bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prolongation of Progression Free Survival In Mantle Cell Lymphoma After RHCVAD: ASCT Consolidation or Rituximab Maintenance
Authors: McQuade Jennifer , Ahmadi Tahamtan , Porter David , Frey Noelle , Loren Alison Wakoff , Goldstein Steven C. , Svoboda Jakub , Stadtmauer Edward , Schuster Stephen J. , Nasta Sunita Dwivedy 
Published: October 2019

Abstract
Abstract 3570
Mantle cell lymphoma (MCL) is a small B cell lymphoma, incurable with standard chemo-immunotherapy. The best initial treatment regimen remains unclear. Although it is generally acknowledged that aggressive approaches using combination chemotherapy and/or high dose chemotherapy can prolong survival, consensus on upfront treatment strategies for advanced MCL is currently lacking without randomized controlled data to guide treatment decisions. We conducted a retrospective cohort analysis to describe and compare the survival experiences of MCL patients at the University of Pennsylvania treated in the first-line setting with R-HCVAD (N=43) with or without autologous stem cell transplant (ASCT) or Rituximab maintenance. The primary study endpoints were PFS and OS as assessed by chart review and confirmed by SSDI database. Median follow up for all pts was 3 years. The median age was 53.7, and 76.7 % (n=33) were stage IV at diagnosis. 15 patients underwent consolidative ASCT. 11 pts received Rituximab maintenance. Comparing patients treated with R-HCVAD vs R-HCVAD + R maintenance vs. R-HCVAD + ASCT, there were no statistical differences in terms of age, ECOG PS, LDH, WBC, beta-2microglobulin, BM or GI involvement, bulky disease or blastoid variant at baseline. Median PFS for all patients was 3.9 years: R-HCVAD alone 2.1 years vs. R-HCVAD+R 3.9 years (P=0.02, HR 3.51, 95%CI: 1.2–10.2) vs R-HCVAD + SCT not reached (p=0.017, HR 3.7, 95%CI: 1.26–10.63). PFS survival rates at 2 years were 50%, 88% and 70%; 33%, 71/% and 63% respectively at 3 years, and 0%, 33% and 33 % at 5 years. 3 year OS for all patients was 84% (95% CI: 65–94) with no significant differences among the three approaches. Notably, only 1/8 patients treated with R-HCVAD + SCT relapsed after 2 years, with a median follow up of 4.8 years for these patients. Our data suggest a further improved PFS when R-HCVAD is consolidated with either Rituximab maintenance or ASCT. While neither of the two consolidative approaches appears superior in our limited data set, both show significant PFS prolongation when compared to R-HCVAD alone. Further prospective investigation of consolidative approaches after RHCVAD in a randomized fashion is warranted.
Figure 1: Figure 1:.

Disclosures:
No relevant conflicts of interest to declare.
Berberine Induces Apoptosis In Acute Lymphoblastic Leukemia Cells by Demethylation of RASSF1A and Self-Ubiquitination of MDM2.
Authors: He Jing , Zhang Hailong , Li Jiansha , Gu Lubing , Zhou Muxiang 
Published: October 2019

Abstract
Abstract 3627
Berberine, an isoquinoline alkaloid derived from a plant used in Chinese herbal medicine, is reported to exhibit anti-proliferative and pro-apoptotic effects on human cancer; however, its mechanism of action is not clearly defined. Herein, we demonstrate that berberine induces apoptosis in acute lymphoblastic leukemia (ALL) cells by demethylation of the tumor suppressor RASSF1A, which results in self-ubiquitination and degradation of the oncoprotein MDM2. In contrast to the conventional chemotherapeutic drug doxorubicin, which directly induced p53 activation and a subsequent upregulation of MDM2, berberine strongly induced persistent downregulation of MDM2 followed by a steady-state activation of p53, which resulted in apoptosis of ALL cells tested, even if they were doxorubicin-resistant. Our investigation discovered that berberine but not doxorubicin can induce demethylation of RASSF1A in RASSF1A-methylated ALL. The result of RASSF1A demethylation was the increased expression of the tumor suppressor, which disrupted the MDM2-DAXX-HAUSP complex, thereby promoting MDM2 self-ubiquitination and subsequent MDM2 degradation. Consistent with this finding, the most potent apoptosis was induced by berberine in wild-type p53 ALL cells with both RASSF1A methylation and MDM2 overexpression, while no pro-apoptotic effect was detected in berberine-treated RASSF1A normal ALL cells that had no or low level of MDM2 expression. Given that ALL cells with RASSF1A methylation and MDM2 overexpression are commonly chemotherapy-resistant our finding that these cells can be potently killed by berberine through effective induction of RASSF1A and inhibition of MDM2 suggests that this naturally-derived agent may have a highly useful role in the treatment of patients with refractory ALL.

Disclosures:
No relevant conflicts of interest to declare.
An Unintended Benefit of Anabolic Steroid Use: Therapy of Hemophilia B Leiden.
Authors: Rimmer Emily K. , Houston Donald Stuart , Seftel Matthew D. 
Published: October 2019

Abstract
Abstract 3661

Introduction:
In Hemophilia B Leiden, administration of testosterone and anabolic steroids in pre-pubertal boys can raise endogenous Factor IX (FIX) to levels sufficient to prevent most hemorrhages. It is unknown whether this androgen responsiveness may occur even in the post-pubertal male. We present a case that unintentionally demonstrated this phenomenon.

Case:
A 29-year-old man with mild Hemophilia B (baseline Factor IX [FIX] level 12%) presented to the emergency department with acute left quadriceps swelling. He received 1,800 units (18 U/kg) of human factor IX concentrate and the following day FIX level was 80%. One day later, he was seen with persistent pain and swelling despite adequate factor replacement. The patient disclosed that he had been using intramuscular injections of the anabolic steroid stanozolol. On suspicion that the thigh swelling may represent abscess rather than hematoma, blood cultures were drawn and grew Staphylococcus aureus. He received appropriate antibacterial therapy and 1.5 liters of purulent material was drained from the thigh. Intriguingly, FIX level one month after discharge was still 61%. One year later, his FIX level had stabilized at 36%.
Given the persistently elevated FIX level compared to baseline, the FIX gene was sequenced. Genotyping revealed a G to A transition at nucleotide -6 within the promoter region of the FIX gene. Mutations within nucleotides -34 to +19 of the 5′ region (Leiden-specific region) of the FIX promoter are associated with the Hemophilia B Leiden phenotype. This rare form of Hemophilia B improves following puberty. There is an androgen-response element (ARE) located at the -26 position of the FIX promoter, within the Leiden-specific region. The mechanism of recovery is not fully understood, but it is postulated that in the presence of androgen binding to the ARE, a normal FIX gene can be transcribed, thus overcoming the mutation causing the hemophilia phenotype prior to puberty. It has been shown that mutations involving the ARE do not resolve after puberty, which suggests an androgen responsive mechanism to the increase in FIX.

Conclusion:
This case raises the possibility of androgen-responsive increases in FIX that persist after puberty. We propose that a further rise in the FIX levels occurred as a result of exogenous exposure to pharmacologic concentrations of androgens in the post-pubertal period. This deliberate use of exogenous androgens carried the unintended benefit of abolishing the Hemophilia B phenotype.

Disclosures:
No relevant conflicts of interest to declare.
Upper Gastrointestinal Bleeding In Patients with Haemophilia In Iran: Prevalence of Helicobacter Pylori Infection.
Authors: Dolatkhah Roya , Khoshbaten Manouchehr , Kermani Iraj Asvadi , Sanaat Zohreh , Bonyadi Mohammad Reza , Ghojazadeh Morteza , Alimoghaddam Kamran , Kermani Touraj Asvadi 
Published: October 2019

Abstract
Abstract 3662

Background:
Upper Gastro Intestinal (UGI) Bleeding is one of the most life-threatening complications, and occurs in up to 25% of persons with Hemophilia (PWH).Recurrent bleeding is common and can be caused by the Helicobacter Pylori (H.Pylori) infection. Our aim was to evaluate the role of H.Pylori infection in UGI bleeding in PWH, using Serology and Stool antigen tests.

Methods:
Ninety patients with hereditary bleeding disorders, 30 patients(group A) with and 60 patients(group B) without a history of UGI bleeding, who were admitted to our Hemophilia Treatment Center, were included in the study. The prevalence of H.Pylori infection was investigated by stool antigen test, as well as serum serologic tests including IgG and anti-CagA.

Results:
Among 90 patients (81 men,16 women, mean age 31 ± 11 years), 66 patients with Hemophilia A,10 patients with Hemophilia B, 6 patients with VWD, and 3 patients with other factor deficiencies were evaluated for H.Pylori infection. About 46.7% of patients in group A, and 23.3% of patients in group B were anti-Cag A – positive in serum (P=0.02), while 76.7% of patients in group A and 51.7% of patients in group B had H.Pylori IgG antibodies in serum (P= 0.02). H.Pylori antigen in stool was positive in 76.7% in group A and 55% in group B (P=0.03). Gastro endoscopy was done to find the bleeding focus and further evaluations, in 19 patients with acute UGI bleeding. No statistically significant difference was found between type and severity of diseases and risk of UGI.

Conclusion:
There was a statistically significant differences in H.Pylori infection between patients with and without UGI bleeding history. So, H.Pylori infection should be considered as an important cause of UGI bleeding in PWH. We would recommend stool antigen test as a new and none-invasive screening test for diagnosis of H.Pylori infection in all patients with hereditary hemorrhagic disorders.

Disclosures:
Dolatkhah: Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Employment, Research Funding. Khoshbaten:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Research Funding. Kermani:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Employment, Membership on an entity's Board of Directors or advisory committees, Research Funding. Sanaat:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Employment, Research Funding. Bonyadi:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Employment, Research Funding. Ghojazadeh:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Employment, Research Funding. Kermani:Hematology and Oncology Research Center, Tabriz University of Medical Sciences: Research Funding.
A Phase II Study to Assess the Safety, Efficacy and Tolerability of Rituximab (Mabthera) In Combination with Plasma Exchange In Patients with Acute Thrombotic Thrombocytopenic Purpura (TTP).
Authors: Scully Marie , Cavenagh James D , Hunt Beverley , McDonald Vicki , Machin Samuel J 
Published: October 2019

Abstract
Abstract 3699
Idiopathic adult TTP is an acute life threatening disorder, in which antibodies, primarily IgG, are detected against ADAMTS 13. We undertook a a phase II trial in 40 patients between 2006-09 of Rituximab, 375mg/m2, weekly for 4 weeks, within 3 days of admission of acute TTP, in conjunction with standard therapy (PEX and steroids). Results have been compared to 40 historical controls (2000-2006), who had not received Rituximab, but had received other immunosuppressive treatments. The female to male ratio was 2:1, age 42 years (21-76), compared to 42 years (15-78) in the historical group. A third of trial patients required ITU admission and 15% were intubated and ventilated at presentation. One patient was withdrawn from the trial. Pre the 2nd Rituximab infusion, 68% had a platelet count >50 ×109/L and 38% >150 ×109/L. Six cases received more than 4 Rituximab infusions (maximum of 8), primarily non-Caucasian, guided by ADAMTS 13 assays. There was a significant reduction in days admitted in hospital in the Rituximab group (median 16.5 days) compared to historic controls (median 20 days) (p=0.04, Spearman correlation), specifically in Caucasian patients (12.5 V's 16 days-Rituximab V's Historical groups) (p=0.0005 Pearson Correlation). There was no overall significant difference in the number of PEX to remission. ADAMTS 13 activity on admission was median <5% (NR 55–126%). Median Anti ADAMTS 13 IgG was 40% (6-162%, NR:<4.2). Following 4 Rituximab infusions, median ADAMTS 13 activity was 43% (7-67%), median Anti-ADAMTS13 IgG was 12% (2-74%). In the historical group, 48% relapsed, median 18 months (3-60 months). In the Rituximab group, follow up 16–40 months, 10% relapsed, median 27 months (17-31). There were no excess infections in the Rituximab group. There were three deaths in the Rituximab cohort at days 11,15 and 25, due to progressive cardiac/neurological disease. In conclusion, Rituximab appears to be a safe and effective therapy given during acute TTP in conjunction with PEX and steroids. No significant difference was seen in the number of PEX to remission, compared to historical controls. However, there was a significant reduction in the number of days in hospital in the Rituximab group. The risk of relapse up to 40 months is significantly reduced in the Rituximab cohort. In patients with acute TTP, Rituximab should be considered in conjunction with standard therapy.

Disclosures:
Scully: MRC, BHF: Research Funding. Off Label Use: Rituximab, in TTP. McDonald:Baxter (uk): Research Funding. Machin:Baxter (UK), Bayer (UK), MRC, BHF: Research Funding.
Human CD8+/TCR−/CD56dim/− facilitating Cells Enhance Engraftment of HSC In NOD/SCID/IL2rγnull Recipients.
Authors: Huang Yiming , Elliott Mary J , Ramsey Deborah M , Miller Thomas , Bozulic Larry D , Ocker Ashley A , King Bradley J , Ildstad Suzanne T. 
Published: October 2019

Abstract
Abstract 3709
CD8+/TCR− graft facilitating cells (FC) enhance engraftment of hematopoietic stem cells (HSC). They are distinct from HSC since they do not generate multilineage engraftment when infused alone, nor do they form colonies in vitro. The function of human FC remains to be defined. Here, we phenotypically characterized the human CD8+/TCR− FC subpopulations and evaluated their function in vivo and in vitro. Human CD8+/TCR− FC comprised 1.1% ± 0.27% of total G-CSF-mobilized peripheral blood mononuclear cells (PBMC). Approximately 55% of FC express CD56dim/−, and the remaining population is CD56bright. In the CD8+/TCR−/CD56bright cell population, approximately 65% of cells express CD11c+ and 67% of cells express CD11b+. In CD8+/TCR−/CD56dim/− cell population, the majority of cells express CD3ε+ (80%), 17% were CD11c+, 16% were CD19+, 14% were CD11b+, 11% were CD123+, and 30% were HLA-DR+. To evaluate whether human CD8+/TCR−/CD56dim/− FC enhance engraftment of human HSC in vivo, we transplanted 100,000 HSC alone or plus 300,000 CD8+/TCR−/CD56dim/− FC into NOD/SCID/IL2rγnull (NSG) recipient mice conditioned with 325 cGy of total body irradiation. At 30 days after transplantation, 8 of 21 (38%) recipients of HSC alone engrafted. In contrast, 81% of recipients (n = 16) receiving HSC plus CD8+/TCR−/CD56dim/− FC engrafted, and donor lymphocyte and monocyte chimerism in PB was 0.53% ± 0.16% and 3.93% ± 1.28%, respectively. At 6 months after transplantation, NSG recipients of HSC alone lost donor chimerism in PB and no donor cells were detected in spleen and BM. In contrast, NSG recipients of HSC + CD8+/TCR−CD56dim/− FC exhibited durable donor chimerism in PB and showed significantly higher levels of donor chimerism in spleen (13%) and BM (9.43%) compared to recipients of HSC alone. To evaluate the function of CD8+/TCR−/CD56dim/− FC in vitro, HSC were incubated with CD8+/TCR−CD56dim/− FC for 18 hrs and then cultured in methylcellulose for 14 days in colony-forming cell assay. HSC plus CD8+/TCR−CD56dim/− FC generated significantly more colonies compared with HSC alone (P = 0.0038), suggesting that human CD8+/TCR−/CD56dim/− FC have a direct effect on the clonogenicity of HSC (Figure). In summary, our data indicate that (1) human CD8+/TCR− FC are a heterogeneous cell population; (2) CD8+/TCR−/CD56dim/− FC enhance engraftment of human HSC in vivo and promote HSC clonogenicity in vitro; and (3) recipients of CD8+/TCR−/CD56dim/− FC maintain a durable donor chimerism in BM and spleen.

Disclosures:
Bozulic: Regenerex, LLC: Employment. King:Regenerex, LLC: Employment, Equity Ownership. Ildstad:Regenerex, LLC: Equity Ownership.
Macrophages Prevent Human Red Blood Cell Reconstitution In NOD/SCID and NOD/SCID/γc−/− Mice.
Authors: Hu Zheng , Yang Yong-Guang 
Published: October 2019

Abstract
Abstract 3723
An animal model supporting human erythropoiesis will be highly valuable for assessing the biological function of human RBCs under physiological and disease settings, and for evaluating protocols of in vitro RBC differentiation from human embryonic stem cells. Although immunodeficient mice on the NOD background have been widely used to study human hematopoietic stem cell function in vivo, the successful use of these mice in the study of human erythropoiesis and RBC function has not been reported. We have previously shown that co-transplantation of human fetal thymic tissue (under renal capsule) and CD34+ fetal liver cells (FLCs; i.v.) in NOD/SCID or NOD/SCID/γc−/− mice results in the development of multilineage human hematopoietic cells. Here, we analyzed human RBC reconstitution in these humanized mice. Although a large number of human erythrocytes, which consisted predominantly of immature nucleated erythrocytes, were detected in the bone marrow of human fetal thymus/CD34+ FLC-grafted mice, human RBCs were undetectable in blood of these mice, even in those with nearly full human chimerism in peripheral blood mononuclear cells (PBMCs). Recipient mouse macrophage-mediated rejection is, at least, one of the major mechanisms responsible for the lack of human RBCs in these mice, as human RBCs became detectable in blood following macrophage depletion and disappeared again after withdrawal of treatment. Furthermore, treatment with human erythropoietin (EPO) and human IL-3 significantly increased human RBC reconstitution in mice that were depleted of macrophages. Like the human RBCs developed in the humanized mice, exogenously injected normal human RBCs were also rapidly rejected by macrophages in NOD/SCID mice. Taken together, our data demonstrate that human RBCs are highly susceptible to rejection by macrophages in immunodeficient mice. Thus, strategies for preventing human RBC rejection by macrophages are required for using immunodeficient mice as an in vivo model to study human erythropoiesis and RBC function.

Disclosures:
No relevant conflicts of interest to declare.
Organ-Specific Protection from CD8+ T Cell-Mediated Acute Skin GVHD by Bortezomib Administration Correlates with Decreased IL-6.
Authors: Sun Kai , Sckisel Gail , Tietze Julia , Li Minghui , Sayers Thomas , Blazar Bruce R. , Murphy William J. 
Published: October 2019

Abstract
Abstract 3735
We have previously demonstrated that while early administration of bortezomib after murine allogeneic bone marrow transplantation (allo-BMT) can prevent acute graft versus host disease (GVHD), delayed administration of bortezomib results in increased GVHD-dependent gastrointestinal toxicity (PNAS 2004, Blood 2005). This was primarily mediated by donor CD4+ T cells in fully MHC-mismatched allo-BMT (Blood 2008). In the current study, we assessed continuous bortezomib administration on CD8+ T cell-mediated acute GVHD. Using an MHC-matched, mHAg-mismatched model (C3.SW donor cells into lethally irradiated C57BL/6 recipients) in which acute GVHD is primarily induced by CD8+ T cells, we confirmed that significant skin and liver GVHD was observed after CD8+ T cell transfer. Histological evaluation of acute liver GVHD displayed severe lymphocyte infiltrates and severe subacute bile duct hepatitis consistent with hepatic GVHD. Severe acute skin GVHD was noted by persistent fur loss and skin ulcerations. Cutaneous histopathological assessment exhibited severe inflammatory infiltrates, epidermal hypertrophy, loss of hair follicles, satellite cell necrosis and dermal fibrosis. Notably, after allo-BMT, 6 weeks or thereafter, a significant increase in serum IL-6 levels was observed in the mice exhibiting marked skin damage, suggesting organ-specific correlation of higher serum IL-6 expression with acute cutaneous GVHD associated injury. We next investigated the effects of continuous treatment with bortezomib in this GVHD model. Surprisingly, we found that continuous bortezomib administration at the conventional dose of 0.75–1 mg/kg twice per week resulted in fractional GVHD- dependent lethal toxicity, exacerbating lymphocyte infiltration in liver, gut and lung as evidenced by pathological examination. However, skin GVHD pathology was markedly improved. Due to the limitations of conventional doses of bortezomib for treatment of acute GVHD, we then performed a low-dose bortezomib treatment at 0.125 mg/kg, twice per week. We found that mice receiving low-dose bortezomib were protected from lethal toxicity and displayed significant amelioration of skin GVHD pathology. Significant decreases in serum IL-6 levels were also consistently observed consistent with the amelioration of skin GVHD. Interestingly, while skin GVHD was markedly improved, there was no difference in liver, gut and lung pathology compared to control GVHD recipients indicating an organ-specific protection. These results demonstrate that continuous bortezomib can be applied in CD8+ T cell-mediated acute GVHD but necessitates lower dosing and organ-specific protection is observed.

Disclosures:
No relevant conflicts of interest to declare.