bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA-Mismatched Stem Cell Microtransplantation Compared to Matched-Sibling Donor Transplantation for Acute Myeloid Leukemia in First Complete Remission
Authors: Liu Limin , Qiu Huiying , Tang XiaoWen , Han Yue , Miao Miao , Fu Chengcheng , Jin Zhengming , Zhu Mingqing , Wu Depei
Published: November 2019

HLA-mismatched stem cell microtransplantation is a new transplantation which has been reported in recent years. We treated 41 patients with intermediate- or high-risk acute myelogenous leukemia (AML) in first complete remission (CR1) undergoing HLA-mismatched stem cell microtransplantation and compared with 56 patients undergoing HLA-matched sibling donor (MSD) transplantation at the same period in our center from July 2012 to August 2015. The estimated leukemia-free survival (LFS) at 2-year was 65.6% ± 9.5% in the MSD group and 27.6% ± 14.0% in the microtransplantation group (P = 0.011). The estimated overall survival (OS) at 2-year was 73.6% ± 8.6% in the MSD group and 46.2% ± 15.6% in the microtransplantation group (P = 0.056). The cumulative incidences of relapse were 22.5% and 58.7% among MSD and microtransplantation groups (P = 0.011). The cumulative incidence of nonrelapse mortality was 7.8% in MSD group, and no nonrelapse mortality in microtransplantation group (P = 0.265). Hematopoietic recovery time was shorter in the microtransplantation group than MSD group (P < 0.05). The infection rate of MSD group was higher than in the microtransplantation group (91.1% vs 75.6%, P = 0.037). The preliminary results suggest that LFS of microtransplantation is inferior to MSD transplantation and a trend toward the OS of microtransplantation was lower than MSD transplantation for intermediate- or high-risk AML in CR1.

Disclosures

No relevant conflicts of interest to declare.
A Comparison Study Between Allogeneic and Autologous Hematopoietic Stem Cell Transplant for High-Risk Peripheral T-Cell Lymphomas (PTCL)
Authors: Huang Haiwen , Wang Qiangli , Jin Zhengming , Tang XiaoWen , Qiu Huiying , Fu Chengcheng , Han Yue , Miao Miao , Chang Huirong , Sun Aining , Wu Depei
Published: November 2019

Objective: To evaluate the efficacy of auto-HSCT and allo-HSCT in the treatment of high risk peripheral T cell lymphoma (PTCL).

Methods: From July 2007 to July 2014, 60 cases of high risk PTCL were analyzed retrospectively.

Results: All 60 patients were at high risk group (carried with IPI≥3), with a median age of 31 (13-55) years old. Of the 60 cases, 22 were PTCL-not otherwise specified (PTCL-NOS), 22 with ALK negative anaplastic large cell lymphoma (ALK-negative ALCL) and 16 with angioimmunoblastic T-cell lymphoma (AITL). Twenty-one (21/60) received allo-HSCT and thirty-nine patients (39/60) received auto-HSCT. Before receiving transplantation, 40/60 patients were in complete remission (CR), 2/60 patients were partial remission (PR) and 18/60 patients were not remission (NR). In the 40 CR patients before transplant, 10 patients received allo-HSCT and 30 patients received auto-HSCT. In the 20 PR/NR patients before transplant, 11 patients received allo-HSCT and 9 patients received auto-HSCT. After a median follow-up of 39 (range 1-96) months, the K-M analysis showed that the 5-year PFS for auto-HSCT and allo-HSCT were 61% and 60% (P = 0.724). The 5-year OS for auto-HSCT and allo-HSCT were 62% and 61% (P = 0.724). There were no statistically significant differences between the auto-HSCT and allo-HSCT. And the cumulative TRM of allo-HSCT and auto-HSCT were 16.5% and 0 (P=0.250) within 5-years after transplantation. At the end of the last follow-up, 7 patients relapsed in auto-HSCT group and 2 patients relapsed in allo-HSCT group, the 5-year cumulative recurrence rates of auto-HSCT and allo-HSCT transplantation were 37.2% and 10.1% (P=0.298), respectively.

Conclusion: There was no significant difference in the long-term survival between auto-HSCT and allo-HSCT for high risk PTCL patients. The effect of allo-HSCT may be better for NR patients.

Disclosures

No relevant conflicts of interest to declare.
Evolving Criteria for Donor Selection for Transplants in Acute Myeloid Leukemia
Authors: Wang Yu , Liu qi-Fa , Wu de-Pei , Huang Xiao-jun
Published: November 2019

Background The best donor for a haematopoietic cell transplant in a person with acute myeloid leukaemia (AML) in 1st remission is controversial. Prior data indicate similar outcomes after transplants from HLA-haplotype-matched relatives, HLA-identical siblings and HLA-matched unrelated donors.

Methods We analyzed data from persons with intermediate- or high-risk AML in 1st complete remission (CR1) treated in a prospective, multi-centre study.

Results 602 consecutive subjects received a transplant from an HLA-haplotype-matched relative (N=328) or an HLA-identical sibling (N=274). 4 year leukaemia-free survivals were 78% (95% confidence interval [CI], 73, 83%) and 80% (75, 85%; P=0.39). In multivariate analyses younger donor/recipient age (donor<30y-receipient<30y) was associated with less transplant-related-mortality (TRM) (hazard ratio [HR)=0.25; [0.06, 1.00]; P=0.05) whereas female-to-male transplants (HR=1.93; [1.10, 3.38; P=0.02) and donor-recipient ABO major-mismatch transplants (HR=1.95 [1.12, 3.38; P=0.02) were correlated with higher TRM. A risk score was generated based on donor/recipient age, gender and ABO-incompatibility. The scoring system predicted TRM and survival. TRMs were 5% (3, 7%), 15% (10, 20%) and 20% (4, 36%) for subjects with scores of 0-1, 2 and 3 (trend comparison P<0.001). 4 year survivals were 85% (81, 89%), 77% (71, 83%), and 75% (59, 91%; trend comparison P=0.04).

Conclusions In contrast to the lack of impact of HLA-disparity on transplant outcomes, selection of the best donor needs to consider D/R age, sex and ABO-incompatibility combined together. Our data suggest choosing young, male, ABO match/minor-mismatch donors is reasonable. Transplants from older female-to-male and from ABO major-mismatched donors should probably be avoided.

Disclosures

No relevant conflicts of interest to declare.
UCBT Using Myeloablative Conditioning Regimen without ATG Has Similar Survival but Better Life Quality Compared with Upbsct for Adults with Primary Acute Leukemia-a Multicenter Study in China
Authors: Tong Juan , Xuan Li , Sun Yanling , Zheng Changcheng , Liu Huilan , Zhu Xiaoyu , Tang Baolin , Liu Qifa , Lin Dongjun , Sun Zimin
Published: November 2019

Objective

Data regarding the outcome of unrelated cord-blood transplantation (UCBT) with myeloablative conditioning regimen without ATG comparing with unrelated peripheral blood stem cell transplantation (UPBSCT) for adults with primary acute leukemia.

Methods

From Jan 2010 to Dec 2014, 175 consecutive adult patients with primary acute leukemia (AL), who had no HLA-compatible related donors who were eligible for a single UCBT (79 patients) or UPBSCT (96 patients). The two groups all received a TBI/CY2 and BU/CY2 based myeloablative conditioning regimen which plus fludarabine (FLU), etoposide (VP-16) or cytarabine (Ara-c). The GVHD prophylaxis regimens of UCBT were cyclosporine A (CsA) and mycophenolate mofetil (MMF) and UPBSCT were CsA, Methotrexate (MTX) and antithymocyte globulin (ATG).

Results

There was no significant difference for the patients characteristics between the UCBT and UPBSCT group except for UCBT patients were younger. Although the UCBT patients received lower doses of nucleated and CD34 positive cells and more patients were HLA mismatched, the two groups acquired the similar percentage of hematopoiesis engraftment rate (97.4% vs 100%, p=0.497) but the granulocyte and platelet recovery was slower in UCBT group. The incidence of bacterial or invasive fungal infection in the UCBT group and the UPBSCT group had no significant difference (46.8% vs 52.6%, P=0.572) may be attributed to the omission of ATG in UCBT group. TRM by three years was 21.0% and 16.9% in the UCBT and UPBSCT cohorts (P=0.46). The 3-year cumulative incidence of relapse was similar between the UCBT group and the UPBSCT group (26.3% vs 27.1%, P=0.75). The grade II to IV acute graft-versus-host disease (aGVHD) and severe aGVHD (grades III and IV) incidence was similar between the two groups (24.7% vs 33.0%, P=0.40), (13.3% vs 8.7%, P=0.28)]. Compared with the UPBSCT cohort, the UCBT cohort had a significantly lower rate of chronic graft-versus-host disease (cGVHD) (20.7% vs 44.0%, p=0.003) or extensive cGVHD (5.6% versus 17.5%, P=0.04). The overall survival (63.2% vs 64.1%, P=0.996) and leukemia free survival (57.5% vs 59.8%, P=0.824) had no significant difference between the two groups. The performance status observed in patients surviving 3 years was measured using the Karnofsky performance score. The UCBT recipients had obviously higher activity scores than UPBSCT recipients (94.5% vs 83.5%, p=0.03).

Conclusion

When use intensive myeloablative conditioning regimen without ATG, UCBT can reach similar survival with UPBSCT and may have better life quality. Primary acute leukemia patients who lack an HLA-matched sibling donor can select UCBT or the UPBSCT of equal status.

Disclosures

No relevant conflicts of interest to declare.
Impact of Disease Status on Outcomes of Allogeneic Hematopoietic Stem Cell Transplantation with Refractory and Relapsed Acute Lymphoblastic Leukemia
Authors: Cao Jing , Zhu Xiaoming , Sun Aining , Qiu Huiying , Jin Zhengming , Miao Miao , Wu Depei , Tang XiaoWen
Published: November 2019

Objective: To evaluate the impact of disease status on the outcomes of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of patients with refractory and relapsed acute lymphoblastic leukemia(ALL).

Methods: 52 patients with refractory and relapsed ALL, including 19 cases in advanced stage (nonremission, NR) and 33 cases in more than or equal to second complete remission(≥CR2), received allo-HSCT after myeloablative conditioning regimen in our department.

Results: 51 patients engrafted successfully. The transplantation-related mortality (TRM) rate of NR and ≥CR2 was 10.5% vs 12.1% (P=0.815). The incidence of aGVHD was 52.6% vs 57.6% (P=0.730), including 42.1% vs 33.3% (P=0.527) with mild (grade I-II) and 10.5% vs 24.3% (P=0.399) with severe (grade III-IV) aGVHD. The incidence of cGVHD was similar also(41.6% vs 57.9% ,P=0.660). With a median follow-up of 12(1.8-44.5) months, the cumulative relapse rate of NR and ≥CR2 was 47% vs 34.3% (P=0.425) respectively. The estimated 2 year overall survival (OS) and 2 year leukemia-free survival (LFS) rate were 42.6% vs 45.7% (P=0.487) and 46.3% vs 46.2% (P=0.571) respectively. Multivariate Analysis results showed that cGVHD was independent favorable risk factor for OS and LFS of R/R ALL. For relapsed patients, OS was significantly better with first CR duration>6 month and time to transplant≤2 months.

Conclusion: AlIo-HSCT is an effective salvage treatment option for patients with refractory and relapsed ALL. Our retrospective analysis showed that R/R ALL with different status prior transplant had similar outcome post transplantation.

Disclosures

No relevant conflicts of interest to declare.
HLA-Haploidentical Donors Seem to be Not Inferior to HLA-Identical Sibling Donors for Hematopoietic Stem Cell Transplantation of Patients with Paroxysmal Nocturnal Hemoglobinuria (PNH)?
Authors: Chen Feng , Wu Depei , Tang XiaoWen , Miao Miao , Fu Chengcheng , Qiu Huiying , Xue Shengli , Ma Xiao , Jin Zhengming
Published: November 2019

Background PNH is an acquired clonal disorder of the hemopoietic stem cells for which the only curative treatment is allogeneic hematopietic stem cell transplantation (allo-HSCT). But allo-HSCT is challenging for those who have no HLA-matched donors.Several recent studies have shown that haploidentical HSCT for patients with hematological malignancy can achieved comparable outcomes with HLA-identical sibling transplantation . There are very few reports on the use of haploidentical HSCT for PNH . Is haploidentical HSCT a valid alternative option for patients with PNH?

Methods 19 PNH patients received allo-HSCT between Dec 2007 and Oct 2015 at our institution. 12 donors were HLA-haploidentical and 7 were HLA-matched siblings. The patients were aged 8 to 54 years (median 28 years) . Of the 12 haploidentical donors, 6 were siblings, 2 fathers,2 mothers,1 son and 1 daughter. 12 patients with haploidentical donors received a myeloablative conditioning regimen consisting of busulfan, cyclophosphamide and ATG (anti-thymocyte globulin), 7 patients with identical siblings were given a reduced intensity conditioning. G-CSF-mobilized bone marrow and peripheral blood stem cells were transplanted as graft. Prophylaxis for graft-versus-host disease (GVHD) consisting of cyclosporine or tacrolimus + short-term methotrexate + mycophenolate mofetil was used for 12 patients with haploidentical donors, and cyclosporine + short-term methotrexate for 7 with identical siblings.

Results All 19 patients were engrafted successfully. The median time of neutrophils (ANC) reached to 0.5×109/L and platelets (PLT) reached to 20×109/L was 12 days and 15 days in haploidentical group , and that to identical group was 11 days and 13 days ,respectively. There were 2 patients developed grade Ⅱ acute GVHD in haploidentical group while 1 patients with grade Ⅳ aGVHD in identical group . Limited chronic GVHD was observed in 2/12 patients in haploidentical group and 1/7 patients in identical group. After a median follow-up time of 22.0 (range 4.0-42.0) months, the 3-year OS probability was 77.8±13.9% and 85.7±13.2% for haploidentical and identical group,respectively (P=0.03). 2 patients died of treatment-related mortality in haploidentical group, including severe pulmonary infection (n=1) and transplant-associated thrombotic microangiopathy (n=1) ,and 1 died of severe aGVHD in identical group. No patients were documented to have a recurrence of a PNH clone after HSCT in both groups.

Conclusion This report seemed that long-term outcomes of HLA- haploidentical HSCT in patients with PNH were comparable to that of HLA- matched donor at our institution . Haploidentical HSCT should be considered as a valid alternative option for PNH patients without HLA- matched donors .

Disclosures

No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation for Multiple Myeloma: a Single Center Experience
Authors: Figueiredo Amarilis , Atkins Harold , Kekre Natasha , Kew Andrea , McCurdy Arleigh
Published: November 2019

Introduction

The use of allogeneic stem cell transplantation (Allo-SCT) in patients with multiple myeloma (MM) remains controversial, but it offers prolonged disease free survival in some patients. It is unclear which patients should undergo Allo-SCT, what conditioning regimen should be used, and what the timing of the transplant should be in the course of the disease. Therefore, we sought to contribute our center's experience to the growing body of literature.

Methods

We performed a retrospective observational cohort study of all patients who underwent Allo-SCT for multiple myeloma at our center between January 1, 1992 and May 31, 2016. Categorical variables were compared using Pearson's chi-square test and the Kaplan-Meier method was used for the overall survival curves.

Results

Thirty-four patients underwent Allo-SCT for multiple myeloma and were included in this analysis. The median age was 40 years and 21 (62%) were male. Nineteen patients (56%) underwent Allo-SCT as upfront therapy, 1 (3%) underwent tandem autologous stem cell transplant (auto-SCT) followed by Allo-SCT, and 14 (41%) had salvage Allo-SCT at the time of relapse. Twenty-four (70.5%) patients had a matched related donor, 1 (3%) had a mis-matched related donor, 8 (23.5%) had matched unrelated donor and matching in 1 (3%) was not available. Myeloablative conditioning was given in 18 patients (52.9%) and non-myeloablative conditioning in 13 (38.2%) with 3 (8.8%) missing. The conditioning regimens included: 5 (15%) Flu-Mel, 7(20.6%) Flu-Bu, 13 (38%) Bu-Cy ± TBI, 6 (17.6%) MelVPTBI, and 3 (8.8%) were missing. Median overall survival (OS) for all patients was 72.5 months from diagnosis (figure 1) and 26.5 months from the time of Allo-SCT. For the 19 patients who had upfront Allo-SCT, median OS from diagnosis was 7.4 years compared to 5.3 years for those having salvage Allo-SCT (figure 2). However, in the upfront group 6 (32%) were alive 10 years post Allo-SCT and the survival curve reaches plateau, whereas in the salvage group, no patient was alive after 8 years post Allo-SCT. There was no difference in median survival between myeloablative and non myeloablative conditioning (2.9 versus 1.33 years, p=0.925). There have been 20 deaths in our cohort (59%); 5 (14.7%) from transplant related mortality within 1 year, 9 (26.5%) from disease progression, and 6 (17.6%) transplanted remotely whose cause of death is unknown.

Conclusions

Our data suggest that Allo-SCT offers prolonged disease free survival in some patients. In our small cohort, a greater proportion of patients undergoing upfront Allo-SCT achieved long term survival, raising the possibility that this group of patients may benefit more from Allo-SCT. Further prospective studies are needed to clarify the role of Allo-SCT in MM.

Disclosures

Kew: Celgene: Honoraria. McCurdy:Celgene: Honoraria.
CXCR3 Upregulation Effect of T Lymphocytic Leukemia Cell Invasion in Vitro
Authors: Feng Xiaoqin , Ding Wenjiao , Li Chunfu
Published: November 2019

Objective

The study was to explore chemokine receptor CXCR3 influence on invasion of T lymphocytic leukemia cell.

Methods

Mouse CXCR3 gene was amplified from T vector containing mouse CXCR3 by PCR, and constructed the mouse CXCR3 overexpressing lentivirus carrying GFP & Puromycin (puro). Jurkat cell line was infected by the constructed CXCR3 overexpression lentivirus. Puromycin was added to screene the successfully infected lymphoblastic leukemia cells. CXCR3 expression on Jurkat cells was analyzed by flow cytometre. Constructed CXCR3-LV5-Jurkat cells were added into the Transwell upper chamber covered with Matrigel. Ligand of CXCR3(CXCL10) was added to the lower chamber. Cell number in the upper chamber and lower chamber which used to calculate the invasion rate was counted by flow cytometry respectively.

Results

GFP+ Jurkat cells was observed less than <10% after lentivirus infection 96h. GFP+Jurkat cells achieved to 90% by Puromycin selection. The expression of CXCR3 on CXCR3-LV5-Jurkat cell group was 90% higher than LV5-Jurkat cell group. CXCR3-LV5-Jurkat cell line stably overexpressed CXCR3 was successfully constructed. The invasion rate of CXCR3-LV5-Jurkat cell lines was significantly higher than LV5-Jurkat control group (12.71 %± 1.03% vs 6.82% ± 0.49%, p <0.0001).

Conclusion

Overexpression of CXCR3 on T lymphocytic leukemia cell line (Jurkat cell) can enhance the invasion of leukemia cells in vitro, CXCR3/CXCL10 path way may induce the T lymphoblastic leukemic cell invading the central nervous system.

Disclosures

No relevant conflicts of interest to declare.
TCR-Ligand Dissociation-Rate, a Reliable and Determining Parameter, Identifying Tumor-Reactive CD8 T Cells of High Avidity and Functional Potency
Authors: Rufer Nathalie , Allard Mathilde , Couturaud Barbara , Carretero Laura , Schmidt Julien , Duong Minh Ngoc , Baumgaertner Petra , Gannon Philippe , Speiser Daniel , Hebeisen Michael
Published: November 2019

Background & Aims: Kinetics of T-cell receptor (TCR) binding to peptide-MHC (pMHC) contribute to T-cell activation and various cellular responses. However, due to technical reasons, the characterization of T-cells is usually only done with functional assays, without assessment of TCR-pMHC binding kinetics, thus limiting the information on the overall quality and clinical relevance of TCR-ligand binding properties. With the aim to broadly evaluate possible correlations between T-cell function and TCR binding kinetics in humans, we undertook a large-scale analysis of combined multiple functional readouts (i.e. CD107a degranulation, cytokine production and proliferation) and off-rate measurements by NTAmers (ref. 1) to characterize tumor- and virus-specific CD8 T-cell clones (n=400) isolated from melanoma patients and healthy donors.

Results: Our results show that, within an antigen-specific repertoire, TCR-ligand off-rates accurately predicted single-functional and poly-functional avidities (i.e. ligand potency) as well as co-receptors expression/modulation of CD8 T-cells. Importantly, the TCR-pMHC dissociation-rate represented a constant and highly reliable bio-physical parameter, contrasting to functional avidity which depended on the activation status of the T-cells following re-stimulation. Remarkably, we found that the TCR-ligand off-rate repertoire depended on the antigenic-specificity, as CD8 T-cells specific for the cancer-testis antigen NY-ESO-1157-165 displayed longer TCR-ligand interactions than T-cells specific for the tumor/differentiation antigen Melan-A26-35, and both these tumor-specific TCRs exhibited significantly lower avidities than those specific for persistent herpes virus antigens (CMV/pp65495-503, EBV/BMFL1259-267).

Conclusions: Our data from a large library of human T-cell clones demonstrate that the TCR-ligand dissociation-rate is a stable/reliable and determining parameter of T-cell responsiveness, arguing that it should be systematically assessed for meaningful monitoring of human T-cell responses. Furthermore, our TCR-ligand off-rate comparisons highlighted strong binding differences between non-self/virus- and self/tumor-specific repertoires. Our data from individual clones suggest that the tumor-specific repertoires contains rare but nevertheless promising TCRs with sufficiently slow off-rate and high functional avidity to potentially confer protective immune responses against cancer.

(ref. 1) Hebeisen et al., Cancer Res, 75(10):1983, 2015

Disclosures

No relevant conflicts of interest to declare.
Dissection of Mechanisms of Realgar Intervening Telomere Protein POT1, TRF1, TRF2 Expression to Regulate Telomere Dynamics in THP-1 Cells
Authors: Xu Rui-Rong , Song Nan-Xin , Wu Xiao-Long , Cui Si-Yuan , Wang Zhen-Zhen , Wang Yan , Wang Xin
Published: November 2019

Objective: To observe the mechanisms of realgar intervening telomere protein POT1, TRF1, TRF2 expression to regulate telomere dynamics in THP-1 cells, and elucidate the experimental and theoretical basis for realgar targeted threapy of AML.

Methods: 1.In vitro: (1)Cultured human acute myeloid leukemia THP-1 cells. (2), The cells were incubated in absence or presence of increasing concentration of realgar for 24/48 h, from which determined the IC50. Cell viability was tested by CCK-8 assay. (3)Apoptotic rate and cell cycle distribution were tested by FCM. (4)Each group of POT1, TRF1, TRF2 protein levels were tested by western-blot analysis. (5)Each group of POT1,TRF1,TRF2 mRNA levels were tested by RT-qPCR analysis. 2.In vivo: (1)Established THP-1 model in NOD/SCID mice, and the mouse were treated by realgar. (2)All the mouse were killed by institutionally approved method after 3 weeks. The apoptotic rate and cell cycle distribution of mice spleen cells were examined by FCM. (3)The changes of POT1, TRF1, TRF2 protein levels in organic tissue were detected by IHC.

Results: 1.In vitro: (1)CCK-8 assay showed that after treated by realgar, THP-1 cells growth were inhibited and the cell viability decreased in a dose- and time- dependent manner, IC50 was 0.023μg/mL. Therefore, we choose a medium concentration was 0.015μg/mL (lower than IC50) at 48h. (2)Our study demonstrated that after treatment for 48h, the apoptotic rate and the G1 arrest of these THP-1 cell increased, compared with control group. (3)As shown by western blot analysis, compared with controls, realgar group POT1, TRF1 protein levels significantly increaesed and TRF2 decreased(P<0.01). (4) As shown by by RT-qPCR analysis, POT1, TRF1, TRF2 mRNA levels were consistent with their protein levels. 2.In vivo: (1) The THP-1/NOD-SCID model was established, and survival curve showed that compared with controls, realgar group mouse had longer life span(P<0.01). (2) The results of FCM showed that after treatment of realgar, the rate of THP-1 cell apoptosis and G1 arrest significantly increased(P<0.01). (3) The results of IHC showed that compared with controls, POT1, TRF1 protein levels significantly increased and TRF2 decreased in realgar group mouse(P<0.01).

Conclusion: In vitro and in vivo studies results indicated that realgar could significantly prolong the life span of the THP-1/NOD-SCID mice, inhibit proliferation and induce apoptosis of THP-1 cell, and regulate telomere dynamics through intervening THP-1 cell telomere protein POT1, TRF1, TRF2 expression. Our studies results provided experimental and theoretical basis for realgar targeted threapy of AML.

Disclosures

No relevant conflicts of interest to declare.
Therapeutic Development of Complement C5 Inhibitor CoversinTM with Extended Half-Life Via PASylation®
Authors: Miles Nunn , Skerra Arne
Published: November 2019

The complement C5 inhibitor Coversin is in clinical development for treatment of paroxysmal nocturnal hemoglobinuria (PNH) and has been shown to be safe and effective in phase 1 single dose and multiple dose trials in healthy volunteers. A single phase 2 PNH patient, resistant to the standard of care eculizumab, has been treated with Coversin for a period of more than 7 months and responded well to treatment.

In contrast to eculizumab, which is intravenously administered twice monthly by a health care professional, Coversin will be self-administered as a once daily subcutaneous injection. While this may provide patient convenience compared to intravenous administration, even longer intervals between subcutaneous dosing would enhance that convenience.

To extend Coversin's half-life and explore the potential for subcutaneous administration at longer dosing intervals Coversin has been modified by the addition of a 600 amino acid proline/alanine/serine (PAS) N-terminal fusion tag to generate PAS-Coversin (68kDa). The unstructured and uncharged PAS polypeptide gives Coversin an apparent molecular size of approximately 720kDa, thus retarding kidney filtration and extending the half-life of PAS-Coversin by 52-fold in mice compared to unmodified Coversin. Furthermore, in vitro complement lytic and C5 binding assays indicate PAS-Coversin inhibits complement C5 as potently as unmodified Coversin.

Recent work has shown that PAS-Coversin can be administered to mice by subcutaneous injection and has very high bioavailability when administered via this route. The bioactive protein can be produced in both (commercially scaleable) bacterial and yeast expression systems. The further development of PAS-Coversin for therapeutic purposes with the potential for weekly dosing will be presented.

Disclosures

Miles: Akari Therapeutics plc: Employment, Equity Ownership. Skerra:XL-protein GmbH: Equity Ownership.
Molecular Mechanism Analysis the Apoptosis of Leukemia Cells Induced By CRM1 Selective Inhibitor KPT-330
Authors: Lu Jun , Hu Shaoyan , DU Weiwei , Pan Jian , Ren Junli , Zhiheng Li
Published: November 2019

Objective: To investigate the effect and mechanisms of KPT-330 on cell apoptosis of human leukemia cells. Methods: CCK-8 assay was used to quantify the growth inhibition of cells after exposure to KPT-330 for 24 hours. Fluorescence microscope was used to observe the morphological change of leukemia cell when treated with KPT330 for 24 hours. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis of leukemia cells. Then Western blot was used to detect the expression level of CRM1 and the activation of apoptosis related protein Caspase-9、Caspase-3 and PARP level. LncRNA microarray was used to analyze the differentially expressed genes from the cluster analyses. Results: The inhibition of cell proliferation of leukemia cells was in a dose-dependent manner and KPT-330 treatment induced expression of CRM1 down-regulated .Abnormal cells were observed under fluorescence microscopy. Cell apoptosis, cell cycle and activation of apoptosis markers demonstrates that KPT-330 induced apoptosis in leukemia cells. LncRNA microarray analysis showed differently expressed mRNAs and lncRNAs in KPT-330 treated HL-60 cells compared with control group. Molecular function analysis showed that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. Conclusion: In this study, KPT-330 treatment resulted in inhibition of cell proliferation and induction of apoptosis in leukemia cells. LncRNA microarray analysis showed differentially expressed genes and lncRNAs in KPT-330 treated HL-60 cells and we demonstrated that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. These results may provide new clue about molecular mechanism of KPT-330 induced apoptosis; however, underlying details will be required to determine further. Taken together, our findings suggest that for the first time that KPT-330 may act as new candidate's drug for leukemia.

Disclosures

No relevant conflicts of interest to declare.
A Fit-for-Purpose Approach to Monitor Circulating Dendritic Cell Subpopulations for Clinical Investigation
Authors: Trampont Paul Charles , Lourenco Elaine , Czechowska-Kusio Kamila , Litwin Virginia
Published: November 2019

Dendritic cells (DC) are a heterogeneous population of antigen presenting cells involved in the capture, processing and surface presentation of antigens to T-cells. Their immune functions serve to connect innate and adaptive immune responses. In human peripheral blood, the two main subpopulations of dendritic cells, myeloid and the plasmacytoid, are of interest as biomarkers for the development of new therapies and clinical indications. A fit-for-purpose approach was applied to the validation of a two related flow cytometric method designed to evaluate both myeloid and the plasmacytoid dendritic cell populations. The iterative approach allowed us to tailor the assays for and the validation for the intended use of the data within the clinical trial. This presentation describes the rationale for the panel design and the technical validation approach.

Disclosures

No relevant conflicts of interest to declare.
Increasing Quality Improvement Capability in a Hemophilia Treatment Center
Authors: Thornburg Courtney , Lane Heidi , Farrow Katharine , Brooks Rosalie , Jahan Mina , Scopelliti Maria
Published: November 2019

Introduction

Persons with hemophilia require complex medical care by a multi-disciplinary team throughout life. There are opportunities for Hemophilia Treatment Centers (HTC) to enhance the care of persons with hemophilia through participation in quality improvement (QI) initiatives. Maintaining comprehensive care and excellent health outcomes are national priorities for Health Resources and Services Administration (HRSA) and the American Thrombosis and Hemostasis Network (ATHN), the National Hemophilia Program Coordinating Center (NHPCC). Rady Children's Hospital San Diego (RCHSD) HTC participated in the NHPCC Dartmouth Improvement Program pilot program with the aim to develop QI capabilities within the HTCs to enhance the care of persons with hemophilia.

Methods

RCHSD participated in the NHPCC Dartmouth Improvement Program starting in December 2015. A QI team was established including the HTC medical director, nurse case managers, pediatric social worker and the social worker of the collaborating adult HTC. The QI team was coached by an expert TDIMA coach and a "coach in training" from another HTC. The team learned QI methods through in person training, web-based training and weekly team meetings with the coaches. The team assessed HTC data (The 5Ps-purpose, patients, professionals, processes, and patterns) to gain system knowledge and insights to determine a QI theme, global aim, specific aims and associated PDSA cycles.

Results

The RCHSD HTC QI team established a QI theme to focus on the transfer from pediatric to adult care. The Global Aim of the team is to improve autonomous communication in RCHSD HTC. The process begins with the 12 year old comprehensive clinic visit and ends with a new patient visit at an adult HTC. By working on the process, we expect patients to have the communication skills to be able to arrange medical insurance, call the home care company to order factor and to communicate their health and personal care needs to adult health care providers. We developed four specific aims which are in various stages of testing in PDSA cycles (Table 1). For Aim 1, we found that patients have only basic knowledge of insurance information. For Aim 2, we found that we needed to adjust our clinic process to make sure that patients receive their "after visit summary" that includes the insurance information prior to leaving clinic.

Conclusions

We developed a QI program within RCHSD HTC focused on improving transfer of care. Once we have completed the specific aim PDSA cycles we will standardize the new and improved procedures. We will continue to develop the program based on internal needs assessment and family input. This training model may be adapted by other HTCs to enhance patient care.

Disclosures

Thornburg: Mast Pharmaceuticals: Research Funding; Bayer Pharmaceuticals: Research Funding; Shire: Consultancy; Biogen Idec: Other: Data Safety Monitoring Board; Bluebird inc: Other: Data Safety Monitoring Board.
Sex-Based Disparities in Venous Thromboembolism Sociodemographics and Outcomes: A National Inpatient Sample (NIS)-Based Analysis
Authors: Marshall Ariela L. , Bartley Adam C , Ashrani Aneel A. , Gonsalves Wilson I , Kapoor Prashant , Hashmi Shahrukh K. , Siddiqui Mustaqeem A. , Durani Urshila , Go Ronald S.
Published: November 2019

Background: Venous thromboembolism (VTE) contributes to significant morbidity, mortality, and socioeconomic burden. Though male sex is a known risk factor for recurrent VTE, there is a paucity of literature regarding sex-based sociodemographic differences as well as differences in VTE outcomes including hospital length of stay and mortality rates.

Methods: We conducted a retrospective analysis from the National Inpatient Sample (NIS) database from 2012-2013. Inclusion criteria were age 18 years and older and primary discharge diagnosis of VTE Sociodemographic features and medical comorbidities were analyzed, as were hospital length of stay and in-hospital mortality rates.

Results: 107,896 patients in the NIS, representing a national estimate of 539,480 patients hospitalized for VTE between 2012 and 2013, met inclusion criteria. 53% were female and 47% were male. Mean age was 63 years and on average women were older than men (65 years versus 62 years, p<0.001). There were significant differences between men and women with respect to race, primary insurance payor, and VTE location as well as medical comorbidities (Table 1). Median overall length of stay (LOS) was 4 days and in-hospital mortality rates were 2.1%. Female sex was associated with a small but significantly longer hospital LOS (RR 1.04, CI 1.03-1.05, p<0.001) which was also seen in subgroup analyses of lower extremity DVT (RR 1.05, CI 1.03-1.06, p<0.001) and pulmonary embolism (RR 1.03, CI 1.02-1.05, p<0.001) but not upper extremity (RR 1.04, CI 0.96-1.12, p=0.36). There were no significant sex-based differences in in-hospital mortality (2.2% in women versus 2.1% in men, p=0.15). In a multivariate model correcting for sociodemographic differences and medical comorbidities, there were no significant differences between women and men with respect to hospital LOS or in-hospital mortality (Table 2).

Conclusion: We used data from the NIS to study a large number of patients hospitalized for VTE, and identified several sex-based disparities in sociodemographic factors and location of VTE. However, in a multivariable analysis correcting for these factors, sex was not associated with significant differences in hospital LOS or inpatient mortality rates.

Disclosures

Kapoor: Celgene: Research Funding; Takeda: Research Funding; Amgen: Research Funding.
Aiming to Reduce Admissions and Also Re-Admissions in Pediatric Sickle Cell Disease: A Single Institutional Experience with Six Interventions
Authors: Onimoe Grace I. , Carlson Aimee , Frazier Rebecca , Bartley Rebecca , Feldman Erin
Published: November 2019

BACKGROUND AND OBJECTIVES

Sickle Cell disease is an inherited blood disorder of the hemoglobin beta-globin protein in the red blood cells which results in significant risk for hospitalization. Since over 100,000 people in the USA have Sickle Cell Disease and recurrent hospitalizations occur; this constitutes a significant disease burden that are not only costly, but also contribute to loss of income, learned helplessness, additional immense morbidities and psychosocial stressors.

Readmission within 30 days after hospitalization for sickle cell crises was developed at the National Association of Children's Hospitals (NACHRI) as a measurable benchmark to improve hospital quality. The NACHRI, multi state study of sickle cell disease care utilization found an overall readmission rate of 33.4% with lower rates in children (12.8% in children age1-9 and 23.4% for patients ages 10-17).

At our institution, we implemented several interventions to reduce the readmission rate for sickle cell pain crises and will share results and insights.

INTERVENTIONS

The following interventions were employed: a) services of a sickle cell coordinator to co-ordinate clinic visits and patient care b) provide additional standardized teaching on sickle cell care via direct interactions with caregivers, provision of educational materials and organization of a sickle cell awareness conference c) verbal discussions to improve understanding and compliance with a written sickle cell action plan, d) obtaining more frequent parental approval for hydroxyurea therapy, e) increasing tracking immunization rates (influenza), and f) 24hr/72hr discharge telephone call.

The sickle cell team comprising of physicians, nurse practitioners, care coordinator, nurses and social worker actively participated in implementing these steps, and majority of the interventions took place in the outpatient setting.

RESULTS

We reviewed sickle cell admissions and readmissions in 2014 and 2015 in patient's age 0-26 years. Mean age of patients was16.8year (N= 14 Standard deviation = 8.04).

Pre-intervention in 2014 there were 91 admissions with 14/91 re-admissions (15.4%). After beginning implementation of interventions in 2015 there were not only 59% fewer admissions (63), but also 58% fewer readmissions (6 re admit/63 discharges = 9.5%). Thus our interventions affected both admission rate and readmission rates.

CONCLUSION

Sickle cell admissions and subsequent readmissions continue to be a major challenge in successful management of young people with sickle cell disease. Our interventions may help abate some of the hospitalizations and re-admissions as shown by an approximately 40% reduction rate. Although there is undoubtedly much room for additional progress in this area, we have made a good start towards improving our sickle cell population's health.

Disclosures

No relevant conflicts of interest to declare.
Limited Benefit of Additional Contrast-Enhanced CT to End-of- Treatment PET-CT Evaluation in Patients with Follicular Lymphoma
Authors: Paone Gaetano , Raditchkova-Sarnelli Mariana , Stathis Anastasios , Giovanella Luca , Zucca Emanuele , Ceriani Luca
Published: November 2019

BACKGROUND: Follicular lymphoma (FL) is generally characterized by a moderate metabolic activity (FDG-avidity) in nodal and extra-nodal sites and by the frequent detection of residual disease in post-treatment PET scans. Since median age at diagnosis is over 60 years, in many FL patients several conditions that affect the elderly often occur in combination with the tumor, thus complicating and limiting the use of conventional imaging for lymphoma staging and response assessment. For this reason, the choice of the imaging technique should take into account clinical needs (preservation of renal function, reduction of diagnostic radiation exposure) as well as the necessity to constrain health care costs.

OBJECTIVE: The aim of the study was to investigate if there is any advantage in the use of contrast-enhanced CT (ceCT) vs. unenhanced low-dose CT (ldCT) in routine protocols for end-of-therapy PET/CT evaluation of patients with FL.

METHODS: Thirty FL patients who underwent end-of-therapy PET/CT protocol with ldCT and ceCT were analyzed retrospectively. Two different observers evaluated PET/ldCT and PET/ceCT in a blinded manner. Number and sites of nodal or extra-nodal disease were compared, using PET/ceCT as gold standard in order to evaluate if the type of CT could result in changes of the DS and therapeutic strategy.

RESULTS: In 26 of 30 patients (87%; 95% confidence interval, 73%-98%), PET/ldCT showed the same number and sites of lesions highlighted by PET/ceCT. The inter-observer concordance and overall concordance between imaging procedures were excellent with a very high Cohen's kappa (respectively 0.82 and 0.83). 97% of lesions (103/107) were found by PET/ldCT and in 4 of 30 patients (13%) PET/ceCT provided additional nodal lesions in the mesenteric and iliac regions (3 mesenteric nodes and 1 iliac node; <1%). In these 4 patients, DS and consequently the therapeutic strategy were not changed after additional ceCT findings. PET/ldCT accuracy, sensibility, specificity, positive predictive value and negative predictive value and NPV were respectively 87%, 83%, 100%,100% and 60%.

CONCLUSION: Our results indicate that the clinical impact of PET/ceCT in assessing end-therapy evaluation in FL is limited. The PET/ldCT could be suggested as primary imaging modality of choice, thus limiting the acquisition of PET/ceCT images only for doubtful cases of residual disease in mesenteric area (represented solely by 13 % in our series). This diagnostic approach would be less expensive, minimize diagnostic radiation exposure, and preserve renal function.

Disclosures

No relevant conflicts of interest to declare.
Validation and Reliability Study for the Korean Version of the Haemophilia Activity List
Authors: Yoon Hwi-Joong , Kim Ji Yoon , Yoo Ki Young , Choi Eun Jin , Kim Soon Ki , Park Sang Gyu , Lee Kun Soo
Published: November 2019

Background: There has been increasing interest in the patients perspective on the outcome of the treatment. The Haemophilia Activity List (HAL) has been developed as a disease-specific questionnaire for haemophilia patients and is a validated self-report measure of function developed according to WHO International Classification of Functioning, Disability and Health. In this study, we performed a cross-cultural adaptation and linguistic validation of the HAL questionnaire to assess the health-related quality of life in Korean hemophilia patients in the future.

Methods: To validate HAL in Korean, the English versions of HAL were translated into Korean using the forward-backward translation method and merged into a final Korean version. Validation was performed against the Korean version of the questionnaires EQ-5D-5L (EQ-5D) as general tool and Routine Assessment of Patient Index Data (RAPID3) as similar disease-specific tool. All processes were done with permission of the developer and according to WHO guidelines.

Results: One hundred patients with severe and moderate forms of haemophilia A and B from Korea Hemophlia Foundation were invited to participate in the study. Spearmans rank correlation test was used for validation and internal consistency of the HAL was calculated with Cronbach alpha.

Eighty-seven patients (87%) (18-62 years old) answered the questionnaires.

The internal consistency of the Korean version of HAL was high, with Cronbachs alpha being 0.80-0.95.

Upper extremity function had the highest consistency and leisure activities and sport had the lowest. The correlation was good between the HAL overall score and EQ-5D overall (r=0.78), EQ-5D usual activity (r=0.79), and RAPID3 physical function (r=0.82).

Conclusion: The Korean version of HAL has reliability from internal consistency and intraclass correlation by test-retest analysis. The Korean version of HAL has validity which correlated with EQ-5D as general tool and RAPID3 as similar disease-specific tool. This questionnaire of Korean version can be useful as a hemophilia disease-specific instrument for evaluation of the health-related quality of life in Korean patients.

Disclosures

No relevant conflicts of interest to declare.
Regional Intra-Arterial Steroid Treatment in 113 Patients with Steroid Resistant or Dependent Graft-Versus-Host Disease
Authors: Shapira Michael Y , Vipul Sheth , Klimov Alexander , Griesaro Sigal , Tsirigotis Panagiotis , Avni Batia Ronit , Or Reuven , Bloom Allan
Published: November 2019

Introduction : Graft-versus-host disease (GVHD) is a major complication of allogeneic bone marrow transplantation (BMT), resulting in death in the majority of steroid-resistant patients.

In the past 15 years, we revolutionized the use of interventional radiology procedures in the field of stem cell transplantation. This study was aimed to assess the efficacy of regional intra-arterial steroid (IAS) treatment in the largest published cohort of patients with resistant/dependent hepatic and/or gastrointestinal (GI) GVHD. In total, 113 patients with steroid resistant or dependent grade 3-4 hepatic (n = 21), gastrointestinal (GI) (n = 62) GVHD or both (n = 30) were given intra-arterial treatment.

Patients and methods: Patients with liver GVHD received IAS in hepatic artery and patients with gastrointestinal GVHD received IAS in the superior and inferior mesenteric as well as the internal iliac arteries. Patients with pronounced upper GI symptoms also received IAS to the gastroduodenal artery. In total 148 procedures were carried out (range 1-3 per patient).

Results : Partial responses were observed in 33 out of 51 (65%) patients with hepatic GVHD, out of which (55%) had complete response. Median time to complete response was 45 days (range - 5 to 300 days).

For GI GVHD complete responses were observed in 69 out of 92 (75%) and partial responses were observed in 76 out of 92 (83%). The median time to complete response was 20 days (range - 3 to 200 days ).

As this is the largest published cohort of regional IAS therapy for GVHD, multivariate and univariate analysis was done in order to identify the patients with highest chance of response.

Conclusions : Regional treatment of severe GVHD with IAS treatment is effective and safe. GI treatment is more effective than intrahepatic treatment. Early administration of isolated intra-arterial therapy in high-risk patients may further improve the outcome and reduce untoward effects of prolonged systemic immunosuppressive treatment.

Figure 1. Figure 1. Figure 2. Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Enhanced MAPK Signaling Constitute Non-Oncogene Addiction in CSF3R Induced Leukemia
Authors: Rohrabaugh Sara , Kesarwani Meenu , Kincaid Zachary , Huber Erika , Grimes H. Leighton , Azam Mohammad
Published: November 2019

Disclosures

No relevant conflicts of interest to declare.
IN VITRO STUDY OF BONE MARROW
Authors: PLUM CLAUS MUNK
Published: November 2019

Two types of apparatus are described for the culture of bone marrow in vitro. The first, a macro method, is suited to the study of large amounts of bone marrow. The second, a micro method, allows observations of the development of individual cells.

Studies of erythropoiesis by these technics has allowed insight into production of erythrocytes. These data will be reported in a subsequent article.
NORMAL BONE MARROW AS OBTAINED BY STERNAL PUNCTURE
Authors: VAUGHAN STUART L. , BROCKMYRE FRANCES
Published: November 2019

A study of aspirated sternal bone marrow of 50 normal human adult volunteer subjects has been presented. Samples of exactly 3 cc. each were collected in a uniform manner. The technic of performing total nucleated cell counts, differential counts, reticulocyte counts, and megakaryocyte counts has been described and the results given. These results have been subjected to statistical analysis and compared with those of other workers.