bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Alterations and Their Clinical Implications in Older Patients with Acute Myeloid Leukemia
Authors: Tsai Cheng-Hong , Hou Hsin-An , Chou Wen-Chien , Lin Chien-Chin , Chen Chien-Yuan , Tang Jih-Luh , Yao Ming , Li Chi-Cheng , Huang Shang-Yi , Ko Bor-Sheng , Hsu Szu-Chun , Lin Chien-Ting , Wu Shang-Ju , Tsay Woei , Tien Hwei-Fang
Published: October 2019

Introduction

Risk-stratification of patients with acute myeloid leukemia (AML) can not only improve treatment response, but also reduce side effects of the treatment, especially in the elderly. A number of patient-specific and leukemia-associated factors are related to the poor outcome in older patients with AML. However, comprehensive studies regarding the impact of genetic alterations in this group of patients are limited.

Methods and Materials

A total of 500 adult patients with newly diagnosed de novo AML who had enough bone marrow cryopreserved cells for analysis at the National Taiwan University Hospital were enrolled consecutively. We compared the clinico-biological features, cytogenetics and molecular gene mutations between patients aged 60 years or older (n=185) and those younger (<60 years, n=315).

Result

Among older patients, those received standard intensive chemotherapy had a longer overall survival (OS) than those treated with palliative care. Compared with younger patients, the elderly had a higher incidence of poor-risk cytogenetic changes, but a lower frequency of favorable-risk cytogenetics. The median number of molecular gene mutations at diagnosis was higher in the elderly than the younger. Older patients had significantly higher incidences of PTPN11, NPM1, RUNX1, ASXL1, TET2, DNMT3A, and P53 mutations but a lower frequency of WT1 mutations. In multivariate analysis for OS among the elderly who received standard intensive chemotherapy, high WBC >50,000/μL at diagnosis, RUNX1 mutations, DNMT3A mutations, and P53 mutations were independent worse prognostic factors, while the presence of NPM1 mutations in the abcence of FLT3/ITD mutations was an independent good prognostic factor. The frequency of acquiring one or more adverse genetic alterations was much higher in older patients than younger ones. Further, the pattern of gene mutations could divide older patients with intermediate cytogenetics into three groups with significantly different complete remission rates, OS, and disease-free survival.

Conclusion

Older AML patients frequently harbored high-risk cytogenetics and gene mutations, and had poorer prognosis. Integration of cytogenetics and molecular alterations could risk-stratify older patients into groups with significant different outcomes. For those patients with poor prognosis under current chemotherapy, novel therapies, such as demethylating agents or other targeted therapies may be indicated.

Disclosures

Tang: Novartis: Consultancy, Honoraria.
CD163+ Tumor Associated Macrophages Predict Inferior Outcome in Patients with Diffuse Large B-Cell Lymphoma Treated with R-CHOP
Authors: Zeng Hong , Wei Yongqiang , Xie Muchen , Yang Lei , Huang Fen , Wei Xiaolei , Hao Xiaoxiao , Wei Qi , Feng Ru
Published: October 2019

Tumor-associated macrophages (TAMs), as the major component in tumor microenvironment, have been reported to correlate with the prognosis in diffuse large B-cell lymphoma (DLBCL). However due to the different subtypes of TAMs and the given therapy regimen, the prognostic value of TAMs in DLBCL patients remains controversial. To explore the prognostic significance of different TAMs subtypes in DLBCL patients treated with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone), we retrospectively analyzed 77 de novo DLBCL patients in present study.

TAM markers including CD68 and CD163 were evaluated by immunohistochemistry. The cutoff value of CD68, CD163 and CD163/CD68 ratio was determined by receiver operating characteristic curve. In a total of 77 patients, CD68 high expression was found in71.4 %( 55/77) DLBCL patients and CD163 high expression in 51.4 %( 40/77 patients. Patients with high CD163 expression were more present with extranodal sites (P =0.049). However other clinical parameters including age, gender, Ann Arbor stage, performance stage, B symptoms, LDH and international prognostic index (IPI) showed no difference in patients with high and low CD68 or CD163 expression (P >0.05). CD68 expression has less effect on overall survival (OS) and event-free survival (EFS) (P =0.852, 0.782). However, high CD163 expression showed poor OS and EFS in DLBCL patients (P =0.006, 0.004) and also high CD163/CD68 ratio (P <0.001, 0.001). The multivariate analysis revealed that the high CD163 expression and CD163/CD68 ratio remained unfavorable factors for both OS (P =0.094, 0.002) and EFS (P =0.044, 0.016) independent of IPI. In conclusion, our date suggest that high CD163+ TAMs in tumor microenvironment may imply poor outcome in DLBCL patients treated with R-CHOP, but not CD68+ TAMs.

Disclosures

No relevant conflicts of interest to declare.
Aspirin-Platelet Sensitivity in Patients with Essential Thrombocythemia: Role βfibrinogen G-455-a Gene Polymorphism
Authors: Cacciola Rossella Rosari , Cacciola Gentilini Elio , Cacciola Emma
Published: October 2019

Essential thrombocythemia (ET) is a myeloid neoplasm characterized by platelet activation and thrombotic risk. Aspirin (ASA) is the standard therapy to normal platelet hyperaggregation and to prevent the thrombosis. It is reported that thrombocythaemic patients are ASA insensitive. It is debated if inherited thrombophilia increases the thrombocythemic platelet activation and, hence, the ASA platelet insensitivity. Therefore, we evaluated βFibrinogen G-455-A gene polymorphism, as thrombophilic molecular mutation associated with increased platelet aggregation, platelet count, β-thromboglobulin (β-TG) and platelet factor 4(PF4) as markers of platelet activation, fibrinogen (Fg), platelet functional activity (PFA), as indicator of ASA platelet sensitivity, clot formation time (CFT) and the maximum clot firmness (MCF), as indicators of aspirinated platelet contribution to clot firmness. We studied 40 patients (24 men, 16 women; mean age 56 years, range 37-77) with ET according to WHO criteria. The mean duration of disease was 11 years. All patients were on ASA 100 mg once daily. The βFibrinogen G455-A genotype was determined using a commercialized polymerase chain reaction kit with sequence-specific primers. Platelets were measured by automated analyzer. β-TG and PF4 were determined by ELISA. PFA, CFT and MCF were measured by Platelet Function Analyzer (PFA-100) and by ROTEM delta, respectively. All patients had heterozygous βFibrinogen G455-A. The mean platelet count was 441±72x109/L. All patients had normal Fg (244±47 mg/dl) high β-TG and PF4 (244±15 IU/ml vs 20±11 IU/ml and 162±56 IU/ml vs 6±2 IU/ml, respectively) (p<.0001 and p<.0001, respectively), prolonged C/EPI closure time (CT, unit: s, n.v. 84-160 s) (252±48 s), normal CFT (CFT, unit: s, n.v. 30-110 s) (50±7s) and MCF (MCF, unit: mm, n.v. 50-72 mm) (71±2 mm). These findings suggest that βFibrinogen G-455-A gene polymorphism does not affect the clonal platelet hyperaggregation in ET.

Disclosures

No relevant conflicts of interest to declare.
A Rapid and Simple Assay for the Determination of Adamts-13 Activity
Authors: Binder Nikolaus B. , Griffiths Margaret , Vetr Helga
Published: October 2019

In recent years the determination of ADAMTS-13 activity in plasma has become an important diagnostics test in the differentiation of Thromobocytopenic Purpura (TTP) patients from those with other forms of Thrombotic micoangiopathies (TMA). Moreover, it is also useful for monitoring the effectiveness of treatment in such patients.

The standard laboratory test for ADAMTS-13 Activity is an ELISA assay, which has an assay which requires 4 hours to process. This assay tends to be performed by specialized laboratories, so samples are usually batch tested. Therefore for a physician to receive a patient ADAMTS-13 activity result can range from 24 hours to as much as one week. This delay in reporting could have a negative impact on patient care.

We have developed a rapid and simple assay which would allow the physician to have an initial reportable result for ADAMTS-13 Activity within 1 hour from the blood draw.

The assay is based on flow through technology, which allows for simple, rapid processing. Plasma samples are pre-incubated with a substrate which is cleaved by ADAMTS-13 present in the plasma. This pre-treated sample is applied to the single test unit which contains a membrane with immobilized antibodies specific for the cleaved substrate. As the sample flows through the membrane into an absorbent pad the cleaved substrate binds to capture antibody. A secondary biotinylated antibody is used to detect the membrane bound complex. Using a Streptavidin-gold conjugate a red colour is produced. The colour intensity is proportional to the level of ADAMTS-13 activity in the plasma sample and compared with a colour standard card, allowing semi-quantitative analysis of the patient sample.

The assay can be performed at room temperature without specialized laboratory equipment and minimal training is needed to perform the test. The assay processing time is under 30 minutes. The test is designed for evaluation of a single patient sample, with a positive control run in parallel.

The designed assay cut-off value of 10% ADAMTS-13 Activity is clinically relevant for differentiation of TTP patients. A 100% correlation was observed when TTP patient samples (n=20) were run in the TECHNOZYM ADATMS-13 ELISA and this assay, as summarized in the figure below.

This rapid and simple assay is versatile and suitable as a cost effective screening assay enabling quick turn-around for ADAMTS-13 activity reporting.

Figure Shows a graphically representation of the method comparison between single test and ELISA results. Figure. Shows a graphically representation of the method comparison between single test and ELISA results.

Disclosures

Binder: Technoclone Herstellung von Diagnostika und Arzneimitteln GmbH: Employment. Griffiths:Technoclone Herstellung von Diagnostika und Arzneimitteln GmbH: Employment. Vetr:Technoclone Herstellung von Diagnostika und Arzneimitteln GmbH: Employment.
The Embt Risk Score Predicts the Outcome of Patients with Leukaemia Receiving Single Umbilical Cord Blood Transplantation
Authors: Zhu Xiaoyu , Tang Baolin , Zheng Changcheng , Liu Huilan , Geng Liangquan , Tong Juan , Zhang Lei , Zhang Xuhan , Yao Wen , Song Kaidi , Wang Xingbing , Ding Kaiyang , Sun Zimin
Published: October 2019

Aim The European Group for Blood and Marrow Transplantation (EBMT) risk score has been implemented as an important tool to predict patient outcomes following allogeneic haematopoietic stem cell transplantation (allo-HSCT). However, to our knowledge, this score has never been applied in cases of single umbilical cord blood transplantation (sUCBT). The aim of this study was to assess the capacity of the EBMT risk score in predicting the outcomes of patients with leukaemia receiving sUCBT. Methods We retrospectively analysed 218 consecutive patients with leukaemia who received sUCBT at our centre between February 2011 and December 2015. Sixty-eight of the 218 patients had AML, 137 had ALL, 10 had CML, 2 had mixed phenotype AL, and only one patient had plasma cell leukaemia. The median age was 12 years (range, 1-46 years), and the median weight was 40 kg (range, 10-100 kg). All of the patients received an intensified myeloablative conditioning regimen with cyclosporine A (CsA) and mycophenolate mofetil (MMF) as graft-versus-host disease (GVHD) prophylaxis. All of the patients were followed until 31 March 2016, with a median follow-up of 10.7 months (range, 0.5-58.9 months). Results The overall survival (OS) and leukaemia-free survival (LFS) of the entire cohort were 66.8% and 55.4%, respectively, whereas the cumulative incidences of relapse rate and non-relapse mortality (NRM) were 22.4% and 18.0%, respectively. In the univariate analysis, a higher EBMT risk score was associated with worse OS, worse LFS, lower NRM and higher relapse rate, ranging from 80.2%, 74.5%,7.5% and 18.0%, respectively, for patients with a score of 1 to 21.8%, 17.0%, 27.3% and 55.7%, respectively, for patients with a score of 5/6. Hazard ratios of the four indexes all steadily increased for each additional score point. Importantly, the prognostic value of the EBMT risk score on OS, LFS, NRM and relapse was maintained in the multivariate analysis. Moreover, considering the importance of allele-level HLA matching, we developed a modified sUCBT-EBMT risk score using HLA matching situations instead of donor type and found that the modified score could also be used as a predictor for patient outcomes following sUCBT. Conclusions The EBMT risk score is a good predictor of outcomes of patients with leukaemia following sUCBT. The modified sUCBT-EBMT risk score can also be used as a pretransplant risk assessment, but this metric still requires further evaluation with a larger cohort.

Disclosures

No relevant conflicts of interest to declare.
Haploidentical Hematopoietic Stem Cell Transplant Compared with Matched Sibling and Unrelated Transplants in Adults with Philadelphia-Negative Acute Lymphoblastic Leukemia in First Complete Remission
Authors: Liu Qifa , Han Lijie , Fan Zhiping , Huang Fen , LI Xuan , Xu Na , Lin Ren , Sun Jing
Published: October 2019

Matched-sibling donor (MSD) and matched unrelated donors (MUD) hematopoietic stem-cell transplantation (HSCT) have been recommended for the adults with acute lymphoblastic leukemia (ALL) in first complete remission (CR1). However, it is not defined for this portion of patients whether HLA haploidentical donor (HID) transplants can achieve equivalent outcomes compared to MSD and MUD transplants. From 2009 to 2014, 379 adult patients with Ph-negative ALL in CR1 underwent HID HSCT (n = 159), or MSD HSCT (n = 135), or MUD HSCT (n = 85). Among HID, MSD and MUD transplants, the 100-day incidences of acute graft-versus-host disease (aGVHD) Grade Ⅲ-IV were 11.0%, 8.4% and 10.6% respectively (P>0.05), and 2-year chronic GVHD (cGVHD) were 38.9%, 28.5% and 35.6% respectively ( P>0.05 ). At 5 years, the incidences of relapse were 19.0%, 24.0% and 23.4% in HID, MSD and MUD transplants respectively (P>0.05). The 5-year transplant related mortality (TRM) of HID was higher than MSD transplants (16.5% vs 10.9%, P = 0.032), and no difference compared with MUD transplants (16.5% vs 18.1%, P > 0.05). However, the 5-year overall survival (OS), disease-free survival (DFS) did not differ among HID, MSD and MUD transplants (OS 70.9%, 72.0%, 65.4% respectively, P > 0.05; DFS 67.3%, 66.0%, 62.0%, respectively, P > 0.05).

Our results indicate that outcomes of adult patients with Ph-negative ALL in CR1 from HID transplants are comparable to MSD and MUD transplants.

Disclosures

Lin: National Natural Science Foundation of China 81400141: Research Funding; National Natural Science Foundation of China 81270647: Research Funding; Science and technology planning project of Guangdong Province 2014B020226004: Research Funding; The project of health collaborative innovation of Guangzhou City 201400000003-4: Research Funding.
Comparison of Disposition and Complications in Geriatric (≥ 65 years) and Non-Geriatric (50-64 years) Cancer Patients Receiving Hematopoietic Stem Cell Transplantation
Authors: Guddati Achuta Kumar , Kumar Gagan , Lewis Robert David , Shapira Iuliana
Published: October 2019

Background:.The disposition and complications of hematopoietic stem cell transplant (HSCT) in the geriatric population has not been extensively studied and compared with younger patients. We have analyzed a database with nationwide representation for outcomes of HSCT in these patients over a 12 year period.

Methods: Data regarding patients who underwent HSCT was extracted from the Nationwide Inpatient Sample (NIS) from 2000 to 2011 using ICD-9-CM codes. HSCT hospitalizations were classified into allogeneic transplantation, autologous transplantation, subsequent hospitalization (s/p transplant) with graft versus host disease (GVHD) and subsequent hospitalization (s/p transplant) with other complications. NIS variables were used to identify in-hospital complications and discharge disposition. Chi square test and Wilcoxon rank test were used to compare categorical and continuous variables respectively.

Results: The proportion of elderly patients (≥ 65 years) receiving autologous transplants and being discharged to nursing homes has increased over the past decade when compared to a younger patient population. Notably, the rate of major complications: mechanical ventilation, tracheostomy and requirement of new dialysis are similar in both the patient populations. The trends from 2000 to 2011 are summarized in Table 1.

Conclusion: Although the rate of transplant-associated complications are similar in geriatric and non-geriatric populations, a higher percentage of geriatric cancer patients who receive HSCT are discharged to nursing homes. These rates have not significantly changed over the past decade (p > 0.05). More medical services and resources will be required to support the growing elderly population undergoing HSCT.

Table 1 Table 1.

Disclosures

No relevant conflicts of interest to declare.
LNK (SH2B3) Deficiency Ameliorates Hematopoietic Stem Cell Defects in Fanconi Anemi
Authors: Balcerek Joanna , Jiang Jing , Jiang Qinqin , Lyu Kaosheng , Greenberg Roger , Tong Wei
Published: October 2019

Fanconi Anemia (FA) is one of the most common inherited bone marrow failure (iBMF) syndromes. Although initially identified over 85 years ago, FA remains a fatal genetic disease. Nineteen FA genes cooperate in a genome stability pathway that is essential for repair of DNA damage and tolerance of replication stress. Cells derived from FA patients are hypersensitive to DNA interstrand crosslink (ICL)-inducing agents, such as Mitomycin C (MMC), and exhibit DNA damage checkpoint and mitosis defects. Mutations in FA genes result in hematopoietic stem cell (HSC) defects, bone marrow failure and cancer predisposition. Importantly, interventions to mitigate HSC defects do not exist, aside from allogeneic bone marrow transplantation (BMT).

HSCs deficient for FancD2, a central component of the FA signaling pathway, are markedly compromised in reconstituting the hematopoietic system in murine BMT models. Remarkably, we found that loss of the adaptor protein Lnk (also called Sh2b3) restores HSC function in FancD2 knockout mice without accelerating neoplastic transformation. LNK negatively regulates HSC self-renewal, in part by attenuating cytokine signaling-activated JAK2 signaling in HSCs (J Clin Invest. 2008;118(8):2832-2844). Fancd2-/-;Lnk-/- (DKO) mice exhibit increased phenotypic HSCs in comparison to wildtype (WT) animals, and DKOHSC function is also largely restored to WT levels in serial transplantation assays. Primary DKOHSC and progenitors (HSPCs) are still sensitive to MMC, indicating that LNK does not play an overt role in ICL repair. Instead, Lnk deficiency notably reduces spontaneous DNA damage and genome instability. This is in agreement with recent studies that reveal a requirement for FA proteins in replication stress, which is a separable function from their role in DNA repair (Cell. 2011;145(4):529-542; Cancer Cell. 2012;22(1):106-116). Strikingly, we demonstrated that Lnk deficiency mitigates replication stress by stabilizing stalled replication forks, and that this effect is dependent upon cytokine signaling.

Together, our data demonstrate that Lnk deficiency ameliorates FA HSPC defects by alleviating replication stress associated DNA damage and genome instability. To our knowledge, this is one of the first examples of in vivo genetic suppression of FA-associated HSPC defects. Our work sheds light on mechanisms underlying the origin of bone marrow failure in FA patients and has implications for new therapeutic strategies to treat FA associated bone marrow failure.

Disclosures

No relevant conflicts of interest to declare.
Osteoclasts Promote Immune Suppressive Microenvironment in Multiple Myeloma: Therapeutic Implication
Authors: An Gang , Acharya Chirag , Feng Xiaoyan , Wen Kenneth , Zhong Mike , Zhang Li , Munshi Nikhil C , Qiu Lugui , Tai Yu-Tzu , Anderson Kenneth C.
Published: October 2019

Purpose: The number and activity of osteoclasts (OCs) are strongly enhanced by myeloma cells, leading to significant bone lesion in patients with multiple myeloma (MM). Mechanisms remain elusive whether myeloma-supporting OCs also induce suppressive immune bone marrow (BM) microenvironment.

Methods: We examined if OCs could modulate T cell-mediated anti-MM immunity; Inhibitory immune molecules expression during osteoclastogenesis; The impact of APRIL on PDL1 expression in MM and the pathway; The impact of Anti-CD38 mAb on osteoclastogenesis and T cell response;

Results: Here, we first show that OCs significantly protect MM cells against T cell-mediated cytotoxicity via direct inhibition of proliferating CD4+and CD8+ T cells. The immune checkpoint molecules PDL1, Galectin-9, HVEM, and CD200, as well as T cell metabolism regulators indoleamine 2, 3-dioxygenase (IDO) and CD38 are significantly up-regulated during osteoclastogenesis. Importantly, levels of these molecules, except CD38, are higher in OCs than MM cells. Anti-PDL1 monoclonal antibody and IDO inhibitor partly overcome OC-inhibited T cell responses against MM cells, confirming their roles in OC-suppressed MM cell lysis by cytotoxic T cells. In addition, Galectin-9 and a proliferation-induced ligand (APRIL), mainly secreted by OCs, are significantly up-regulated during osteoclastogenesis. Galectin-9 specifically induces apoptosis of T cells while sparing monocytes and MM cells. APRIL induces PDL1 expression in MM cells, providing additional immune inhibition by OCs. Moreover, CD38 is significantly up-regulated during osteoclastogenesis. When targeted by an anti-CD38 monoclonal antibody, suppressive T cell function by OCs is alleviated, associated with down-regulation of HVEM and IDO.

Conclusion: Taken together, these results define expression of multiple immune proteins and cytokines in OCs essential for suppressive MM BM milieu. These results further support combination of targeting these molecules to improve anti-MM immunity.

Disclosures

Anderson: Celgene: Membership on an entity's Board of Directors or advisory committees; C4 Therapeutics: Equity Ownership; Gilead: Membership on an entity's Board of Directors or advisory committees; Acetylon: Equity Ownership; Oncoprep: Equity Ownership; Millennuim: Membership on an entity's Board of Directors or advisory committees; Oncoprep: Equity Ownership; Gilead: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Acetylon: Equity Ownership; Millennuim: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; C4 Therapeutics: Equity Ownership; Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees.
Feasibility of a Smartphone-Based Health Coaching Intervention for Patient Self-Management of Nutrition in the Post-Chemotherapy Setting
Authors: Chen Connie E , Mao Alice Y , Murray Jontrice G , Oran Betul , Qazilbash Muzaffar H. , Shah Nina , Ahmed Sairah , Bashir Qaiser , Hosing Chitra , Shpall Elizabeth J. , Steele Joseph R , Champlin Richard E. , Parmar Simrit
Published: October 2019

Introduction. While quality of life (QoL) has become an increasingly important outcome measure following hematopoietic stem cell transplant (HSCT), ongoing support services available to patients post-HSCT patients are limited due to high cost and limited reimbursement. Smartphone devices offer a novel means by which to offer low cost, longitudinal support through the use of virtual health coaches. However there is little existing research on the acceptability and effectiveness of using smartphones to deliver health coaching for patients with malignant diseases after HSCT.

Methods. Patients were recruited in the outpatient setting in the stem cell transplant clinic at MD Anderson Cancer Center to enroll in digital health coaching. Upon enrollment, patients were asked to download a smartphone application (Vida Health), which allowed participants to communicate in real time with a health coach through live video, phone, or text messages. Coaches provided health education, offered ongoing encouragement, and helped patients set small weekly goals to enhance nutrition, physical activity, stress management, and medication adherence. Patients completed the Functional Assessment of Cancer Therapy - Bone Marrow Transplant (FACT-BMT) survey at baseline and 3-month follow up.

Results. A total of 62 patients completed at least 3 months of Vida health coaching. The median age was 56 years (45-72 years) and 41% were women. Time elapsed since HSCT ranged from 3 months to 9 months with primary diagnoses being: Myeloma 50%, Lymphoma 20%, Other Hematologic Malignancy 30%. Primary goals as stated by patients were: 72% optimizing nutrition, 60% stress management and emotional support, 60% remembering to take medications, 45% improving sleep, 20% staying physically active. 68% of patients sent at least one message to coach with an average 29 messages sent from patient to coach during the 3 month coaching period. FACT-BMT scores improved significantly after coaching with a 27% improvement in physical wellbeing (pre-coaching mean 6.1, post-coaching 4.7) and a 53% improvement in functional wellbeing (pre-coaching mean 13.8, post-coaching 21.1). Emotional wellbeing also increased 40% (from mean of 10.3 to 6.2).

Conclusion. These preliminary findings suggest digital health coaching may help post-HSCT patients improve QoL, particularly among physical, functional, and emotional domains. We believe that utilizing novel tools such as smartphone-enabled health coaching may offer an important means by which to continue to support patients' wellbeing outside of the clinic.

Disclosures

Chen: Vida Health: Employment, Equity Ownership. Mao:Vida Health: Employment.
Better Transplant Outcomes with Umbilical Cord Stem Cells in Patients with Dyskeratosis Congenita
Authors: Gupta Ashish O. , Shein Steven L , Dalal Jignesh D
Published: October 2019

Background:Dyskeratosis congenita (DKC) is a rare genetic bone marrow failure (BMF) syndrome of unknown etiology with a multisystem involvement and predisposition for cancer. It presents as a classic triad of dystrophic nails, oral leukoplakia and skin hyperpigmentation. Hematopoietic stem cell transplant (HSCT) has been tried as a treatment option but there is limited data on outcomes in these patients.

Methods:As the disease is rare with high mortality, there have been no prospective studies. Retrospective database analysis is an effective means to evaluate outcomes in this patient population. Our study utilized Pediatric Health Information Systems (PHIS) database, which is an administrative quality-controlled database from 43 not-for-profit children's hospital, to analyze healthcare outcomes in pediatric patients with DKC. ICD-9 code 757.39 was used to identify the disease classification and as the only condition with an indication for stem cell transplant, we isolated the cases that underwent HSCT with that diagnostic code.

Results: A total of 40 patients with DKC were identified who underwent transplant. A higher incidence was noted in Caucasian males with no significant difference in mortality across gender and ethnicity. The median length of hospitalization was 48 days (range 8.5 to 160.5 days) with a mean age of hospital admission at 93 months (95% CI: 70, 117). All the transplants were allogeneic with almost 20% umbilical cord (UCT) stem cell transplants. There was 15% mortality noted in this patient population, with all of deaths occurring prior to 2012. The complication rate (including acute and chronic GVHD as well as graft failure) was noted to be 37.5%, with acute GVHD being most common (20%). Fludarabine, Alemtuzumab, Cyclophosphamide, and Melphalan based conditioning regimen were most commonly used. Cyclosporine (65%), steroids (80%), Mycophenolate mofetil (50%) and Tacrolimus (40%) were commonly used for GVHD prophylaxis. Despite small numbers, significantly lower mortality was noted in UCT as compared to the other allogeneic stem cell transplants (p<0.01).

Conclusion: Limited literature is available on pediatric transplant outcome data in DKC. UCT has better transplant outcomes in DKC patients due to longer telomere lengths of donor stem cells. Larger studies are needed for to compare transplant outcomes from different sources.

Disclosures

No relevant conflicts of interest to declare.
Erythroferrone Regulates Hepcidin Expression Independently of Matriptase 2
Authors: Kautz Leon , Aschemeyer Sharraya , Gabayan Victoria , Ganz Tomas , Nemeth Elizabeta
Published: October 2019

Introduction: The iron-regulatory hormone hepcidin regulates the body iron stores and its expression is repressed when erythropoietic activity intensifies to meet the iron requirements for erythropoiesis (e.g. during anemia). Under the influence of erythropoietin (EPO), the hormone erythroferrone (ERFE) is secreted by erythroid precursors in the bone marrow and the spleen, and suppresses hepcidin synthesis to facilitate the recovery from anemia. However, the mechanism by which ERFE suppresses hepcidin is unknown. In contrast with forms of anemia in which hepcidin is suppressed, patients with mutations in transmembrane serine protease 6 (TMPRSS6) have iron-refractory iron deficiency anemia (IRIDA) but increased hepcidin production despite a severe anemia and elevated EPO levels. Recently, it has been suggested that matriptase-2 activity facilitates ERFE-mediated suppression of hepcidin. We therefore investigated the potential crosstalk between ERFE and Matriptase 2.

Methods: We first measured serum ERFE concentration in Tmprss6-/- mice. To assess the contribution of ERFE to the phenotype of Tmprss6-/-mice, we next generated Tmprss6-/-mice with disrupted Erfe (Erfe+/- Tmprss6-/-; Erfe-/- Tmprss6-/- and Erfe+/+ Tmprss6-/-). To determine whether ERFE requires TMPRSS6 to regulate hepcidin production, we treated freshly isolated hepatocytes from wild-type (WT) or Tmprss6-/- mice with conditioned medium from cells expressing recombinant ERFE or not.

Results: While wild-type mice have undetectable plasma ERFE (below the 500 pg/ml limit of detection), plasma ERFE concentration was elevated in Tmprss6-/- to levels comparable to those of WT animals 24 hours after phlebotomy (~3 ng/ml) but was lower than ERFE levels in thalassemic mice (~10 ng/ml). Ablation of Erfe in Tmprss6-/- mice did not result in any change in hematological parameters, hepcidin expression and iron levels compared to Tmprss6-/- animals at 6 weeks of age. However, treatment of WT and Tmprss6-/-hepatocytes with ERFE resulted in a comparable suppression of hepcidin mRNA expression.

Conclusion: Although matriptase-2 may dampen the BMP signaling under the influence of EPO, it is not part of the ERFE signaling pathway.

Disclosures

Ganz: Intrinsic Lifesciences: Other: shareholder and scientific advisor; Merganser Biotech: Other: shareholder and scientific advisor; Silarus therapeutics: Other: shareholder and scientific advisor; Keryx Biopharmaceuticals: Consultancy. Nemeth:Intrinsic Lifesciences: Other: shareholder and scientific advisor; Merganser Biotech: Other: shareholder and scientific advisor; Silarus therapeutics: Other: shareholder and scientific advisor.
Differentiating Thrombotic Microangiopathies Based on Laboratory Tests Other Than ADAMTS13 Using Machine Learning Technology
Authors: Koh Youngil , Lee SuYeon , Yun Hong-Seok , Yoon Sung-Soo , Kim Inho , Kim Byung-Su , Oh Doyeun
Published: October 2019

Introduction: ADAMTS13 activity level is crucial for differentiating thrombotic microangiopathies. However, ADAMTS13 testing is not readily available at site in many parts of the world. Hence, we developed an innovative algorithm that allow differentiation of thrombotic thrombocytopenic purpura (TTP) from other TMA's based on laboratory results other than ADAMTS13 using machine learning.

Methods: Two hundred- eight adult patients with either TTP (N=64) or TMA other than TTP (N=144) (ADAMTS13 cutoff level of 10%) were classified using three machine learning techniques (decision tree, random forest, and neural network), using a set of easily measured 19 clinical variables such as fever, Hb, ALT and so on. Basically, each clinical variable is not correlated with TTP (Absolute values of correlation coefficients are lower than 0.5), so we applied machine learning algorithms. First, we divided patient data into three parts, train, test and validation set. And then, we applied these 3 machine learning techniques, decision tree, random forest and neural network. Principal component analysis was also performed.

Results: As a single variable, platelet count, BUN and total bilirubin were the most important three variables that are predictive of differentiating TTP from other TMA's with accuracy of 82%. Random forest method increased accuracy to 85% and precision, and recall statistic is 0.828, and 0.832, respectively. Neural network did not do better without optimization than random forest method.

Conclusion: Machine learning technology seems promising in differentiating TTP from other TMA's if ADAMTS13 value is not available. These algorithms could support the physician in tailoring the management of TMA.

Correlation coefficient in our study Correlation coefficient in our study Scheme of Random Forest method used in our study Scheme of Random Forest method used in our study

Disclosures

Lee: SamsungSDS: Employment. Yun:Samsung SDS: Employment.
The Transcription Factor PU.1 Controls a Reversible Differentiation Program in Acute Myeloid Leukemia
Authors: McKenzie Mark D , Ghisi Margherita , Cimmino Luisa , Erlichster Michael , Oxley Ethan P , Liu Cynthia , Witkowski Matthew T , Liu Grace , Dakic Aleksandar , Simankowicz Emilia , DiRago Ladina , Metcalf Donald , Nutt Stephen L , Wall Meaghan , Ritchie Matthew E , Zuber Johannes , Dickins Ross A
Published: October 2019

Background: Acute myeloid leukemia (AML) is an aggressive malignancy characterized by clonal expansion of transformed myeloid precursors that fail to differentiate into mature cells. Since myeloid lineage maturation curbs self-renewal and is considered irreversible, engaging this process in AML is an attractive therapeutic strategy.

Results: Normal myeloid differentiation requires the transcription factor PU.1 (SPI1), which is functionally compromised in several AML subtypes and is directly inhibited by the recurrent fusion oncoproteins AML1-ETO and PML-RARA. To examine the importance of PU.1 suppression in AML maintenance in vivo, we have combined RNAi-mediated PU.1 inhibition with p53 deficiency to drive highly aggressive AML in mice. Using these models we find that restoring endogenous PU.1 activity in established AML in vivo is sufficient to trigger robust transcriptional, immunophenotypic, and morphological differentiation of leukemic blasts, yielding polymorphonuclear, neutrophil-like cells. Maturation of AML is associated with significant loss of cell viability and yields sustained disease clearance in vivo. Although PU.1 restoration is potently anti-leukemic, remarkably we find that subsequent suppression of PU.1 in mature neutrophil-like cells reverts them to a transformed state within several days. While mature AML-derived cells are slower to form blast colonies in methylcellulose cultures, their clonogenic frequency is only reduced four-fold relative to AML blasts suggesting highly efficient de-differentiation.

Conclusions: These results demonstrate that triggering myeloid differentiation can effectively resolve a p53-deficient model of treatment resistant AML, but also identify a previously unrecognised ability of AML cells to bidirectionally transition between transformed and differentiated states based on the activity of a single transcription factor. Our findings challenge the concept of 'terminal differentiation' in AML and highlight the importance of therapeutically eradicating leukemia cells at all stages of myeloid lineage maturation.

Disclosures

No relevant conflicts of interest to declare.
Blood Transfusion in Adult Patients with Sickle Cell Disease: Incidence of Alloimmunization
Authors: Master Samip , Ong Menchu , Mansour Richard Preston
Published: October 2019

Background:

Chronic red blood cell transfusion has been proven to be effective in prevention of strokes, silent cerebral infarcts, acute chest syndrome, recurrent priapism and in pregnancy. The use of regular transfusions to mitigate other morbidities of sickle cell disease (SCD) is evolving. In the silent infarct transfusion (SIT) trial in children, chronic transfusion lead to a significant improvement in quality of life. Some of the common reasons patient with SCD do not get chronic transfusion is fear of alloimmunization, iron over load and risk of viral infections. We did a retrospective analysis of adult patients with SCD who need chronic blood transfusion to determine the incidence of alloimmunization. At our institute all pediatric sickle cell patients needing chronic transfusion are placed on protocol, receive C, E, and K matched blood, and remain on the protocol until they become adults.

Methods:

We electronically collected data from 180 adult SCD patients who need chronic transfusions and analyzed the data for the number of transfusions received, incidence of allo- immunization and most common antibodies identified.

Results:

A total of 3967 red blood cell transfusions were administered on 180 adult sickle cell disease patients. Twenty five patients developed antibodies (13.8 %). Fifteen out of the 25 had multiple antibodies (60%). The alloantibodies identified were : anti- K(11), anti- E(12), anti- Fya(5), anti-C (4), anti-V (4), anti- S (3), anti-D (2), anti- Jkb (1), anti-Jsa(1) , and anti- Lutherana (3). Two patients had cold and 5 patients had warm autoantibodies.

Conclusion:

The policy to place patients with SCD needing chronic transfusion on protocol to receive C, E, and K matched red blood cells has decreased the alloimmunization rate to 13.8 %. We conclude that, fear of alloimmunization should not preclude physicians from using chronic red cell transfusions to prevent complications in sickle cell disease.

Disclosures

No relevant conflicts of interest to declare.
Prevalence of Chronic Hepatitis B, Hepatitis C and HIV in Adult Patients with Sickle Cell Disease
Authors: Master Samip , Patan Shajadi , Cingam Shashank , Mansour Richard Preston
Published: October 2019

Introduction:

Previous studies have shown that general prevalence rate of Hepatitis B is 0.3%, Hepatitis C is 1.3% and HIV is 0.39% in United States and that these infections are more common in low income groups and African Americans.

We did retrospective analysis of 458 adult patients with SCD to estimate the prevalence of these infections in our large population.

Methods: We retrospectively collected data of 458 adult patients with SCD to calculate prevalence of Hepatitis C, Hepatitis B and HIV. We also collected data of type of SCD and ethnicity.

Results: Out of total 458 adult patient with SCD, 25/458(5.4%) had Hepatitis C, 10/458(2.1%) had Hepatitis B and 4/458(0.87%) had HIV.

In SC type of SCD, prevalence of Hepatitis B was 1/114(0.87%), Hepatitis C was 4/114(3.5%) and HIV was 1/114(0.87%).

In SS type of SCD, prevalence of Hepatitis B was 7/263(2.66%), Hepatitis C was 19/263(7.22%) and HIV 3/263(1.14%).

In Sickle beta thal group, prevalence of Hepatitis B was 1/75(1.33%), Hepatitis C was 2/75(2.66%). No cases of HIV were fund in this group.

Conclusion:

The prevalence of HIV, hepatitis C and hepatitis B is higher in adult patient with SCD compared to general US population. At our institute we screen all patients with SCD for these infections, especially Hepatitis C and refer infected patients for the appropriate treatment.

Disclosures

No relevant conflicts of interest to declare.
Education, Employment, Social Support and Insurance Coverage in Adult Patients with Sickle Cell Disease
Authors: Master Samip , Arnold Connie , Davis Terry , Mansour Richard Preston
Published: October 2019

Introduction:

The average life expectancy of patients with sickle cell disease (SCD) has increased from early 20s well over 50 now. With improving pediatric healthcare, nearly all SCD patients become chronically ill adults, however little is known about these adult patients. This pilot study assessed adult SCD patients' age, education, literacy, employment, marital status, social support, and insurance status.

Methods:

A convenience sample of 100 sickle cell patients 18 and older cared for at academic medical system hematology clinic were enrolled in the study. A research assistant administered a structured interview which included demographic questions (insurance, education, employment, social support, marital status and persons taking care of them at home). PROMIS (Patient-Reported outcomes measurement information system) surveys assessed depression, anxiety and pain.Literacy was assessed using the Rapid Estimate of Adult Literacy in Medicine (REALM-SF) and cognitive ability, using the Callahan 6-Item Screener.

Results: Patients ranged in age from 19 to 66 years (mean age - 33), 100% were African Americans, 52% females. Education was varied - over one third ( 37%) did not graduate from high school, 26% were high school graduates, 14% had some college, and 12% were college graduates. Literacy was extremely low. No patient read at or above a 9th grade level, 99% < 6th grade level and 1% 7th- 8th grade. Only 16% were employed; 45% were married; 31% lived alone and 63% live with family. Almost all patients (98%) had insurance - 61% had Medicaid, 25% Medicare and 12% private insurance. Mean score of 5.77 on Callahan cognitive screener indicating patients did not need to be screened further for cognitive ability.

Conclusion:

Adult SCD patients in our study had very low education, literacy and employment levels. Many patients had inadequate social support. Our findings indicate that despite the fact that almost all patients were insured and theoretically had access to care, they still needed literacy and culturally appropriate information and on- going support to be able to understand and use health information and services to make appropriate health decisions and properly care for themselves.

Disclosures

No relevant conflicts of interest to declare.
Genome-Scale Screen Reveals Genes Required for Lenalidomide-Mediated Degradation of Aiolos By CRL4-CRBN
Authors: Sievers Quinlan , Gasser Jessica , Cowley Glenn , Doench John G , Fischer Eric , Ebert Benjamin L
Published: October 2019

Lenalidomide exerts its therapeutic effects in the malignancy multiple myeloma by facilitating the degradation of the transcription factors Ikaros (IKZF1) and Aiolos (IKZF3) by the CRL4-CRBN E3 ubiquitin ligase. In the following study we utilized a positive selection, genome-scale CRISPR-Cas9 screen in the lenalidomide-sensitive myeloma cell line, MM1S, to further our understanding of the molecular machinery which regulates and is required for lenalidomide-mediated modulation of CRL4-CRBN. The gRNAs demonstrating the greatest enrichment following selection with lenalidomide belong to cereblon, the molecular target of lenalidomide and substrate receptor for the CRL4-CRBN ubiquitin ligase. Additionally, 20 of the top 30 genes highlighted by the screen are implicated in both the positive and negative regulation of cullin-ring ligases (CRLs), emphasizing both the importance of cereblon's E3 ligase function in mediating cell death as well as the equilibrium of forces required for proper function of CRLs. Among these genes were the ubiquitin-donor E2 enzymes UBE2D3 and UBE2G1; despite the relevance of E2s to E3 ligase biology it has been difficult to determine via traditional methods which of the ~35 E2 enzymes are utilized by a given E3 ligase. Here we demonstrate that G1 and D3 fulfill distinct functional roles and cooperate with the CRL4CRBN E3 ligase to ubiquitinate an aiolos reprorter with lysine-48 linkages. In aggregate these findings confirm the power of CRISPR-Cas9 positive selection screening to reveal drug mechanism-of-action and provides a paradigm for the identification and functional characterization of E2-E3 pairings.

Disclosures

Fischer: Novartis: Consultancy.
Detection of MYD88 L265P and Whim-like CXCR4 Mutation in Patients with IgM Monoclonal Gammopathy
Authors: Cao Xinxin , Meng Qi , Cai Hao , Su Wei , Sun Jian , Zhou Daobin , Li Jian
Published: October 2019

Objectives The presence of IgM monoclonal gammopathy can occur in a broad spectrum of diseases, including Waldenstrom macroglobulinemia (WM), various B cell non-Hodgkin's lymphoma (NHL), monoclonal gammopathy of undetermined significance (MGUS), and so on. MYD88 L265P and WHIM-like CXCR4 mutation are highly prevalent in WM. However, the data is lacking in other IgM monoclonal gammopathy related diseases (IgM-RD). Therefore we investigated MYD88 L265P and WHIM-like CXCR4 mutation in various IgM-RD.

Methods Patients with sIFE confirmed IgM monoclonal gammopathy and had enough material for DNA extraction between January 2008 and May 2016 at Peking Union Medical College Hospital were enrolled in this cohort. Bone marrow samples or paraffin-embedded tumor tissues were collected. We performed Real-time allele-specific-polymerase chain reaction (AS-PCR), AS-PCR and Sanger sequencing to explore the presence of MYD88 L265P and WHIM-like CXCR4 mutation.

Results Ninety-three patients (51 male and 42 female patients) were included in this retrospective study. Median age at diagnosis was 62 years (range, 30-84 years). As for the type of light chain, 65 patients were IgM kappa (69.9%), 28 patients were IgM lambda (30.1%). The median level of serum IgM was 7.9g/L (range, 0.6-104.0g/L). There were 32 patients (34.4%) diagnosed with WM, 5 patients (5.4%) diagnosed with smoldering WM (sWM), 38 patients (40.9%) diagnosed with B cell NHL and 9 patients (9.7%) were diagnosed with MGUS. We outlined the main diagnosis, clinical characteristics and MYD88 L265P, WHIM-like CXCR4 mutation of the 93 patients in Table 1. MYD88 L265P mutation was detected in 30/32 (93.8%) patients with WM, 2/5 (40%) with sWM, 4/9 (44.4%) with IgM-MGUS, 6/14 (42.9%) with diffuse large B cell lymphoma (DLBCL), 2/6 (33.3%) with marginal zone lymphoma (MZL), 3/5 (60%) with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL), 1/7 (4.3%) with mucosa-associated lymphoid tissue lymphoma (MALT), 1/2 (50%) with mantle cell lymphoma (MCL), 1/1 patients with primary amyloidosis (AL) and 1/1 patients with Castleman disease (CD). WHIM-like CXCR4 mutation was detected 7/32 (21.9%) patients with WM, 1/5 (20%) with sWM and 1/14 (7.1%) with DLBCL.

Conclusion IgM monoclonal gammopathy is most frequently found in WM, but it contains a broad spectrum of diseases. MYD88 L265P mutation is highly prevalent in WM and IgM MGUS, and the prevalence of MYD88 mutation in IgM related NHL is higher than that in not specified NHL reported in the literature. WHIM-like CXCR4 mutation is common in WM, but rare in other IgM-RD.

Disclosures

No relevant conflicts of interest to declare.