD200 treated mixed lymphocyte reaction (p=0.04). In summary, CD200 blockade may provide therapeutic benefits in CLL by

enhancing T-cell expansion, augmenting an antigen-specific T cell response with suppression of regulatory T cells.

CD200 seems to be an important immunosuppressive molecule in CLL: by CD200 blockade immune suppression can be overcome by altering tolerance to tumor antigens and deregulation of regulatory T cells. This combination of an immune induction paralleled by a disruption of immunosuppressive factors makes anti-CD200 mAb a powerful tool for future treatment of CLL, possibly in combination with other B cell cytotoxic or immunostimulatory approaches.
Payer Costs of Autologous Stem Cell Transplant: Results from a U.S. Claims Data Analysis
Authors: Pelletier Elise M , Smith Paula J , Dembek Carole J
Published: April 2020

Objectives: Autologous stem cell transplant (ASCT) includes costly pre-transplant care which must be considered in the overall cost of treatment. The objective of this study was to evaluate total costs of an ASTC, from mobilization/apheresis through 100 days following transplant, in U.S. managed care plans.

Methods: Patients 18+ years of age with evidence of an ASCT between January 1, 2000 and December 31, 2006 were identified from a nationally-representative database of medical and pharmacy claims from over 90 U.S. managed care health plans. Patients were required to have a claim for apheresis within 60 days prior to the ASCT and have a history of multiple myeloma, non-Hodgkin’s lymphoma, or Hodgkin’s disease. Patients were included in the study only if they were continuously enrolled in one health plan for a minimum of 90 days before to 100 days following the ASCT; patients with evidence of a prior ASCT were excluded. Costs were calculated for 4 phases of ASCT:

Mobilization/Apheresis: 5 days prior to the first G-CSF administration through the last apheresis claim, Conditioning: 1 day after the last apheresis claim to 1 day prior to ASCT, Transplant: ASCT through 30 days post-transplant, and Follow-up: 31 to 100 days following transplant.

Paid claims (payer perspective) were used as a proxy for medical costs and were expressed in 2006 US$, adjusted as necessary using the medical care component of the U.S. Consumer Price Index.

Results: A total of 249 ASCT patients were identified; the mean age was 50 years and 57% were male. Total costs from mobilization to 100 days post-transplant averaged $146,890 per patient (median $134,878). Mobilization and apheresis costs comprised 27% of the total costs (mean $38,803), while conditioning therapy represented 43% of the total costs (mean $63,369). Transplant costs (mean $22,032) and follow-up costs (mean $22,686) each accounted for approximately 15% of the total costs.

Conclusion: ASCTs are costly procedures with mean payer costs of almost $150,000. This study reviewed recent U.S. managed care health plan claims data to determine the component costs of ASCT. Conditioning therapy and mobilization/apheresis were identified as the main cost drivers, comprising 43% and 27% of the total costs, respectively. Delineating these ASCT-related costs is important in evaluating the overall economic burden of ASCT on the healthcare system and in estimating the benefit of new therapies that could provide greater efficiency and reduce cost in these areas.

Total Medical Costs from Mobilization through 100 Days Post-ASCT in ASCT Patients (N=249).

Mean (SD) % of Total Costs Total Costs from Mobilization through 100 Days Post-ASCT $146,890 (87,628) -- Mobilization/Apheresis $38,803 (31,060) 26.5% Conditioning $63,369 (70,836) 43.1% Transplant $22,032 (30,960) 15.0% Follow-up $22,686 (38,129) 15.4%
nPKC Epsilon Negatively Regulates Platelet Functional Responses
Authors: Bynagari Yamini Saraswathy , Nagy Bela , Bhavaraju Kamala , Woulfe Donna , Kim Soochong , Kunapuli Satya P.
Published: April 2020

Protein Kinase C (PKC) are family of serine threonine kinases, known to regulate various platelet functional responses. Among them novel class of PKC isoforms (nPKC) including delta(δ), theta(𝛉), eta(η), and epsilon(ε) are expressed in platelets. Although, the role of nPKC ε and η in platelets is fairly understood, not much is known about nPKC ε and η in platelets. In this study, we investigated the role of nPKC ε in platelet functional responses using ADP-induced signaling as our stereotype. ADP causes platelet activation via Gq-coupled P2Y1 receptor and Gi-coupled P2Y12 receptor. Thus, we primarily studied the role of P2Y1 receptor in nPKC ε activation. ADP activated nPKC ε in time- and concentration- dependent manner. In the presence of P2Y1 receptor antagonist MRS-2179 and in P2Y1 knockout (KO) murine platelets ADP failed to activate nPKC ε, suggesting that ADP activates nPKC ε via P2Y1 receptor. We further investigated the functional role of nPKC ε using specific nPKC ε inhibitory RACK peptide (ε V1-2). ε V1-2 is a peptide designed to compete with native nPKC ε to bind ε-Receptors for activated C Kinase (ε-RACK) and thereby inhibits nPKC ε catalytic activity due to decreased substrate accessibility. ADP-induced thromboxane generation in human platelets pretreated with ε V1-2 peptide was more compared to the platelets pretreated with control peptide. Similarly, ADP-induced thromboxane generation in platelets derived from nPKC ε KO mouse was more compared to the wild type (WT) littermates. However, ADP- induced alpha granule secretion and aggregation in aspirin treated platelets derived from PKC ε KO mice was not significantly different from platelets derived from wild type littermates. These data suggest that nPKC e regulates an unknown pathway, which primarily regulates thromboxane generation with minimal effects on aggregation and alpha granule secretion. Furthermore, we also investigated the role of nPKC ε in PAR- and GPVI- mediated platelet aggregation and dense granule secretion. Interestingly, in both aspirin-treated and non-aspirin-treated platelets PAR- and GPVI- mediated platelet aggregation and dense granule secretion were potentiated. Consistent with ex vivo studies, FeCl3-induced arterial thrombosis was enhanced in nPKC ε KO mice compared to WT littermates.
The Frequency, Manifestations and Duration of Prolonged Cytopenias after First Line Fludarabine-Combination Chemotherapy for Chronic Lymphocytic Leukemia and Non-Hodgkin Lymphoma
Authors: Gill Saar I. , Carney Dennis , Ritchie David S. , Wolf Max , Westerman David , Prince H. Miles , Januszewicz Henry , Seymour John F.
Published: April 2020

The combination of fludarabine with other cytotoxic agents or rituximab is established in the treatment of lymphoid malignancies, with well-recognised efficacy and short-term toxicity. However, the presence and significance of prolonged cytopenias after completion of treatment have not been thoroughly described. In addition, recent guidelines on the management of chronic lymphocytic leukemia (CLL) highlight the importance of delineating the cause of post-treatment cytopenias. In order to address this important and under-reported aspect of treatment outcome, we evaluated 61 patients with indolent non-Hodgkin lymphoma or chronic lymphocytic leukemia treated in our institution with fludarabine with or without other agents as their first-line cytotoxic therapy. Post-treatment cytopenias (defined as hemoglobin < 130 g/L, neutrophils < 2.0 × 109/L or platelets < 140 × 109/L) persisting longer than 3 months were found in 43% of patients. Increasing patient age was found to be the only predictive factor (p= 0.02), whereas the presence of pre-treatment cytopenias, dose delivered, and requirement for dose delay during treatment were not. Patients failing to achieve complete count recovery by 3 months were more likely to experience post-treatment complications (infection, transfusion requirement, and bleeding) (p=0.03). The median time to resolution of anaemia, neutropenia and thrombocytopenia was 7, 9, and 10 months respectively, and the median hemoglobin, neutrophil count and platelet count were 94 g/L, 0.94 × 109/L and 86 × 109/L, respectively. Persistent cytopenias at 3 months did not impact on overall survival at 36 months. Bone marrow biopsies revealed residual disease infiltration and autoimmune aetiology in only 4% and 11% of patients, respectively, and there was no typical morphological appearance in the remaining 85%. After the completion of first-line fludarabine-based combination therapy for CLL and NHL, persistent cytopenias are frequent and clinically significant. Although cytopenias tend to resolve over time, treating physicians should be aware of these factors when considering fludarabine combination chemotherapy and when documenting post-treatment response status in CLL.
Reprogrammed Murine Fibroblasts Differentiated into Hematopoietic Progenitors Are Able to Successfully Engraft and Repopulate the Bone Marrow
Authors: Alipio Zaida , Xu Dan , Yang Jianchang , Fink Louis M. , Xu Wilson , Ward David C , Ma Yupo
Published: April 2020

Cellular therapy using embryonic stem cells has always been an area of great interest due to the pluripotent characteristics of stem cells. In 2006, Takahashi and Yamanaka (Cell 126, 663–676) demonstrated that somatic cells can be reprogrammed into a stem cell-like state, termed induced pluripotent stem (iPS) cells, by ectopic expression of Oct4, Sox2, Klf4 and c Myc. A later report (Nakagawa et al. Nat. Biotechnol.26:101–106, 2008) showed that iPS cells can be produced in the absence of the c Myc oncogene. We have used this latter strategy to successfully reprogram somatic cells derived from C57BL/6 mouse tail fibroblast to iPS cells. Retrovirus infected fibroblasts exhibited stem cell-like morphology by 14 days post infection. These iPS cells were then infected with a retrovirus that expressed HOXB4. Recombinant leukemia inhibitor factor (LIF) supplement was removed from media at this time and the cells allowed to differentiate into embryoid bodies. These cells were screened for specific differentiation stem cell markers, such as Oct4, Nanog, Sall4 and SSEA-1. iPS cells were converted into embryonic bodies and then infected with retroviruses expressing HOXB4. Embryoid bodies stably expressing HOXB4 were induced to hematopoietic differentiation by treatment of thrombopoietin (TPO), stem cell factor (SCF), vascular endothelial growth factor (VEGF), interferon gamma (IFNg) and fms-like tyrosine kinase (FLT3 ligand). Evaluation of iPS-derived hematopoietic cells on smears show strikingly similarity in morphology to the W4 mouse embryonic stem (ES) cells differentiated into hematopoietic cells as a control. Flow cytometry analysis of iPS-derived hematopoietic cells after 1 week exposure to cytokines revealed 7% B220+ cells (B cells), 11% Ter119+ cells (erythroid), and 13% Gr-1+ cells (granulocytes) similar to W4 ES cells. The iPS-derived hematopoietic cells were transplanted into irradiated immunodeficient mice via lateral tail vein injection. Transplantation of these iPS-derived hematopoietic progenitors tagged with GFP into irradiated SCID mice revealed that the hematopoietic progenitors were able to home to the bone marrow after 1 week of transplantation. Importantly, after 1 month, GFP+ engrafted cells remained in the bone marrow suggesting a long-term engraftment. This long term engraftment of the iPS-derived hematopoietic cells to the bone marrow constitutes an important step toward potential therapy of numerous patient-specific blood based diseases.
Splenic Marginal Zone Lymphoma (SMZL): Clinical Course after Splenectomy. A Single Institution Experience
Authors: Gaykazyan Susanna S , Janakiraman Nalini , Kuriakose Philip , Maeda Koichi , Hammour Tareq
Published: April 2020

SMZL is an indolent B-cell malignancy accounting for 1–2% of chronic lymphoid leukemia found on bone marrow examination and up to 25% of low-grade B-cell neoplasms in splenectomy patients. Aggressive transformation of SMZL rarely occurs. It usually presents as an incidental finding or with symptoms of splenomegaly and anemia. There is still no reliable clinical or biological scoring system for prognostic stratification. We reviewed pathology reports of 41 splenectomized patients at HFHS from 1994 to 2007 and identified 14 patients with splenic marginal zone lymphoma (SMZL). The reasons for splenectomy were symptoms of splenomegaly in all 14 patients, anemia in 13 patients, thrombocytopenia in 12 patients, AIHA in 4 patients, splenic laceration in one patient. We report here the demographics, clinical course and pathology review of these patients. The median age of patients was 77.8 years. There were 7 male and 7 female patients. ECOG performance status was 0–1 in 12(86%), and 2 in 2(14%). Of the 14 patients, 8(57%) were at Ann Arbor stage IV, 1(7%) was at stage III, 4(29%) were at stage II, and 1(7%) at stage I. LDH was above normal in 9(64%) patients B-symptoms were observed in 1(7%).

Bone marrow involvement was documented in 8(57%) of the patients. Anemia in 13(93%), thrombocytopenia in 12(86%), AIHA in 4(29%). IPI score was 1–2 in 5(36%), and score 3–4 in 9(64%) of the patients. Median weight of the spleen was 1235 gm. Bone marrow cytogenetics were abnormal in 4(29%) cases. Following splenectomy, cytopenias resolved completely or partially (CR/PR) in 13(93%) patients. Bacterial infections were observed in 4(29%) patients and 2(14%) died of infectious complications. Progressive disease requiring additional systemic therapy was documented in 5(36%) patients. Total of 5(36%) patients died. One secondary to NSLC, 1(7%) of urothelial carcinoma, 1(7%) secondary to hypercalcemia, 2(14%) due to bacterial sepsis. Patients were followed up to 139 months (with median follow-up time of 42 months). The estimated median overall survival (OS) for this group was 116.5 months (9.7 years), the median progression-free survival (PFS) was 91 months (7.6 years). The Kaplan Meier method was used to calculate these estimates. A simple median was calculated for the sample median. In summary, we report the course of 14 patients with SMZL who underwent splenectomy for symptomatic disease. Only 5(36%) required systemic therapy following splenectomy. No death was attributed to progressive SMZL. Overall course was indolent even after splenectomy. Estimated OS was 116.5 months (9.7 years), PFS - 91 month (7.6 years).
HLA-Identical Umbilical Cord Blood Transplantation from a Sibling Donor in Juvenile myelomonocytic Leukemia
Authors: de Vries Andrica , Bredius Robbert , Lankester Arjan , Bierings Marc , Trebo Monika , Stary Jan , Zecca Marco , Locatelli Franco , Niemeyer Charlotte , Van Den Heuvel Marry
Published: April 2020

Background: Juvenile myelomonocytic leukemia (JMML) is a rare malignant myeloid disorder in childhood, for which stem cell transplantation is currently the only curative treatment option. Although unrelated donor umbilical cord blood transplantations (UCBTs) have been described, series of HLA-identical sibling donor UCBTs in JMML are not available.

Methods: From the European Working Group on Childhood MDS (EWOG MDS) registry the 5 JMML patients who underwent a fully HLA matched sibling UCBT are described.

Results: The median age at diagnosis was 18 months (range 15–30 months). In one case no mutation analysis was performed, in one case a PTPN11 mutation was found. In the other cases no mutation in RAS or PTPN11 was found. None of the patients had clinical signs of NF1. The conditioning consisted of busulfan, cyclophosphamide and melphalan in four cases and of cyclophosphamide, etoposide and total body irradiation in one case. All patients engrafted slowly. In 3 patients mild acute graft versus host was noticed (grade 1 and 2), no chronic graft versus host was reported. Two patients relapsed after the initianal transplantation and underwent a second transplant with marrow of the initianal donor. One of them is in second complete remission and the other died after a second relapse. One patient developed increased donor chimerism from day 42 without any clinical sign of relapse. She was treated with DLI, 6 mercaptopurine and 13-cis retinoic acid respectively. She is still in complete remission 5 years after transplantation.

Conclusion: This EWOG serie illustrates that, although this needs to be confirmed in larger series, relatively immunologically naive HLA-matched sibling UCBT is feasible in selected cases of JMML.
Autologous Peripheral Blood Stem Cell Transplantation for POEMS Syndrome
Authors: Ohwada Chikako , Nakaseko Chiaki , Takeda Yusuke , Abe Daijiro , Takeuchi Masahiro , Sakaida Emiko , Masuda Shinichi , Shimizu Naomi , Cho Ryuko , Nishimura Miki , Kanai Kazuaki , Misawa Sonoko , Kuwabara Satoshi
Published: April 2020

Background: POEMS syndrome is a rare plasma cell disorder characterized by polyneuropathy, organomegaly, endocrinopathy, M-protein, and skin changes associated with multiorgan involvement and monoclonal plasma cell dyscrasia. The pathogenesis of POEMS syndrome is not well understood, but overproduction of vascular endothelial growth factor (VEGF) probably secreted by plasma cells is considered to be responsible. High-dose melphalan (Mel) with autologous stem cell transplantation (ASCT) has been recently considered to be an effective strategy, but its treatment related mortality is considered to be high. Here, we report the results of ASCT for POEMS syndrome.

Methods: Between January 2004 and June 2008, twelve patients with POEMS syndrome underwent ASCT at Chiba University Hospital and were retrospectively analyzed in the study. Conditioning treatment for ASCT was Mel 200 mg/m2, while Mel dose was reduced to 140 mg/m2 if a patient had poor performance status (PS).

Results: Ten out of twelve patients were male and their median age was 48 years (range 34–61). Median time from first symptom and diagnosis to ASCT was 21 and 10 months, respectively. Seven patients received induction therapy before preparation of ASCT, thalidomide-dexamethasone in four, intravenous Mel in two, and bevacizumab followed by thalidomide in one patient. Peripheral blood stem cells were mobilized by cyclophosphamide/G-CSF in nine cases and G-CSF alone in three. Median serum VEGF level before ASCT was 2835 pg/ml (range; 334–5300). Median number of infused CD34+ cells was 2.3 × 106/kg (range; 2.1–4.3 × 106/kg). Eleven patients were conditioned with Mel 200 mg/m2, while one patient with poor PS (Karnofsky score 30%) received Mel 140 mg/m2. Median time to neutrophil recovery (>500/mm3) was 13 days post transplant (range 9–17 days). Only two patients developed respiratory distress during neutrophil engraftment period. Both cases were successfully treated with corticosteroid therapy, none of them required intubation. During a median follow up period of 16.5 months (range 2–54), there was no transplant related death. Neurologic improvement was significantly observed in all patients and median post transplantation serum VEGF level was decreased to 330 pg/ml (range 87–3250). Only one patient experienced relapse of polyneuropathy at 43 months post transplant following reincrease of serum VEGF level.

Conclusions: ASCT is an effective therapy for POEMS syndrome. Despite previous published reports arousing potential risk of life-threatening engraftment syndrome, treatment related mortality and periengraftment pulmonary complications were significantly low in our patient series. We speculate that stabilization of serum VEGF level by induction therapy before ASCT could play a potential role for reducing the risk of peritransplant complications.
Efficacy of Nilotinib (formerly AMN107) in Patients (Pts) with Newly Diagnosed, Previously Untreated Philadelphia Chromosome (Ph)-Positive Chronic Myelogenous Leukemia in Early Chronic Phase (CML-CP)
Authors: Cortes Jorge , O’Brien Susan , Jones Dan , Ferrajoli Alessandra , Konopleva Marina , Borthakur Gautam , Garcia-Manero Guillermo , Letvak Laurie A. , Kantarjian Hagop
Published: April 2020

Background: Nilotinib is an oral tyrosine kinase inhibitor with high selectivity towards Bcr-Abl and approximately 30-fold more potent than imatinib, and is effective in patients with CML after imatinib failure. We initiated a phase II study to evaluate the efficacy of nilotinib as 1st line therapy in pts with newly diagnosed CML-CP.

Aims: To investigate the efficacy and safety of nilotinib as initial therapy for patients with CML-CP.

Methods: The primary objective was to estimate the proportion of pts attaining major molecular response (MMR) at 12 months (mo). Pts with untreated CML-CP (or with <1 months of therapy with imatinib) were eligible and received nilotinib 400mg twice daily. A cohort of patients with previously untreated CML in accelerated phase (AP) was also included.

Results: Forty-nine pts have been treated for a median of 13 months (mo). The median age was 47 years (yrs) (range, 21 to 81); 69% are Sokal low risk. Eight (16%) had received imatinib for <1 months. Overall, 46/48 (96%) of evaluable CP pts achieved a complete cytogenetic response [CCyR]. The rate of CCyR at 3, 6 and 12 mo for pts in CP compares favorably to those observed in historical controls treated with imatinib 400mg or 800 mg daily:

Percent with CCyR (No. evaluable) Months on therapy Nilotinib Imatinib 400mg Imatinib 800mg P value 3 93 (45) 37 (49) 62 (202) < 0.0001 6 100 (36) 54 (48) 82 (199) < 0.0001 12 96 (27) 65 (48) 86 (197) 0.0003 18 92 (12) 68 (38) 89 (179) 0.0042 24 91 (11) 70 (40) 88 (173) 0.0151

MMR was observed in 45% at 6 mo and 52% at 12 mo. Two of 44 (5%) evaluable pts have achieved confirmed complete molecular response, and 3 others unconfirmed (ie, only achieved on their last assessment). Grade 3–4 hematologic toxicity (transient) included thrombocytopenia in 10%, neutropenia in 12%, and anemia in 2%. Grade 3–4 non-hematologic adverse events (regardless of causality) included elevation of bilirubin in 8% and lipase in 6%. 19 (36%) pts had transient treatment interruptions and 17 (32%) had dose reductions. The actual median dose is 800mg daily. Three pts have come off study: 1 pt’s choice and 2 because of toxicity (1 liver, 1 pericardial effusion). One of them (liver toxicity) transformed to blast phase shortly after coming off study. Estimated 24 month EFS (event = loss of CHR, loss of MCyR, AP/BP, death, or off because of toxicity) is 95%.

Conclusion: Nilotinib 400 mg twice daily induces a CCyR in nearly all patients as early as 3 months after the start of therapy with a favorable toxicity profile. Accrual is ongoing.
A Novel Mutation (Q694X) in the ITGB3 Gene Causing Glanzmann’s Thrombasthenia from the Sultanate of Oman
Authors: Albalushi Talal , AlZadjali Shoaib , AlHaddabi Hamood , Dennison J. David , Alkindi Salam , Muralitharan Shanmugaonkar , Arinami Tadao , Pathare Anil
Published: April 2020

Background : Glanzmann Thrombasthenia (GT) results from mutations in the genes ITGA2B and ITGB3, located on chromosome 17q21–23 which encodes the platelet glycoprotein αIIbβ3 complex, namely GPIIb (αIIb) and GPIIIa (β3), the fibrinogen receptors on platelets, which play an important role in platelet aggregation. Patients with GT can require frequent hospitalization and can be a burden on the nation’s health resources. The possibility that GT could be cured by gene replacement therapy makes it essential to study the molecular basis of the GT patients in a particular family or kindred.

Objectives : Our aim was to identify the underlying mutations responsible for GT in Omani patients in order to establish a strategy for genetic counseling and carrier detection to prevent the occurrence of the homozygous state by prenatal diagnosis.

Methods: GT was diagnosed in a 17 year old Omani female at the Sultan Qaboos University Hospital. The diagnosis of GT was based on clinical features, platelet aggregometry and biochemical studies. Platelet surface expression of GPIIb/IIIa was also studied by flowcytometry. Molecular studies performed at Medical Genetics Department, Tsukuba University, Japan, include DNA sequencing of all exons and exon-intron junctions of ITGA2B and ITGB3 of the two genes by the ABI 3100 Genetic Analyzer®. [Applied Biosystems, Foster City, CA, USA]. Genomic DNA was also analyzed by Illumina Human-1 Bead Chip Illumina® (Illumina Inc., San Diego, CA, USA) to exclude the whole region of the two genes that could produce an apparent homozygous state.

Results : We have identified a novel nonsense causative mutation (Q694X) by sequencing the ITGB3 gene. [Figure 1a & b]. In addition, sequencing ITGB3 gene also revealed 2 SNPs (rs 3809863; IVS14+9C/T, rs 3809865; 3383T/A). The Micro-Array assay using Illumina Human-1 Bead chip excluded the possibility of deletion of these genes in chromosome 17 in this patient.

Summary/Conclusion: A stop codon was found in exon 13 of ITGB3 gene causing the translated protein to be abnormally shortened. It is hypothesized that the altered form of ITGB3 gene is both extremely unstable and rapidly degraded after its biosynthesis, leading to a loss of function of the protein. Further RNA expression studies, transfection tests and cDNA sequencing are ongoing to elucidate the molecular mechanisms responsible for GT.

Fig. 1 a] Structure of αIIbβ3 complex. The blue box indicates the position of the nonsense mutation in ITGB3 gene, which is clearly before the transmembrane (Red Box) AA sequence. 1 b] Electropherogram showing a nonsense mutation in exon 13 of ITGB3. Fig. 1. a] Structure of αIIbβ3 complex. The blue box indicates the position of the nonsense mutation in ITGB3 gene, which is clearly before the transmembrane (Red Box) AA sequence. 1 b] Electropherogram showing a nonsense mutation in exon 13 of ITGB3.
Selective Serotonin Reuptake Inhibitors Influence Agonist-Induced Platelet Aggregation. Preliminary Results from Comorbidity of Depression and Cardiovascular Disease Study
Authors: Iqbal Omer , Adiguzel Cafer , Fareed Jawed , Hoppensteadt Debra , Litinas Evangalos , Zhu He , Halaris Angelos
Published: April 2020

Selective serotonin reuptake inhibitors (SSRI) are a first-line treatment option for depressive illnesses, anxiety and obsessive-compulsive disorders with a favorable risk benefit ratio and side effects. They selectively inhibit neuronal reuptake of serotonin and result in depleted serotonin stores in the dense bodies of platelets. It is hypothesized that depressive illnesses cause platelet activation and endothelial dysfunction, which could be modulated by the use of SSRIs. In order to validate this hypothesis an institutional-based co morbidity of depression and cardiovascular study is undertaken. Patients with depression (n=25) and healthy control (n=25) are recruited in this study. Males and females between the ages of 20 and 65 years meeting the DSM-IV criteria for major depressive disorder served as the inclusion criteria for the study. The exclusion criteria included patients with heart disease, diabetes, lipid disorders, history of smoking, pregnant and lactating women, psychosis, schizoaffective illnesses and presence of other Axis I diagnosis with the exception of generalized anxiety disorder. Blood samples were collected at baseline and then 4 and 8 weeks following treatment with Escitaloprim, an SSRI. Preliminary results from the agonist-induced platelet aggregation showed varying degrees of inhibition of collagen, ADP, arachidonic acid and epinephrine-induced platelet aggregation as shown in the table below.

Agonist induced platelet aggregation profile Averaged % aggregation Time Collagen AA ADP Epinephrine Saline Baseline 71.5± 22.4 64.1± 35.1 48.15± 30.9 55.65± 29.2 6.9± 4.5 Week 4 63.4± 19.5 67.1± 25.1 33± 26.7 37.375± 25.8 9.375± 7.8 Week 8 74.7± 21.3 55.71± 36.1 52.4± 31 50.4± 32.3 5± 3.9

While a number of patients showed an inhibition of ADP (2.5μM) and epinephrine (10μg/ml) and collagen (380μg/ml), however, other patients showed an inhibition of collagen-induced platelet aggregation. These results indicate that patients treated with SSRIs may have inhibition of platelet activation and aggregation, which may have an impact in the prevention of atherothrombotic cardiovascular disease. SSRIs might increase the bleeding tendency in some patients by inhibiting platelet aggregation. However, when the SSRIs are given in combination with other drugs patients may have a propensity for bleeding complications due to drug-drug interactions. Further large-scale trials are warranted to validate these results.
Delayed Onset of T Cell-Mediated Xenogeneic Disease in Rag2−/− γc−/− mice after Co- Transplantation of in Vitro Expanded Human CD4+CD25highCD45RA+ Regulatory T Cells
Authors: Boeld Tina J , Obermann Ellen , Andreesen Reinhard , Hoffmann Petra , Edinger Matthias
Published: April 2020

In murine disease models, the adoptive transfer of donor CD4+CD25+ regulatory T cells (Treg) prevents lethal graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation. We recently described methods for the isolation and in vitro expansion of human Treg from adult peripheral blood and found that only CD45RA+ but not CD45RA− CD4+CD25high Treg (RA+ and RA− Treg, respectively) maintain FOXP3 expression as well as suppressive activity after prolonged culture (Hoffmann et al. Blood108:4260; 2006). Here, we compared the immunoregulatory capacity of these two different Treg subpopulations in vivo in a xenogeneic GVHD (xGVHD) model. Transplantation of 20x106 human peripheral blood mononuclear cells (huPBMCs) into immunodeficient BALB/c Rag2−/−γc−/− mice led to engraftment and extensive proliferation of human CD45+ cells (huCD45+), which was accompanied by clinical signs of xGVHD. Histological and flow cytometric evaluation revealed that mainly CD8+ T cells within PBMC caused xGVHD and that main targets were liver, lung and spleen as well as mouse hematopoiesis. When expanded CD45RA+ Treg were co-transplanted at a 1:1 ratio with CD4+ and CD8+ T cells contained in huPBMC, the frequency of huCD45+ cells in peripheral blood (PB) at d14 was reduced to 12 ± 4.9 % (n=15), while animals that received expanded RA− Treg had 30 ± 7.5 % (n=15) and animals without co-transfer of Treg 40 ± 8.3 % (n=14) huCD45+ cells. The expansion of conventional T cells, however, was only delayed but not prevented, as by d21 the percentage of huCD45+ cells in PB increased to 35 ± 9.2 % (n=15) when RA+ Treg were co-transplanted as compared to 40 ± 9.3 % (n=15) with RA− Treg and 50 ± 6.4 % (n=14) without Treg administration. Subsequently, all mice developed clinical signs of xGVHD and 73.3 % of mice co-transplanted with RA+ Treg, 86.7 % co-transplanted with RA− Treg and 78.6 % of mice transplanted with huPBMCs alone died within 100 days. Since Treg mainly inhibited early expansion of huPBMC, we examined the splenic cellularity of xeno-transplanted animals at d10 after cell transfer. While spleens of mice that received only huPBMC contained 20 ± 3x106 huCD45+ cells (n=10), co-transfer of RA− Treg reduced huCD45+ cells to 14 ± 4.8×106 (n=9) whereas animals that received RA+ Treg contained only 5 ± 1.1×106 huCD45+ cells (n=11) at that time. Furthermore, IL-2 and IFN-γ production of conventional T cells re-isolated from the spleen on d10 was reduced 5fold in mice co-transplanted with RA+ Treg as compared to recipients of only huPBMCs or additional RA− Treg. Thus, in vitro expanded human RA+ Treg suppress the proliferation and cytokine production of conventional T cells in early phases of xGVHD, but ultimately do not prevent disease onset and mortality.