bluuebunny/crossref_metadata_2025_split huggingface dataset:
Therapeutic Efficacy and Cure Of Sensitive and T315I Pan-Resistant Human Ph+ Leukemia In Mice Using a TCR-Like Antibody To WT1/HLA-A0201 Alone, Or In Combination With Tyrosine Kinase Inhibitors
Authors: Dubrovsky Leonid , Brea Elliott , Pankov Dmitry , Veomett Nicholas , Dao Tao , Yan Su , Liu Cheng , Scheinberg David A. 
Published: October 2019

Abstract
Acute and chronic leukemias, including CD34+ CML stem cells, overexpress the Wilms tumor gene 1 (WT1) protein, making WT1 an attractive therapeutic target. ESKM is a fully human IgG1 antibody that targets a 9 amino acid sequence (RMF) of the protein WT1 in the context of HLA-A0201, allowing it to target an undruggable, widely expressed, intracellular oncogene product. BV173 is an HLA-A0201+, human Ph+ ALL cell line that expresses WT1, and tagged by our lab with luciferase. We engineered a tyrosine kinase inhibitor (TKI) resistant BV173-R cell line by transducing BV173 with the resistant T315I Bcr-Abl plasmid. Antibody-dependent cellular cytotoxicity (ADCC) was evaluated in vitro by chromium release assay, utilizing human PBMC effectors. Tumor growth in vivo was assessed in NOD/SCID gamma (NSG) mice with bioluminescence imaging (BLI). RT-PCR was used to evaluate minimal residual disease in mice with negative BLI signal at the end of therapy. Imatinib, dasatinib, and ponatinib were used at up to maximally tolerated doses, given IP once daily. ESKM was administered at 100 µg twice weekly IP. ESKM mediated ADCC against both BV173 and BV173-R cell lines in vitro. In a BV173 engrafted human leukemia xenograft model, ESKM was more potent than imatinib, with median tumor growth reduction of 78% vs 52%. Combination of imatinib and ESKM therapy resulted in a 94% reduction in leukemic growth. High dose dasatinib (40 mg/kg daily) was more potent than ESKM, but discontinuation of therapy due to dasatinib toxicity resulted in relapse. Combination with ESKM therapy with dasatinib resulted in cure in 75% of mice, confirmed by bone marrow RT-PCR three weeks after termination of therapy. For mice cytoreduced with dasatinib followed by consolidation therapy with ESKM, delayed relapse was observed, but no cures. ESKM was highly superior to imatinib and dasatinib against the T315I BV173-R leukemia in vivo. Cures were not achieved with combination therapy of ESKM and either first or second generation TKIs against resistant T315I leukemia. Ponatinib at 10 mg/kg had higher efficacy than ESKM alone against BV173-R, but mice treated with combination of ESKM and ponatinib had superior tumor reduction. CONCLUSION: ESKM is an effective therapeutic antibody for sensitive and T315I Ph+ ALL. Resistant T315I Ph+ leukemic growth is inhibited more effectively by ESKM therapy compared to imatinib and dasatinib, and combination therapy with ESKM is superior to ponatinib.
Supported by the Leukemia and Lymphoma Society, NIH R01CA55349, P01 23766 and T32CA62948-18.

Disclosures:
Yan: Eureka Therapeutics: Employment. Liu:Eureka Therapeutics: Employment, Equity Ownership.
Human Reticulocytes Extrude Inside-Out, Phosphatidylserine-Exposed, Autophagic Vesicles During The Final Step In The Formation Of Erythrocytes
Authors: Mankelow Tosti J. , Griffiths Rebecca E. , Flatt Joanna F. , Anstee David J. 
Published: October 2019

Abstract
During maturation to an erythrocyte, a reticulocyte must degrade or eliminate its residual cytosolic organelles and reduce its surface area. Using confocal microscopy, we show that in late reticulocytes produced from an in vitro culture system, this is achieved through a novel form of exocytosis whereby large (∼1.4um) intact, inside-out phosphatidylserine-exposed vesicles are expelled from the maturing reticulocyte. The vesicles contain organelle marker proteins and numerous erythroid membrane proteins, notably CD71, CD147 and stomatin. The presence of ubiquitin within these vesicles suggests a recognised mechanism for the targeting of proteins for extracellular export or degradation. The exocytosed vesicles are identical to intracellular GPA-decorated autophagic vesicles previously identified in cultured reticulocytes (Griffiths et al 2012). GPA-decorated vesicles are also seen in some cells in peripheral blood. Proteins involved in vesicle trafficking, SNARE (VAMP7), ESCRT (CHMP4B) and myosin, locate to defined positions at the point of vesicle extrusion. We hypothesize that the exposed “eat me” phosphatidylserine signal ensures that released autophagic vesicles are rapidly removed from circulation by professional phagocytic cells within the spleen thus ensuring maturation to erythrocytes without the release of potentially toxic material into circulation. Our results describe a previously unrecognised mode of exocytosis which may have significance beyond erythropoiesis particularly with respect to apoptosis and autophagy and reveal the final step in the formation of human erythrocytes.
Reference
Griffiths RE, Kupzig S, Cogan N, Mankelow TJ, Betin VM, Trakarnsanga K, Massey EJ, Lane JD, Parsons SF, Anstee DJ.Maturing reticulocytes internalize plasma membrane in glycophorin A-containing vesicles that fuse with autophagosomes before exocytosis. Blood. 2012 Jun 28;119(26):6296-306.

Disclosures:
No relevant conflicts of interest to declare.
How Does CD47-SIRPα ‘Don’t Eat Me Signal’ Physically Signal Self
Authors: Nisha Sosale , Discher Dennis E. 
Published: October 2019

Abstract
Resident macrophages in spleen and liver are particularly adept at recognizing foreign pathogens through recognition of ‘non-self’ proteins on the pathogen surface but also through the absence of ‘self’ proteins that are highly displayed on circulating blood cells. Red blood cells display a ‘marker of self’ protein CD47 which increases the in vivo half-life and decreases red-pulp splenic macrophage uptake of mouse RBC (Oldenborg et al, Science 2000) and also of particles displaying human-CD47 in recent studies by our group (Rodriguez et al, Science 2013). CD47 signals self through its counter receptor SIRPa, which is highly expressed on the surfaces of myeloid cells but also highly polymorphic. The CD47 protein functions in vitro as a marker of self toward human SIRPa on human macrophages and monocytes, inhibiting accumulation of myosin II motor protein to the phagocytic synapse (Tsai 2008). The work here aims to clarify when and how CD47-SIRPa inhibition physically signals ‘self’ during macrophage phagocytosis uptake. While it is clear that CD47 reduces the number of uptake events, here we use time-lapse and confocal microscopy to examine the forces of distortion imparted by phagocytes on opsonized red blood cell targets during uptake. Glutaraldehyde-fixed RBC are also used as a model to assess the affects of cell rigidity in this self-recognition process, since rigidity is relevant to processes as diverse as RBC aging and sickle RBC to malaria but also because adhesion (by macrophages) is expected to activate the myosin-II contractility system and oppose CD47 signaling. Through blocking and pharmacological approaches, we parse the pathways between foreign, self, and rigidity sensing.

Disclosures:
No relevant conflicts of interest to declare.
Consolidation Therapy Is Associated with Significantly Lower Bleeding Risk Compared to Induction Therapy in Patients with Acute Myeloid Leukemia
Authors: Schaefer-Eckart Kerstin , Wendelin Knut , Pilz Bettina , Kramer Michael , Ehninger Gerhard , Wilhelm Martin , Wandt Hannes 
Published: October 2019

Abstract
Background: Between 2005 und 2010 we conducted a multicenter randomized study comparing a therapeutic and a prophylactic (morning platelet (ptl) trigger < 10/nl) platelet transfusion strategy in patients with hematological disorders. (The Lancet Vol 380, No 9850, pp 1309-16). Briefly, we could show that a therapeutic plt transfusion strategy, where platelets are transfused in clinical stable patients (pts) only if bleeding ≥ WHO grade II occurs is safe in patients after autologous transplantation. In patients with acute myeloid leukemia (AML) we observed significantly more severe bleedings (WHO IV°) with the therapeutic regimen. To proof, if there is a difference in bleeding risk related to the remission status of the patients, we conducted a post-hoc analysis to compare the risk of severe bleeding (WHO III° and IV°) in induction with consolidation therapy.
Patients and Methods: We analyzed 175 pts with 175 cycles of induction therapy, 90 with a prophylactic transfusion regimen and 85 cycles with a therapeutic regimen. 131 pts received 268 cycles of consolidation therapy, 155 with a prophylactic and 113 with a therapeutic transfusion strategy.
Results: Bleedings WHO III° were neither different between the two strategies nor between induction and consolidation therapy. In contrast, there were significantly more bleedings WHO IV° in induction (7,4%) compared to consolidation therapy (1,5%; p=0,01). In addition, there were significantly more bleedings WHO IV° with the therapeutic regimen (11,8%) compared to the prophylactic strategy (3,3%) in induction therapy (p=0,012). This difference was less pronounced in consolidation therapy. But even in the prophylactic arm most WHO IV° bleedings occurred in pts with more than 10/nl ptl, which shows, that the morning platelet count should not be the only trigger for a platelet transfusion.
Conclusions: In consolidation therapy the risk of bleeding is significantly less compared to induction therapy, even with a therapeutic platelet transfusion strategy. During induction therapy the prophylactic strategy should remain the standard of care. The therapeutic transfusion strategy in consolidation therapy will be proven prospectively in an ongoing multicenter study.

Disclosures
No relevant conflicts of interest to declare.
Day 21 Chimerism Is a Critical Prognostic Factor for Graft Failure and Survival in Recipients of Myeloablative Conditioning and Umbilical Cord Blood (UCB) Transplantation
Authors: Bejanyan Nelli , Brunstein Claudio G , Cao Qing , Thyagarajan Bharat , Sandhu Karamjeet S. , Arora Mukta , Weisdorf Daniel J. , Wagner John E 
Published: October 2019

Abstract

Compared to other hematopoietic stem cell sources, UCB carries an increased risk of graft failure (GF), a life-threatening complication of transplant. Timely recognition of GF could help plan potentially curative salvage therapy, such as second grafting [Ruggeri et al, Haematologica 2014]. This study was aimed at identifying patients at high risk of GF. We hypothesized that early chimerism results might be a useful prognosticator. Therefore, we examined the results of day 21 bone marrow (BM) and peripheral blood (PB) chimerism testing in 327 (123 single UCB and 204 double UCB) patients receiving myeloablative conditioning (2000-2013) for various hematological malignancies. Recursive partitioning analysis was used to determine the optimal cut off of day 21 chimerism for neutrophil engraftment. Chimierism ≥85% was observed in 283 and <85% in 44 patients. For all patients surviving >21 days without evidence of disease relapse the incidence of neutrophil recovery was 92% (95% CI 89-95%) by day 42. However, for those with <85% chimerism, the rate of GF was 42% (n=20) as compared to only 3% (n=9) among patients with ≥85% chimerism at day 21 (p<0.01). As expected, 1-year overall survival (OS) was very poor in the <85% chimerism (21% with complications of GF as the most common cause of death). In multiple regression analysis, <85% chimerism was associated with 3.7-fold less likelihood of achieving neutrophil recovery by day 42 (HR 0.27, 95% CI 0.18-0.41; p<0.01) after adjustment for number of UCB units (p=0.04), HLA disparity (p=0.15) and year of transplant (p=0.10). Chimerism was also an independent risk factor for 1-year treatment related mortality (TRM, 19% vs. 39%; p<0.01), disease-free survival (63% vs. 42%; p=0.03), and OS (69% vs. 42%; p<0.01) among engrafted subset of patients with chimerism ≥85% versus <85%, respectively. In multivariable analysis, <85% chimerism remained prognostic for increased 1-year TRM (HR 2.14, 95% CI 1.06-4.31; p=0.03), treatment failure (HR 1.85, 95% CI 1.05-3.25; p=0.03) and mortality (HR 2.19, 95% CI 1.23-3.87; p<0.01) for all engrafted patients with myeloablative UCB transplant. Chimerism, however, was not an independent predictor of grade II-IV acute GVHD at 100 days (p=0.41), 1-year relapse (p=0.78) or chronic GVHD (p=0.41). Our data indicate that chimerism <85% at day 21 is critically important prognostic factor for graft failure and survival after UCB transplant. These results argue for early evaluation in order to intervene at an earlier time point to enhance survival.

Disclosures
No relevant conflicts of interest to declare.
Efficacy of Mesenchymal Stem Cells for the Treatment of Steroid-Refractory aGVHD
Authors: Watakabe Kyoko , Miyamura Koichi , Ozawa Yukiyasu , Okada Masaya , Yamashita Takuya , Murata Makoto , Ishikawa Takayuki , Uike Naokuni , Hidaka Michihiro , Kobayashi Ryoji , Imamura Masahiro , Tanaka Junji , Ohashi Kazuteru , Taniguchi Shuichi , Ikeda Takashi , Eto Tetsuya , Mori Masaki , Muroi Kazuo , Ozawa Keiya 
Published: October 2019

Abstract
Background: Steroid-refractory acute graft vs. host disease (SR-aGVHD) remains a significant complication of allogenic hematopoietic stem cell transplantation (allo-HSCT). In this phase II-III multicenter clinical study we evaluated the efficacy and safety of MSCs(JR-031; JCR Pharmaceuticals Co., Ltd., Japan) for SR-aGVHD.
Method: 25 patients (age: 5-66 years, median 33years; male 15, female 10) who developed SR-aGVHD (grade III 22, grade IV 3) after allo-HSCT (Nov. 2011-Sept. 2012) were enrolled and given 8 biweekly infusions of 2x106 cells/kg of JR-031 for 4 weeks, with an additional 4 infusions weekly after 28 days in patients with partial response (PR). They were followed up to 24 weeks.
Results: At 4 weeks after the first dose, 6 (24%) patients showed complete response (CR), 9 (36%) patients showed PR, 4 (16%) patients showed mixed response (MR) and 1(4%)patient showed no change (NC). During 24 weeks of observation period, 12 (48%) patients achieved durable CR (CR >=28days). As for response by organ, 80% (16/20) of GI, 66.7% (8/12) of skin and 66.7%(4/6) of liver GVHD completely responded (stage 0). 15 (60%) patients survived up to 24 weeks after the first dose.All patients experienced at least one adverse event(AE). Common AEs were leukopenia (12 patients), thrombopenia (9 patients), sepsis, anemia, TMA and hepatic dysfunction (6 patients). Among 10 death, causal relationship with JR-031 were not completely ruled out in 4 cases (TTP, pneumonia, sepsis and relapse of underlying malignancy), however there was no case in which the causality was strongly suggested.
Conclusion: It was suggested that JR-031 is a safe and effective therapy in the treatment of patients with SR-aGVHD.

Disclosures
Miyamura: Novartis: Honoraria, Speakers Bureau.
Androgen Function in Long-Term Survivors of Haematopoietic Stem Cell Transplantation
Authors: Topping Jonathon E , Todd Jeannie , Apperley Jane F. , Salooja Nina 
Published: October 2019

Abstract

Endocrinopathies are well known complications of haematopoietic stem cell transplants (HSCT) and their onset may occur many years after the transplant process. Infertility in men is common and occurs soon after exposure to high doses of radiation/chemotherapy as a result of Sertoli cell damage. Despite this testosterone levels are usually normal. We have investigated testosterone levels in male patients surviving a minimum of 10 years following of HSCT to see whether there are long term effects of chemoradiotherapy on testosterone production.
We have investigated testosterone levels in 46 long-term survivors of allogeneic HSCT attending a dedicated late effects clinic over a 12 month period. Data were compared to age-matched non-transplant controls in a paired case-control analysis. The controls were being investigated for fatigue but had no specific underlying illness. Where testosterone levels were low, repeat measurements were taken with LH and SHBG.
The median age at HSCT was 30 years (range 12-53) and the median duration of follow up was 19 years (range 10-34). 44/46 received cyclophosphamide/TBI conditioning. 32 patients had CML, 5 had ALL, 7 had AML, 1 had aplastic anaemia and 1 had lymphoma.
26/46 (57%) patients had a low testosterone reading at some point compared to15% in the control group (p < 0.0001). Two of these patients had a low SHBG and a normal free androgen index. 16/46 patients had persistently low levels which for 8 was primary testosterone deficiency with elevated LH and for 8 was secondary with inappropriate low gonadotrophins. In four patients the testosterone fluctuated around the normal level and for four it was an isolated low reading. The transplanted patient group had a significantly lower mean testosterone level at 10.8 nmol/l (range 3.2-20.3) compared to 15.2 nmol/l (range 7.5-28.1) in the control group (p = 0.00009). The probability of developing true hypogonadism (defined as consistently low testosterone levels after repeat measurements) increased from 0.087 at 10 years to 0.554 after 25 years.
We have shown that true hypogonadism is a common finding between 10 and 25 years post transplant. Implications include reduced energy levels, decreased libido, decreased bone density and sexual dysfunction. Our data indicate that for male patients surviving 10 years or more after HSCT annual biochemical surveillance is required.

Disclosures
Apperley: Ariad: Research Funding; Bristol Myers Squibb: Research Funding; Novartis: Research Funding; Pfizer: Research Funding.
Safety and Efficacy of Total Dose Infusion of Low Molecular Weight (LMW) Iron Dextran in a Large Population of Anemic Patients Across a Broad Spectrum of Diagnoses Associated with Iron Lack
Authors: Mehmood Tahir , Swe Khine , Das Gopimohan , Gozu Aysegul , Auerbach Michael , Aung Sein 
Published: October 2019

Abstract

Background: Physicians are reluctant to recommend high dose intravenous (IV) iron infusion for iron deficient (ID) patients due to concerns of adverse events (AEs) including anaphylaxis. This retrospective analysis evaluates efficacy and AEs with LMW iron dextran (ID) total dose infusion (TDI) in patients with iron deficiency.
Methods: We retrospectively evaluated the Electronic Medical Records of 468 consecutive patients with iron deficient anemia (IDA) who received IV LMW ID between 01/2008 and 07/2011 in an ambulatory infusion center. Diagnostic parameters for IDA included hemoglobin [Hgb] <11 g/dL, transferrin saturation [TSAT] 19%, ferritin <100 ng/mL. The majority had documented intolerance of or inadequate response to oral iron. All received TDI of LMW ID (1130 ± 217 mg) over 1 hour after a test dose of 25 mg. Pre-medication was not used. Patients were closely monitored for AEs during and immediately after infusions. Laboratory data for baseline and repeat hematological parameters within 8 weeks following infusions were analyzed. End points were safety and efficacy. Safety data was available for all 468 patients. Follow-up data at 8 weeks was available on 315.
Figure 1 Figure 1.
Results: There were 103 males and 365 female patients. Most common diagnoses were GI bleed, chemotherapy, menorrhagia and chronic kidney disease. At baseline, mean Hgb was 9.8 g/dL, ferritin 92.08ng/mL, and TSAT 12.37%. At 8 weeks the mean hemoglobin increased to 10.97 g/dL (p =0.0001), Ferritin 279.63 ng/mL (p =0.0001), and TSAT 21.85% (p=0.0001). All tolerated the infusion well. There were no incidents of fever, chills, wheezing, hypotension, stridor, periorbital edema, anaphylaxis or death. One complained of chest tightness, but completed the infusion after several minutes and the administration of diphenhydramine and steroids. One developed a rash. Fatigue and myalgia were reported in 17.31% and 8.33% of patients respectively.
Conclusions: This study demonstrates TDI of LMW ID can be given safely and conveniently over 1 hour in an outpatient setting without premedication to ID patients. Easily manageable minor reactions did not affect the therapeutic plan. No allergic or anaphylactic reactions were observed. This study demonstrated TDI of LMW ID improved hemoglobin levels more than 1 g/dL within 8 weeks. Consistent with the preponderance of published evidence TDI of LMW ID should be considered an important therapeutic option for iron replacement in patients with IDA.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Myeloproliferative Syndrome with Leukemia Preceding Polycythemia Vera: A Case Report
Authors: CALABRESI PAUL 
Published: October 2019

Abstract
A case of chronic erythroleukemia is reported in which a picture of chronic granulocytic leukemia preceded the erythremic proliferation by one to three years. The patient was followed throughout his entire 10-year course and the clinical impressions were confirmed initially by biopsy and eventually at necropsy.
The rare examples of this occurrence usually cited in the literature are reviewed and briefly discussed. In every case the development of erythremia was preceded by radiation therapy for the leukemic dyscrasia.
A set of criteria for evaluating such cases in order to provide definite evidence for this sequence of events is suggested.
The transformation of chronic granulocytic leukemia into polycythemia vera or chronic erythroleukemia lends further support to the concept of fundamental similarity and close inter-relation of these myeloproliferative disorders.
Hematologic Findings in Human Beings Given Therapeutic Doses of Gallium-72
Authors: WOLINS W. , BOND V. P. 
Published: October 2019

Abstract
The hematologic effects of Ga72 administered intravenously to patients with diffuse bony metastases or primary bone malignancy were studied.
1. The effects of this internally administered isotope were, in general, similar to those resulting from total-body external radiation. The degree of effect was greater than that anticipated from estimates of the total-body dose received from the isotope. Localization in the bone may have been responsible for this finding.
2. The depth and duration of platelet count depression probably represented the best indices of the degree of marrow damage after radiation, and may be indicative also of the total damage sustained by the marrow from previous exposure.
3. The effects of large doses of radiation on the marrow appeared to be cumulative to a point of no return, beyond which regeneration may not be possible.
4. It was found that a total white count below 1,000 and a platelet count below 25,000 could be tolerated for weeks without infection or gross bleeding and with ultimate recovery. Therapy because of these findings alone did not appear to be indicated.
Folate Deficiency in Chronic Liver Disease
Authors: KLIPSTEIN FREDERICK A. , LINDENBAUM JOHN 
Published: October 2019

Abstract
Fifty-five patients with liver disease of varied etiology and severity have been studied. Serum folate concentrations were subnormal and folic acid clearances rapid, when studied, in 19 actively imbibing alcoholic cirrhotics who had a megaloblastic anemia. Eleven patients, from both the nonalcoholic and alcoholic groups, had rapid folic acid clearances, with subnormal serum folate levels in seven, in the absence of morphologic evidence of folate deficiency. Serum B12 concentrations were uniformly normal or elevated.
Dietary deficiency appeared to be the major cause of folate deficiency; all 19 patients who had megaloblastic changes were considered to have an inadequate dietary intake. Increased requirement for folate due to hyperactivity of the bone marrow secondary to gastrointestinal bleeding. hypersplenism, or hemolysis appeared to contribute to the development of abnormal folic acid determinations in many patients in both the alcoholic and nonalcoholic groups. Two of 10 patients studied had malabsorption of folic acid. Such factors as the presence of ascites, an expanded plasma volume, a Patent portacaval shunting procedure, the type of alcoholic beverage imbibed, and the severity of impairment of liver function did not appear to be of significance in the development of folate deficiency.