bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of Autonomic Function in Patients with Sickle Cell Disease in Relation to Nighttime Hypoxemia
Authors: Erin McGuire , Kato Roberta Miyeko , Bush Adam M , Zmyewski Heather , Detterich Jon , Gardner Daniel , Khoo Michael , Meiselman Herbert J , Coates Thomas D. , Wood John C
Published: October 2019

Abstract 4764

Background:

Sickle cell disease (SCD) is a genetic condition resulting from a homozygous hemoglobin S mutation of the beta globin gene. HbS polymerizes in the deoxygenated state, causing the red cell membrane to become rigid and obstruct blood flow. Ischemia results leading to early organ damage, impaired quality of life and short life expectancy. Patients with SCD die from many causes including infection, splenic sequestration, stroke, and pulmonary emboli, however; a number of deaths in this population are acute and unexplained. The increased risk of sudden death is hypothesized to be associated with autonomic dysfunction. Heart rate variability (HRV) is a measurement of autonomic function and predictor of risk of sudden death. Measures of HRV can be compared to other factors associated with autonomic dysfunction. Individuals with obstructive sleep apnea have increased sympathetic tone. We hypothesized that SCD subjects that experienced nighttime hypoxemia will have autonomic dysfunction.

Methods:

Subjects were fitted with an ambulatory electrocardiogram device (Acitheart, CamNTech, Inc) and monitored for 24 hours. High frequency power (HFP), a measure of cardiac parasympathetic tone, and low:high ratio (LHR), a measure of sympathovagal balance, were calculated from the HRV by the product software. During sleep, baseline oxyhemoglobin saturation (SpO2) and number of desaturations below 88% was monitored using an ambulatory pulse oximeter (WristOX, Respironics, Inc). Bivariate analysis of the median HFP and LHR values over a 24 hour period were compared to baseline SpO2and number of desaturation events. Sex, age and hematologic values including hemoglobin, CRP and lactate dehydrogenase values were also considered.

Results:

30 SCD subjects (mean age= 26.1 years+10.96, 36% male) were evaluated. Baseline nighttime SpO2 was 95.5+2.8% with 1 subject with a baseline SpO2 < 90%. Baseline SpO2 did not correlate with HF with R2 = 0.03 but did correlate with LHR with R2 = 0.46. There was a mean number of desaturation events of 16.6+38.3 with 4 subjects greater than 25 desaturation events. The number of desaturation events did not correlate with HF with R2 = 0.027 but did correlate with LHR with R2= 0.63. Stepwise multivariate analysis was performed including the baseline SpO2, number of desaturation events, sex, age and hematologic parameters showed that the number of desaturation events was the best predictor of LHR.

Conclusions:

SCD subjects that experienced nighttime hypoxemia and increased number of desaturation events during sleep have higher LHR ratios consistent with increased sympathovagal tone.

Disclosures:

Coates: Novartis: Speakers Bureau; Apopharma: Consultancy. Wood:Ferrokin Biosciences: Consultancy; Shire: Consultancy; Apotex: Consultancy, Honoraria; Novartis: Honoraria, Research Funding.
Cerebral Venous Thrombosis in Adolescents and Young Adults with Classical Hodgkin Lymphoma
Authors: Kumar Poornima , Ahmed Rebekah , Riat Renu , Ardeshna Kirit M , Daw Stephen
Published: October 2019

Abstract 4784

Background

Patients with classical Hodgkin Lymphoma (cHL) have a relatively high risk of venous thrombo-embolism (VTE); reported incidence 4.6–7% in adults and up to 11.5% in children and adolescents. Most VTE episodes are peripheral or related to central venous catheters, with very limited data on central or life-threatening thromboses in adolescents. There is only 1 reported case series on cerebral venous thrombosis (CVT) in adolescents. We report 4 cases of CVT from our centre, all treated with chemo-radiotherapy. Chemotherapy comprised OEPA (vincristine, prednisolone, doxorubicin, etoposide) and COPP/COPDAC (cyclophosphamide, vincristine, prednisolone, procarbazine/dacarbazine respectively).

Results

All patients received involved field radiotherapy (IFRT) 19.8 – 30Gy on completing chemotherapy. All were female, aged 12–23. All received norethisterone contraception. All had indwelling central venous catheters (PICC). Patient 4 alone had a raised body mass index. All were exposed to steroids; Patient 4 completed steroid therapy several weeks before developing CVT symptoms. Patients 2 and 4 received treatment dose low molecular weight heparin (LMWH) for 6 weeks after diagnosis of PICC-associated thrombosis, and were not on anticoagulation or thromboprophylaxis when CVT was diagnosed. Regarding other risk factors, 3/4 had no documented prothrombotic tendency. Patient 4 was found to have a moderately positive IgM anti beta 2 glycoprotein antibody present 12 weeks apart, consistent with antiphospholipid syndrome.

All patients were therapeutically anticoagulated for 6 months to 1 year. LMWH of choice at our centre was dalteparin. Patient 1 was switched to warfarin upon completion of chemo-radiotherapy, and Patient 4 was commenced on warfarin with dalteparin cover at diagnosis of CVT as she had completed treatment.

Patients 1 and 2 had raised intracranial pressure on lumbar puncture, and required therapeutic lumbar punctures and acetazolamide.

Patients 2 and 3 both required anticonvulsant therapy for 1 year. Patient 2 was initially treated with phenytoin, and switched to carbamazepine. Patient 3 was also initially managed with phenytoin, and switched to levetiracetam. Neither patient had any subsequent seizures.

All 4 patients have recovered completely from CVT with no residual neurological deficits or further thromboses.

Conclusion

CVT is a rare and potentially life threatening complication in adolescents and young adults with cHL with paucity of data. The risk factors are unclear however all patients in our series were female, received steroids and were on norethisterone. Only 1 patient had a prothrombotic tendency detected on thrombophila screening. CVT is treatable, and complete resolution of signs and symptoms can be expected. More studies are required to elucidate risk factors which may help develop thromboprophylaxis guidance in this group of patients.

Disclosures:

No relevant conflicts of interest to declare.
Clinical Characterization and Prognostic Factors of 144 Adult Acute Monocytic Leukemia
Authors: Wu Yingying , Ye Xiujin , Li Li , Zhu Jingjing , Xie Wanzhuo , Zhang Jie , He Jingsong , Cai Zhen , Huang He
Published: October 2019

Abstract 4798

Background:

Acute monocytic leukemia (M5) is one kind of acute leukemia with poor prognosis, which has the characterization of high number of white blood cells, high risk of extramedullary metastasis and abnormal chromosome karyotype.

Objective:

To analyze clinical characterization and prognostic factors of adult acute monocytic leukemia.

Methods:

Clinical data of 144 patients diagnosed between Jan 1st 2006 and Dec 31 2010 were collected. Clinical manifestation, cytogenetics, immunophenotyping and minimal residual disease (MRD) were analyzed respectively. The numeration data was tested by χ2 test, while the measurement data by Levene test to variance homogeneity and by T test to the mean difference. The data were analyzed by SPSS 17.0.

Results:

81.9% of 144 patients reached complete remission. 27.5% patients were abnormal in cytogenetics, including +8, −5/del (5q), −7/del (7q). Patients were divided into three groups according to chromosome karyotype. Intermediate prognostic group has better OS and DFS than poor prognostic group (P<0.05). 26 patients treated with SCT.These patients have higher OS rate for one year and low relapse rate than those who did not receive SCT.CR rate for old patients(≥55) and young patients (<55) were 69.4% and 86.1% respectively. Old patients had shorter OS and DFS than young patients, but no singnificant difference in FAB type, number of white blood cell and immunophenotyping. Old patients occur chromosome abnormalities more frequently than young patients. Relapse rate is 20.7% in MRD+ group versus 11.5% in MRD- group. Multivariate COX analysis showed that chromosome karyotype and MRD had most significant correlation with relapse.

Conclusion:

Chromosome karyotype, age, therapy, MRD were all prognostic factors of M5. Chromosome karyotype and MRD had most important significance in prognosis.

Disclosures:

No relevant conflicts of interest to declare.
High WT1 Expression in 2nd CR of AML Is Predictive for Early Relapse (prospective study)
Authors: Girshova Larisa , Romanova Ekaterina , Kholopova Irina , Nikulina Tatiana , Mirolubova Julia , Gorunova Elena , Motorin Dmitry , Zaritskey Andrey
Published: October 2019

Abstract 4812

Backgound:

The Wilms' tumor gene 1 (WT1), which is over-expressed in more than 90% of AML, is a useful marker for monitoring MRD. Quantification of WT1 transcript level in peripheral blood (PB) and bone marrow (BM) at recovery from chemotherapy reliably discriminates patients at different risks of relapse. It is known that WT1 expression may be a marker of future relapse in 1 CR AML patients, after allogenic BMT and WT1 reduction also is a prognostic during induction of the first episode of AML. But there no evidence of WT1 reduction value in AML relapse.

Patients and methods:

In our prospective study were 10 patients with the first AML relapse and 2nd bone marrow complete remission were studied. High level of WT1 at relapse and achieving 2nd remission were the main inclusion criteria. Median WT1 level at relapse was 4302,2 (range 540–13184) copies WT1/104 Abl. Monitoring of WT1 level were estimated in patients with median age 36,6 (range from 20 to 64). Median time from the first CR to the relapse was 16,47 months (range from 3 to 51 mo), so there were 6 patients with early relapse and 4 patients with late relapse respectively. Chemotherapy with high dose cytarabine (HiDAC, FLAG) used for 2nd CR induction.

Results:

Only 66,7% (6 patients) demonstrated WT1 normalization. All of patients (n=4) with high WT1 expression in bone marrow remission developed second relapse in a few months, median time to relapse was 4,45 months (range from 2,8 to 7,4 mo). While patients with normal WT1 level in 2nd remission had relapse free survival 83.3% at 20 months (p=0.048, Fisher exact test, two tailed). Only one of 6 patients developed an extramedullary relapse in 6 months. The groups of patients were comparable to cytogenetic and molecular risks.

Conclusion:

We have shown that high WT1 level in 2nd remission is predictive for early relapse. Thus, high WT1 level in 2nd remission is an indication for very early stem cell transplantation.

Disclosures:

No relevant conflicts of interest to declare.
Eph-A4 Plays a Important Role in Metastasis and Invasive Activity of Myeloid Leukemia Cells
Authors: Xiaoli Liu , Zhou Xuan , Zuo Yuan , Xu Lulu , Zhang Jinfang , Xu Na , Huang Bintao , Xiao Xiaozhen , Huang Yuanlu , Du Qingfeng
Published: October 2019

Abstract 4819

The Eph receptors are found in a wide range of cancers and correlate with metastasis. However, their precise role in cancer has only started to be addressed. In this study, we investigated the role of Eph-A4 receptor in metastasis and invasive activity of myeloid leukemia cells. We fisr tested the expression of Eph-A4 in eight primary myeloid leukemias imclulding four with extramedullary metastasis and four without it, and leukemia cell line K562 by Real-time PCR and western blotting, then found that Eph- A4 was wildly expressed in myeloid Leukemia cells, especially in myeloid leukemia cells with high invasive activity. To further clarified the question, the stable over-expressing Eph-A4 cell line (K562-EphA4) based the wild K562 cell and pGC lentivirus vector were established to declare the metastasis and invasive activity in myeloid leukemia cells in vitro by trans-well migration assay. The results indicated that the mRNA level and protein expression of Eph-A4 were significantly increased in myeloid leukemias with extramedullary metastasis and K562 cells compared to those without it (P<0.05).After we successfully established the stable over-expressing Eph-A4 cell line, we verified its mRNA level and protein expression were both significantly increased ((P<0.05).Furthermore, RhoA and Rac1/cdc42, which are important adhesion molecules and related to metastasis and invasive activity were both highly expressed in K562-EphA4 cells compared to wild K562 cells (P<0.05). Moreover, the trans-well migration assay showed that cells that migrated to lower chambers and matrigel hydrogel were both increased in K562-EphA4 cells compared to wild K562 cells (5.22±2.11*104/ml vs 13.56±2.70*104 /ml P<0.000;18.07±3.15/cm2 vs 28.53±2.50/cm2P<0.000 respectively). Our findings suggest that Eph- A4 is likely to play an important role in the regulation the of myeloid Leukemia through the control of RhoA and Rac1/cdc42 associated signaling, migration and invasive activity, and therefore may represent a novel target for cancer treatment.

Disclosures:

No relevant conflicts of interest to declare.
More Sun Light Exposure May Be Associated with Improved Overall Survival in Diffuse Large B-Cell Lymphoma. A Population Based Swedish Lymphoma Registry Study
Authors: Jerkeman Mats , Székely Elisabeth , Hagberg Oskar , Linden Ola
Published: October 2019

Abstract 4847

Background

Low levels of circulating 25-hydroxyvitamin D [25(OH)D] has been associated with inferior EFS and OS in DLBCL[1]. Ultraviolet B radiation from the sun contributes strongly to the vitamin D status in humans. In Scandinavia, the difference in sun light exposure is highly dependent on season. Consequently, vitamin D levels in Norway have been found to be 15–50 % lower in winter compared to summer[2]. In this study we tested the hypothesis that diagnosis of DLBCL during the lighter period of the year may be related to superior overall survival in DLBCL.

Patients and materials

The study population was collected from the Swedish Lymphoma Registry 2000–2010.

A Cox regression model was analyzed for overall survival. Light exposure was defined as a continuous variable - cos((month of diagnosis -6)/12 × 2π) – varying between -1 in December and +1 in June. In addition, age, stage, year of diagnosis, performance status, number of extranodal sites, and LDH were included as cofactors.

Results

During this period, 5268 adult patients with DLBCL were identified.

In multivariate analysis, including all factors above, overall survival was significantly superior for patients diagnosed during the lighter part of the year, p = 0.032.

The hazard ratio for death due to all causes for patients diagnosed in June compared to December was 0.90. Monthly

Discussion

On a population level, seasonal differences in sun light exposure may contribute to differences in overall survival in DLBCL. However, to prove if normalizing vitamin D levels in this group of patients would improve survival; this will require testing in randomized trials.

Disclosures:

No relevant conflicts of interest to declare.
Bone Turnover In Multiple Myeloma: Impact Of Bortezomib-Based Induction Therapy, High-Dose Melphalan, and Lenalidomide Consolidation On Osteoblast and Osteoclast Function Within The GMMG-MM5 Trial
Authors: Seckinger Anja , Meißner Tobias , Bertsch Uta , Salwender Hans , Dürig Jan , Hänel Mathias , Blau Igor W. , Scheid Christof , Weisel Katja , Klein Bernard , Goldschmidt Hartmut , Hose Dirk
Published: October 2019

Introduction

In multiple myeloma, initially, there are increased numbers of osteoclasts showing increased activity, but bone formation by osteoblasts is keeping step. In later stages, parts of the bone remodeling compartments are disrupted by the interaction with myeloma cells leading to increased bone resorption which can no longer be compensated (myeloma bone disease, uncoupling of bone formation and bone resorption). Lenalidomide and bortezomib have been shown to target both, myeloma cells and the microenvironment: lenalidomide inhibits osteoclastogenesis, bortezomib is also able to stimulate osteoblast differentiation leading to increased bone formation. Aim of this study is to evaluate the impact of bortezomib-based induction treatment, high-dose therapy, and lenalidomide consolidation on alterations of bone turnover, i.e. surrogates of osteoblast- (osteocalcin, OC) and osteoclast- (collagen type I fragments, CTX-I) function, and their induction by myeloma cells (DKK1-level).

Methods

Serum was collected during routine sampling within the GMMG-MM5 trial (EudraCT 2010-019173-16), and levels of CTX-I, OC, and DKK1 were assessed by ELISA in triplicates using commercially available assays according to the manufacturer’s instructions (RnD Systems and Immunodiagnostic Systems). The following time points were assessed: at inclusion (n=365), after induction therapy with either PAd (n=88) or VCD (n=84), stem cell mobilization using CAD (n=69), high-dose melphalan (n=92), and 2 months lenalidomide consolidation (n=92). Up to now, serum samples of 69 patients were measured sequentially at five time points in line with the GMMG-MM5 trial. DKK1 levels were correlated with the expression in CD138-purified myeloma cells (Affymetrix microarrays, n=365).

Results

Prior to treatment, CTX-I levels are increased, those of OC decreased compared to healthy donors (uncoupled bone turnover). DKK1 protein levels are increased and correlate with DKK1-expression in myeloma cells. After induction therapy, osteoclast activity (CTX-I) is decreased below normal values. PAd unlike VCD further decreases osteoblast activity (OC-levels); DKK1-levels are normalized. Subsequent treatment further decreases DKK1-levels below normal values and blocks osteoclast function. After 2 months lenalidomide consolidation, no normalization of osteoblast activity is found.

Conclusion

The main impact on bone turnover by bortezomib-based induction treatment is a reduction of osteoclast activity alongside a decrease in DKK1-levels. During the reported period, no normalization of decreased osteoblast function was observed.

Disclosures:

Seckinger: Novartis Pharma: Research Funding. Goldschmidt:Novartis Pharma: Research Funding. Hose:Novartis Pharma: Research Funding.
Responsiveness Of Cytogenetically Discrete Human Myeloma Cell Lines To Lenalidomide: Lack Of Correlation With Cereblon and Interferon Regulatory Factor 4 Expression Levels
Authors: Greenberg Alexandra J , Walters Denise K , Arendt Bonnie Kaye , Tschumper Renee C. , Kumar Shaji K. , Rajkumar S. Vincent , Jelinek Diane F.
Published: October 2019

The introduction of novel immunomodulatory drugs (IMiDs), such as lenalidomide, has drastically improved the survival of patients with multiple myeloma (MM). While it has been shown that patients with specific cytogenetic subtypes, namely t(4;14), have the best outcomes when treated with bortezomib-based regimens, the relationship between cytogenetic subtypes and response to IMiDs remains unclear. Using DNA synthesis and apoptosis assays, we investigated the relationship between cytogenetic subtype and lenalidomide response in a representative panel of human myeloma cell lines (HMCLs). Additionally, we examined HMCL protein expression levels of the lenalidomide target cereblon (CRBN) and its putative downstream target interferon regulatory factor 4 (IRF4), both of which have previously been shown to be predictive of lenalidomide response in HMCLs. Our results reveal that response to lenalidomide did not correlate with specific cytogenetic translocations. There were, however, distinct groups of lenalidomide-responsive and non-responsive HMCLs, as defined by inhibition of cellular proliferation; notably, all of the hyperdiploid HMCLs fell into the latter category. Repeated dosing of lenalidomide significantly lowered the IC50 of the responsive HMCL ALMC-1 (IC50 = 2.6 μM versus 0.005 μM, p=0.004), but did not have an effect on the IC50 of the non-responsive HMCL DP-6 (p>0.05). Moreover, no association was found between lenalidomide responsiveness and CRBN or IRF4 expression levels. In summary, our data indicate that lenalidomide sensitivity is independent of cytogenetic subtype in HMCLs. Additionally, while CRBN and IRF-4 have been shown to be associated with response to lenalidomide in patients, these findings do not translate back to HMCLs, which could be attributable to factors present in the bone marrow microenvironment. These results suggest that the development of a new treatment paradigm based on cytogenetic subtype of MM alone may be insufficient, and that additional markers may need to be incorporated. Furthermore, the disconnect between our findings of expression of CRBN and IRF-4 in lenalidomide-treated HMCLs and patient data highlights the need for further investigation into the role of the bone marrow microenvironment on the regulation of these previously demonstrated markers of lenalidomide response.

Disclosures:

No relevant conflicts of interest to declare.
The Choice Of Regimens Based On Bortezomib For Patients With Newly Diagnosed Multiple Myeloma
Authors: Yang Li , He Jingsong , Han Xiaoyan , Zheng Gaofeng , Zheng Weiyan , Wei Guoqing , Wu Wenjun , Ye Xiujin , Shi Jimin , Xie Wanzhuo , Li Li , Zhang Jie , Huang Weijia , Zhao Yi , Huang He , Zhang Xuejin , Fu Jiaping , Cai Zhen
Published: October 2019

Bortezomib significantly improved response rates in the treatment of patients with multiple myeloma (MM). However, the process of selecting an optimal bortezomib based regimen as the initial therapy of MM remains ambiguous due to a lack of modern clinical trials demonstrating the efficacy of various bortezomib based treatments. Here, we report four bortezomib-based regimens for treatment of MM patients from three hematological treatment centers in China.

Methods

Newly diagnosed MM patients in three hematological centers in China between February 1, 2006 and May 31, 2013 were treated with combination therapies including bortezomib plus dexamethasone (PD), or triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) every 28 days.

Results

The overall response rate (≥ partial response, ORR) of all the 215 eligible patients was 90.2%. The ORR for PCD, PAD, PTD and PD were 97.4%, 93.2%, 85.3% and 77.8% respectively, while the effects with VGPR or better were 63.7%, 62.7%, 44.2% and 37.8% respectively. The effect of ORR, VGPR and CR/nCR for PCD regimen was significantly better than PD scheme (P = 0.009, 0.011, 0.005 ). The median PFS of all the patients was 29.0 months with significant differences observed between groups (P =0.047). The median OS of all the patients was not reached, but triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) were more efficient in treatment of MM patients compared to PD (P =0.005). The frequently observed toxicities were neutropenia, thrombocytopenia, fatigue, infection, herpes zoster, and peripheral neuropathy. Incidence of peripheral neuropathy (PN) in PTD group was significantly higher than other three groups, especially grade 2-3 PN. Treatment with anti-viral agent acyclovir significantly reduced the incidence of herpes zoster.

Conclusions

Our study demonstrated that bortezomib-based regimens were active and well-tolerated in the Chinese MM patients, and triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) were more efficient for treatment of MM patient, and the patients received PCD or PAD demonstrated significant higher ORR compared to PD.

Disclosures:

No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation - A Long-Term Curative Approach In Patients ≥ 60 Years With Advanced Disease AML/MDS, - The Freiburg Experience Of 250 Consecutive Patients
Authors: Bertz Hartmut , Lübbert Michael , Ohneberg Kristin , Wäsch Ralph , Zeiser Robert , Marks Reinhard , Finke Jürgen
Published: October 2019

Since the introduction of reduced-toxicity conditioning prior to allogeneic hematopoietic stem cell transplantation (alloSCT) we transplanted from 1999 to 2012, 250 consecutive patients (pts) with myeloid malignancies (AML, MDS) aged ≥ 60 years (yrs).

The 144 male and 106 female pts with a median age of 66 yrs (range 60-77) were transplanted for de novo AML (n=95), s/tAML (n=104) and MDS (n=51) with 89% unfavorable cytogenetics (CALGB). Since 2004 pts received a prospective fitness assessment (Deschler et al., Haematologica 2013). In 74% the donor was matched/mismatched unrelated and in 26% related. Only 16% were transplanted in CR1/2, 84 % with advanced or untreated disease. The conditioning regimen was the FBM protocol (fludarabine, carmustine, melphalan; Bertz et al., JCO 2003) in 98%, and 97% of the pts received PBSC. For GVHD prophylaxis in 91% a combination of cyclosporine plus alemtuzumab or ATG-F™ was used.

At day +30, 94% of the pts had achieved CR by standard measures. With a median follow up of 57 months (3-157) 37% of the pts are alive; main causes of death were relapse (n=62), infection (n=35) and age-related diseases (n=13). The probability of OS/DFS was at 1yr 61%/49%, at 2 yrs 49%/41% and at 5 yrs 37%/34%, respectively. The probability for NRM at 1 yr is 24%. Nineteen known prognostic factors for outcome were evaluated: e.g. patient and donor age, graft size, days between diagnosis and alloHCT, CMV, early/advanced disease, cytogenetics, Sorror and Gratwohl score, donor type, HLA-identity. In the multivariate analysis a better OS (factors with p<0.1; table) was seen with a matched donor; a better DFS with a related donor, and high CD34+ graft content; in contrast, a mismatched donor is a risk factor for reduced DFS.TableMultivariate analysis of prognostic factors* for OS and DFSvariablevalueHazard Ratio95% CI lower limit95% CI upper limitP valueOverall survivalRemission at alloHCTadvanced1.370.862.160.1825HLA mismatchyes1.401.011.960.0463HCT-CI (Sorror)>= 21.311.011.960.1007Peripheral blood blastsyes1.210.841.760.3034Disease-free survivalRemission at alloHCTadvanced1.290.722.300.3946Donorrelated0.640.430.950.0258HLA mismatchyes1.440.992.090.0561CD34+ cells> median0.760.551.040.0867Bone marrow blasts> 5%1.210.781.880.3915*in univariate analysis p<0.157 (AIC criterion; Sauerbrei W, 1999 Applied Statistics,48:313-329.70.)

In conclusion, this unique large cohort of older pts with AML/MDS with mainly advanced disease and unfavorable cytogenetics shows a high feasibility, safety and efficacy of alloHCT after the FBM protocol. AML/MDS pts in their 7th and 8th decade of life fit for transplant should be evaluated for alloHCT as very important long-term curative option.

Disclosures:

No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Stem Cell Transplantation From Haploidentical Donor After Reduced Intensity Conditioning Regimen Allows Promising Disease Control In Patients With Hematological Diseases
Authors: Devillier Raynier , Castagna Luca , Crocchiolo Roberto , Furst Sabine , Harbi Samia , El Cheikh Jean , Granata Angela , Mohty Bilal , Faucher Catherine , Blaise Didier
Published: October 2019

Introduction

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) from haploidentical donors (HID) as alternative donor is currently increasing for patients with hematological diseases. In this setting, post Allo-HSCT high dose Cyclophosphamide (Cy) allows the use of T-repleted graft from HID by decreasing early alloreactive T-cell, leading to low incidences of graft-versus-host disease (GVHD) and non-relapse mortality. However, the efficacy of this procedure is still a matter of debate. The aims of this study were to evaluate the efficacy of HID Allo-HSCT in a comparison with our historical control cohort of patients transplanted from matched related or unrelated donors.

Patients and methods

We report here the experience of HID Allo-HSCT prepared by reduced intensity conditioning (RIC) regimen and post infusion Cy (HID group; n=60) and compared outcome to that of historical control cohort of patients with hematological diseases (n=220) who underwent RIC Allo-HSCT from matched related donors (MRD group; n=110) or unrelated donors (UD group; n=110). The control cohort received RIC regimen based on Fludarabine (150 mg/m2 total dose over 5 days), intravenous Busulfan (6.4 mg/kg total dose over 2 days) and antithymocyte globulin (ATG; Thymoglobulin; 5 mg/kg total dose over 2 days). To better compare the results between these two heterogeneous cohorts of patients including different hematological diseases, we used the disease risk index (DRI) score recently published (Armand at al., Blood, 2012).

Patient characteristics are illustrated in the Table 1. Although HID and MRD/UD groups were significantly different regarding age and graft source, they presented with similar disease risk profile when classified according to the DRI and comorbidities according to the HCT-CI.

Results

In the HID group, day-100 acute GVHD, overall incidence of chronic GVHD and 2-year NRM were 21%, 9% and 15% respectively. Regarding the disease control, with a median follow up of 17 months (range: 5-50), the 2-year cumulative incidence of relapse (CIR) was 23%, which is similar to the MRD and UD control groups (2-year CIR: MRD 31%; UD: 18%). The 2-year PFS and OS were 62% and 68% respectively in the HID group, 52% and 66% respectively in the MRD group and 55% and 56% respectively in the UD group.

Conclusions

These results suggest that RIC Allo-HSCT from HID allows promising anti-tumor effect that is comparable to our previous experience of Allo-HSCT from MRD or UD. We also confirm the low toxicity profile after HID Allo-HSCT. Taken together, these observations suggest that HID is a valuable alternative for patients with hematological diseases but no available MRD or UD.

Disclosures:

No relevant conflicts of interest to declare.
Lower Relapse and Better PFS Among Chemosensitive Patients Undergoing Allogeneic Transplantation By Haploidentical Compared With HLA-Identical DONOR: Results On A Cohort Of 94 Patients With Hodgkin’S Lymphoma
Authors: Castagna Luca , Crocchiolo Roberto , Bramanti Stefania , Furst Sabine , Sarina Barbara , El Cheikh Jean , Granata Angela , Devillier Raynier , Garciaz Sylvain , Bouabdallah Reda , Santoro Armando , Blaise Didier
Published: October 2019

Introduction

haploidentical stem cell transplantation (SCT) represents an alternative option for those patients lacking a HLA-identical donor. Patients with relapsed or refractory Hodgkin's lymphoma may benefit from allogeneic SCT and, in particular, patients without a HLA-identical donor may receive SCT from a haploidentical one. The aim of this retrospective analysis is to compare outcome of HL patients undergoing allogeneic SCT from haploidentical vs. HLA-id. Donor.

Methods

patients with diagnosis of HL who received allogeneic SCT at Istituto Clinico Humanitas (Milan, Italy) and Institut Paoli-Calmettes (Marseille, France) were included in the analysis. Outcome was estimated according to SC source (HLA-id. vs. haplo) and disease status at transplant: CR, PR or SP/PD.

Results

a total of 58 and 36 patients received SCT from a HLA-id. and haploidentical donor, respectively. SCT were performed from February 1999 to December 2012. Ninety-two patients received reduced-intensity conditioning; all but one patient in haplo group received post-transplant cyclophosphamide as GVHD prophylaxis. Median follow-up was 63 months (range: 6-160) in HLA-id. group and 20 months (range: 6-50) in haplo group. In HLA-id. group, 2-year PFS was 62%, 33% and 17% among patients in CR, PR or SD/PD before allogeneic SCT, respectively; 2-year PFS in haplo patients was 86%, 53% and 0% among CR, PR and SD/PD patients. Results are shown in Table 1. Notably, a lower number of relapse events were observed in chemosensitive patients in haplo group compared with HLA-id.: 1/21 vs. 7/28 among CR patients and 2/9 vs. 10/18 among those in PR (see Table 1); this translates into better PFS after haplo-SCT. Conversely, outcome of refractory patients confirmed to be poor in both groups. At last follow-up, among the 30 chemosensitive patients receiving haplo-SCT, last relapse occurred at day +392 post-SCT.

Conclusions

present analysis showed better PFS in haplo group among high-risk, chemosensitive (CR + PR) patients compared with HLA-id. and better OS among patients who were in PR before SCT; this was due to lower relapse rate among chemosensitive patients after haplo-SCT vs. HLA-id. In conclusion, haplo-SCT showed high antitumor activity, leading to promising outcome of chemosensitive patients compared with HLA-id transplants. Present data may challenge donor choice in this subset of patients, who may benefit from graft-versus-lymphoma effect, potentially enhanced by haplo disparity.

Disclosures:

No relevant conflicts of interest to declare.
Platelet-Like-Particles Sheared From Myosin-II-Inhibited Megakaryocytes Highlights The Elevated Thrombocrit Of May-Hegglin Anomaly
Authors: Spinler Kyle R , Shin Jae-Won , Discher Dennis E
Published: October 2019

Megakaryocytes (MKs) in the marrow extend projections into blood flow and generate platelets under shear. Understanding MK differentiation and platelet production is of broad clinical importance and extends a need to augment platelet numbers in patients. Reversible but sustained inhibition of non-muscle myosin-II (NMM-II) with the drug blebbistatin increases MK polyploidization, proplatelet formation, and membrane flexibility, thereby increasing platelet generation under shear. Using a cone and plate rheometer to apply fluid shear to drug-treated MKs in bulk, platelet-like-particles (PLPs) that are collagen-I responsive can be generated with intermediate shear. The MKs naturally down-regulate NMM-IIA activity through phosphorylation of S1943, but this site proves shear sensitive, consistent with results for human platelets. Using micropipette aspiration of MKs, inhibition of NMM-IIA is found necessary to generate CD41+ fragments that approximate the size of human platelets. Localization of NMM-IIA to the fragments is modulated by S1943 as seen by unique distribution patterns resulting from specific S1943 mutations that can be abrogated by addition of blebbistatin. The approach is extended to clinically relevant mutations associated with May-Hegglin anomaly (MHA) co-expressed with wild type protein to mimic heterozygotes. As with blebbistatin inhibition of myosin, May-Hegglin mutants result in a higher frequency of fragmentation during micropipette aspiration, indicating a dominant negative effect. Immunofluorescence documents abnormal myosin aggregation in cells transfected with May-Hegglin myosin mutations compared to wild type constructs. Finally, peripheral blood from a patient with a D1414N May-Hegglin mutation is cultured to produce megakaryocytes used to support both the micropipette and immunofluorescence results. These findings reveal a phospho-switch in NMM-II, from inactive to active in the terminal stages of platelet-poiesis, and that proper myosin activity is critical to fragment size and number. Disruption of normal activity enhances fragment generation suggesting a novel mechanism in MHA: in particular, MHA thrombocytopenia results in an increased thrombocrit due to abnormally large platelets, which overcompensates for the reduction in platelet number.

Disclosures:

No relevant conflicts of interest to declare.
Morphological Distinction Unravels Mechanisms Of Platelet Biogenesis From Bone Marrow Megakaryocytes
Authors: Nishimura Satoshi , Eto Koji , Nagai Ryozo
Published: October 2019

Blood platelets are generated in the bone marrow (BM) from their precursors, megakaryocytes (MK). Although we know that MKs produce platelets throughout life, precisely how platelets are produced in vivo remains uncertain, largely because of the rarity of MKs in the BM and the lack an adequate visualization technique. In the present study, we were able to visualize MK dynamics leading to platelet release in living animals at high resolution.

To clearly understand the nature of thrombopoiesis in BM MKs, we optimized an in vivo imaging technique based on two-photon microscopy that enabled us to visualize living BM in CAG- enhanced green fluorescent protein (eGFP) mice. By visualizing living bone marrow in vivo, we observed that two modes (fragmentation and proplatelet formation) can be ongoing simultaneously in the same mouse. We observed that these two modes detectable by different morphological behavior can be ongoing simultaneously in the same BM of mouse, and are regulated by specific cytokines. Short proplatelets from megakaryocytes predominated at steady state, and more elongated proplatelets were accelerated by thrombopoietin (TPO) with responding to chronic platelet needs including recovery form BM transplantations. In contrast, acute platelet needs by blood loss, 5-FU administration or pritoneal acute inflammation increased cytoplasmic fragmentation following rapid ‘rupture’. Observed two modes are both dependent on tubulin reorganization on platelet biogenesis. In addition, platelet increase at acute phase is independent of proliferation by MK progenitors and this factor might exert apoptosis machinery on already reserved mature type of MKs.

This humoral factor was identified by combination of in vitro screening systems and in vivo MK visualization analysis. Factor serum levels were reduced independently of the thrombopoietin level in human subjects with low platelet counts. It thus appears the cytokine balance dynamically regulates the mode of thrombopoiesis and the cellular programming of MKs. Thus, these novel factor may be a novel therapeutic target in thrombocytopenic situations, especially when associated with acute loss of platelets or when platelet transfusion is limited or unsuccessful.

Disclosures:

No relevant conflicts of interest to declare.
RUNX1C-Mediated Reprogramming Of Human Bone Marrow Stromal Cells into Early Blood Progenitors
Authors: Yin Weihong , Porada Christopher D , Walker Stephen , Bishop Colin , Almeida-Porada Graca
Published: October 2019

Somatic cell reprogramming to the hematopoietic lineage, either through a pluripotent state or directly, opens the possibility of production of a ready source of autologous hematopoietic stem cells (HSC) that can be used to treat/cure a wide variety of blood disorders. While it has previously been shown that dermal fibroblasts (HFF) can be directly reprogrammed to the hematopoietic lineage, the efficiency was relatively low and the resultant hematopoietic cells lacked multilineage differentiative potential. Stro1(+) isolated stromal progenitors (SIPs) can easily be isolated from the bone marrow (BM) and expanded ex-vivo to obtain clinically significant numbers of cells. In similarity to HSC, SIPs are derived from the mesoderm, and are intimately linked with HSC specification during ontogeny. As such, they are likely to be epigenetically closer to HSC than HFF, and therefore good candidates for reprogramming into hematopoietic cells. To verify the uniqueness of SIPs for reprogramming, we transduced SIPs and HFF with OCT4 and/or RUNX1C, a master transcription factor (TF) that triggers the developmental onset of definitive hematopoiesis, in the following combinations: 1) OCT4 alone; 2) RUNX1C alone; or 3) OCT4+RUNX1C. We then performed a timeline of gene/cell surface marker expression (using microarray, qRT-PCR, and flow cytometry) from day 3-16 post-transduction. Visual inspection of the cultures showed that, while reprogrammed colonies began to appear in SIPs cultures at day 9, no colonies were seen during this time period in HFF cultures. Flow cytometry and molecular analyses of colonies obtained from OCT4+RUNX1C combination demonstrated that expression of CD41, the earliest marker of commitment to the hematopoietic lineage, commenced within only 3-4 days and peaked at day 5-6, by which time ∼20% of SIPs expressed this marker. This peak in CD41 expression coincided with commencement of expression of CD34 and CD45, and maximal induction of several hematopoiesis-specific TFs and phenotypic markers such as PU.1, HOXB4, GATA2, MIXL, WNT3, KDR, CDX4, which occurred at 1-3 logs higher levels in SIPs than HFF. Further studies demonstrated that the chromatin remodeling function of OCT4 could be replaced with the histone methyltransferase inhibitor Bix-01294, with the combination of RUNX1C and Bix-01294 inducing levels of CD34 and CD41 expression by day 5 that were similar to those achieved with RUNX1C plus OCT4. The present studies thus take several important steps towards making the promise of producing autologous hematopoietic cells for transplantation via direct reprogramming a reality.

Disclosures:

No relevant conflicts of interest to declare.
Phase II Randomized Study Of Lenalidomide Or Lenalidomide and Rituximab As Maintenance Therapy Following Standard Chemotherapy For Patients With Intermediate-High/High Risk Diffuse Large B-Cell Lymphoma (DLBCL)
Authors: Reddy Nishitha M. , Morgan David S. , Park Steven I. , Greer John P , Richards Kristy L.
Published: October 2019

Background

DLBCL patients (pts) with an intermediate/high to high risk international prognostic index (IPI) are at an increased risk of disease relapse rate in the first year after completion of standard therapy with R-CHOP. Lenalidomide (len), an immunomodulatory drug, has activity in relapsed diffuse large B cell Lymphoma. Len enhances the natural-killer cell mediated antibody-dependant cellular cytotoxicity of rituximab in lymphoma cell lines and inhibits angiogenesis as well as alters cytokine production. Lenalidomide received FDA approval based on its clinical activity in relapsed mantle cell lymphoma.

Methods

Intermediate-high/high risk IPI pts with DLBCL were randomized to len (arm A) alone or len and rituximab (arm B). The primary endpoint of the study was to assess the one year disease-free survival. We expected that a 25% difference of relapse will have clinical significance when compared with current standard therapy. Pts in arm A received len at a dose of 25 mg daily for 21 days of 28 days. Patients on arm B received len at a dose of 20mg daily for 21 days of 28 days along with rituximab on day 8 of even cycles. Treatment on both arms was continued for one year. Treatment was discontinued for disease progression. Len dose adjustments are incorporated in the protocol.

Results

Forty three pts, 21 arm A/ 22 arm B, 21 female/22 male, with a median age of 59 yrs were enrolled. The median IPI was 4 for pts over the age of 60 and the median aa-IPI was 3. Three pts received XRT to areas of bulky disease. At a median follow up of 27 months, the 2 yr DFS and OS was 88%. For patients in arm A and arm B the 2 yr DFS was 90% vs. 86% (figure 1) and the 2 yr OS was 95.2% vs. 81% (figure 2), respectively (P=NS). Two pts discontinued treatment due to adverse events. Grade 3-4 toxicities include neutropenia (28%), fatigue (16%), diarrhea (6%), DVT (3%), rash (3%), febrile neutropenia (3%). Related grade 1-2 toxicities include hypothyroidism (15%) and rash (54%).

Conclusions

Len as maintenance therapy demonstrates clinical activity following standard chemotherapy and improves DFS and OS in DLBCL patients with high risk prognostic features as compared with historical controls. This trial is registered with NCI- #NCT00765245

Disclosures:

Reddy: Celgene: Research Funding. Off Label Use: lenalidomide in diffuse large B cell lymphoma. Park:TEVA: Research Funding; Seattle Genetics, Inc.: Research Funding.
Multiple Myeloma Cells Express IL-17A Creating An Autocrine Loop: An Attractive Therapeutic Target
Authors: Prabhala Rao H. , Fulcinitti Mariateresa , Pelluru Dheeraj , Prabhala Harsha K. , Rashid Naim , Sperling Adam , Nanjappa Puru , Lee Saem , Negroiu Andreea , Lazo-Kallanian Suzan , Munshi Aditya , Richardson Paul G. , Daley John , Anderson Kenneth C. , Munshi Nikhil C.
Published: October 2019

We have previously demonstrated that Th17 cells, which produce IL-17A, are significantly elevated in peripheral blood and bone marrow (BM) of patients with Multiple Myeloma (MM) and IL-17A promotes MM cell growth and survival, both in vitro and in vivo via IL-17A receptor. We have recently evaluated and observed that anti-IL-17A monoclonal antibody (mAb) significantly inhibited MM cell growth in vitro, while IL-17A induced proliferation of MM cells compared to control. We have also observed significant down-regulation of IL-6 production by anti-IL-17A mAb in MM-BMSC co-culture. Importantly, the administration of anti-IL-17A mAb weekly for 4 weeks in the SCIDhu model of human myeloma, where MM cells grow within the human microenvironment in mice led to a significant inhibition of tumor growth compared to the control mice. This remarkable activity of anti-IL17 mAb raised the question of whether the myeloma cells themselves are a possible source of IL-17. In this study, we used transcriptome sequencing (RNA-Seq) data to evaluate the expression of IL-17A in primary CD138+ myeloma cells (N=17) compared to normal plasma cells (NPC) (N=5). Whereas none of the NPCs expressed IL-17A, it was significantly over-expressed in majority of MM cells. In addition, these data also showed that the expression of other IL-17 family members (IL-7B, C, D, E & F) and Th17-associated pro-inflammatory cytokines (IL-21, IL-22 & IL-23) were not significantly elevated in primary myeloma cells compared to normal donor plasma cells. We further validated these observations by IL-17 immunoblot showing IL17 expression in all MM cell lines and 10 out of 14 primary patient MM cells; confirmed IL-17 expression in MM cells by quantitative RT-PCR, and flow cytometry and by immuno-histochemistry and confocal microscopy. We observed that IL-17 knock down by IL-17-specific siRNA inhibited MM cell growth as well as their ability to induce IL-6 production in co-cultures with BMSC. Finally, expression profile data from 172 uniformly treated patients showed that patients with lower IL-17A expression had superior overall survival compared to those with higher expression. These data confirms that MM cells express IL-17 and targeting it with a mAb will abrogate the autocrine loop making it an attractive therapeutic target.

Disclosures:

No relevant conflicts of interest to declare.