bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential diagnosis of monoclonal gammopathy of undetermined significance
Authors: Merlini Giampaolo , Palladini Giovanni
Published: October 2019

Monoclonal gammopathy of undetermined significance (MGUS) is an asymptomatic plasma cell disorder occurring in 4.2% of adults > 50 years of age, which can progress into symptomatic diseases either through proliferation of the plasma cell clone, giving rise to multiple myeloma and other lymphoplasmacellular neoplasms, or through organ damage caused by the monoclonal protein, as seen in light-chain amyloidosis and related conditions. Differential diagnosis of asymptomatic and symptomatic monoclonal gammopathies is the determinant for starting therapy. The criteria for determining end-organ damage should include markers of organ injury caused by the monoclonal protein. Patient assessment and optimal follow-up are now performed using risk stratification models that should also take into account the risk of developing AL amyloidosis. Patients with low-risk MGUS (approximately 40% of all MGUS patients) need limited assessment and very infrequent follow-up. The ongoing development of novel molecular biomarkers and advanced imaging techniques will improve the identification of high-risk patients who may benefit from early therapeutic intervention through innovative clinical trials.
Catheter-related thrombosis: lifeline or a pain in the neck?
Authors: Kamphuisen Pieter W. , Lee Agnes Y. Y.
Published: October 2019

Central venous catheters (CVCs) are used extensively in cancer patients for the administration of therapy and phlebotomy. An important complication of CVCs is the development of catheter-related thrombosis (CRT), which becomes symptomatic in approximately 5% of the patients. Several factors, such as insertion location and position of the catheter tip, increase the risk of CRT. Prevention of CRT with systemic anticoagulant prophylaxis has largely been ineffective. In addition, the optimal diagnostic strategy and anticoagulant treatment are unclear due to the lack of well-designed studies. The most recent American College of Chest Physicians guidelines recommend (color) Doppler ultrasound more than venography as the initial diagnostic test in patients with suspected arm thrombosis. Only if the ultrasound is negative and clinical suspicion is high is further testing with D-dimer, serial ultrasound, or venography advocated. In case of CRT, removal of the catheter is not necessary if it is functional and needed for chemotherapy. Anticoagulant treatment of CRT consists of treatment with low-molecular-weight heparin (LMWH) followed by vitamin K antagonists for at least 3 months. Whether long-term treatment with LMWH is more effective than vitamin K antagonists in cancer patients with CRT is unknown, but LMWH may be advocated following the recommendations in lower limb thrombosis and cancer. In addition, the effect of new anticoagulants in CRT has not been studied.
B- and T-cell prolymphocytic leukemia: antibody approaches
Authors: Dearden Claire
Published: October 2019

B- and T-cell subtypes of prolymphocytic leukemia (PLL) are rare, aggressive lymphoid malignancies with characteristic morphologic, immunophenotypic, cytogenetic, and molecular features. Prognosis for these patients remains poor, with short survival times and no curative therapy. The advent of mAbs has improved treatment options. In B-PLL, rituximab-based combination chemoimmunotherapy is effective in fitter patients. TP53 abnormalities are common and, as for chronic lymphocytic leukemia, these patients should generally be managed using an alemtuzumab-based therapy. Currently, the best treatment for T-PLL is IV alemtuzumab, which has resulted in very high response rates of more than 90% when given as frontline treatment and a significant improvement in survival. Consolidation of remissions with autologous or allogeneic stem cell transplantation further prolongs survival times, and the latter may offer potential cure. The role of allogeneic transplantation with nonmyeloablative conditioning needs to be explored further in both T- and B-PLL to broaden the patient eligibility for what may be a curative treatment.
Recent insights into the role of the contact pathway in thrombo-inflammatory disorders
Authors: van Montfoort Maurits L. , Meijers Joost C.M.
Published: October 2019

The contact pathway of coagulation consists of the proteins factor XI, factor XII, prekallikrein, and high-molecular-weight kininogen. Activation of the contact system leads to procoagulant and proinflammatory reactions. The contact system is essential for surface-initiated coagulation, as exemplified by aPTT, but there is probably no role for the contact system in initiating physiologic in vivo coagulation. However, over the last few years, there has been renewed interest, especially because of experimental evidence suggesting that the contact system contributes to thrombosis. Knockout mice deficient in one of the contact proteins were protected against artificially induced thrombosis. Furthermore, inhibiting agents such as monoclonal antibodies, antisense oligonucleotides, and small molecules were found to prevent thrombosis in rodents and primates in both venous and arterial vascular beds. Although it remains to be established whether targeting the contact system will be effective in humans and which of the contact factors is the best target for anticoagulation, it would constitute a promising approach for future effective and safe antithrombotic therapy.
Evidence for Janus kinase (JAK) inhibitors for the prevention of major morbid events in patients with myeloproliferative neoplasms (MPNs)
Authors: Pemmaraju Naveen , Mesa Ruben
Published: October 2019

A 69-year-old man presents with a Dynamic International Prognostic Scoring System (DIPSS) intermediate-risk 2 post polycythemia vera myelofibrosis with significant splenomegaly, 30 pound weight loss, constitutional symptoms, and 2% peripheral blood blasts. He has no other significant past medical history and no other major comorbid conditions. You are asked by the hematology fellow you are supervising whether or not treating this patient with a Janus kinase (JAK) inhibitor will decrease major morbid events.
STANDARDIZATION OF THE QUICK PROTHROMBIN TEST
Authors: AGGELER PAUL M. , HOWARD JOAN , LUCIA S. P. , CLARK WILLIAM , ASTAFF ALEXIS
Published: October 2019

A large specimen of human brain, obtained at autopsy, was dehydrated and stored in an evacuated desiccator. Portions of the material were removed when needed and the desiccator was re-evacuated. Under these conditions there was no loss of thromboplastic potency over a period of two years.

See PDF for Table

See PDF for Table

The dry oxalate mixture (6 mg. of ammonium oxalate and 4 mg. of potassium oxalate) was used as an anticoagulant in order to obtain plasma for the prothrombin tests.

With the specimen of thromboplastin used in these experiments, normal standards for the Quick prothrombin test were found as shown in table 8. These results do not necessarily mean that the normal range of prothrombin concentrations is from 75 to 140 per cent of the actual amount of prothrombin in the plasma of the average normal subject. They mean only that with the methods here employed, values within this range may be expected to occur.

There was no significant difference between the prothrombin values obtained in a group of normal subjects by methods using diluted or undiluted plasma. In repeated tests on the same subject there appeared to be slightly greater accuracy when the tests were performed on diluted plasma, but there was no significant difference between the results obtained with plasma diluted to 50 or to 25 per cent of its original concentration.

Repeated tests on diluted and undiluted plasma from the same normal subject produced the range of normal variation about the various prothrombin concentrations shown in table 9.

The normal variation that may be expected to occur at any given prothrombin level must be considered in interpreting the variations that appear after administration of vitamin K or dicumarol.

The use of a single specimen of blood from one random normal subject as a normal standard may lead to considerable error, because of unavoidable technical variables and because of differences in the prothrombin concentrations of different normal subjects. To establish reliable normal standards, the blood of at least 5 normal subjects should be individually tested with the same specimen of thromboplastin and the results averaged.
CELLULAR CHANGES PRODUCED BY EXTRACTS OF HUMAN ORGANS
Authors: ERF L. A. , TURNER D. L. , MILLER F. R.
Published: October 2019

By the injection of lipoid extracts of organs (liver, spleen, and lymph nodes) of 5 patients with leukemia, 1 with Hodgkin's disease, and of 4 "control" cases (3 of heart disease and 1 of cerebral tumor) into 28 guinea-pigs, it was observed that:

a. extracts from human myeloid leukemic organs produced "myeloid reactions" or infiltrations in guinea-pigs;

b. extracts from lymphoid leukemic organs produced "lymphoid reactions";

c. extracts from organs involved with Hodgkin's disease produced "Hodgkin's reactions"; and

d. extracts from "normal" organs produced slight lymphoid or monocytoid infiltrations.
Evidence that Platelet Accelerator (Platelet Factor 1) Is Adsorbed Plasma Proaccelerin
Authors: HJORT PETER , RAPAPORT SAMUEL I. , OWREN PAUL A.
Published: October 2019

1. Human platelets possess an accelerator activity equal to about six per cent of the proaccelerin activity of normal citrated plasma. This accelerator activity is only slightly reduced by 10 washings.

2. Thrombin increases platelet accelerator activity as much as ten fold. This means that platelet accelerator activity behaves like proaccelerin (plasma Acglobulin) rather than accelerin (serum Ac-globulin).

3. Platelets from a patient with parahemophilia (congenital proaccelerin deficiency) possess only a trace of accelerator activity. They acquire a normal amount of accelerator activity after contact with normal platelet-poor plasma.

4. Trypsin destroys 90 per cent or more of the platelet accelerator activity of normal platelets without altering their appearance. Trypsinized platelets regain normal accelerator activity upon incubation with normal platelet-poor plasma.

5. These findings strongly suggest that the platelet accelerator (platelet factor 1) is adsorbed plasma proaccelerin.
The Cytophotometric Estimation of the Desoxyribonucleic Acid (DNA) Content of Individual Plasma Cells from Multiple Myeloma and Nonspecific Plasmacytoses
Authors: PETRAKIS NICHOLAS L. , FOLSTAD LOIS J.
Published: October 2019

The cytophotometric technic employing the Feulgen reagent was used to determine the relative amount of DNA in the nuclei of individual myeloma and plasma cells aspirated from the bone marrow. Individual myeloma cells were found to contain markedly elevated amounts of DNA as compared to that in the plasma cells of non-specific plasmacytoses. Myeloma cells were found to contain geometric multiples (2, 4, 8) of the amount of DNA present in normal lymphocytes, and the degree of DNA "ploidy" was related to the morphologic stage of immaturity. Polyploid DNA values were not found in the non-specific plasmacytoses. The possible mechanisms of formation were discussed. The findings are interpreted as suggesting the presence of a basic alteration in the quantitative DNA-chromosome relationship in myeloma cells, and they offer additional support for the separation of the myeloma plasmacytoses from nonspecific plasmacytoses.
Hemolytic Transfusion Reaction Due to Anti-Lea
Authors: MATSON G. ALBIN , COE JOHN , SWANSON JANE
Published: October 2019

A case is reported in which a transfusion of whole blood of group Le (a+b-) produced a severe hemolytic transfusion reaction in a patient of group Le (a-b-). The anti-Lea in the patient's serum had apparently been stimulated by previous transfusions. No other antibody was demonstrable by us in this case.
Differences in Susceptibility of Polymorphonuclear Leukocytes from Several Species to Alteration by Systemic Lupus Erythematosus Serum: Application to a More Sensitive L. E. Phenomenon Test
Authors: CARRERA ANA E. , REID MAY VIRGINIA , KURNICK N. B.
Published: October 2019

In the article by Carrera, Reid, and Kurnick: Differences in Susceptibility of Polymorphonuclear Leukocytes from Several Species to Alteration by Systemic Lupus Erythematosus Serum: Application to a More Sensitive L. E. Phenomenon Test (Blood 9: 1165, 1954), the following footnote, pertaining to Dr. Carrera, was inadvertently omitted: Fellow, American Cancer Society, recommended by the Committee on Growth, National Research Council.
Electron Microscope Observations of the Platelet-Fibrin Relationship in Blood Clotting
Authors: DE ROBERTIS E.
Published: October 2019

The behavior of stretched out human platelets in different stages of blood clotting has been observed with the electron microscope. In the early stages of coagulation the morphologically intact platelets apparently act as centers of orientation and condensation of the fibrin fibrils. This effect is not found in platelets which have previously undergone extensive damage or disintegration. In more advanced stages of clotting, many platelets remain normal, showing their attachment to the fibrin, while others have disintegrated and appear as residual dense bodies.

The probable action of platelets in the retraction and syneresis of the blood clot is discussed. It is suggested that at the surface of the morphologically intact platelets, adhesion forces develop which hold and orient the molecular chains of fibrin increasing the number of interacting junctions and producing syneresis. On the contrary, the platelets which have suffered an injury which alters these surface adhesive properties do not interact with the fibrin fibrils and, as a result, the mechanism of clot retraction is impaired.
Hematologic Observations of the Course of Erythroblastosis Fetalis
Authors: DESFORGES JANE F. , O’CONNELL LIAM C.
Published: October 2019

Serial studies revealed characteristic changes in the peripheral blood of infants with erythroblastosis fetalis. In the presence of elevated nucleated red cells and reticulocytosis, replacement transfusion brought about a momentary decline to normal levels followed by a second rise during the succeeding three days before the values returned to low levels. Platelet counts diminished within a day after replacement and only gradually rose toward normal after approximately three days. Such a picture was not seen in the mildly affected babies who did not receive replacement therapy, in whom the platelet values varied.

Within the first three months in both groups anemia developed in the presence of little evidence of compensating marrow activity. After a decline of the hemoglobin to an average level of 7.6 Gm. per cent, erythropoiesis became adequate for return to normal levels. This hypoplastic defect could not be correlated with any other parameter of the disease.

Abnormalities were noted in the osmotic and mechanical fragility of the red cells in this disease. The major population of cells was found to be more susceptible to osmotic stress while the limits of hemolysis were within the normal range. The osmotic curves of the blood of those patients who were not transfused returned to normal within 4 weeks. Mechanical fragility of the red cells was increased in a significant number, and this abnormality was observed until the time when minimal hemoglobin values were reached. There was no correlation of these variations in fragility with other laboratory or clinical manifestations of the disease.
Unique Evidence for Autosomal Dominant Inheritance of Severe Congenital Neutropenia.
Authors: Boxer Laurence A. , Stein Steven , Buckley Danielle , Subramanian Sandhya , Bolyard Audrey Anna , Olson Maynard V. , Dale David C.
Published: October 2019

Severe congenital neutropenia was originally described as an autosomal recessive disorder characterized by severe neutropenia and recurrent bacterial infections from birth, but other reports indicate that autosomal dominant inheritance occurs. Several report now indicate that most, but not all, patients with severe congential neutropenia have heterozygous mutation in the gene for neutrophil elastase (NE or ELA2). We recently investigated an unusual series of cases of severe congenital neutropenia occurring when the same sperm donor was utilitized by four different families to impregnate mothers by either artificial insemination or in vitro fertilization. Three sets of twins were conceived and a single child in the four families. Collectively one set of twins and one child each from the other two families conceiving twins and the single child were all found to have severe congenital neutropenia. The mean for their median blood neutrophils (ANC) at diagnosis was 84 +/−32 x109/L (range 0–166 x 109 /L); and their other blood counts were remarkably similar. On treatment with G-CSF (mean dose 6.7+/− 1.3 SEM mcg/kg/day) the mean for the median ANC increased to 3290 +/−1410 x109/L( range 420–8600 x 109 /L )

To determine whether the sperm donor was responsible for transmitting severe congenital neutropenia, geonomic DNA from peripheral blood leukocytes was used to sequence the gene for neutrophil elastase in the affected children and their mothers using standard techniques. None of the mothers had a mutation in their NE gene but all five affected children had the same mutation effecting the fourth exon at site S97L. Since it was not possible to perform genetic analysis on biological samples from the donor, more detailed genetic linkage analysis was performed on the affected children and their mothers. Linkage mapping analysis of 22 microsatellite markers on chromosome 14 and 19 confirmed that all affected children possessed one of the two paternal alleles. For an 11 centiMorgan region immediately adjacent to the NE gene on chromosome 19, all affected children shared a single paternal allele, and 4 of 5 affected children shared a single paternal allele for a 20 centiMorgan region adjacent to the NE gene. The studies indicate that the father of the affected children provided consistent haplotypes that led to the expression of severe congenital neutropenia in all of the affected children. This investigation strongly implicates that autosomal dominant inheritance for severe congenital neutropenia. It also raises important questions for the evaluation of subjects in sperm donor programs.
Uric Acid Induces High Mobility Group Box1 Protein Release in Monocytes/Macrophages through P38 MAPK, ERK1/2, JNK and AP-1 Activation.
Authors: Biswas Kamal K. , Oyama Yoko , Abeyama Kazuhiro , Hashiguchi Teruto , Maruyama Ikuro
Published: October 2019

Uric acid, a final metabolite of purine metabolism has gained attention for its pathogenic role in the development of hypertension, vascular disease, renal disease, and cardiovascular events. Recent studies have shown that soluble uric acid induces proliferation as well as synthesis of proinflammatory chemokine, monocyte chemoattractant protein 1 in rat vascular smooth muscle cells. It also stimulates human monolayer cells to produce IL-1B. IL-6 and TNF alpha. However, high mobility group box 1(HMGB1), a non histone DNA binding protein which is recently identified as a proinflammatory cytokine and can be released from cells of the macrophage/monocyte lineage by proinflammatory stimuli or by cells undergoing necrotic cell death pathway. In the present study, we hypothesized that soluble uric acid may stimulates monocytes/macrophages to release HMGB1. Here we demonstrate that crystal and endotoxin free uric acid triggers the release of HMGB1 in a time and dose dependant fashion in mouse macrophage cells, Raw 264.7, human leukemic promonocytes(THP-1 cells) as well as in macrophages but not in fibroblast obtained from synovial fluid of rheumatoid arthritis patients. Moreover, translocation of HMGB1 to the cytoplasm before secreted in the extracellular milieu was significantly detected within 6h of Uric acid exposure. No loss of cell viability was observed in our experimental condition as judged by trypan blue exclusion, MTT reduction and LDH release assays, indicating that HMGB1 release was not due to cell death. Uric acid treatment resulted in the activation of p38 mitogen activated protein kinase(MAPK), c-Jun N terminal kinase(JNK) and extracellular regulatroy kinase (ERK1/2). Inhibition of each of p38 MAPK, ERK1/2 and JNK by using potent and selective inhibitors, SB203580, U0126 and SP60012 respectively, almost completely blocked uric acid-induced HMGB1 release. Furthermore, Curcumin, a potent inhibitor of the transcription factor, activator protein 1(AP-1) strongly suppressed HMGB1 release, indicating that these signaling patways are essential for the induction of HMGB1 release in response to uric acid. In conclusion, our data suggest that uric acid may regulate critical proinflammatory pathways at least in part through its action as a monocyte/macrophage derived potent proinflammatory cytokine, HMGB1 release. This may play pathologic role for hyperuricemia-induced morbidities including arthritis, atherosclerosis, renal disease and so on.
Minimal Residual Disease (MRD) Kinetics after Autologous Stem Cell Transplantation for Chronic Lymphocytic Leukemia (CLL) Correlate to IgH VH Mutational Status and Predict Post Transplant Outcome: Real Time ASO-IgH PCR Results of the GCLLSG CLL3 Protocol.
Authors: Ritgen Matthias , Dreger Peter , Stilgenbauer Stephan , von Neuhoff Nils , Greinix Hildegard T. , Hertenstein Bernd , Schmitz Norbert , Emmerich Berthold , Hallek Michael , Döhner Hartmut , Kneba Michael
Published: October 2019

The purpose of the present study was (1) to assess the quantitative impact of myeloablative therapy with autologous stem cell transplantation (SCT) on MRD levels in patients with CLL; (2) to assess the predictive value of post transplant MRD kinetics for clinical outcome; and (3) to study the influence of the IgH VH mutational status on MRD kinetics. Blood and bone marrow (BM) samples were obtained from patients from a prospective multicenter trial of the German CLL Study Group (GCLLSG CLL3 protocol) after informed consent. MRD monitoring was performed by real-time quantitative IgH PCR using allele-specific primers (ASO RQ-PCR).

Results: As of July 2004, 476 blood samples from 46 patients who had an informative primer and repeated follow-up material available were analyzed by ASO RQ-PCR with a minimum sensitivity of 1E-4. In 145 cases, BM samples obtained on the same occasion were analyzed in parallel. Blood and BM showed highly significant correlating MRD values (p=.0001; slope 0.9, r<.0001), with a tendency to higher MRD levels and higher sensitivity in BM. Myeloablative treatment with SCT resulted in a strong reduction of the CLL load (median MRD level 6.6E-3 pretransplant vs. 1.0E-4 at 90–180 days post transplant; p<.0001) with no significant difference between patients with mutated and unmutated VH, respectively. Whereas MRD levels had no prognostic impact during the first 6 months after SCT, stable or decreasing MRD kinetics below 1E-4 between 6 and 12 months after SCT were strongly predictive for a favourable outcome (4-year progression-free survival (PFS) 100%). In contrast, PFS of those patients who had increasing MRD levels at this time was significantly poorer (4-year PFS 37%; p 0.02; n = 25). Patients with increasing MRD levels early post transplant almost exclusively belonged to the subgroup with unmutated VH.

Conclusions: Myeloablative SCT can lead to substantial additional reduction of the tumor load in patients with CLL in remission. The cause of the dismal post transplant outcome of patients with unmutated VH is faster regrowth of the leukemic clone rather than less effective reduction of MRD levels immediately after SCT.
Activation of VEGFR-2 in Hematopoietic Progenitor Cells Leads to Accumulation of Myeloid Cells through Up-Regulation of GM-CSF.
Authors: Larrivee Bruno , Pollet Ingrid , Karsan Aly
Published: October 2019

VEGF is a secreted growth factor that mediates its biological effects by binding to two transmembrane tyrosine kinase receptors, VEGFR-1 and VEGFR-2. VEGF has previously been shown to be critical in the establishment of cells of the hematopoietic and endothelial lineages. VEGF and its receptors are expressed on subsets of hematopoietic cells, and are also expressed in some hematopoietic malignancies. However, the requirement for each of the VEGF receptors, and whether the effects of VEGF are due to autocrine or paracrine signals remains unclear. We have designed a strategy that allows us to specifically study the unique effects of VEGFR-2 signaling in hematopoietic cells in vivo. By using a VEGFR-2 cytoplasmic domain fusion protein that can be dimerized with a synthetic drug, we were able to specifically study the effects of VEGFR-2 signaling in isolation. Using mice transplanted with bone marrow transduced with the inducible VEGFR-2 fusion construct, we demonstrate an expansion of myeloid progenitors by CFC assay, as well as increases in Gr-1+ and CD11b+ cells in the bone marrow. VEGFR-2 activation did not affect the proportion of lymphoid and erythroid cells. We show, by RT-PCR and ELISA, that VEGFR-2 activation can up-regulate granulocyte/macrophage-colony stimulating factor (GM-CSF) in bone marrow cells. Using activation and blocking experiments, we show that the VEGFR-2 activity is due to autocrine effects as well paracrine effects through the induction of GM-CSF in bone marrow cells.
Honokiol Overcomes Conventional Drug Resistance in Human Multiple Myeloma.
Authors: Ishitsuka Kenji , Hideshima Teru , Hamasaki Makoto , Noopur Raje , Shaji Kumar , Hideshima Hiromasa , Shiraishi Norihiko , Richardson Paul , Munshi Nikhil C. , Chauhan Dharminder , Arbiser Jack , Anderson Kenneth C.
Published: October 2019

Honokiol is an active component isolated and purified from Magnolia, a plant used in traditional Chinese medicine. It is an anti-oxidant, and inhibits both xanthine oxidase and angiogenesis. In this study, we first examined the direct toxicity of honokiol against human multiple myeloma (MM) cell lines in vitro. Honokiol significantly inhibited growth of MM cell lines (RPMI8226, U266 and MM.1S) via induction of G1 growth arrest, followed by apoptosis, with IC50 values at 48h of 5 to 10 μg/ml. Moreover, honokiol similarly inhibited growth of doxorubicin (Dox)-resistant (RPMI-Dox40), melphalan resistant (RPMI-LR5), and dexamethasone (Dex)-resistant (MM.1R) cell lines. Furthermore, flow cytometric analysis demonstrated that honokiol (6–10 μg/ml, 48h) induced death of CD38+CD138+ tumor cells isolated from 5 patients with relapsed refractory MM. In contrast, no toxicity was observed in normal peripheral blood mononuclear cells or long term-cultured bone marrow stromal cells (BMSCs) treated with honokiol (≤20 mg/ml). Neither culture of MM cells with BMSCs nor interleukin-6 (IL-6) and insulin like growth factor-1 (IGF-1) protected against honokiol-induced cytotoxicity in MM.1S cells. We next delineated the mechanism of honokiol-triggered cytotoxicity. Honokiol triggered increased expression of Bax and Bad; down regulated Mcl-1 protein expression, followed by caspase-8/9/3 cleavage. Importantly, the pan-caspase inhibitor z-VAD-fmk only partially inhibited honokiol-induced apoptosis in MM.1S cells. Furthermore, honokiol induced apoptosis even in SU-DHL4 cells, which express low level of caspase-8 and -3 and are resistant to both conventional (doxorubicin, melphalan, dexamethason) and novel (bortezomib, revimid) drugs. These results suggest that honokiol may induce apoptosis via both caspase-dependent and -independent pathways. Finally, honokiol inhibited IL-6-induced phosphorylation of ERK1/2, STAT3, and Akt, known to mediate growth, survival, and drug resistance, respectively. Taken together, our results suggest that providing the rationale for clinical evaluation of honokiol to improve patient outcome in MM.
Genetic Polymorphisms Associated with Clinical Outcome in the Intergroup Trial S9321, Comparing High Dose Therapy with Standard Dose Therapy for Myelomaon, on Behalf of ECOG, SWOG, CALGB, and the Bank on a Cure.
Authors: Santana-Davila Rafael , Crowley John , Durie Brian , Barlogie Bart , Greipp Philip , Van Ness Brian
Published: October 2019

Genetic variations in patient populations likely contribute to disease progression and therapeutic outcomes. We have begun a systematic approach to examine the association of genetic variations (ie. functional, single nucleotide polymorhisms, SNPs) involved in myeloma growth promotion, metabolic events, drug responses, and DNA repair on clinical outcome in the intergroup trial S9321. This trial tested a single high dose regimen with autologous stem cell support against a conventional dose regimen, with further randomization of responders to maintenance with interferon or not, in newly diagnosed patients with multiple myeloma. ECOG, CALGB and SWOG enrolled 899 patients with newly diagnosed MM to receive VAD induction x 4 cycles followed by randomization to PBSC-supported high dose therapy (HDT) versus standard dose therapy (SDT) of VBMCP, using CTX 4.5 g/m2 + G-CSF for PBSC mobilization in all patients. Responders to VBMCP or HDT were randomized to IFN or no maintenance. Specimens were distributed through ECOG for biologic correlative studies, including candidate SNP analysis. SNP assays have been developed using the Sequenom Mass-extend platform for functional SNPs in IL-6, IL-1, IL-RA, IL-10, TNF, Lta, TGFb, MDR1, MPO, CYP3A4, GST (M, P, T), ERCC2 and XRCC1, and are being evaluated on 803 DNA samples prepared from patients enrolled in S9321. Preliminary findings (n=135) demonstrate functional genetic variants of IL-10 (position -1082), IL-1 (position +3953), TGFb (postion -509), and TNFa (position -308) are showing trends associated with differences in progression free survival; and variants in IL-6 (position -174) are associated with response. Median survival of the IL-10 variants was 31 months for A/A low producer alleles versus 19 months for the G/G high producer alleles (p=.5). For TNFa, 2 cases with the high producer A/A alleles died within a year, while the median survival for the lower producer G/G alleles was 2 years (p=.04). For patients with the high producer C/C/ allele of IL-1 (n=67), median survival was 2 years, versus 5 patients with the T/T low producer alleles that had a median survival greater than 5 years (preliminary p=.29). While these preliminary results are now only suggestive of trends in genetic polymorphisms associated with clinical outcome, completion of the full SNP panel on the entire sample base should provide a extensive association study, and analysis of potential differences in therapy arms of the trial. The full panel and association studies will be presented.ααββααα
Prognostic Diversity of Cytogenetic Lesions in Myelofibrosis with Myeloid Metaplasia: A Prospective Study of 88 Patients.
Authors: Dingli David , Arora Brijesh , Sirhan Shireen , Gordon Dewald W. , Ayalew Tefferi
Published: October 2019

Background : Thirty to 50% of patients with myelofibrosis with myeloid metaplasia display detectable bone marrow cytogenetic abnormalities at diagnosis with some(Demory et al. Blood1988;72:855, Dupriez et al. Blood1996;88:1013, Reilly et al. BJH1997;98:96) but not other studies (Cervantes et al. BJH1998;102:684, Okamura et al. Int J Hematol2001;73:194) suggesting a detrimental effect on survival.

Methods : The current prospective study was conducted as a follow-up of the observations from an antecedent retrospective study that identified sole abnormalities of either 13q- or 20q- as ‘favorable’ to survival (Tefferi et al. BJH2001;113:763). All clinical and laboratory measurements, including bone marrow examination and karyotype analysis, were performed concurrently in consecutive cases of histologically confirmed MMM regardless of either duration of disease or presence of active therapy. Survival, leukemic transformation rate, and clinical progression were determined from the time of cytogenetic analysis.

Results : Karyotypic analysis was successfully performed in a total of 88 patients (median age 61 years; 31 females) and the results were normal in 49 (56%) and abnormal in the remaining 39 patients with 9 (10%) displaying ‘favorable’, and 30 (34%) ‘other’ clonal abnormalities. At the time of study entry, 57 patients were not recieving any form of therapy, 18 were newly diagnosed, and the Dupriez prognostic score distributions were ‘0’ in 33 patients, ‘1’ in 35 and ‘2’ in 20 (Dupriez et al. Blood1996;88:1013). All patients were followed for a minimum of 3 years and a median of 42 months. The 3 cytogenetic categories were similar in age and gender distribution. Survival in the favorable cytogenetic group was similar to that of the normal group and significantly better than the group with other cytogenetic abnormalities (p=0.02).

Figure Figure

Similarly, no patient (0%) in the ‘favorable’ group transformed into acute leukemia while 6 of 30 (20%) in the ‘other’ and 5 of 49 (10%) in the normal groups did (p=0.21). In a Cox regression analysis, cytogenetic lesions other than the ‘favorable’ kind, hemoglobin below 10 g/dL, leukocytosis, hypercatabolic symptoms, increased blood blast %, blood CD 34 count above 100 x 106/L were individually associated with shortened survival. In a multivariate analysis, only hemoglobin level and leukocyte count retained their significance.

Conclusion : Single chromosomal abnormalities of either 13q- or 20q- should be considered as ‘favorable’ cytogenetic lesions in MMM while other abnormalities are associated with an adverse prognosis. The current observation explains the discrepancy in the literature regarding the prognostic relevance of cytogenetic findings in MMM and warrants additional studies to determine the value of incorporating the specific information in prognostic scoring systems.
A Potential New Application of Mobilization/Leukapheresis for Enrichment of Peripheral Blood in Circulating Non-Hematopoietic CXCR4+CD45− Tissue-Committed Stem Cells (TCSC) for Organ/Tissue Regeneration.
Authors: Kucia Magda , Reca Ryan , Wysoczynski Marcin , Gozdzik Jolanta , Ratajczak Janina , Janowska-Wieczorek Anna , Ratajczak Mariusz Z.
Published: October 2019

During mobilization hematopoietic stem cells (HSC) egress from the bone marrow (BM) into peripheral blood (PB) where they temporarily circulate and can be collected by leukapheresis. However, recently we demonstrated that BM, in addition to HSC, contains heterogeneous populations of CXCR4+ tissue-committed stem cells (TCSC) and we contend that the contribution of these cells to organ/tissue regeneration after transplantation of BM cells has been misinterpreted as evidence for “plasticity” or “trans-dedifferentiation” of HSC (Leukemia2004:18;29–40). To determine whether TCSC could also be mobilized into PB we i) evaluated the presence of these cells in the PB of G-CSF-mobilized patients (n=11), ii) attempted to increase mobilization of the TCSC in a murine model by combining G-CSF with T140 (a CXCR4 antagonist) or SB290157 (the C3a complement fragment receptor antagonist which we have found to desensitize the responsiveness of HSC to SDF-1), and iii) tested the hypothesis that TCSC could also be mobilized into PB during stress related to tissue/organ injury, e.g., heart infarct, stroke, partial body irradiation. The presence of mobilized/circulating TCSC in PB was evaluated by i) real-time RT-PCR, ii) immunohistochemical staining for TCSC markers and iii) demonstrating the potential of mobilized TCSC to grow neurospheres or to form myotubes in vitro. We present evidence for the first time that i) G-CSF efficiently enhances the release into PB not only of HSC but also of TCSC expressing markers for early skeletal muscle, myocardium, neural tissue, pancreas and liver, ii) a combination of G-CSF with T140 or SB290157 is 25–30 times more effective in selectively mobilizing TCSC compared to G-CSF alone, and iii) TCSC are also mobilized to PB during stress related to heart infarct, stroke or partial body irradiation. Furthermore, we observed that TCSC, like HSC, express CXCR4 and Sca-1 but do not radioprotect lethally irradiated mice and, unlike HSC, are CD45-negative. Based on these findings we postulate that mobilization/leukapheresis procedures may find a new application for obtaining TCSC for use in tissue/organ regeneration. We are currently testing this hypothesis in murine models.
Pilot Study of Reduced-Intensity Conditioning Followed by Allogeneic Stem Cell Transplantation in Patients with Myelofibrosis.
Authors: Kroeger Nicolaus , Zabelina Tatjana , Schieder Heike , Panse Jens , Ayuk Francis , Fehse Natalja , Fehse Boris , Waschke Olga , Kvasnicka Hans M. , Thiele Jurgen , Zander Axel
Published: October 2019

We evaluated in a prospective pilot- study the effect of reduced intensity conditioning with busulfan (10 mg/kg), fludarabine (150 mg/m²) and anti-thymocyte globulin followed by allogeneic stem cell transplantation from related and unrelated donors in 14 patients with myelofibrosis. Study objectives were engraftment, chimerism, treatment- related mortality (TRM) and response. The median age of the patients was 51 (range, 32 – 63) years. According the Lille score there were low risk (n=4), intermediare risk (n= 7) and high risk (n=3) The median time until leukocyte (> 1.0 x 109/l) and platelet (> 20 x 109/l) engraftment was 16 (range, 11 – 26) days and 26 (range, 9 – 139) days, respectively. No graft failure occurred. Complete donor chimerism on day 100 was seen in 13 patients (93%). Acute graft-versus host disease (GvHD) grade II-IV occurred in 50% and grade III/IV 29 % of the patients. The incidence of chronic GvHD was 54 %. Two patients died due to treatment complications, resulting in a TRM at one year of 15% (95% CI: 0–35%). Hematological response after allogeneic transplantation was seen in all patients and complete histopathological remission was observed in 90%of patients. After a median follow-up of 13 (range, 3 – 48) months, the 3-years estimated overall and disease-free survival is 85 % (95 % CI: 65–100 %).
Prognostic Factors in Primary Chronic Myelofibrosis in Patients Aged Less Than 70 Years: A Report on 207 Patients with the Description of a Scoring System and Its Validation on 100 Other Patients.
Authors: Tanimoto Tetsuya E. , Shimoda Kazuya , Yamaguchi Takuhiro , Okamura Takashi , Mizoguchi Hideaki , Omine Mitsuhiro , Niho Yoshiyuki , Harada Mine
Published: October 2019

Primary chronic myelofibrosis (PCMF) is a rare myeloproliferative disorder among young individuals. Conventional therapies are often ineffective and only palliative. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) using myeloablative or reduced-intensity conditioning offers the only possible curative treatment. However, the course of PCMF is highly variable from only a few months to more than a decade. To identify PCMF patients for allo-HSCT candidates, we analyzed a Japanese cohort aged less than 70 years and established a new prognostic system based on simple clinical parameters. In a training series, we used 207 patients with complete data, whose diagnoses were made over a period of 10 years (1988 to 1997) (Okamura T et al. Int J Hematol 2001). Patients who received allo-HSCT were not included. Using the Cox proportional hazard regression model, four initial variables were independently associated with shorter survival: Male sex (hazard ratio [HR], 2.26; 95% confidence interval [C.I.], 1.35–3.46), Hemoglobin < 10 g/dL (HR, 3.01; 95% C.I., 1.83–4.93), the presence of constitutional symptoms (fever, sweats and weight loss) (HR, 2.33; 95% C.I., 1.33–4.06), and circulating blasts ≥ 1% (HR, 1.53; 95% C.I., 1.00–2.33). Based on the above criteria, two groups were identified; a low-risk goup with up to one adverse prognostic factor and a high-risk group with two or more adverse prognostic factors. With the median follow-up time of 83 months, the median survival was 292 months in the low-risk group, and 66 months in the high-risk group, respectively (P <.0001, Figure). An external data set of 100 patients who diagnosed after 1999 was used for validation. The 4 prognostic systems separated the validation series in groups with significantly different survival rates despite the median follow-up time of 12 months. A simple prognostic model using the combination of sex, Hb level, constitutional symptoms and circulating blasts was developed and validated. This model might help to select patients with PCMF for treatment and to evaluate therapeutic results including allo-HSCT.

Figure Figure
Increased Granulocyte CD11b and Monocyte Tissues Factor Expression in Patients with Essential Thrombocythemia and Thrombosis.
Authors: Arellano-Rodrigo Eduardo , Alvarez-Larran Alberto , Cervantes Francisco , Reverter Juan-Carlos , Villamor Neus , Montserrat Emili
Published: October 2019

The mechanisms accounting for the increased risk of thrombosis in patients with essential thrombocythemia (ET) are not well known. Recently, a possible role for the granulocytes in the development of thrombosis in ET has been suggested. To analyze the possible relationship between granulocyte activation and a history of thrombosis in ET, granulocyte activation was studied in 53 ET patients (26 with a previous history of thrombosis and 27 without) and 26 healthy controls. Neutrophil and monocyte CD11b expression, monocyte tissue factor (mTF) expression, and platelet-neutrophil (PNC) and platelet-monocyte (PMC) complexes were studied using whole blood cytometry and the results were expressed as percentages and in MESF (molecules of equivalent soluble fluorochrome) units. Granulocyte CD11b and mTF expression were measured at baseline and after activation with lipopolysaccharide (LPS). The results are as follows: 1) ET patients had significantly higher baseline percentages of circulating PNC and PMC as well as neutrophil CD11b MESF expression than the controls (p=0.0001); 2) monocyte CD11b expression was higher in patients with ET and thrombosis (mean 151241 MESF) than in those without (127191 MESF) and in the controls (65717 MESF) (p= 0.0001); 3) mTF baseline surface expression was increased in ET patients as compared with the controls (15635 vs 9385 MESF, p= 0.0001); 4) following LPS activation, patients with ET and previous thrombosis had significantly higher mean percentage of mTF expression than those without thrombosis and the controls (48%, 36% and 25%, respectively) (p= 0.0001). No differences were noted when the results were analysed according to the treatment that the patients were receiving at the time of study. In conclusion, patients with ET and a history of thrombosis show a marked increase in granulocyte activation, especially in monocyte activation, and therefore such alteration might play a role in the pathogenesis in these patients.
Sub-Cellular Localization Determines the Delicate Balance between the Anti and Proapoptotic Activity of Livin.
Authors: Ben-Yehuda Dina , Nachmias Boaz , Ashhab Yaqoub , Lazar Itay , Perlman Rebeca
Published: October 2019

Inhibitor of Apoptosis proteins (IAP) comprise a family of intracellular proteins that play an essential role in the regulation of apoptosis. IAP family members are defined by one or more repeats of a highly conserved anti-apoptotic domain termed the baculovirus IAP repeat (BIR), located at the amino-terminus. In addition, some IAPs contain a conserved domain termed RING finger at the carboxy-terminus. In a previous study, we identified an IAP family member termed Livin, which contains a single BIR domain and a carobxy-terminal RING finger motif. Livin inhibits apoptosis induced by a variety of stimuli. We further demonstrated that following apoptotic stimuli, Livin is cleaved by effector caspases 3 and 7, to produce a truncated form with paradoxical proapoptotic activity. In our current study, we reveal that while the full-length Livin shows a cytoplasmic distribution, the truncated form is readily detected in the nucleus. Using mutation analysis, we identified two domains outside the BIR sequence that are strongly engaged in determining the subcellular localization as well as the proapoptotic activity of Livin. These two domains are the N-terminus post-cleavage exposed area and the C-terminus RING domain. Remarkably, a single substitution of the first exposed amino acid Glycine, is sufficient to significantly diminish nuclear localization as well as the proapoptotic activity, thus linking the nuclear accumulation of the cleavage fragment with the proapoptotic activity. We demonstrate that the cytoplasmic localization of Livin is mediated through the RING domain, as a mutation that disables Livin from forming the zinc finger results in both a cytoplasmic and nuclear localization. Moreover, RING-mutated Livin subunit also loses its proapoptotic activity. Thus we suggest a model in which the exposed N-terminal region counteracts the cytoplasmic localization signal of the RING domain, promoting nuclear localization of the truncated form. The change in sub-cellular localization allows the RING domain to mediate the proapoptotic effect. To the best of our knowledge, this is the first example of such a regulatory mechanism in IAP family members.
Enforced BCL6 Expression Inhibits B Cell Development in Vivo.
Authors: Robbiani Davide F. , Colon Kaity , Naik Kruti , Nickerson Helen , Affer Maurizio , Chesi Marta , Bergsagel Peter Leif
Published: October 2019

The B-Cell Lymphoma 6 (BCL6) gene encodes for a zinc finger motifs containing transcriptional repressor that is frequently dysregulated by chromosomal translocations in germinal center lymphomas. A putative protooncogene, its transforming ability in vivo was reported in I-mu-HA-BCL6 knock-in mice by Cattoretti et al last year. We also tested this assumption in transgenic mice expressing BCL6 in B cells under the control of kappa light chain regulatory elements. We replaced the murine C-kappa locus with the 16kb human BCL6 genomic locus in a construct containing the murine kappa light chain regulatory elements (Vk, EiK, 3′RR).

While control transgenics were readily obtained (5/32 founders), only 3/68 founders were positive for the BCL6 transgene, of which only one (bearing a single copy of the transgene) was able to transmit the transgene to its progeny, thus suggesting embryonal toxicity of exogenous BCL6.

In the bone marrow, flow cytometry revealed a nearly complete block of B cell development at the pro-B to pre-B transition. This was also the stage at which we first detected expression of EGFP in control reporter mice that were generated in parallel. Spleens of transgenic mice weighed about 50% of control spleens and less than 5% of splenocytes were CD19+ B cells. These were IgM high, IgD intermediate, corresponding to an immature B cell phenotype. Lymph nodes were smaller and B cells barely detected. Peyers’ patches were not visible.

Combined, our analysis of 6–8 weeks old VkHABCL6 transgenic mice reveals that enforced expression of BCL6 early in development results in a profound block of B lymphocyte differentiation. How transgenic BCL6 modulates this effect at the transcriptional level remains to be investigated. To test the oncogenic potential of BCL6 in B cells, it will be interesting to precisely turn on this gene in the germinal center.