bluuebunny/crossref_metadata_2025 huggingface dataset:
Hodgkin’s Disease Presenting with Osseous Involvement.
Authors: Chandra Divay , Ewton April , Baker Kelty R.
Published: October 2019

A 51-year-old Caucasian female with a past history of hypertension and rheumatoid arthritis presented with intermittent fevers and fatigue for one year and left hip pain for six months. During the month prior to presentation she noted temperatures to 102° F at home and was having difficulty bearing weight on her left hip. She denied any weight loss, skin rash, morning stiffness and pain or swelling in other joints. Treatment with NSAIDS and a course of steroids had provided minimal relief. Physical exam was remarkable for the presence of pallor, two 1.5 cm, firm, non tender, mobile lymph nodes in the left posterior superior cervical chain and one 3 cm tender, fixed lymph node in the left inguinal chain. She had limited range of motion with both active and passive movement of the left hip and the joint was tender to heel percussion. Her lab work was significant for a moderate anemia of chronic disease, thrombocytosis to 614,000/μl, alkaline phosphatase of 400 U/L, lactate dehydrogenase of 204 U/L, erythrocyte sedimentation rate of 95 and β2-microglobulin of 3.6 mg/L. Magnetic resonance imaging of bilateral hips showed an aggressive marrow replacing process in the left iliac bone that had eroded posteriorly into the adjoining gluteal musculature with multiple enlarged lymph nodes along the left pelvic sidewall and bilateral inguinal chains. PET/CT revealed increased tracer uptake in the left iliac bone, adjacent soft tissues and in multiple lymph nodes in the neck, medistinum and pelvis and the liver and spleen. Excisional biopsy of an enlarged left inguinal lymph node revealed infiltration by nodular tissue with thick fibrous bands and classic Reed-Sternberg cells which were positive for CD 15 and CD 30. The patient was diagnosed with stage IV B Hodgkin’s Disease. Planned treatment consisted of 8 cycles of ABVD, followed by consolidative radiation to the left iliac bone. Patients with Hodgkin’s disease usually present with painless lymphadenopathy typically in the cervical region. Although radiologically detectable bone involvement is not unusual in disseminated disease, osseous lesions are seldom the primary manifestations. Less than 20 cases of Hodgkin’s disease presenting with osseous involvement have been described. A majority of these are of the nodular sclerosing subtype and involve the axial skeleton, most commonly the vertebrae, femur, and iliac bones as was seen in our patient.
Dose Adjusted EPOCH Chemotherapy Plus Rituximab Combination (DA-R-EPOCH), an Effective Upfront Therapy for Patients with Poor-Risk Aggressive B-Cell Lymphomas. Results of a Pilot Study in a Single Center.
Authors: López Andrés , Gallur Laura , Palacios Andrés , Navarrete Mayda , Ruiz-Marcellán Carmen , Castellví Josep , Cajal Santiago Ramón y , Juliá Antonio
Published: October 2019

Poor risk aggressive non-Hodgkin’s lymphomas have a short EFS and OS when treated with conventional chemotherapy schedules. High dose therapy followed by stem cell rescue has been attempted with disparate results. Based on Wilson WH et al. schedule (Wilson WH, Grossbard ML, Pitaluga S et al. Blood2002; 99: 2685–93), a pilot study has been conducted in order to assess the efficacy and toxicity of DA-R-EPOCH in patients with Intermediate-High/High (IH/H) IPI score diffuse large B-cell (DLBCL) and follicular large (grade 3b) B-cell (G3bFL) lymphomas. No restrictions were done regarding the performance status.

Methods. From August 2002 to May 2005, 28 untreated patients (pts) were enrolled in the study. Basically the regimen consisted on 6 cycles every 21 days of Rituximab (day 1) followed by a 96 hours infusion of etoposide, vincristine and doxorubicine and bolus cyclophosphamide on day 5 plus prednisone for 5 days. The doxorubicine, etoposide and cyclophosphamide doses were adjusted according to the hematological toxicity as reported by Wilson WH et al (Wilson WH, Gutierrez M, O’Connor P et al. 2002; 29 (Suppl. 2):41–7). Response rate and survivals were calculated in intention to treat. Pts not reaching a PR ≥ 70% after 3 cycles or a CR/CRu after completion of therapy were assigned as failure in the time to treatment failure (TTF) calculation.

Results. Patients characteristics: median age 62,5 years (range:26–72); male/female: 11/17; DLBCL 21 (75%) and G3bFL 7 (25%); 19 (68%) had IV clinical stage; 13 (46%) B symptoms; PS (ECOG) > 2 in 12 pts (42%); 21 pts (75%) had extranodal and 10 (36%) bone marrow involvement; 32% had serum albumin < 3 g/dL; Hb < 100 g/L in 13 (46%); 46% had high b2microglobulin levels, and 100% elevated LDH. High age-adjusted IPI score in 15 pts (54%). Dose adjustment: Dose escalation could be done in most patients (18 out of 21 receiving all 6 cycles). There were an inverse correlation between the mean dose of doxurubicin given and age (r=−0.555; P2=0.009) and IPI (r=−0.507; P2=0.019). Response and survival: 17 out of 23 pts evaluated for response (74%) achieved a CR/CRu. There were 2 early deaths. At a median follow up of 14 months (range: 2–35) the estimated 2-year TTF and OS were 58% and 73% respectively. Toxicity: After 143 cycles administered grade 3–4 mucositis was observed in 9 courses (6%), grade 3–4 nueropathy in 1 (<1%) and neutropenic fever in 20 (14%).

Conclusion. DA-R-EPOCH is a feasible approach and as effective as other more intensive chemotherapy schedules as upfront therapy in patients with intermediate-high or high risk IPI score aggressive large B-cell lymphomas (DLBCL and G3bFL) with acceptable toxicity. Updated data will be presented at the meeting.
Role of Plasmin and uPAR in Bone Marrow HSC/HPC Retention and Mobilization.
Authors: Tjwa Marc , Moons Lieve , Theunissen Koen , Moura Rute , Blasi Francesco , Dewerchin Mieke , Sidenius Nicolai , Collen Desire , Carmeliet Peter
Published: October 2019

We previously identified the plasmin protease family as a critical determinant of the mobilization of hematopoietic stem and progenitor cells (HSC/HPC), but the role of the urokinase receptor uPAR remained unclear. uPAR is a membrane-anchored glycoprotein, which not only localizes its ligand urokinase (uPA) to the cell surface via its GPI-anchor but also regulates β1-integrin dependent cell adhesion and migration.

Following 5-FU myeloablation or G-CSF treatment, mice lacking uPAR (uPAR−/−) had impaired hematopoietic recovery and HSC/HPC mobilization as compared to wild type (WT) mice. However, this phenotype was not mimicked in mice lacking uPA, suggesting a role of uPAR in mobilization independent of uPA-mediated proteolysis.

The impaired mobilization in uPAR−/− mice was reversed upon pre-transplantation with WT BM cells (BMC), suggesting functional expression of uPAR on transplantable BMCs. Conversely, loss or inhibition of uPAR on transplanted BMCs impaired homing to the BM but not to the spleen, and compromised survival of myeloablated WT recipients. In vitro experiments revealed that loss or inhibition of uPAR impaired BMC adhesion to stromal cells and fibronectin. Anti-α4-β1 antibodies blocked adhesion of WT but not uPAR−/− BMCs. Thus, uPAR appears to regulate BM homing and α4-β1 dependent retention of transplantable BMCs, possibly HSC/HPCs.

If uPAR mediates retention of HSC/HPCs, then this signal should be inactivated upon mobilization. Indeed, in 5-FU or G-CSF-treated WT mice, we found increased uPAR cleavage, and elevated levels of soluble uPAR (suPAR) in BM plasma. These processes failed to occur in mice lacking plasminogen, suggesting that plasmin cleaves uPAR during mobilization. Cleavage of uPAR appeared critical as the inactivation of the retention signals membrane-bound Kit ligand and SDF-1α was normal in uPAR−/− mice. Moreover, the generated suPAR may also affect the BM, as administration of recombinant suPAR in WT mice enhanced hematopoietic recovery and HSC/HPC mobilization after 5-FU or G-CSF. In vitro and transplantation experiments revealed that suPAR blocked α4-β1 dependent adhesion.

Thus, in steady state, membrane-anchored uPAR appears to function as a BM retention signal for transplantable BMCs, possibly HSC/HPCs. In conditions of mobilization, the uPAR retention signal is cleaved, which weakens α4-β1 dependent adhesion and allows mobilization out of the BM. Soluble uPAR may then additionally amplify mobilization, in part by further attenuating α4-β1 dependent adhesion to the BM. Currently, we are investigating the role of uPAR on subsets of HSC/HPCs, and in the different BM niches. We are also performing long-term competitive repopulation experiments to further delineate the therapeutic potential of uPAR.
Fludarabine-Based Regimens in the Treatment of Indolent Non-Hodgkin’s Lymphoma Patients: A Clinical Study of the Fludarabine Cooperation Group in 16 Hospitals in Eastern China.
Authors: Li Junmin
Published: October 2019

Objective: We performed this prospective, multi-center study to evaluate the efficacy and safety of fludarabine-based regimes in the treatment of previously untreated and refractory, or relapsed, indolent non-Hodgkin’s lymphoma (NHL).

Methods: Two fludarabine-based regimens were used in the clinical study. The FC regimen which consisted of fludarabine 50 mg/day, and cyclophosphamide 200 mg/day, intravenously, for 3 consecutive days, was given to the previously untreated patients. The FMD regimen which consisted of fludarabine 50 mg/day, intravenously, for 3 consecutive days, mitoxantrone 10 mg/m2, intravenously, on the first day, and dexamethasone 20 mg/m2/day, intravenously, for 5 consecutive days, was given to the relapsed and refractory patients.

Results: Forty-seven indolent NHL patients (29 male and 18 female) were enrolled in this study from February 2003 to May 2005. The main disease subtypes were: chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL; n = 25), follicular lymphoma (FL; n = 18), lymphoplasmic lymphoma (LPL; n = 3), marginal zone lymphoma (MZL; n =1). Overall response (OR) rate was 93.62%, and 44 of 47 patients achieved a response. The complete response (CR) rate was 63.83%, and 30 of 47 patients achieved a response. In the 16 previously untreated patients, the CR rate was 75.0% (n =12); the OR rate was 93.8% (n = 15). In the 31 relapsed and refractory patients, the CR rate was 61.3% (n =19) and OR rate was 93.5% (n =29). Median patient follow-up was 12 months (range, 1–27 months). The 12-month progression free survival (PFS) rates of previously untreated patients and relapsed or refractory patients were 93.3±6.4% and 91.04±6.1%, respectively; and the 18-month overall survival (OS) rates were 90.9±8.67% and 90.32±6.90%, respectively. Both regimens were well tolerated and the major toxicities were bone marrow suppression and gastrointestinal response.

Conclusion: The FC and FMD regimens are effective and safe for indolent NHL patients.
Primary Mediastinal Large B-Cell Lymphoma: Treatment Outcomes in 43 Patients.
Authors: Mello Celso L. , Lima Vladimir C. , Valadares Christiane , Fernando Soares A. , Lopes Carvalho A. , Olavo Feher
Published: October 2019

Objectives: Primary mediastinal B-cell Lymphoma (PMBL) is recognized as a separate entity in the WHO classification. Treatment for PMBL is based on a combination of conventional dose chemotherapy, high dose chemotherapy and radiation therapy. The best strategy is still undefined. We conducted a retrospective analysis of patients with PBML to identify clinical prognostic factors.

Materials and Methods: Between 1989 and 2004, 43 consecutives patients treated at Hospital do Cancer AC Camargo, São Paulo, Brazil were identified and clinical and histological data were reviewed. All patients had previous diagnosis of diffuse large B-cell lymphoma, with positive CD20 on neoplastic cells. For patients with extrathoracic involvement at presentation a predominant anterior mediastinal lesion should be present. Induction chemotherapy regimens were grouped in first generation (CHOP or CHOP-like), third generation (PromaceCytabon or MACOP) and other (pediatric regimens, COP).

Results: Age ranged from 16 to 82 years-old, 30 females and 13 male. Patients aged < 35 yo had better outcome compared with those older than 35 yo (5 years OS - 56% x 34%, p=0.048). Among clinical variables, female gender, stage IA-IIB, IPI 0-1, normal LDH, absence of mediastinal bulky disease were associated with better prognosis, although not statistically significant. Response rate to first generation regimens was: 37% CR (11/29), 24% PR (7/29) and 24% PD (7/29). Four patients were treated with third generation regimens with 2 CR and 2 PR. 20 out of 25 patients with PR or CR to first line chemotherapy received mediastinal radiation therapy. More than 65% of patients had a minimum time of follow up 5 years. With a median follow up of 22.3 months, projected 5 year OS was 47% and for the responders the median PFS was 8,4 months. No difference in OS and PFS was observed among the three chemotherapy groups.

Conclusion: Our analysis showed that response rate to first line regimens was around 60% and 25% of patients were primarily refractory to CHOP regimen. Age younger than 35 years old was associated with a better prognosis. 5 years overall survival was 45 % and is in accordance with the literature. Although recent studies have demonstrated biological similarities between PMBL and Hodgkin’s Lymphoma, the prognosis of PMBL is less favorable than HL. Better understanding of the disease will help in developing more appropriate therapeutic strategies for PMBL.
Generation of a Fully Human High Affinity Neutralizing Antibody Against MT-SP1/Matriptase and Its Potential Role for the Treatment of B Cell Lymphoma.
Authors: Foltz Ian N. , Morkowski Stan K. , Scatena Caroline D. , Ling Peter , Li Hao , Liu Ying , King Chadwick T. , Kang Jaspal , Manchulenko Kathy , Perkins Lisa , Kang Sukhbir , Chen Minchao , Godfrey Wayne , Keyt Bruce , Richardson Jennifer H.
Published: October 2019

Membrane-type serine protease 1 (MT-SP1)/Matriptase is a type II transmembrane serine protease that is primarily expressed in epithelial cells and is over-expressed in several cancers. MT-SP1 has been implicated in the processes of tumor cell invasion and metastasis, primarily due to its ability to degrade extracellular matrix proteins and to activate the latent forms of hepatocyte growth factor (HGF), urokinase type plasminogen activator and protease-activated receptor 2. To further explore the role of this protein in human cancer, we developed a fully human IgG1, high affinity, neutralizing antibody against MT-SP1 (95/96) using XenoMax® technology. Using this antibody, we confirmed the expression of MT-SP1 on epithelial cells. In addition, we observed MT-SP1 expression on human peripheral blood B lymphocytes and monocytes. Furthermore, expression was also observed on several B cell lymphomas including Ramos, Raji and Daudi cell lines where, in contrast to epithelial cells, MT-SP1 was found in the absence of its inhibitor and cognate receptor, HGF activator inhibitor -1 (HAI-1). As 95/96 is a potent inhibitor of MT-SP1 enzymatic activity, we used the antibody to examine the functional role of MT-SP1 in B cell lymphoma. Using a tripeptide fluorogenic substrate of MT-SP1, we detected cell surface enzymatic activity on Ramos cells that was completely inhibited by 95/96, implying that the MT-SP1 expressed on Ramos cells is catalytically active. The ability of 95/96 to modulate the invasive properties of Ramos lymphoma cells was evaluated using a rodent hind-limb paralysis model. The median survival of SCID mice systemically inoculated with Ramos lymphoma cells, and receiving 95/96, was significantly extended compared to the animals receiving an isotype-matched control antibody (35 days vs. 26 days, p < 0.01). 95/96 had no effect on the in vitro proliferation or invasion of Ramos cells and was unable to induce cell death in a whole blood assay or by complement-dependent cytolysis. Collectively, these data suggest a role for MT-SP1 catalytic activity in the invasion or metastasis of Ramos B cell lymphoma.
Autoregulation of RUNX1 Activity by Homodimerization.
Authors: Sinha Kislay K. , Li Donglan , Hay Maher A. , Nucifora Giuseppina
Published: October 2019

RUNX1 also known as AML1 is a transcription factor essential for normal hematopoiesis, platelet production and thymocyte development. RUNX1 is the most frequent target of chromosomal translocations and acquired or inherited point mutations associated with human leukemia. RUNX1 is a DNA-binding transcription factor that can act both as an activator and a repressor of gene expression depending probably on the association of RUNX1 with co-activator or corepressors in large transcription complexes at promoter sites. The C-terminus of RUNX1 contains an inhibitory region, ID, which represses positive regulation of RUNX1-dependent genes. Thus, this region could potentially act as a switch and co-operate with RUNX1-interacting transcription factors in the choice between gene activation or gene repression. Here we have examined the role of ID in gene regulation by RUNX1. We found that this region is a homo-dimerization motif that controls RUNX1-RUNX1 interaction in vitro and in vivo. The association of RUNX1 with itself through this domain appears to reduce the positive transactivating potential of RUNX1 and, if provided in trans, this domain can repress by itself the activity of RUNX1 in reporter gene assays. Our studies suggest that RUNX1 autoregulates itself through its ability to form a homodimer. Data will be shown on the effects of the ID region in hematopoietic differentiation of cell lines. The results indicate that the ID region plays critical role in RUNX1 activity and is essential to control the correct execution of hematopoietic programs. It is of importance that aside from the t(12;21), all chromosomal translocations and virtually all point mutations associated with leukemia profoundly affect the integrity of the C-terminus including the ID region. These studies suggest a novel pathway involved in RUNX1 leukemogenesis and provide new targets for the management of RUNX1-leukemia. Kislay Sinha and Donglan Li contributed equally to this study.
Prevention of Red Cell Alloimmunization by CD25 Regulatory T Cells.
Authors: Yazdanbakhsh Karina , Zheng Xiaoying , Yu Jin , Heck Susanne
Published: October 2019

BACKGROUND: Alloimmunization after exposure to donor red cell (RBC) antigens remains a major clinical problem in transfusion medicine, particularly for patients in need of chronic transfusions. The identification of T regulatory cells (Tregs) among the CD4+CD25+ T cell subset as key regulators of peripheral tolerance in mice as well as humans has opened an exciting era in the prevention and treatment of autoimmune disease and for improving organ transplantation. However, their potential in inducing transfusion tolerance so as to prevent or blunt the alloimmune response to allogeneic red cells has not been explored.

METHODS AND RESULTS: To determine a role for Tregs in control of alloimmune responses, we have manipulated Treg numbers by either depletion of CD25-expressing cells or transfer of purified CD4+CD25+ Treg cells in two different mouse models of RBC immunization. Using a mouse model of RBC immunization in which weekly injection of rat RBCs results in the development of antibodies to rat red cells, we previously showed that depletion with anti-CD25 increased the antibody response to rat RBCs (Mqadmi et al., 2005, Blood, 105, 3746). We now found that adoptive transfer of purified populations of CD4+CD25+ cells from non-primed C57/Bl6 mice into naïve syngeneic recipients (n=9) completely inhibited the anti-Ig response to rat RBCs in 8 out of 9 mice even up to 5 weeks post-immunization but transfer of control CD4+CD25- cells did not (p=0.029). As an experimental system to study regulation of antibody production following allogeneic RBC transfusion, we used red cells (equivalent to either 1 or 2 packed red cell units) from mice transgenic for human glycophorin A (GPA) blood group antigen as donor cells and transfused wild type C57/Bl6 mice to induce alloantibodies. We found that depletion with anti-CD25 enhanced the alloantibody (IgG and IgM) production (p=0.042), indicating that CD25 Tregs play an important role in regulation of alloantibody responses. More importantly, adoptive transfer of purified splenic population of CD4+CD25+ from naïve mice prevented the induction of IgG and IgM alloantibody production in transfusion recipients (n=8).

CONCLUSIONS: Altogether, our results demonstrate that Tregs control transfusion-associated alloantibody responses. Our data indicate that Treg immunotherapy by enhancement of Treg function may be exploited for future therapeutic approaches for suppressing transfusion immunization events.
Bioluminescence Imaging of Spontaneous Lymphomas and Contribution of Distinct NFκB Pathways.
Authors: Ratner Lee , Bernal-Mizrachi Leon , Rauch Daniel , Harding John , Gross Shimon , Piwnica-Worms David
Published: October 2019

The nuclear factor κB (NFκB) pathways have been implicated in tumorigenesis in several lymphoid malignancies, but the responsible mechanisms remain to be fully defined. EBV and HTLV-1 immortalized lymphoma cell lines with constitutively active NFκB were found to be resistant to inducers of the extrinsic and intrinsic apoptosis pathways. Several NFκB-target genes were over-expressed in these cell lines, including Bcl-xL, Fas-associated death domain-like IL-1β converting enzyme inhibitor protein (FLIP), and inhibitors of apoptosis (CIAP, XIAP, and survivin). Inhibition of the canonical or non-canonical NFκB pathways with small interfering RNAs (siRNA) or adenovirus expressing a stable form of inhibitor of NFκB (IκB) enhanced sensitivity to apoptosis inducers, and resulted in lower levels of expression these NFκB target genes. Repression of XIAP with siRNA was particularly potent in enhancing apoptosis sensitivity. These findings demonstrate an important role of both NFκB pathways in mediating resistance to apoptosis, and distinctive anti-apoptotic downstream target gene profiles responsible for this effect. In order to utilize HTLV-1 Tax activity as an in vivo biomarker for tumorigenesis, we generated transgenic mice with firefly luciferase driven by the HTLV-1 long terminal repeat (LTRLUC) and bred them with the GzBTax animals to produce double transgenic LTRLUC/GzBTax (LLT) mice which develop large granular lymphocytic leukemia, splenomegaly, localized bone destruction, and solid tumors of the ears, legs, nose, and tail. We found that the bioluminescence of the LLT mice

serves as a reporter for spontaneous tumorigenesis in vivo, is transferable as a useful tumor marker in cell culture and allograft models derived from primary tumors, is inducible by immunomodulatory agents, and is a sensitive indicator for even the earliest events in tumor development.
Genomic Profiling of Follicular Lymphoma Identifies Novel High-Frequency Copy-Neutral Loss of Heterozygosity on Chromosomes 1p and 6p and Identifies Candidate Genes Involved in FL Biology.
Authors: Malek Sami N. , Ouillette Peter D. , Saddler Chris M. , Shedden Kerby A. , Ross Charles W.
Published: October 2019

Follicular lymphoma (FL) constitutes the second most common non-Hodgkin’s lymphoma in the Western world. The clinical course is variable and only in part explained by known tumor-intrinsic (genetic) or -extrinsic (infiltrating immune cells, infiltrating macrophage content) factors. FL carries the hallmark chromosomal translocation t(14;18), deregulating expression of Bcl-2, but this is not sufficient to explain either FL biology or clinical behavior. We have employed genomic profiling technology using the Affymetrix 50K-Xba1 oligonucleotide single nucleotide polymorphism (SNP) chip platform to interrogate the genomes of 58 FL specimens for loss of heterozygosity (LOH) and chromosomal copy number changes. Where available, these high-density genomic data were correlated with published FL gene expression data. Using this integrated approach, we provide data that suggest that i) Bcl11A is a likely target gene for the amplification on chromosome 2p15–16 in FL, ii) that the amplification of der(18)t(14;18) does not include Bcl-2, iii) high-frequency copy-neutral LOH exists at 1p36 (50%) and 6p (33%), and that 6p LOH possibly deregulates IRF4 and, iv) deletion 6q is non-overlapping, complex and possibly involves four or more gene loci of importance. In addition, high-frequency LOH (with and without chromosomal copy loss) exists on chromosomes 9p, 10q and 16p. These data should facilitate analysis of specific genes in FL and should allow for development of a SNP -and genomics-based assay for FL risk stratification and outcome prognostication.
Pharmacokinetics and Pharmacodynamics from a First-in-Human Phase 1 Dose Escalation Study with Antagonist Anti-CD40 Antibody, HCD122 (Formerly CHIR-12.12), in Patients with Relapsed and Refractory Chronic Lymphocytic Leukemia.
Authors: Byrd John C. , Flinn Ian W. , Khan Khuda D. , Kipps Thomas J. , Aukerman Lea , Fox Judith , Girish Sandhya , Guzy Serge , Bilic Sanela , Solinger Alan , Dort Sherri , Wang Yongyu , Hurst Deborah , O’Brien Susan
Published: October 2019

The CD40 antigen is expressed in most B-cell malignancies, including chronic lymphocytic leukemia (CLL), and represents an attractive target for antibody therapy. HCD122 is a high affinity, fully human IgG1 antagonistic anti-CD40 monoclonal antibody designed to exert antitumor activity as an inhibitor of CD40-ligand-mediated survival signals and as a potent mediator of antibody-dependent cellular cytotoxicity (ADCC). In this phase 1 study to determine maximum tolerated dose (MTD) in CLL, eligible patients (pts) include those with relapsed or refractory disease after prior fludarabine treatment and are assigned to receive one cycle of 4 weekly infusions of HCD122 at doses ranging from 0.3 to 10 mg/kg depending upon the dose cohort. A standard phase I design is used with each dose cohort enrolling 3–6 pts for evaluation of MTD, toxicity, pharmacokinetics (PK), and pharmacodynamics (PD). To date, 14 pts have been treated at 3 dose levels: 3 pts at 0.3 mg/kg, 4 pts at 1 mg/kg, and 7 pts at 3 mg/kg. Median patient age was 65 yrs (41–74 yrs); median number of prior therapies was 3 (2–11); median WBC at enrollment was 38.8 K/μ L (3.4–284 K/μ L). No dose limiting toxicity (DLT) occurred at the 0.3 and 1 mg/kg dose levels. At 3 mg/kg, streptococcal sepsis was reported in 1 pt after 2 study infusions and was considered DLT. Transient asymptomatic grade 3 or 4 elevation of amylase and/or lipase occurred in 2 subjects. In 9 pts with available data, infusions were associated with manageable grade 1–2 toxicity, primarily chills (7 pts), nausea (4 pts), and fever (3 pts) that was most predominant with the first infusion. PK analysis showed rapid clearance of HCD122 at the 0.3 and 1mg/kg dose levels, with no detectable levels at 2 and 7 days after infusion, respectively. PK data were available from 6 of 7 patients receiving 3mg/kg. These data indicated that some accumulation of HCD122 occurred at this dose level. Complete antigen saturation data were available for 2 of these subjects. Both showed sustained 100% saturation of antigen on peripheral CD5+CD19+ cells throughout the treatment period. Peripheral CD40+ CLL cells dropped transiently with a mean of 31% during each infusion in the majority of patients but the decreases were not maintained week-to-week. There were no substantive changes in NK or T-cell populations during therapy. In summary, HCD122 was safe and well tolerated up to the 3mg/kg dose level that is currently under evaluation. PK analysis to date suggests that levels higher than 3 mg/kg will be necessary to sustain levels of HCD122 in the expected therapeutic range, and dose escalation will continue to the maximum tolerated dose.
Thromboprophylaxis of Adult Medical Inpatients in St. John’s Newfoundland.
Authors: Hickey Joanne M. , Grewal Kuljit , Duggan Peter
Published: October 2019

Introduction: Venothromboembolic disease (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE), are causes of significant morbidity and mortality in hospitalized patients. Medical inpatients often have multiple risk factors for developing VTE and as a population have been shown to have significant rates of VTE ranging from 5–15% with approximately 75% of in hospital VTE occurring in non-surgical patients. Recent large randomized controlled trials have demonstrated the safety and efficacy of low molecular weight heparins (LMWH) in the prevention of VTE in this population. Despite the evidence of the effectiveness of this prophylactic treatment, numerous studies have demonstrated its underutilization in many at risk medical inpatient populations. We aim to establish rates of VTE prophylaxis in the population of at risk general medical inpatients in two acute care adult hospitals in the St. John’s area to assess if similar underutilization exists and if there is a need for future interventions to improve outcomes.

Methods: We used a modification of the inclusion/exclusion criteria of the MEDENOX trial to retrospectively identify consecutive medical inpatients at The General Hospital/St. Clare’s Mercy Hospital who were appropriate to receive VTE prophylaxis during January and February of 2005. Data was collected on demographics, VTE risk factors, and the use of any duration of low dose subcutaneous unfractionated heparin or low dose LMWH during their hospitalization. Two tailed t-tests were performed to identify any significant differences in demographics or VTE risk factors between the prophylaxis and non-prophylaxis groups.

Results: 114 medical inpatients were identified as being appropriate to receive pharmacologic VTE prophylaxis between Jan.1, 2005 and Feb. 28, 2005. 26 patients (23%) received some form of pharmacologic VTE prophylaxis. Of those who received prophylaxis, 19 patients (73%) received subcutaneous unfractionated heparin and 7 patients (27%) received LMWH. The only significant difference between the prophylaxis and non prophylaxis group was a higher proportion of patients with malignancy (27% vs 11% p=0.05) in the prophylaxis group.

Conclusion: This data demonstrates a significant underutilization of VTE prophylaxis in all at risk general medical patients in the two major adult acute care centres of the Eastern Health Board of Newfoundland. The data is consistent with the growing body of literature that exhibits that VTE prophylaxis in medical inpatients is deficient, with previously reported prophylaxis rates of only 20%–50%. Thus future work needs be done in translating “best practice” into “usual practice” as it relates to VTE prophylaxis.
Novel Murine Model To Study Modulation of Genes and Molecular Pathways Induced Following In Vivo Interaction between Multiple Myeloma Cells and Human BM Milieu.
Authors: Neri Paola , Tassone Pierfrancesco , Shammas Masood , Fulciniti Mariateresa , Tai Yu-Tzu , Blotta Simona , Prabhala Rao , Di Martino Mariateresa , Catley Laurence , Tagliaferri Pierosandro , Venuta Salvatore , Carrasco Ruben , Anderson Kenneth C. , Munshi Nikhil C.
Published: October 2019

Interaction between multiple myeloma (MM) cells and the bone marrow (BM) microenvironment plays a critical role in promoting MM cell growth, survival, migration and development of drug resistance. This interaction within the bone marrow milieu is unique and its understanding is important in evaluating effects of novel agents in vitro and in vivo. We here describe a novel murine model that allows us to study the expression changes in vivo in MM cells within the human BM milieu. In this model, the green fluorescent protein (INA-6 GFP+) transduced IL-6-dependent human MM cell line, INA-6, was injected in human bone chip implanted into SCID mice. At different time points the bone chip was retrieved, cells flushed out and GFP+ MM cells were purified by CD138 MACS microbeads. Similar isolation process was used on INA-6 GFP+ cells cultured in vitro and used as control. Total RNA was isolated from these cells and gene expression profile analyzed using the HG-U133 array chip (Affymetrix) and DChip analyzer program. We have identified significant changes in expression of several genes following in vivo interaction between INA-6 and the BM microenvironment. Specifically, we observed up-regulation of genes associated with cytokines (IL-4, IL-8, IGFB 2–5) and chemokines (CCL2, 5, 6, 18, 24, CCR1, 2, 4), implicated in cell-cell signalling. Moreover genes implicated in DNA transcription (V-Fos, V-Jun, V-kit), adhesion (Integrin alpha 2b, 7, cadherin 1 and 11) and cell growth (CDC14, Cyclin G2, ADRA1A) were also up-regulated and genes involved in apoptosis and cell death (p-57, BCL2, TNF1a) were down-regulated. Using the Ingenuity Pathway Analysis the most relevant pathways modulated by the in vivo interaction between MM cells and BMSCs were IL-6, IGF1, TGF-beta and ERK/MAPK-mediated pathways as well as cell-cycle regulation and chemokine signalling. These results are consistent with previously observed in vitro cell signalling studies. Taken together these results highlight the ability of BM microenvironment to modulate the gene expression profile of the MM cells and our ability to in vivo monitor the changes. This model thus provides us with an ability to study in vivo effects of novel agents on expression profile of MM cells in BM milieu, to pre-clinically characterize their activity.
Detection of Chromosomal Abnormalities in Chronic Lymphocytic Leukemia Increased by Interphase Fluorescence In Situ Hybridization in TPA-Stimulated Peripheral Blood Cells.
Authors: Aventin Anna , Sanchez Jana
Published: October 2019

Chromosomal abnormalities, namely deletion 11q-,13q-, 17p- and trisomy 12, have prognostic significance for patients with chronic lymphocytic leukemia (CLL). Several studies have demonstrated that the interphase fluorescence in situ hybridization technique (I-FISH) in CLL identifies such genomic aberrations in a higher frequency than classical karyotyping, including stimulated cultures using B-cell specific mitogens. However, there appears to be no information in the literature comparing I-FISH on non-cultured and cultured cells in CLL. A total of 56 samples from 49 patients with CLL were studied using commercially available probes for chromosomes 11q22.3(ATM), 13q14(13S272), 17p13(p53) and 12 centromere(D12Z3). We compared the results obtained by I-FISH-PBMC and those by interphase fluorescence in situ hybridization on TPA-stimulated peripheral blood cells (I-FISH-TPA) used for conventional cytogenetics in order to evaluate the usefulness of I-FISH-TPA. The proportion of abnormal nuclei obtained with the I-FISH-TPA was higher than that found with I-FISH-PBMC (P<0.001). Consequently, 15 cases with a negative or borderline result by I-FISH-PBMC became positive by I-FISH-TPA for deletion 11q- (n=2), 13q- (n= 9) and trisomy 12 (n=4). In all but one of these, chromosomal abnormalities were reconfirmed by either metaphase-FISH or conventional G-banding. Disease detection thus increased from 51% with I-FISH-PBMC to 78% with I-FISH-TPA. Interestingly, all 15 cases which reached the diagnostic thresholds for deletion 11q-,13q- and trisomy 12 had a slight lymphocytosis. An absolute lymphocyte count <8.7×109/l was found to be the critical threshold (P=0.037) below which I-FISH-TPA should be performed rather than I-FISH-PBMC. We have shown that I-FISH-TPA can not only detect a higher proportion of abnormal interphase nuclei but can also identify abnormal CLL cases which may be overlooked by I-FISH-PBMC, especially those with low absolute lymphocyte counts. I-FISH-TPA is thus a reliable technique for clinical diagnostics in CLL.
Vaccination with Lymphoma and Dendritic Cell Hybrids Intended as an Adjuvant Therapy for Lymphoma Patients - A Feasibility Study.
Authors: Markowicz Sergiusz , Jakubowska-Mucka Anna , Biernacka Marzena , Rymkiewicz Grzegorz , Borawska Anna , Federowicz Irena , Rozewska Danuta , Sawadro-Rochowska Maria , Polowniak-Pracka Hanna , Szymczyk Michal , Skurzak Henryk , Pojda Zygmunt , Walewski Jan
Published: October 2019

A feasibility of hybrid cell vaccination performed in remission following a standard chemotherapy (CHT) is currently being studied in patients with indolent B cell lymphomas at our institution under approval of the Ethics Committee. Adequate samples of fresh tumor tissue were collected from 16 patients. Seven patients were vaccinated with irradiated lymphoma/dendritic cell (DC) hybrids, and whenever the amount of collected viable lymphoma cells was not sufficient to complete vaccination with the use of cell hybrids, booster injections were alternatively performed with the use autologous DCs pulsed with tumor lysate. Autologous lymphoma cells were electrofused with autologous and/or allogeneic DCs, depending on accessibility of autologous DCs. Monocyte-derived DCs were basically generated from the peripheral blood adherent cells stimulated with GM-CSF and IL-4, and if available, from the bone marrow (BM) adherent cells. If only possible, autologous DCs were also generated from the BM non-adherent cells or from a sample of leukapheresis product collected for transplantation cultured with GM-CSF, TNF-α, SCF and FLT3-L to induce differentiation of DC progenitors. DC hybrids and/or DCs preincubated with tumor lysate were administered to uninvolved lymph nodes under USG guidance. Four patients failed to achieve a complete remission after initial therapy and the vaccination was discontinued due to disease progression (after 2, 2, 6, and 5 cycles). Three patients were vaccinated during remission following CHT. At present - 16, 18, and 10 months after starting the immunization, and after 8, 9, and after 7 cycles of vaccination, one patient with mantle cell lymphoma (MCL) is symptomless with persistent bone marrow (BM) involvement, one patient with MCL presenting with IgM paraproteinemia is disease-free with residual BM involvement (Fig 1), and one patient with follicular lymphoma is disease-free. Cutaneous delayed type hypersensitivity (DTH) response to DC/lymphoma hybrids exceeded the response to DCs alone in 3 of 7 vaccinated patients, and interferon-γ producing CD8+ cells specific to autologous lymphoma cells were demonstrated in one patient. In another patient, cutaneous DTH reaction was accompanied by transient pruritus and swelling of the injected lymph node two days after the sixth booster injection. No other adverse reactions to vaccinations were observed. Vaccination using the DC/lymphoma hybrids appears feasible, safe, and capable of inducing lymphoma cell specific immune responses. Further evaluation of efficacy in terms of disease control is mandatory.

Fig 1. Disease and treatment course of a representative case Fig 1. Disease and treatment course of a representative case
Novel Model of Stimulation-Responsive Splicing for Generation of Immunogenic Isoforms of Tumor Antigens and Autoantigens.
Authors: Yang Fan , Yan Yan , Xiong Zeyu , Chen Irene H. , Wang Hong , Yang Xiao-Feng
Published: October 2019

Alternative splicing is a process that removes introns and alters exons to generate multiple isoforms from a single pre-mRNA transcript. Alternative splicing is the major mechanism by which a small number of human genes (6 × 104) can encode the larger complexity of the human proteome (1 × 106 proteins). Previously we demonstrated that alternative splicing of apoptosis-regulatory protein transcripts regulates immune responses by modulating lymphocyte survival (Immunity, 1997; Mol. Immunol. 2002; J Exp Med, 2002; Oncogene 2005; Biochem J, 2005). To examine the hypothesis that alternative splicing plays a role in selection of nonmutated self-protein isoforms for tumor antigens and autoantigens, recently, we showed that alternative splicing is a major mechanism in regulation of the immunogenicity of tumor antigen CML66 (J. Immunol. 2004). In addition, we found that alternative splicing occurs in 100% of the autoantigen transcripts. This is significantly higher than the approximately 42% rate of alternative splicing observed in the 10,000 randomly selected human gene transcripts (p<0.001) [J. Allergy Clin. Immunol., 2004 (cover article)]. Here, we report that essential alternative splicing factor ASF/SF2 expression in samples from patients with chronic inflammation is lower than that of the healthy controls (p<0.05). In addition, TNF-a significantly downregulates ASF/SF2 expression (7 folds) in cultured cells in comparison to the expression variations of b-actin control. These findings demonstrate that ASF/SF2, presumably affecting splicing of self-antigen transcripts, is downregulated in autoimmune inflammatory disease potentially via a TNF-a-mediated pathway. Collectively, we propose for the first time a novel model of “stimulation-responsive splicing”, which emphasizes that stimulation-responsive splicing plays a critical role in selection of nonmutated self-protein isoforms to become tumor antigens and autoantigens (Clin. Immunol. Invited Review, in press, 2006). The new model for the definition of immunogenic isoforms of tumor antigens and autoantigens is significant in facilitating the development of:

immunogenic antigen isoform microarrays for disease diagnosis and prognosis; autoantigen-tolerizing therapy and splicing-redirection therapy for autoimmune diseases; and immunogenic antigen isoforms-based immunotherapy for tumors.
Low-Dose Cytosine Arabinoside and Aclarubicin in Combination with Granulocyte Colony-Stimulating Factor for Patients with Refractory and Relapsed Adult Acute Lymphocytic Leukemia.
Authors: Sun Aining , Ji Chengfu , Wu De Pei , He Guangshen
Published: October 2019

Objective To explore the efficiency and safety of Low-dose Cytosine arabinoside and aclarubicin in combination with granulocyte colony-stimulating factor (CAG regimen) for patients with refractory and relapsed adult acute lymphocytic leukemia (ALL).

Methods We treated 10 patients of refractory and relapsed adult ALL with CAG regimen (median age 28 years, range 17–58, male 5, female 5). The regimen consisted of low-dose cytosine arabinoside (10mg/m2 q12h, day1to14), aclarubicin (10mg/day, day1to8), and granulocyte colony-stimulating factor (G-CSF) (200 μg/m2/day).

Result 8/10 patients (80%) achieved complete remission (CR) after one course, including 4 out of 5 refractory and 4 out of 5 relapsed patients. 1 patient who relapsed after the first course, then achieved CR again after the second course. 6 of the 8 complete remission received consolidation therapy with the CAG regimen or other regimens. Toxic effects were very rare and mainly consisted of neutropenia and thrombocytopenia due to myelosuppression; 8 of all patients had neutropenia or thrombocytopenia that exceeded WHO grade II. However, severe non-hematologic toxicity included nausea and vomiting (WHO grade ≥III) was characteristically rare.

Conclusion CAG regimen is an effective and safety in the treatment of refractory and relapsed ALL.
Treatment with Erythropoietin and G-CSF Improves Survival in MDS Patients with Low Transfusion Need.
Authors: Jädersten Martin , Malcovati Luca , Dybedal Ingunn , Porta Matteo Giovanni Della , Invernizzi Rosangela , Montgomery Scott M. , Pascutto Cristiana , Porwit-MacDonald Anna , Cazzola Mario , Hellström-Lindberg Eva
Published: October 2019

Erythropoietin (Epo) and G-CSF alleviates anemia and increases quality of life in myelodysplastic syndrome (MDS). In patients with low transfusion need and low S-Epo level the erythroid response rate reaches 70% with a median response duration of two years, while patients with high transfusion need and high S-Epo level respond poorly and do not benefit from treatment. A recent study comparing Epo and G-CSF treated MDS patients with untreated historical controls indicates that the treatment can be given safely as chronic administration. To investigate the affects of growth factor treatment on outcome in patients with different probability of response, we compared treated patients included in three Nordic MDS Group studies (n=123) with untreated patients from Pavia, Italy (n=240). In both cohorts data on level of transfusion need and S-Epo level was recorded at baseline. All patients were transfusion dependent (n=176) or anemic with a hemoglobin level below 10 g/dl (n=187). An erythroid response, defined as at least abolition of transfusion need, was observed in 41% of treated patients, with a median response duration of 23 months (range 3–116+). Multivariate Cox regression analysis showed that treated patients with transfusion need of less than 2 units of RBC per month had a survival benefit, with HR 0.57 (P=0.015), while no difference in survival was observed in patients with higher transfusion need (P=0.36). There was no significant impact on risk of leukemic transformation in patients with the low (P=0.75) or high (P=0.21) transfusion need. This is the first study implicating that treatment of anemia in MDS with Epo and G-CSF in selected patients has a positive impact on outcome. Whether this effect is mediated by limiting the iron overload or counteracting other negative effects chronic anemia remains to be investigated.
Non-Intensive AML/MDS Treatment with Low-Dose Decitabine Prior to Reduced-Intensity Conditioning (RIC) and Allogeneic Blood Stem Cell Transplantation of Older Patients.
Authors: Lübbert Michael , Bertz Hartmut , Rüter Björn H. , Mertelsmann Roland H. , Finke Jürgen
Published: October 2019

Patients (pts) undergoing allogeneic stem cell transplantation for AML or high-risk MDS usually receive standard induction chemotherapy first, with the goal of achieving a complete remission (CR) or at least best response to conventional chemotherapy. This disease control is often necessary to bridge the time needed for identifying an unrelated donor in case no sibling donor is available, although CR is no absolute prerequisite for successful allografting following RIC (Bertz et al., J Clin Onc21:1480, 2003). Recently, non-intensive treatment with low-dose azanucleosides has been developed for older pts with high-risk MDS. Low-dose Decitabine (DAC) shows a particularly interesting activity in inducing cytogenetic remissions (38%) in MDS pts with poor-risk cytogenetics. The role of DAC prior to allografting has not yet been determined. Here we report our single-center experience of 10 consecutive pts treated with low-dose DAC for MDS (n=5), CMML (n=2) or AML (n=3). The median number of DAC courses given was 3 (range 1–13), with CR and PR as best response in 4 and 1 pt respectively, stable or persistent disease in 4 and 1 pt, resp. Allografting was either done as consolidation at time of best response (n=4) or as salvage for relapsed/refractory disease (n=6). Median age at time of allografting was 65 years (range 63–71). After conditioning with either FBM (n=8) or Flu/2 Gy (n=2), pts were transplanted from either matched sibling (n=3) or unrelated donor (n=7). One pt died before engraftment on day +13 due to infection. The other 9 pts engrafted, and no unexpected toxicities were noted. 6/9 pts have died either due to relapse (n=3) or infection in CR (n=3). Three pts are alive in CR at the time of this report (+1, +10, +26 months following transplant). Prior to transplantation two of them were in PR and CR following 2 and 8 courses of DAC, respectively, one had progression of CMML after 5 courses. In conclusion, we show that RIC and allografting in older pts with myeloid neoplasia following DAC treatment is feasible, and no unexpected toxicities have occurred. DAC pretreatment can induce improvement or even remission with negligible nonhematopoietic toxicity, thus bridging the time period of donor search prior to allografting. The allogeneic option in these pts should be explored more often and response to DAC and transplant risk-assessment (comorbidities, performance status etc.) will likely be major determinants of optimal timing of allografting in this setting.