bluuebunny/crossref_metadata_2025 huggingface dataset:
Successful Engrafment After HLA Identical Granulocyte Transfusion As Support Therapy in a Septic Shock Patient with Acute Lymphoblastic Leukemia
Authors: Ovilla Roberto , Barrera-Carmona Claudia , Guzman-Bouilloud Nicolas , Buganza-Torio Elizabeth , Jimenez-Alvarado Rosa , Monreal-Alcantara Claudia
Published: October 2019

Abstract 4507

A 53 years old male started in February 2010 with ecchymosis, petechiae and spontaneous gum bleeding. He was diagnosed with acute lymphoblastic leukemia, and was started on HYPER-CVAD, in combination with anti-tumoral lysis syndrome measures and antimicrobial, antiviral y antifungal prophylaxis. After finishing HYPER-CVAD phase A, he developed severe myelosuppression even with the use of G-CSF, 72 hours later he presented with abdominal cramping, fever up to 38.2°C and hypotension, with a high clinical suspicion of neutropenic colitis; his mean arterial pressure average was 45 mmHg, he was started on intravenous colloids, dobutamine and norepinephrine drips.

Because of severe myelosuppression, septic shock and myocardial depression, a granulocyte transfusion without previous mobilization was performed with an identical sibling donor with concomitant use of granulocytic colony stimulation factor. Severe myelosuppression was maintained during 7 days, however shock state was reversed some hours after performing granulocyte transfusion, without infectious signs, 36 hours later a mononuclear cell infusion was performed from the same donor, with previous 2 days G-CSF mobilization. A gradual increase in leukocyte number appeared, with 400, 900 and finally 1900.

A new bone marrow aspiration was performed, where hematopoietic recovery was confirmed from his sister's cells with confirmation by karyotype and microsatellites. He was then considered bone marrow grafted HLA compatible.

On April 2010 he presented with acute diarrheic syndrome secondary to a CMV infection, he was started on ganciclovir and intravenous immunoglobulin. On May he presented an Aspergillus pneumonia that was treated both clinically with antifungal therapy and surgically with thoracoscopy to remove the fungi lesion. On October 2010 he started with graft versus host manifestations on the skin and in the liver. He started immunosuppressive treatment with Prednisone and Sirolimus. On November he developed anogenital herpes zoster infection. After this complication, every GVHD manifestation ceded. Now, 28 months post-bone marrow transplant he presents full remission with no evidence of leukemia.

Granulocyte transfusion is an uncommon technique. Preferably it should be done with and identical donor. In this case report, the justification of the procedure was to be able to achieve a remission from a potentially irreversible septic shock; this was successfully done, in an unexpected clinical scenario with severe life threatening, and in a casual manner, the patient achieved a successfully bone marrow transplant, and now 30 months post-granulocyte infusion the patient is free from any evidence of disease.

Disclosures:

No relevant conflicts of interest to declare.
Comparison of Lengths of Stay for Inpatients with Immune Thrombocytopenia Treated with Anti-Rh(D), Intravenous Immunoglobulin, and Steroids
Authors: Melinscak Hrvoje , Shapira Ilan , Varma Mala
Published: October 2019

Abstract 4646

Rationale:

Anti-Rh(D) is an effective treatment for acute immune thrombocytopenia [ITP]. It has a faster onset of action (1 day) vis a vis corticoids (3 days) and intravenous immunoglobulin [IVIG] (4 days). A direct comparison of length of stay for adult inpatients receiving these therapies has hitherto not been performed. We hypothesized that the length of stay would be shortest for patients treated with anti-Rh(D).

Methods:

A retrospective chart review was conducted to assess length of stay in relation to treatments proffered for ITP. The defining diagnosis of ITP [coded 287.31] was rendered from the computerised record at St. Luke's-Roosevelt Hospital Center and identified 303 patients, of which 147 received treatments for active ITP within a period spanning 01SEP2005 through 29FEB2012. Treatments consisted of prednisone alone, dexamethasone alone, anti-Rh(D) alone, IVIG alone, and combinations of corticoids and the latter two. An average length of stay was tabulated for each treatment regimen. Age and gender were also recorded.

Results:

A total of 147 hospitalisations for ITP were noted and the analysis of variance statistical calculation applied thus. The median age was 48 years and the male:female ratio was 1.1:1. Eleven groups were delineated and the means for length of hospital stay with confidence intervals derived. The groups were as follows: Prednisone, Dexamethasone, Methylprednisolone, Anti-Rh(D), IVIG, Prednisone and Anti-Rh(D), Dexamethasone and Anti-Rh(D), Prednisone and IVIG, Dexamethasone and IVIG, Methylprednisolone and IVIG, and Anti-Rh(D) and IVIG or Dexamethasone, Anti-Rh(D), and IVIG. The overall p-value for length of stay was 0.0016 (Table 1). The shortest stays were recorded for the corticoid alone groups; however, the mean stay for anti-Rh(D) was shorter than that of IVIG, both in sole and combined modality treatments.

Conclusion:

Anti-Rh(D) is favourable with respect to hospital stay duration. Although corticoids result in still shorter lengths of stay, anti-Rh(D) demonstrated a shorter length of stay compared with IVIG. Combined with its single dosing and relative cost savings, anti-Rh(D) is an excellent alternative to IVIG.

Disclosures:

No relevant conflicts of interest to declare.
Platelet Apoptosis in Uremic Patients
Authors: Li Ming , Ma Tongling , Lu Guoyuan , Deng Kun , Yan Rong , Dai Kesheng
Published: October 2019

Abstract 4647

Introduction:

Apoptosis, which serves for the regulation of cell lifespan and is long attributed exclusively to nucleated cells, has been well-documented in anucleate platelets. Diverse cell-external chemical and physical stimuli have been reported to induce transformation of resting platelets to apoptotic state. Uremia, a clinical syndrome associated with retention of various solutes that would normally be excreted by the kidneys, is frequently accompanied by bleeding tendency, and the mechanism remains unclear. The aim of the present study is to investigate whether platelet apoptosis occurs in uremia patients.

Methods:

Venous blood was drawed from 13 patients with end stage renal disease (ESRD) who exhibited uremia syndrome and 13 health controls. Platelet-rich plasma (PRP) was prepared and then was detected for apoptotic events including depolarization of mitochondrial inner membrane potential (ΔΨm), phosphatidylserine (PS ) exposure, variations of apoptotic Bcl-2 family proteins, activation of caspases-3 by Flow cytometry or Western-blot. Furthermore, normal washed platelets were incubated with the poor- platelet plasma (PPP) from uremic patients, and then were detected for apoptotic events. The comparisons of the data were made using paired Student's t-test.

Results:

Compared with controls, ΔΨm significantly depolarizated in platelets from uremic patients (P<0.05). Furthermore, Bax was up-regulated, Bcl-2 and Bcl-XL were down-regulated, and caspase-3 was activated in platelets from uremic patients. However, there was not significant difference in PS exposure and P-selectin expression between the platelets from uremic patients and health contols (P>0.05). Next, PPP from uremic patients was incubated with normal platelets, and the platelets presented ΔΨm depolarization, up-regulation of Bax, down-regulation of Bcl-2 and Bcl-XL, and caspase-3 activation.

Conclusion:

The data demonstrate that platelets are incurred apoptosis in uremia patients. The finding might suggest a novel pathogenic mechanism for bleeding tendency in uremic patients.

Disclosures:

No relevant conflicts of interest to declare.
Surface Expressions of Platelet Glycoprotein Ibα, GpIIb/IIIa, and P-Selectin Are Elevated in Lung Cancer Patients
Authors: Zhang Jiansheng , Zhao Lili , Huang Jianan , Zhang Pingping , Dai Kesheng
Published: October 2019

Abstract 4655

Objective:

Thromboembolic events are common symptoms in patients with cancer, and the mechanisms are not totally understood. The platelet adhesion moleculars play important roles in the process of thrombus formation. The aim of the current study is to investigate the surface expression of platelet glycoprotein (GP) Ibα, GPIIb/IIIa, and P-selectin in patients with lung cancer.

Method:

Platelets were isolated from patients with lung cancer and normal volunteers, and then the surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were measured by Flow cytometry. The glycocalicin (GC) fragments in the peripheral blood of patients with lung cancer were tested by Western blot. The surface expression of GP Ibα on normal volunteers' platelets incubated with the platelet-poor plasma (PPP) of patients with lung cancer was measured by Flow cytometry and Western blot.

Results:

The surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were obviously increased in patients with lung cancer compared with that of normal volunteers, and the GC fragments in PPP were also increased in patients with lung cancer. Incubation of control platelets with the PPP from cancer patients resulted in the elevation of the surface expression of GP Ibα in control platelets. Furthermore, the expressions of GP Ibα, GPIIb/IIIa, and P-selectin on platelets from patients with advanced stage of lung cancer (III-IV stage) were significantly higher than those with early stage (I-II stage) (P<0.05). However, there were not significantly difference in the surface expressions of GP Ibα, GPIIb/IIIa, and P-selectin among the pathological types of lung cancer (squamous cell carcinoma, adenocarcinoma, small cell lung carcinoma, large cell lung carcinoma) (P>0.05).

Conclusion:

The surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were obviously elevated in patients with lung cancer, and the plasma from cancer patients is responsible for the elevations. These findings provide a new view to understand the pathogenic mechanisms of thromboembolic events in the patients with lung cancer.

Disclosures:

No relevant conflicts of interest to declare.
Pulmonary Endothelial Cell Irradiation Damage Signaling Initiates Late Fibrosis
Authors: Kalash Ronny , Houghton Frank , Zhang Xichen , Wang Hong , Epperly Michael W. , Greenberger Joel S.
Published: October 2019

Abstract 4682

Thoracic irradiation of C57BL/6NHsd mice produces an acute lung inflammatory period (0 to 30 days after irradiation), followed by a stable latent period (30 to 100 days), followed by a delayed fibrosis reaction (100 –150 days). Fibrosis involves migration of bone marrow derived mesenchymal stem cells (fibroblast progenitors) into the lung. The molecular mechanisms which initiate migration and homing of fibroblast progenitors are unknown. Increased pulmonary TGF-β expression has been correlated with the onset of fibrosis. To identify the cells and genetic pathways involved, C57BL/6NHsd female mice were irradiated with 20 Gy to the thoracic cavity and sacrificed at serial time points including days 0, 2, 14, 28, 50, 75, 100, 110, 125, 150, and 200, lungs were removed and RNA extracted from either whole lung or purified cell populations. Real Time Polymerase Chain Reaction (RT-PCR) was used to quantitate gene expression.

In whole lung tissue, when compared to un-irradiated lungs, a significant elevation in expression of multiple inflammation and chemotaxis associated genes (including TGF-β, TNF-α,and MnSOD) and stress response genes (including NFkB, AP-1, SP-1,and Nrf2) was detected both during the acute inflammatory phase (p= 0.0112, 0.0037, 0.0245 and p= 0.0001, < 0.0001, =0.0010, =0.0181 respectively) and late-onset fibrosis phase (p= 0.0079, 0.0459, 0.0001, and p < 0.0001, = 0.0006, =0.0128, < 0.0001 respectively), but were either not elevated or significantly decreased during the latent period ( p< 0.0001, = 0.3785, 0.8123, and p=.0243, < 0.0001, < 0.0001, < 0.0001 respectively). In contrast, increased expression of endothelial cell associated genes including: vWF, VEGF, CTGF, FGF1 and CCL13 was detected during the latent period, as well as during the acute and chronic reaction phases. Purified lung endothelial, but not epithelial cells, showed stable vascular gene elevations of vWF, VEGF, CTGF, FGF1 and CCL13 during the entire time course including day 7 (p < 0.0001, < 0.0001, =0.0035, =0.0009, =0.0056), day 75 (p=.00263, 0.0104, 0.0284, 0.0038, and 0.0007), and day 150 (p < 0.0001, < 0.0001, =0.0005, =0.0039, =0.0014 respectively). The data supports a significant role of vascular gene expression in the activation and recruitment of cells involved in radiation induced late pulmonary fibrosis.

Further analysis of lung endothelial cells may elucidate the mechanisms involved in the late effects of thoracic irradiation, and identify new targets for drug development to prevent irradiation pulmonary fibrosis.

Acknowledgments:

This project was supported by NIH-2R01CA119927-08A1.

Disclosures:

No relevant conflicts of interest to declare.
Rapid Increase of CD8+ T Cell Count in Peripheral Blood of Pediatric Patients After HLA-Identical Stem Cell Transplantation for African Sickle Cell Anemia
Authors: Isgrò Antonella , Marziali Marco , Sodani Pietro , Gaziev Javid , Fraboni Daniela , Paciaroni Katia , Simone Maria Domenica , De Angelis Gioia , Alfieri Cecilia , Gallucci Cristiano , Roveda Andrea , Torelli Fabio , Cardarelli Luisa , Ribersani Michela , Lucarelli Guido
Published: October 2019

Abstract 4769

Background.

Sickle Cell anemia (SCA) remain a disease with high risk of morbidity and early death, especially in African patients. Allogeneic haematopoietic stem cell transplantation (HSCT) is the only curative treatment for SCA. To analyze immunohematological reconstitution after transplant, we report our experience concerning 12 geno-identical HSCT for SCA-patients prepared with the same myeloablative conditioning regiment consisting of Busulfan and Cyclophosphamide.

Patients and Methods.

Twelve patients with a median age of 12 years (range, 2–16), affected by sickle cell anemia (SCA), received hematopoietic stem cell transplantations from HLA-identical, related donors following a myeloablative conditioning regimen. To analyze the mechanisms involved in immunological reconstitution post transplant, we analyzed T cell subsets by flow cytometry at + 60 post transplant.

Results.

All patients had sustained engraftment and remained free of any SCA-related events after transplantation. Sixty days after the transplant, the patients had significantly lower CD4+ T cells in comparison to the controls (15.6 ± 5.9 % vs. 47.5 ± 6% respectively), whereas CD8+ T cells were the first lymphocytes to repopulate the peripheral blood with up to 45% of these cells being CD8+ T cells (in mean 48.5 ± 14.3 % vs. 20 ± 7%). All patients displayed reduced numbers of B cells versus normal value, and 10/12 patients had only 0% to 1% of control levels of CD19+ cells. CD3-CD56+bright NK cells were 18.6 ± 12.2 %, whereas CD3-CD16+ (with cytotoxic functions) were 16.7 ± 11.8 %.

Conclusion.

Our primary finding include the following: 1) rapid increase of lymphocytes in peripheral blood after transplant; 2) rapid expansion of CD8+ T cell but not CD4 T cell counts. Probably reactivation of cytomegalovirus (CMV) infection, observed in 11/12 patients on the early stages of T-lymphocyte recovery after transplant, might induce a dramatic increase in CD8 but not in CD4 T-cell counts.

Disclosures:

No relevant conflicts of interest to declare.
The Revised High-Dose Regimen of Beacop in the Primary Treatment of the Advanced Hodgkin's Lymphoma: A Interim Report
Authors: LIU Peng , Shi Yuankai , Wang Zhao
Published: October 2019

Abstract 4783

Objective

In the study, we compared the efficacy between the revised high-dose and standard-dose of the revised BEACOP regimen (Bleomycin, Etoposide, Epidoxorubicin, Cyclophophamide, Vincristine, Prednisione) in the treatment of the advanced Hodgkin's lymphoma (HL).

Patients and methods

Patients with HL of Stage III/IV were 1:1 randomized into either the standard-dose BEACOP regimen (the same to the BEACOPP regimen in GHSG-HD9 study except procarbazine) in the control arm or the revised high-dose BEACOP regimen (different from BEACOPPesc regimen in GHSG-HD9 study, excluding procarbazine, increasing the dose of cyclophophamide and doxorubicin, with the standard-dose of etoposide, and without the preventive G-CSF support) in the experimental arm. Patients with large tumor or residual tumor after 6 to 8 cycles chemotherapy would be received local radiotherapy. All patients were evaluated by imaging examination every 2 cycles during treatment and were followed up every 3 months from the end of the treatment.

Results

33 cases with advanced HL completed whole cycles chemotherapy and were followed-up a medium time of 12 months. The interim analysis showed that, patients received revised high-dose BEACOP had better response than those with standard-dose BEACOP. After 2 cycles chemotherapy, 2/17 cases had complete response (CR), and 8/17 cases had reduced more than 75% in the tumor size in the experimental arm; while none of 16 cases in the control arm achieved the similar response. After 6 cycles chemotherapy, 13/17 cases in the experimental arm achieved CR/CRu, compared with 8/16 cases in the control arm. Simultaneously, adverse drug reactions, such as neutropenia, nausea and vomiting, were observed significantly increased in the experimental arm. However, no related death and SAE occurred. All the patients showed good tolerance in the study.

Conclusions

In the HD9 study, asian patients received with BEACOPPesc were not tolerated enough to complete the whole chemotherapy, because of the serious toxicity such as long-term bone marrow suppression. In our study, we demonstrated that revised high-dose BEACOP in the primary treatment of the advanced HL would achieved the more high rate of CR/CRu compared with standard-dose BEACOP, and patients with revised high-dose BEACOP showed good tolerance and controllable acute hematotoxicity, even without the preventive use of G-CSF. Therefore, the revised high-dose BEACOP regimen might be a potential choice for Asian patients with advanced HL. However, more cases and long-term follow-up should be needed in the future to evaluate the long-term overall survival (OS) rate, efficacy and toxicity.

Disclosures:

LIU: Research Project of Cancer Hospital, CAMS: Research Funding.
Prognostic Significance of Minimal Residual Disease in Adult Patients with B-Lineage Acute Lymphoblastic Leukemia by 8-Color Flow Cytometry
Authors: Weng Xiang-Qin , Shen Yang , Sheng Yan , Chen Bing , Wang Jing-han , Li Jun-ming , Zhu Yong-Mei , Chen Sai-Juan
Published: October 2019

Abstract 4789

Monitoring of minimal residual disease (MRD) in patients with acute lymphoblastic leukemia (ALL) by immunophenotyping and/or molecular techniques provides a way to precisely evaluate early treatment response and predict relapse. In this study, we have investigated the prognostic significance of MRD in adult patients with B-lineage acute lymphoblastic leukemia (B-ALL) by 8-color flow cytometry. A cohort of 106 patients with B-ALL who had achieved a complete remission (CR) and at least 1 LAIP characteristics were enrolled to perform MRD assessment at the end of induction and 1 cycle of consolidation. LAIPs were identifiable in 96% of the patients by 8-color flow cytometric assay, in which, most cases (90.6%) containing 2 or more LAIPs had a sensitivity as high as identifying 1 leukemic blast among 1×105 BM nucleated cells. MRD negative status could clearly predict a favorable 1 year relapse free survival (RFS) and 2 year overall survival (OS) when a cut-off level of 0.01% was used to define MRD positivity at the point of achieving CR (P=0.000 and 0.000, respectively) and after 1 cycle of consolidation (P=0.000 and 0.000, respectively), respectively. In multivariate analysis including cytogenetic abnormalities, clinical factors and MRD status, late CR (P=0.046), MRD status at the points of obtaining CR (P=0.016) and 1 consolidation (P=0.007) were associated with RFS independently, while only MRD status after 1 course of consolidation was independent prognostic factor for OS (P=0.000). Of note, in exploring the fewer patients with MRD negative status experienced recent relapse, we have identified that most of such patients had a MRD level of 10−4−10−5 comparing to undetectable MRD level. Furthermore, our evidences showed that MRD assessed by flow cytometry and by RQ-PCR assay targeting to BCR-ABL fusion gene yielded concordant results in the vast majority of cases (90%). In conclusion, immunophenotypic evaluation of MRD by 8-color flow cytometry could work as an important tool to assess the treatment response and prognosis precisely in adult B-ALL.

Disclosures:

No relevant conflicts of interest to declare.
A Novel Non-Genetic Photostable Approach to Labelling Acute Myeloid Leukemia and Bone Marrow Cells with Quantum Dots
Authors: Zheng Yanwen , Mao Zhengwei , Yin Bin
Published: October 2019

Abstract 4818

Acute myeloid leukemia (AML) is a detrimental disease with difficult diagnosis and treatment. Understanding the biology of AML at the molecular and cellular levels would be essential to successful management of the disease. However, the notoriously known difficulty in manipulation of leukemia cells has long hindered the dissection of AML pathogenesis. The advent of CdSe/ZnS quantum dots (QDs) represents an important advancement in the research field of nanotechnology, which have recently also been applied for imaging of live cells. Here, we have introduced a non-genetic approach of marking blood cells, by taking advantage of QD technology. We compared QDs complexed with different vehicles, including a peptide Tat (QDs-Tat), cationic polymer Turbofect (QDs-Tf) and liposome Lipofectamine 2000 (QDs-Lip), in their abilities to mark cells. QDs-Tat showed the highest efficiency in delivery into hematopoietic cells, among the three vehicles. We then examined QDs-Tat labelling of leukemia cell lines, and found that QDs-Tat could label 293T, bone marrow (BM) cells, THP-1, MEG-01 and HL-60 with a decreasing efficiency. The efficiency of QDs-Tat delivery was dependent on the concentration of QDs-Tat applied, but not the length of incubation time. In addition, more uniform intracellular distributions of QDs in 293T and leukemia cells were obtained with QDs-Tat, compared with the granule-like formation obtained with QDs-Lip. Clearly, QD fluorescence was sharp and tolerant to repetitive photo excitations, and could be detected in 293T for up to one week following labelling. In summary, our results suggest that QDs have provided a photostable, non-genetic and transient approach that labels normal and malignant hematopoietic cells in a cell type-, vehicle-, and QD concentration-dependent manner. We expect for potentially wide applications of QDs as an easy and fast tool assisting investigations of various types of blood cells in the near future.

Disclosures:

No relevant conflicts of interest to declare.
Peripheral Regulatory B Cell Phenotype in Multiple Sclerosis Patients
Authors: Habib Jakob , Deng Jiusheng , Pennati Andrea , Lava Neil , Galipeau Jacques
Published: October 2019

Abstract 4843

Regulatory B cells (Bregs) are a unique CD5+/CD1d+/IL10+ B cell subtype which has novel immunosuppressive capabilities. It remains to be determined if subjects suffering from autoimmune ailments such as multiple sclerosis (MS) display a defect in number or function of Bregs in association with their disease. Our hypothesis is that total B cell number, and Breg subset in particular, is distinct when comparing peripheral blood B cell populations from MS to healthy controls (HC). This study herein focuses on establishing a phenotypic baseline of the peripheral B cell profile. The total number of CD19+ B cells in HC is 892±816 cells/μL (average ±SD, n=28), of which CD5+/CD1d+ Breg subset is 4.6±5.6 cells/μL. We found a greater number of B cells in MS subjects, 1441±1011 cells/μL (n=22, p<0.05, Student's t-test). However, the number of Bregs in MS patients is not significantly different from HC, 7.5±7.6 cells/μL (p=0.13). We further interrogated the phenotype of B cells in MS subjects and found that there is an increased number of CD27−IgD+ naïve/intermediate phenotype in MS, 1041±794 cells/μL, versus HC, 624±606 cells/μL (p<0.05). Our findings demonstrate that MS is not associated with a deficiency in the absolute number of CD5+/CD1d+ Bregs but rather a 60% increase in CD27−IgD+ B cells. We hypothesize that altered B cell distribution may be functionally associated to the underlying immune disease process of MS. This finding may inform the design of clinical trials examining B cell depletion strategies for treatment of MS.

Disclosures:

No relevant conflicts of interest to declare.
Clinical Features, Treatment and Prognosis of Extranodal NK/T Cell Lymphoma, Nasal Type: A Retrospective Analysis of 88 Cases From a Single Institution in China
Authors: Xuemei Wang , Xu Juan , JIngjing Wen , Zhibin Liu
Published: October 2019

Abstract 4848

Background and Objective:

Extranodal NK/T Cell Lymphoma, nasal type (NKTCL), a rare malignancy in western countries, behaves distinct clinicopathologic features. Patients are generally refractory to conventional chemotherapy. No standard therapy regimen has been established yet. Prognosis of this highly aggressive disease is usually dismal. As it is relatively prevalent in the southwest China, the analysis will provide useful information on the clinical characteristics, treatment options and survival.

Methods:

Hospitalized patients newly diagnosed as NKTCL according to the WHO classification in our institution from January 2000 to December 2010 were surveyed. 88 cases were retrospectively analyzed. Patients were treated by different regimens: chemotherapy-alone, radiotherapy-alone or radio-chemotherapy. All patients had thorough baseline and follow-up assessments. The median observation time was 28 months (range 3∼96).

Results:

Conclusions:

NKTCL occurs most often in young or middle aged adults with a male predominance and has a strong association with Epstein-Barr virus (EBV) infection. Patients with advanced stage disease usually have a poor prognosis. Better response and higher survival chances are expected by radiotherapy or combined radio-chemotherapy.

Disclosures:

No relevant conflicts of interest to declare.
Predictors Of Survival Outcomes In Phase I Relapsed Or Refractory Multiple Myeloma Patients
Authors: Harvey R. Donald , Barbee Meagan S. , Nooka Ajay K. , Kim Sungjin , Chen Zhengjia , Heffner Leonard T. , Kaufman Jonathan L. , Lonial Sagar
Published: October 2019

Objectives

Categorization of response criteria for multiple myeloma (MM) is based on magnitude of change in serum and urine paraprotein values and normalization of free light chain ratio (rFLC). However, the association between improvements in these surrogate markers and patient outcomes is not validated in the phase I setting. Early measures of response would be beneficial for patients and agents to identify those likely to have prolonged disease-free intervals and to validate agent activity for rapid movement to subsequent development.

Methods

We identified 31 trials that met enrollment criteria of phase I, relapsed or refractory disease, and non-transplant study population. Clinical and demographic data collected included age, sex, race, ECOG performance status (PS) at entry, myeloma subtype and isotype, prior therapy, cytogenetics at study entry, date of progression, and date of expiration. Patients with t(4;14), del13, del17p, t(14;16), or t(14;20) were considered to have non-standard risk cytogenetics. Evaluation of the relationship between progression free survival (PFS) and change in plasma cell activity by the rFLC and magnitude of response in serum/urine paraprotein per IMWG criteria was performed. Landmark analyses occurred at cycle 2 and 4, 8, and 12 months. Progression free survival (PFS) at 12 months was the primary outcome of interest.

Results

Among 87 patients; 47 (54%) were female; 56 (64%) white, 29 (33%) black; 27 (31%) non-standard risk cytogenetics; ECOG PS 1 in 73 (84%); and median prior lines 5 (1-11). Eighty were evaluable for paraprotein changes, 71 for rFLC. Normalization of rFLC at 4 months conferred a PFS advantage (11.3 v 2.8 months, p = 0.038) (Table 1). Normalization of rFLC by 12 months was found to predict PFS (6.1 vs. 2.8 months, p = 0.015) and a longer OS (45 vs. 17.4 months, p = 0.002). Magnitude of response in paraprotein was found to predict and correlate linearly with PFS at all time landmarks (r2 = 0.769 to 0.952). Analysis of PFS by IMWG criteria and by quartiles of 50% changes were both linear (p < 0.001) (Figure 1).

Conclusion

These findings suggest that normalization of the rFLC and magnitude of paraprotein response are viable surrogate disease endpoints in phase I clinical trials of novel agents and combinations. The use of current IMWG criteria in the phase I setting is valuable, but the addition of time of response and alterations of response boundaries should be further evaluated in the setting of improved treatments.

Disclosures:

Kaufman: Onyx: Consultancy; Celgene: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Janssen: Consultancy; Millenium: Consultancy; Merck: Research Funding. Lonial:Millennium: Consultancy; Celgene: Consultancy; Novartis: Consultancy; BMS: Consultancy; Sanofi: Consultancy; Onyx: Consultancy.
Recovery Of Natural Killer Cells and Monocyte Subsets Following Autologous Peripheral Blood Stem Cell Transplantation Predicts Longer Progression Free Survival Among Multiple Myeloma Patients
Authors: Callander Natalie S , Rathouz Paul J , Asimakopoulos Fotis , Juckett Mark B , Coe Christopher L , Sheerar Dagna , Ersland Karen , Hematti Peiman , Costanzo Erin S
Published: October 2019

Prior research has shown that a timely immune recovery following peripheral blood stem cell transplantation (PBSCT) may improve progression free and overall survival for lymphoma and leukemia patients (pts). However, this has not been studied in multiple myeloma (MM) pts. We conducted an exploratory investigation of the extent to which the recovery of Natural Killer (NK) cell and monocyte (Mo) subsets predicted progression free survival (PFS) among MM pts following autologous PBSCT. Patients (N = 37) were treated with 200mg/m2 of melphalan, followed by infusion of >2 x106 CD34 cells/kg, and were followed prospectively for 3 to 36 months. Immunophenotyping and multicolor flow cytometry were used to identify NK and Mo subsets at Day 100 post-PBSCT. Disease progression or relapse was defined using IMWG criteria. Cox proportional hazard regression models were used to examine relationships between cell subpopulation counts and time from PBSCT to MM progression. All models were adjusted for ISS stage and risk (high, defined as plasma cell leukemia; loss of p53, myc rearrangement, t(14;16), t(14:20) by FISH; minus 13 by cytogenetics; versus standard) at diagnosis, and hazard ratios were standardized to correspond to a 2 standard deviation difference in the logarithm of NK or Mo cell counts. Compared to a healthy reference sample, MM patients showed good recovery of all Mo subsets with normal counts by Day 100, but CD56+CD3- NK cell numbers remained low, with the subset of NK cells expressing high CD16 particularly suppressed. During the follow-up period, 25 patients (67.6%) progressed or relapsed. Pts with higher Day 100 CD56+CD16hi NK cell counts had a longer PFS (hazard ratio = .351, p = .033). A similar effect was seen for CD56+CD16lo NK cells (hazard ratio = .321, p = .079). Higher CD14+ Mo counts were also associated with longer PFS (hazard ratio = .230, p = .007). An examination of Mo subpopulations showed that higher numbers of Mo expressing low or no CD16 predicted a longer time to disease progression (hazard ratio = .207, p = .011), but the inflammatory subsets with moderate or high CD16 expression were not significantly associated with PFS. Results suggest that MM patients who have lower NK cell and CD14+ Mo counts may be at risk for a poorer outcome following autologous PBSCT, with these subsets adding prognostic significance above and beyond cytogenetic risk and ISS stage. Subpopulations with greater regulatory and less inflammatory function had the most salient effects. These NK cell and Mo subsets may inhibit residual MM growth and progression and could serve as novel markers of the clinical outcome following PBSCT for MM.

Disclosures:

No relevant conflicts of interest to declare.
A Comparative Study Of The Outcome Of Isolated Chromosome 13q and Clonal Progression Detected By I-FISH Do Additional Cytogenetic Abnormalities Impact Survival In Multiple Myeloma
Authors: Peres Edward , Farhan Shatha , Kuriakose Philip , Michalowski Susan , Sitarik Alexandra , Janakiraman Suseela M , Jessica Sanchez , Janakiraman Nalini
Published: October 2019

Background

Chromosomal abnormalities detected by interphase fluorescence in situ hybridization (I-FISH) are an important prognostic marker in patients with multiple myeloma (MM). Isolated chromosome 13q has been considered standard risk when identified by I-FISH and high risk by conventional cytogenetics. The impact of additional cytogenetic abnormalities with chromosome 13q identified by I-FISH in regards to prognosis has not been fully defined. In this report, we describe the outcome of patient’s with multiple myeloma with isolated chromosome 13q and 13q+ (additional cytogenetic abnormalities) identified by I-FISH at our institution between January 2003 and January 2013 and had I-FISH analysis prior to treatment.

Methods

The primary objective was to compare patient’s outcomes in regards to response, time to progression, and overall survival between patients who had an isolated 13q and 13q+ identified by I-FISH in the bone marrow plasma cells. Kaplan & Meier curves were generated to calculate overall survival (OS) between the two groups.

Results

Between January 2003 and January 2013, we identified 76 patients by I-FISH who had either an isolated 13q or 13q+ in patients with multiple myeloma (Patient characteristics Table 1). Of the patients with an isolated 13q abnormality 33% received a bortezomib-based regimen and 38% in the 13q+ group. Of the patient’s with a isolated 13q 38% went onto receive high dose chemotherapy followed by autologous stem cell transplant (ASCT) while 20% with a 13q+ received ASCT. African American patients with 13q consisted of 65% and 60% with 13q+ in our patient population. For the 13q or 13q+ who underwent high dose chemotherapy followed by autologous stem cell transplant OS was 85% compared to the non-transplant group 45% (p=0.01) (Figure 2). On follow up at a median of 2.5 years mortality occurred in 31% of the 13q patients compared to 62% in the 13q+ group. The overall survival at 5 years was 25% in the 13q+ group compared to 65% in the patient’s with an isolated 13q, With the 13q+ group having an overall poor OS (p=0.03)

Conclusion

Patients who harbor the 13q and additional cytogenetic abnormalities identified by I-FISH have a significant worse outcome compared to patients with an isolated 13q. These patients should be considered high risk and consideration for treatment with novel agents and autologous stem cell transplant followed by post-transplant maintenance therapy should be considered.

Disclosures:

No relevant conflicts of interest to declare.
Chromosome Instability Underlies Stem Cell Dysfunction and Neoplasia In Widespread Hematologic Disease Induced By Deregulated Cyclin E
Authors: Siu Ka Tat , Xu Yanfei , Bhattacharyya Mitra , Gurbuxani Sandeep , Hua Youjia , Minella Alexander C.
Published: October 2019

The Fbw7 ubiquitin ligase controls the expression of a number of oncoprotein substrates including cyclin E, Notch, c-Jun, and c-Myc. Using a knock-in mouse model (cyclin ET74AT393A), in which mutations were introduced into the cyclin E1 allele (Ccne1) to disrupt Fbw7-mediated ubiquitination specifically, we previously found that cyclin E dysregulation in vivo induces anemia with defects in erythroid differentiation and morphologic dysplasia of erythroid progenitors. We also found that cyclin ET74AT393A mice have fewer hematopoietic stem cells (HSCs) during steady-state hematopoiesis compared to wild-type counterparts. We performed serial transplantation experiments to assay comprehensively the self-renewal and multi-lineage reconstitution capacities of cyclin ET74AT393A HSCs. Contrary to our expectations, cyclin ET74AT393A HSC self-renewal appears normal after three rounds of serial transplantation; however, we identified defects in their multi-lineage reconstitution function. In cyclin ET74A T393A bone marrow erythroid cells, induction of a p53-dependent DNA damage response pathway appears to promote compensated erythropoiesis. In cyclin ET74A T393A HSCs, we similarly observed induced expression of canonical p53 target genes. We studied the effect of p53-loss on cyclin ET74A T393A HSCs and found that p53-null; cyclin ET74A T393A HSCs exhibit defects in both self-renewal and multi-lineage reconstitution. By enumerating chromosomes in metaphase spreads, we found p53-null; cyclin ET74A T393A hematopoietic stem and progenitor cells (HSPCs) demonstrate significant chromosomal instability (CIN). Importantly, we can recapitulate the self-renewal defects and CIN of cyclin ET74A T393A HSPCs with intact p53 by treating recipient animals with a single dose of 5-fluorouracil (5-FU). Thus, chromosomal stability is a key determinant for the maintenance of HSC self-renewal, and hematologic stress appears to unmask the potential for impaired Fbw7-dependent cyclin E ubiquitination to engender CIN in the presence of intact p53. Moreover, CIN is a characteristic feature of fatal T-cell malignancies that ultimately develop in recipients of cyclin ET74A T393A; p53-null HSCs. In pre-malignant thymocytes isolated from recipients of cyclin ET74A T393A; p53-null HSCs, aneuploidy is associated with the marked potentiation of cyclin E kinase activity in these cells by p53-loss. In malignant thymocytes, comparative genome hybridization analysis demonstrates clonal CIN associated with deregulated cyclin E expression combined with p53-loss. In toto, our data demonstrate the functional importance of cyclin E regulation by the Fbw7 ubiquitin ligase to the hematopoietic system and highlight CIN as a key mechanism underlying HSC dysfunction and malignancy induced by deregulated cyclin E in vivo.

Disclosures:

No relevant conflicts of interest to declare.
Hydroxyurea Therapy Reduces The Hypercoagulability State In a Sickle Cell Mouse Model
Authors: Colella Marina Pereira , Almeida Camila Bononi , Conran Nicola , Pallis Flavia Rubia , Franco-Penteado Carla Fernanda , Annichino-Bizzacchi Joyce Maria , Costa Fernando Ferreira , Vinicius de Paula Erich , Olalla-Saad Sara Terezinha , Traina Fabiola
Published: October 2019

Several lines of evidence show that sickle cell anemia (SCA) is characterized by a hypercoagulable state. SCA patients present an elevated rate of thrombotic complications and increased biological markers of coagulation activation. In a previous study we demonstrated that therapy with hydroxyurea (HU) in SCA patients was associated with reductions in hypercoagulability markers, in addition to reductions in hemolysis, inflammation and endothelial activation markers (Colella et al Journal of Thrombosis and Haemostasis 2012). In the present study, we attempted to further investigate whether the effect of HU on hemostatic activation is dependent on fetal haemoglobin (HbF) expression, hemolysis and/or inflammation. To do so, we used the BERK murine model of SCA, a homozygous model of SCA, incapable of expressing HbF. In this BERK model, treatment with HU results in no improvement in hemolysis, due to the lack of HbF induction (Lebensburger et al Haematologica 2010). The murine SCA model was generated by transplantation of nucleated bone marrow cells from BERK mice into lethally-irradiated C57BL6 mice. Only mice expressing > 97% of human hemoglobin S (HbS) at 10 weeks after transplantation were used in the study. HU therapy at the dose of 50mg/kg (HU group) or saline alone (control mice) was administered through intraperitoneal injections 5 times per week, initiated at 11 weeks after transplantation (approximate time of sickle marrow engraftment). After a treatment period of 16 weeks, blood samples drawn from the inferior cava vein were collected and submitted to evaluation of plasma levels of thrombin-antithrombin III (TAT), a final marker of thrombin generation, in both animal groups. We also evaluated plasma levels of the inflammation marker interleukin 6 (IL6), and the endothelial marker soluble vascular cell adhesion molecule-1 (sVCAM-1). TAT, IL6 and sVCAM-1 were all measured by commercially-available ELISA kits. Statistical analyses were performed using Mann-Whitney’s U test. Treatment with HU resulted in a significant reduction of TAT plasma levels (106.7 μg/L vs 138.5 μg/L; P = 0.05). Plasma levels of IL6 (7.1pg/mL vs 12.4pg/mL; P = 0.18) and sVCAM-1 (765.8 ng/mL vs 848.4 ng/mL; P= 0.43) presented non-significant reductions with HU treatment. Our results show that, in this murine SCA model, long-term HU treatment results in an improvement in the hypercoagulability state, even in the absence of HbF induction, or of a significant improvement of inflammation or endothelial activation. This indicates that HU may have a direct effect on inhibiting the activation of coagulation in SCA.

Disclosures:

No relevant conflicts of interest to declare.
Neurologic Complications In Pediatric Sickle Cell Patients Hospitalized For Priapism: An Analysis Of The Pediatric Health Information System (PHIS) Database
Authors: Jones Gary , Lee Brian R , Woods Gerald M , Dalal Jignesh , Kalpatthi Ram
Published: October 2019

Background

Priapism is a known complication of sickle cell disease (SCD). Though, blood transfusion is often used in the management of SCD-associated priapism (SAP), its safety and efficacy is not known. Transfusion has been associated with neurologic events including ASPEN syndrome (Association of Sickle cell disease, Priapism, Exchange transfusion, and Neurologic events) [Siegel et al. J Urol 1993]. A recent systematic review suggests that transfusion does not appear to be effective in SAP as determined by time to detumescense (Merritt et al. Can J Emerg Med 2006). Our objective is to examine the relationship between blood transfusions and neurologic complications (NCs) in the management of SAP.

Methods

We reviewed the data from 33 children’s hospitals within the Pediatric Health Information System (PHIS) database from 2000-2011. Patients ≤ 21 years of age with SCD were included for the analysis. SCD, priapism and all related conditions were identified by ICD-9 codes. We abstracted data on demographics, hospitalizations, transfusion, urologic procedures and neurologic changes (including stroke, seizure, headache, weakness, transient-ischemic attacks (TIAs), intracranial hemorrhage, and mental status changes). Fisher’s exact test was used to compare the prevalence of NCs. A p-value <.05 was considered statistically significant.

Results

From the year 2000 to 2011, a total of 7,929 unique male pediatric patients with SCD were identified. Among these 465 (5.9%) patients with 1,069 hospitalizations were identified as having SAP. The majority of these patients were HbSS (n=447; 96%). The frequency of SAP hospitalizations at various age groups were; 4.7% (0-2 years of age), 11.2% (3-5 years), 38% (6-12 years) and 46% (13-21 years). In the majority of SAP hospitalizations (63%, n=673) neither transfusions nor urologic procedures were performed, whereas blood transfusions, urologic procedures or both were performed in 25.2% (n=269), 6.6% (n=71) and 5.2% (n=56) of SAP hospitalizations respectively. There was no significant difference in the incidence of NCs between the patients who received transfusions and those who did not (4.1% vs. 4.7%, p = NS). Both bivariate (Table 1) and multivariate regression analysis (Table 2) revealed no statistically significant associations between NCs and blood transfusion, urologic procedure, age at hospitalization for priapism, or other SCD co-morbidities (VOC and ACS).

Conclusion

In our large pediatric sickle cell cohort, NCs associated with priapism were rare. Moreover, there was no significant association between the neurologic complications and blood transfusions or urologic procedures in SAP hospitalizations. Further studies are needed to understand the mechanism behind these neurologic complications and any associated risk factors.

Disclosures:

No relevant conflicts of interest to declare.
Aln-TMP: A Subcutaneously Administered RNAi Therapeutic Targeting Tmprss6 For The Treatment Of β-Thalassemia
Authors: Butler James , Fishman Shannon , Racie Tim , Hettinger Julia , Bettencourt Brian R , Charisse Klaus , Fitzgerald Kevin
Published: October 2019

The b-Thalassemias are a group of hereditary blood disorders resulting from insufficient beta globin production, ultimately giving rise to the signature clinical sequelae associated with β-Thalassemia which includes anemia, ineffective erythropoiesis, and secondary iron overload. Previously, we demonstrated that intravenous administration of an siRNA targeting hepatic Tmprss6 expression significantly ameliorated the disease phenotype in the Hbbth3/+ mouse model of Thalassemia Intermedia (Blood. 2013; 14;121(7):1200-8). The Tmprss6 gene encodes for the protein Matriptase-2 which negatively regulates Hepcidin gene expression by cleaving the Hepcidin regulatory protein Hemojuvelin; RNAi-mediated suppression of Tmprss6 removes this negative regulator, ultimately leading to an increase in Hepcidin expression. Increased Hepcidin expression leads to a significant decrease in serum iron concentration and Transferrin Saturation (TfSat), which in the β-Thalassemia disease setting, corrects ineffective erythropoiesis, ameliorates anemia, and mitigates secondary iron overload. The role of Tmprss6 in iron metabolism has been extensively characterized in animal and human studies and, together with the observation in the Hbbth3/+ mouse, represents an attractive therapeutic target for the treatment of β-Thalassemia.

To this end, we developed ALN-TMP, a subcutaneous RNAi therapeutic targeting hepatic Tmprss6 for the treatment of β-Thalassemia. ALN-TMP employs the GalNAc conjugate siRNA delivery platform that safely and effectively delivers siRNA to the liver for hepatic gene silencing. Preclinical animal data demonstrate ALN-TMP exhibits robust and durable dose-dependent gene suppression as single dose administration of ALN-TMP leads to > 80% Tmprss6 gene suppression for up to 3 weeks post-dose. This leads to concomitant increases in Hepcidin gene expression (>2x baseline levels) and subsequent decreases in total serum iron and TfSat (>50% decrease from baseline). The degree to which ALN-TMP modulates Hepcidin and serum iron mobilization is nearly identical to that observed in the previous Hbbth3/+ mouse studies and suggests ALN-TMP is a potent RNAi therapeutic with the potential of producing disease modifying effects in β-Thalassemia.

Disclosures:

Butler: Alnylam: Employment. Fishman:Alnylam: Employment. Racie:Alnylam Pharmaceutical, Inc: Employment. Hettinger:Alnylam Pharmaceuticals: Employment. Bettencourt:Alnylam Pharmaceuticals: Employment. Charisse:Alnylam Pharmaceuticals: Employment. Fitzgerald:Alnylam: Employment.