bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clonal Hematopoiesis in Patients with Diamond Blackfan Anemia
Authors: Nash Michelle , Vlachos Adrianna , Wlodarski Marcin W. , Lipton Jeffrey Michael 
Published: November 2020

Background: Diamond Blackfan anemia (DBA) is a rare inherited bone marrow failure syndrome characterized by anemia, congenital anomalies and a predisposition to cancer. Patients usually present during infancy or early childhood, but can also be diagnosed as adults. In the vast majority of cases DBA is due to a mutation in a gene encoding a small or large subunit-associated ribosomal protein (RP) leading to RP haploinsufficiency. In a study of 702 patients enrolled in the DBA Registry (DBAR), the observed to expected ratio for acute myeloid leukemia (AML) was 28.8 and for myelodysplastic syndrome (MDS), 352.1 (Vlachos et al, Blood, 2018). The average age of onset for MDS in the DBA cohort was 26 years, compared to 60-70 years in the general population. Evolving clonal hematopoiesis (CH) with age has been observed as a precursor to MDS, with CH rarely observed in individuals younger than 40 years of age. Thus we hypothesized that the young age at the development of MDS in DBA would be presaged by evolving CH.
Objective: The primary objective was to perform whole exome sequencing (WES) specifically screening for previously reported somatic mutations in 56 genes associated with CH (Jaiswal et al, NEJM, 2014).
Design/Method: A total of 69 samples were analyzed from 65 patients, mostly targeting patients older than 18 years (median age 30 years). Multiple samples were run on patients who had available samples in the DBAR Biorepository to determine rate of acquisition of mutations. 468 age- and sex-matched healthy controls were made available from GeneDx who performed the WES for the study. We used a threshold for variant calling of minimum 5% with a minimum of 2 variant reads.
Results: Three of the 65 DBA patients (5%) were found to have somatic mutations in STAG1, U2AF1, SF3B1, and DNMT3A at 8, 20, 41, and 70 years, respectively (Table 1). The patient who was 20 years of age had a sample in the DBAR biorepository from when he was age 8 years which was found to have a different somatic mutation (STAG1) than was found at present (U2AF1). This patient did go on to develop MDS at the age of 21 years. In comparison, of the 468 controls, 4 (0.8 %) had a somatic mutation in SF3B1, LUC7L2, DNMT3A, and LUC7L2 at ages 12, 31, 33 and 40 years, respectively.
Conclusion: Patients with DBA show more somatic mutations as compared to controls (p&lt;0.05). This early acquisition of mutations may be the driving force for their developing MDS at an earlier age than that of the general population. Further studies with more sensitive methods are warranted to accurately determine the prevalence of somatic CH mutations and their potential association with the development of myelodysplastic syndrome in these patients.

Disclosures
No relevant conflicts of interest to declare.
This Is a Title: Single-Center Data Report of Clhag Regimen for R/R-AML Patients
Authors: Jiang Chuanyan , Wang Yu , Zou Mengying , Wang Yijing , Wang Jun , Wu Jiafei , Yang Xi , LI Hui 
Published: November 2020

Objectives: To explore the synergistic effect of CLAG combination with HHT in cell experiments and to evaluate the effectiveness and safety of the CLHAG regimen for patients with relapsed/refractory acute myeloid leukemia(R/R-AML).
Methods:In thecell experiments, the Kasumi-1 cell line was treated with a single drug or CLAG combined with HHT. MTT colorimetric method was used to determine the cell growth inhibition rate(GIR). CompuSyn drug cooperation software was used to assess the synergistic effect.In the clinical study,eligible R/ R-AML patients (n=24) were included and CLHAG regimen (cladribine 5mg/m2, d1-5, arabinoside cytosine 2g/m2 q12h, d1-5, homoharringtonine 2mg/m2, d1-d5, GCSF 300μg, d0-d5, WBC&gt;20×109/L disuse) was performed.The efficacy was evaluated by CR, PR, ORR, PFS, OS, and ED. The safety were evaluated by hematological toxicity and non-hematological toxicity.
Results: The GIR value with a combined treatmentregimenwas higher than the sum that with the single drug treatment, showing thata synergistic effect. Twenty-four patients with R/R-AML were included in the clinical study, including 2 cases of sAML (MDS transformation), 8 cases of primary refractory, and 14 cases of relapse.The total ORR was 70.8%; among them,14 cases achieved complete remission, and 3 cases achieved partial remission. Besides, seven patients underwentallo-HSCTsuccessfully. In this study, all patients were well tolerated, and early death was not observed. The IV grade hematological toxicity was the major adverse reaction, and no IV grade non-hematological toxicity occurred.
Conclusions:TheCLHAG regimen is well tolerated and has a high remission rate in R/R-AML patients, and it can be used as a choice of salvage treatment for R/R-AML patients.

Disclosures
No relevant conflicts of interest to declare.
The Role of 5-HT on Proplatelet Formation and Thrombopoietin Production
Authors: Yang Mo , Liang Enyu , Ye Jieyu , Chong Beng H , Li Liang 
Published: November 2020

Background: Our previous work confirmed that serotonin (5-HT) promotes the proliferation of hemopoietic stem cells and megakaryocytes (Yang M et al, Stem Cells, 2007; 2014). However, the mechanisms remain indefinite.
Methods: Q-PCR, Flow Cytometry, Western Blot, or Immunofluorescence microscope were used in the receptor and TPO study. MTT/CCK-8, Proplatelet assay, and Flow Cytometry were also used in cell proliferation and apoptosis study. The relationship between 5-HT and TPO was studied in a traumatic stress mice model.
Results: In-vitro study, there was a stimulating effect of 5-HT on proplatelet formation in human bone marrow megakaryocytes. Human BM MK progenitors cultured in serum-free medium with either 5-HT (200nM) or TPO (100 ng/ml) had more proplatelet bearing MKs than the control group (5-HT (12.3 ± 5.0)% vs. Control (6.2 ± 3.5)%, P=0.025; TPO (15.6 ± 2.5)% vs. Control, P=0.04; n=4). The 5-HT treatment group showed more mature and more in the final stage MK cells as compared to the TPO group. 5-HT2A, 2B, 2C receptors were detected in the surface of megakaryocytes. The effect of 5-HT on proplatelet formation in MK cells was via 5-HT2 receptors and this effect was reduced by 5-HT2 receptor inhibitor ketanserin. 5-HT acted on cytoskeleton reorganization in MKs via 5-HT2 receptors and ERK1/2 pathway. Using an immunofluorescence microscope with F-actin specific binder rhodamine-phalloidin staining, the polymerized actin level was lower in the control group than the 5-HT group and actin distributed diffusely throughout the cytoplasm. In contrast, the polymerization actin level was higher in the 5-HT group. Adding ketanserin and ERK1/2 inhibitor PD98059 to 5-HT treatment, the fluorescence intensity was correspondingly reduced. Our data also demonstrated that ERK1/2 was activated in MKs treated with 5-HT for 30 minutes. In a traumatic stress mice model, both of 5-HT and TPO were increased, but the increasing of TPO is posterior to 5-HT. After added LX1606, the synthesis inhibitor of 5-HT, 5-HT was reduced markedly, as well as TPO. The expression of TPO mRNA and the production of TPO protein were increased as compared with the control in this model.
Conclusions: This study suggests that 5-HT promotes thrombopoiesis from two aspects: one is the direct effect on megakaryocytes. 5-HT could promote the proplatelet formation from megakaryocytes. The second is the indirect effect by promoting the production of TPO, which is a paracrine secretion to influence thrombopoiesis.

Disclosures
No relevant conflicts of interest to declare.
Chronic Myeloid Leukemia (CML) in Adolescents & Young Adults (AYA): Single Institution 10 Years Experience with 1st Line Imatinib Mesylate (IM)
Authors: Malhotra Hemant , Yadav Ajay , Gupta Naveen , Soni Satyajeet , Singh Jitender Kumar , Malhotra Bharti 
Published: November 2020

Introduction and Aims:
CML is the commonest adult leukemia in India and the disease is diagnosed commonly in the 3rd or 4th decade of life - at least 10 years earlier than in the west, making the disease an important issue in AYA cancers.
The present study reports our 10 year experience (all patients registered after 2005 with minimum of 3 years follow up) with the use of 1st line IM in patients of CML Chronic Phase (CML-CP) in patients aged 15 to 29 years.
Methods:
Patients of CML-CP receiving 1st line IM, aged 15 to 29 were included in the study. Standard demographics; hematological response, molecular responses (RQ-PCR on International Scale) q 6 monthly and toxicity of IM were assessed.
Results
A total of 337 patients of CML-CP were registered, 89 (28.7%) of which were aged between 15 to 29 and were on 1st line IM. These are reported in this study.
Age distribution: 15 to 19 years: 17 (19.1%); 20 to 24: 26 (29.2%); 24 to 29: 46 (51.7%). Male female ratio: 53 (59.5%) to 36 (40.5%).
Sixty one (68.5%) received innovator IM (Glivec) through the GIPAP (Glivec International Patient Assistance Program), 28 (40.5%) generic IM.
At 10 years f/u, 6 (6.7%) patients were lost to f/u, 15 (16.8%) off imatinib (12 sub-optimal molecular response/relapse, and 3 b/o toxicity - 2 hepatitis, 1 renal). Seventy four (74) patients continue on IM, 56 on 400 mg/day and 18 on &gt; 400mg/day. Fifty nine are in deep molecular response, 15 have a bcr/abl between 0.1 to 1.0 but are in complete hematological remission.
Musculo-skeletal toxicity, grade I to II: 11/74 (14.8%), skin toxicity grade I: 27/74 (36.5%) and GI grade I to II: 16/74 (21.6%) were seen.
Discussion and Conclusion
CML is a common cancer in AYA in developing countries. IM, including generic IM, remains the standard 1st line drug for the majority and is effective, well tolerated in most patients. Issues related to treatment-free-remission (TFR), compliance, long-term drug toxicity and fertility need to be studied in this young population.

Disclosures
No relevant conflicts of interest to declare.
Comprehensive RNA Editome Reveals Azin1 As a Functional Regulator in Hematopoietic Stem and Progenitor Cells
Authors: Wang Fengjiao , He Jiahuan , Ma Yanni , Hao Sha , Liu Siqi , Gao Ai , Cheng Hui , Yu Jia , Cheng Tao 
Published: November 2020

RNA editing, adenosine (A)-to-inosine (I), plays a vital role in many biological processes. Our previous study has demonstrated that the hematopoietic stem and progenitor cells (HSPCs) deficient in adenosine deaminase acting on RNA 1 (Adar1), an RNA-editing enzyme, cannot reconstitute the irradiated recipients in vivo and form colonies in vitro (Xufeng R et al, PNAS 2009). However, the overall profile of RNA editome in hematopoiesis has not been established and the underlying mechanism how RNA editing governs the function of HSPCs is poorly defined. In this study, we sorted 12 murine adult hematopoietic cell populations and performed RNA sequencing. We depicted the landscape of RNA editome in hematopoietic cells and identified 30,796 editing sites in total. The dynamic landscape of RNA editome comprised of stage/group-specific as well as house-keeping editing patterns. Notably, antizyme inhibitor 1 (Azin1) was uncovered to be highly edited in HSPCs. To understand whether edited Azin1 was required for functioning of HSPCs, we transduced c-Kit+ HSPCs with the lentivirus carrying Azin1 cDNAs with distinct editing frequencies. c-Kit+ cells transduced with fully edited Azin1 showed enhanced reconstitution, compared to that transduced with partially edited or non-edited Azin1. Specifically, inability of RNA editing in Azin1 blocked the differentiation of hematopoietic stem cells (HSCs) in vivo. Moreover, a similar finding was obtained when Azin1 was knocked down. In conclusion, RNA editing of Azin1 (i) results in amino acid change to induce AZIN1 translocation to the nucleus, (ii) enhances AZIN1 binding affinity for DEAD box polypeptide 1 (DDX1) to alter the DDX1 chromatin distribution, and (iii) changes the expression of multiple hematopoietic regulators to ultimately promote HSPC differentiation. This work provides a valuable resource for studying RNA editing and delineates an essential role of Azin1 RNA editing in HSPCs.

Disclosures
No relevant conflicts of interest to declare.
Anlotinib Shows Potent Antileukemia Effects in B-Cell Acute Lymphocytic Leukemia through the Blockage of Angiogenic Related Pathways
Authors: Chen Qiuling , Jiang Yuelong , Chen Qinwei , Liu Long , Xu Bing 
Published: November 2020

Acute lymphoblastic leukemia (ALL) derives from the malignant transformation of lymphoid progenitor cells with ~85% being originated from B-cell progenitors (B-ALL). Despite fairly good prognoses for most pediatric B-ALL patients, the outcome is fatal in over 50% of adult patients who have a recurrent or progressive disease and lack of effective therapeutic approaches. Therefore, novel treatment strategies with high efficacy and low toxicity are an unmet need for B-ALL patients, especially those with relapsed or refractory status.
Angiogenesis is a process of new vessel formation that requires the participation of multiple proangiogenic factors (e.g., VEGF, PDGF, and FGF) and their corresponding receptors (e.g., VEGFR, PDGFR, and FGFR). Angiogenesis, a well-established feature of solid tumors, also contributes to leukemia progression and correlates with the involvement of specific sanctuary sites in ALL, highlighting that the perturbation of angiogenesis would be an attractive approach for ALL treatment. Anlotinib is an oral tyrosine kinase (TKI) inhibitor with a broad range of antitumor effects via the suppression of VEGFR, PDGFR and FGFR. Of importance, anlotinib has been approved for the treatment of advanced lung cancer in China. Here, we evaluated the antileukemia activity of anlotinib in preclinical B-ALL models and its underlying molecular mechanisms.
In this study, we observed that anlotinib significantly blunted the capability of cell proliferation and arrested cell cycle at G2 phase in B-ALL cell lines. Subsequently, we found that anlotinib resulted in remarkably enhanced apoptosis in B-ALL in vitro. To assess the in vivo antileukemia potential, we established a B-ALL patient-derived xenograft (PDX) mouse model and then treated the B-ALL PDX model with anlotinib. As a result, oral administration of anlotinib pronouncedly delayed in vivo B-ALL cell growth and reduced leukemia burden with acceptable safety profiles in this model. As for the mechanism of action, the antileukemia effect of anlotinib was associated with the disruption of the role of VEGFR2, PDGFRb, and FGFR3. Moreover, we revealed that this drug blocked the PI3K/AKT/mTOR/ signaling, a pathway that is linked with angiogenesis and its proangiogenic regulators, including VEGFR2, PDGFRb, and FGFR3.
In aggregate, these results indicate that anlotinib is a potent antitumor agent for the treatment of B-ALL via the inhibition of angiogenic relevant pathways, which provide a novel potential treatment intervention for patients with B-ALL who have little effective therapy options.

Disclosures
No relevant conflicts of interest to declare.

OffLabel Disclosure:
Anlotinib originally designed by China is a novel orally active multitarget inhibitor that is evaluating in clinical trials against multiple solid tumors.
Efficacy and Safety of Lenalidomide, Rituximab Combined with Second-Line Chemotherapy (R2-chemo) in Patients with Relapsed/Refractory Diffuse Large-B Cell Lymphoma
Authors: Xiao Fei , Huang Honghui , Hua Yanni , Fang Yi , Zhang Minyue , Hou Jian 
Published: November 2020

Objective: Outcome of patients with relapsed/refractory (r/r) diffuse large B-cell lymphoma (DLBCL) remains poor, highlighting the need for novel treatment regimens. Lenalidomide is an immunomodulatory drug that could reverse rituximab refractoriness in lymphoma patients. This study aimed to explore the efficacy and safety of lenalidomide, rituximab combined with second-line chemotherapy (R2-chemo) in patients with r/r DLBCL.
Methods: In this study, r/r DLBCL patients received R2-chemo as second-line salvage therapy. The treatment plan was as follows: lenalidomide 10mg/day, day 1-14; rituximab 375mg/m2, day 1; chemo-regimen in combination with R2 as per Investigator's discretion. The overall response rate (ORR), complete remission rate (CR rate), duration of remission (DOR), disease-free survival (PFS) and overall survival (OS) were evaluated.
Results: As of Aug 1, 2020, 15 patients with r/r DLBCL were enrolled in this study with median age 65 years (range, 49 to 74 years), and 60% female. 20% (3/15) was transformed DLBCL, 33.3% (5/15) was relapse and 46.7% (7/15) had refractory disease. All received at least 1 prior line of immune-chemotherapy including rituximab, and 1 case was recurrence after autologous hematopoietic stem cell transplantation. 6.7% (1/15) had double-hit lymphoma, 66.7% (10/15) had MYC or BCL-2 high expression by IHC, of whom 26.7% (4/15) had co-expression of the MYC and BCL2. Cell of origin features using the Hans algorithm: 26.7% (4/15) were GCB, and 73.3% (11/15) were non-GCB. The chemo-regimens in combination with R2 were as follows: 5 cases of GEMOX or GCD, 7 cases of DICE or ICE, 2 cases of DHAP, 1 case of CHOPE regimen. As of the cutoff date, 13 cases were evaluable for efficacy. The ORR (CR+PR) was 61.5%, with 53.8% achieving a CR, and the median DOR was 9.7m (range, 2.9 to 23.7m). With a median time to follow-up of 15.3m (95%CI, 12.2 to 18.4m), median PFS and OS had not yet reached. The estimated 2-year PFS and OS rates were 53.8% and 75.2%, respectively. The most common treatment-related adverse event was hematological toxicity, and no treatment-related deaths occurred.
Conclusion: The R2 combined with second-line salvage chemotherapy regimen has a significant efficacy in the treatment of patients with r/r DLBCL, and can be well- tolerated with no significant additive toxicities. The efficacy and safety of this regimen and the choice of chemo-regimen as an optimal combination partner for R2 need to be further verified by prospective clinical studies with a larger sample size.
Figure

Disclosures
No relevant conflicts of interest to declare.
Crispra: A Promising Tool for β-Thalassemia Treatment
Authors: Jia Xiaoqing , Yao Qi , Li Hui , Chen Jieping 
Published: November 2020

Induction of hemoglobin γ expression is a reliable strategy to treat β-thalassemia. Gene editing using CRISPR/Cas9 technology has been widely used. However, application in vivo is limited due to the uncertainty on genomic cleavages of Cas9. In contrast, CRISPR/Cas9-based gene activation (CRISPRa) can only locate genomic locus but not interrupt sequence. Here, we use SAM system of CRISPRa to locate and activate HBG1 and HBG2, exploring the great potential of CRISPRa for β-thalassemia treatment.
WWe designed 8 single-guide RNAs (sgRNAs) online and cloned into vector SAM V2, which fused dCas9 and VP64. To test the over-expression efficiency, vector containing sgRNA and MPH (fused HSF1, p65 and MS2) were transfected into 293T cell. After 72h transfection, 293T cells were collected. Q-PCR data showed that two sgRNAs were excellent on activating HBG expression with over 1000-fold increase.
WTo test the activating function in hematological cell and the persistence of hemoglobin γexpression, two screened sgRNA were transfected into NB4 cells using lentivirus system. We harvested NB4 cells at different time-point (3 day, 1 week and 2 weeks), and implemented q-PCR assay. HBG expression were increased 50-hold and 1000-hold, respectively. However, the expression were reducing over time and the intrinsic mechanism is unknown.
WThis study set out to increase HBG without interrupt genome using CRISPRa system. This study has found two sgRNA to activate the expression of HBG in 293T cell and NB4 cell. Further research is required to vertify the efficiency of sgRNA in hematopoietic stem cells and prolong the expression time.
Figure

Disclosures
No relevant conflicts of interest to declare.
Impact of Pulmonary Hypertension on in-hospital outcomes amongst Sickle Cell Disease hospitalizations
Authors: Jhaveri Khushali , Patel Raj , Barnett Christopher , Smith Hedy 
Published: November 2020

Introduction: Pulmonary hypertension (PH) is a common and severe complication of Sickle Cell Disease (SCD), and an independent risk factor for mortality. While there is a clear association between SCD and PH, the predictors of PH in SCD and the impact of PH on in-hospital outcomes of SCD hospitalizations remains unknown. In our study, we sought to assess the in-hospital prevalence, predictors, and the impact of PH in SCD hospitalizations.
Methods:
We used the 2016 and 2017 National Inpatient Sample (NIS) to identify all adult hospitalizations with a primary discharge diagnosis of SCD. The sample was then stratified based on the presence or absence of PH. We used the Pearson chi-square test and weighted Student's t-test to analyze categorical and continuous variables, respectively. Multivariate logistic regression analysis was performed to calculate the adjusted odds ratio for various clinical outcomes. SAS was used for the analysis, and the p-value was defined as &lt;0.05.
Results:
We identified n=191,080 weighted hospitalizations for SCD, of which, 5.54% (n=10590) had concomitant PH. Female gender and comorbidities including hypertension, obesity, illicit drug use, hepatic cirrhosis, renal failure, prior venous-thromboembolism, valvular, and congenital heart disease were identified as significant predictors of PH in SCD. PH was associated with increased in-hospital mortality (1.04% vs 0.22%, AOR=2.14, 95% CI 1.15-3.98, p=0.0158). PH in SCD hospitalizations also increased the odds of - acute kidney injury (AKI), need for dialysis, acute respiratory failure (ARF), and need for mechanical ventilation for &gt; 96 hours. The adjusted odds ratio for venous thromboembolism, shock-state, and the need for cardiac catheterization (both right and bilateral) were also higher in patients with PH. Overall hospitalization cost and length of stay increased (7.06±0.16 vs 1.82±0.02 days) in patients with SCD and PH (see table 1).
Conclusion:
In sickle cell disease hospitalizations, PH is independently associated with increased in-hospital morbidity and mortality, with an increased need for in-hospital catheterizations thereby, prolonging the length of stay and overall health care costs. Identifying and treating PH in the SCD population would improve in-hospital outcomes.

Disclosures
No relevant conflicts of interest to declare.