bluuebunny/crossref_metadata_2025 huggingface dataset:
Ixazomib Based Maintenance Therapy for the Patients with Newly Diagnosed Multiple Myeloma in Real-World Practice
Authors: Zhong Liye , Wei Yongqiang , Ping Baohong , Li Xin , Peng Hongling , Li Yongqain , Tian Yuyang , Liu Zelin , Luo Jun , Su Yongzhong , Chen Haifei , He Yanjuan , Li Hongliang , Wu Congming , Sun Zhiqiang , Shi Keqian , Li Xiaoliang , Zhang Qing , Hu Guoyu , Huang Gang , Huang Zhenqian , Wang Weida , Xia Zhongjun
Published: November 2021

Background

The Phase III TOURMALINE-MM4 Trial demonstrated that the oral proteasome inhibitor, Ixazomib maintenance significantly improved progression-free survival (PFS) in patients with newly diagnosed multiple myeloma (NDMM) not undergoing ASCT. However, patient recruitment for clinical trials is highly selective so the extent to which study populations represent real-world patients is unclear. Currently, data validating the use of ixazomib maintenance in clinical practice in Chinese landscape is scarce.

Methods

we conducted a retrospective, observational study of NDMM patients meeting International Myeloma Working Group (IMWG) criteria, who received ixazomib-based maintenance therapy. Data was collected from 26 medical centers in southern China. Enrollment was stratified by risk stage, transplant status and maintenance regimens. The characteristics, toxicities and survival outcomes were recorded.

Results

We included 105 NDMM patients who received ixazomib-based maintenance treatment. Mean age at diagnosis was 59.82 ± 9.80 years, with 62.9% male patients. MM International Staging System (ISS), stage 3 was the most common with 49.04% of patients; 62.86% had an Eastern Cooperative Oncology Group performance score ≥2, and 81.9% with concomitant diseases. By the end of over 70 months follow-up since initial diagnosis, median PFS was 69.45 (95%CI: 31.42-107.48) months, and median OS was not achieved yet. We explored potential influencing factors of progression at the post-maintenance evaluation with a crude model using logistic regression. Difference in PFS was not statistically significant by status at pre-maintenance response, transplantation, maintenance therapy regimens (single/two/three-agent) and dexamethasone in maintenance regimens (p >0.05). Eight patients (7.62%) had grade≥3 treatment-emergent adverse events (TEAEs); 5.7% discontinued treatment because of TEAEs. Common any-grade TEAEs included fatigue (18.1%), diarrhea (16.19%), and peripheral neuropathy (11.43%).

Conclusion

Although there are certain differences in maintenance treatment options for newly diagnosed multiple myeloma patients in real-world practice, the results of our retrospective study support that the ixazomib-based maintenance therapy was effective with acceptable toxicity and single-agent ixazomib maintenance provided enough clinical benefit compared to two or three agents.

Keywords: multiple myeloma, ixazomib, maintenance therapy, real-world practice

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Clinical and Molecular Biology Analysis of Patients with Mantle Cell Lymphoma Resistant to BTK Inhibitor
Authors: Wang Lingli , Yang Ping , Zhang Weilong , Wang Jing , Jing Hongmei
Published: November 2021

Background. BTK inhibitor(BTKi) has transformed the therapeutic landscape for mantle cell lymphoma(MCL). Results of clinical trials demonstrate the efficacy of BTKi to salvage patients with MCL progression, however, mechanism of MCL patients resistant to BTKi is still lack of study.

Aim In order to describe the clinical and molecular characteristics of BTK inhibitor resistance and guide the clinical rational application of BTK inhibitor.

MethodsWe retrospectively evaluated the 19 consecutive patients(relapsed/refractory MCL ,n=15;Primary MCL,n=4) who received a BTKi(Ibrutinib, n = 13; Zanubrutinib, n = 2; Orelabrutinib, n=4;Acalabrutinib,n=1) ,while the MCL still progressed.Fully detailed information of patients, disease characteristics, treatments and the next generation sequencing collected.

Results Median time of taking BTKi and median time of BTKi efficiency were 10 months and 5 months, respectively. Median progression-free survival (PFS) and median overall survival after BTKi initiation were 14 months and 21 months, respectively. Using deep targeted next generation sequencing, we performed somatic mutation profiling from 19 MCL patients, top 10 genes including ATM(47%)、TP53(47%)、KMT2D(32%)、NOTCH2(26%)、BTK(16%)、NSD2(16%)、BIRC3(11%)、CCND1(11%)、NOTCH1(11%)、KMT2C (11%).B cell activation, histone modification, inflammatory response to antigenic stimulus genes were increased in patients with mantle cell lymphoma resistant to BTKi according to Kyoto Encyclopedia of genes and Genomes pathways on gene-set enrichment analysis.

Conclusions Demonstration of ATM and TP53 genes mutations and related pathways opens the door for future investigations in BTKi resistance in MCL patients.

Disclosures

No relevant conflicts of interest to declare.
Response Kinetics of Daratumumab-Based Regimens in Patients with Newly Diagnosed or Refractory/Relapsed Multiple Myeloma
Authors: Wang Jiasheng , Arroyo-Suarez Raul , Tse William
Published: November 2021

Background: It is controversial in multiple myeloma (MM) whether early and late responders to therapies have similar clinical outcomes. Daratumumab (DARA) is a human anti-CD38 antibody that has been increasingly used in newly diagnosed MM (NDMM) and relapsed/refractory MM (RRMM). The association between response kinetics to DARA and clinical outcomes remains unexplored.

Methods: Individual-participant data were obtained from phase 3 trials: POLLUX (Dimopoulos, NEJM, 2016), CASTOR (Palumbo, NEJM, 2016), and MAIA (Facon, NEJM, 2019). Patients were divided into early and late response groups based on the median time to the response of interest. Modified PFS (mPFS) and OS (mOS) were calculated from the time of first response of interest. Minimal residual disease (MRD) negativity was defined as less than 10 5 tumor cells by NGS assays.

Results: After a median follow up of 16.1 months, 670 patients achieved a response of very good partial response (VGPR) or better, and 213 achieved MRD negativity. The median time to achieving VGPR or better was similar between NDMM and RRMM (86 vs. 84 days, respectively), while the median time to MRD negativity was longer among NDMM than RRMM (407 vs. 197 days, respectively). Among patients achieving VGPR or better, there was no significant difference of mPFS (HR 1.00, 95%CI 0.69 to 1.44) (fig. a), duration of response (DOR) (HR 1.02, 95%CI 0.68 to 1.53) (fig. b), or mOS (HR 0.98, 95%CI 0.54 to 1.75) (fig. c) between early and late responders. In the subgroup analysis, no significant difference of DOR was observed across all prespecified groups, including sex, age, cytogenetic risk groups, lines of previous therapy, types of measurable disease, NDMM vs. RRMM, prior treatment with autologous hematopoietic cell transplant, immunomodulatory drugs, or proteasome inhibitors. Among patients with NDMM achieving MRD negativity, there was no significant difference of mPFS (p=0.66) (fig. d), DOR (p=0.21) (fig. e) or mOS (p=0.87) (fig. f) between early and late responders. However, among patients with RRMM achieving MRD negativity, late responders had significantly longer mPFS (p=0.038) (fig. g) and DOR (p=0.043) (fig. h) than early responders; mOS was not significantly different (fig. i). In the multivariable Cox analysis among patients achieving MRD negativity, only lower baseline LDH level, NDMM, and IgG type MM were independently associated with later response.

Conclusions: For patients with NDMM or RRMM achieving VGPR or better, early and late responders had similar survival; for patients with RRMM achieving MRD negativity, late responders had significantly longer mPFS and DOR. Our data support that in patients who failed to achieve an early or deep response to daratumumab based regimens, therapies should be continued with the goal of achieving ongoing and stepwise improvement of response.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
A Real World Study of Venetoclax Combined with Azacitidine in 64 Chinese Patients Newly Diagnosed Acute Myeloid Leukemia
Authors: Qian Jiejing , Xu Jieyu , Hong Qing , Lou Yinjun , Mao Liping , Xu Weilai , Yang Min , Yu Wenjuan , Meng Haitao , Mai Wenyuan , Ye Xiujing , Zhu Honghu , Jin Jie
Published: November 2021

Background: Venetoclax combined with azacitidine has been demonstrated to have a favorable overall response rate and tolerable safety in acute myeloid leukemia (AML) patients who are unfit for intensive chemotherapy.

Methods: This study retrospectively recruited 64 Adults (≥18 years) with newly diagnosed AML ineligible for intensive chemotherapy who had received at least one cycle of treatment with venetoclax plus azacitidine. The primary endpoint was overall survival (OS). Secondary endpoints were remission rates. Safety was evaluated based on the Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0.

Results: The median age of the enrolled patients was 69 years. The median OS for the cohort was 15.5 months, with 21.2 months for complete responders as opposed to 13.3 months for those who did not achieve a complete response. 39 (60.9%) patients achieved composite complete remission (complete

remission (CR) + CR with incomplete count recovery (CRi)) and 7/64 (10.9%) achieved morphologic leukemia-free state (MLFS) with a median follow-up time of 14.7 months. The median CR+CRi duration was 10.9 months. It is noteworthy that 43/64 (67.2%) patients achieved the best response after one cycle with a median time of 1.2 months. Normal karyotype (P=0.015) was significantly associated with extended OS in multivariate analysis, while higher white blood cell count (P=0.032), lower platelet count (p=0.018) and antifungal drug combination (P=0.013) were predictors of shortened OS in univariate analysis. Common grade 3/4 Adverse Events (AEs) included infection (68%) and hematological AEs consisting of neutropenia (93%), anemia (90%), leukopenia (86%), thrombocytopenia (77%) and febrile neutropenia (52%). The median neutropenia duration was 16 days.

Conclusions: The novel venetoclax-azacitidine combination regimen showed prolonged survival period, promising efficacy, sound and rapid response, and superior tolerance in Chinese patients newly diagnosed AML.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Clinicopathologic Characteristics and Genomic Profiling of De Novo CD5 Positive Diffuse Large B-Cell Lymphoma: A Multicenter Retrospective Study in China
Authors: Sang Wei , Ma Yuhan , Wang Xiangmin , Shen Ziyuan , Xu Linyan , Qiao Jianlin , Xu Kailin , Liu Hui
Published: November 2021

De novo CD5 positive diffuse large B cell lymphoma (CD5+ DLBCL) has poor survival in the era of immunochemotherapy. We conducted a multi-center retrospective study to explore the clinicopathologic characteristics, genomic profiling and prognostic elements of 61 CD5+ DLBCL and 60 CD5- DLBCL patients in China. In contrast with CD5- DLBCL, elderly onset, advanced stage, central nervous system (CNS) involvement, as well as MYC/BCL-2 and P53 overexpression were more prevalent in CD5+ DLBCL. In addition, most of CD5+ DLBCL patients were of non-germinal center B-cell-like (non-GCB) and activated B-cell-like (ABC) subtype according to immunohistochemistry and Lymph2Cx assay. Genetic analysis by next generation sequencing (NGS) showed the proportion of MCD subtype in CD5+ DLBCL was higher than that of CD5- DLBCL (50% vs 5%, p = 0.0007). Compared with CD5- cohort, CD5+ DLBCL patients showed poorer 5-year overall survival (OS) (70.9% vs 39.0%, p<0.001), independent of cell-of-origin and MYC/BCL-2, P53 and BCL-6 status. Multivariate analysis showed that age >76 years, advanced stage, CNS involvement and hypoalbuminemia were independent factors associated with poor prognosis in CD5+ DLBCL. In conclusion, CD5+ DLBCL conveyed poor prognosis and showed distinctive clinicopathologic characteristics and predominant genetic features of ABC and MCD subtypes.

Disclosures

No relevant conflicts of interest to declare.
G-CSF Upregulates the Expression of Aquaporin-9 through C/EBP-Beta to Enhance the Cytotoxic Activity of Arsenic Trioxide to Acute Myeloid Leukemia Cells
Authors: Fu Wanbin , Xu Lan , Liu Jia , Han Xiaofeng , Wang Junying , Wang Xinpeng , Zhao Huanbin , Hou Jian , Hu Jiong , Zhong Hua
Published: November 2021

Arsenic trioxide (ATO) is the first-line drug for the treatment of acute promyelocytic leukemia (APL). Aquaporin-9 (AQP9), a transmembrane transporter, is required for leukemia cells to uptake ATO. APL cells express high levels of AQP9, while other types of acute myeloid leukemia (AML) cells express much lower levels of AQP9, which limits the transportation and the cytotoxic activity of ATO in those types of leukemia. Recently, we found that granulocyte colony stimulating factor (G-CSF) can significantly upregulate the expression of AQP9 in AML cell lines (THP-1 and HL-60), and can significantly enhance the intracellular concentrations of ATO, when compared with the treatment with ATO alone. We also found that the combination of ATO and G-CSF inhibited the cell proliferation and induced cell apoptosis more significantly than the treatment with ATO or G-CSF alone. The overexpression of AQP9 with lentivirus in HL-60 or THP-1 cells significantly enhanced the cytotoxic activity of ATO, while the knock-down of AQP9 gene with small interfering RNA (siRNA) significantly attenuated the combination effect of G-CSF and ATO. Moreover, we found that the upregulation of AQP9 by G-CSF depends on the upregulation of C/EBP-beta, the transcription factor downstream of G-CSF. In conclusion, our study suggests that the pretreatment of G-CSF could significantly enhance the cytotoxic activity of ATO in AML cells through the upregulation of AQP9, and that the combination of G-CSF and ATO would be a potential therapeutic strategy for AML patients.

Disclosures

No relevant conflicts of interest to declare.
The Serine Threonine Kinase Inhibitor Ots-167 Improves Erythropoiesis through Suppression of Nlk Activity in Diamond Blackfan Anemia Models
Authors: Shibuya Aya , Wilkes Mark C , Serrano Manuel , Flygare Johan , Glader Bert , Narla Anupama , Sakamoto Kathleen M.
Published: November 2021

Diamond Blackfan Anemia (DBA) is associated with a hypoproliferative anemia, congenital abnormalities, and cancer. The disease typically presents within the first year of life with the majority of patients carrying mutations in one of at least 17 ribosomal proteins, with RPS19 being the most common. Current therapies for DBA have undesirable side effects, including iron overload from repeated red cell transfusions, chronic effects from long term corticosteroid use, or complications from stem cell transplantation.

The serine threonine kinase Nemo-like Kinase (NLK) is an atypical member of the MAP kinase family of enzymes and has been shown to be chronically hyper-activated in RPS19- and RPL11-haploinsufficient murine and human models of DBA, as well as in erythroid progenitors from DBA patients. In RPS19-insufficient human hematopoietic stem and progenitor cells, genetic silencing of NLK by shRNA increased erythroid expansion by 220.3% (SD = 6.6%), indicating that aberrant NLK activation may contribute to the pathogenesis of the disease and is a potential target for DBA therapy.

A number of clinically approved or advanced compounds have been developed to inhibit MAP kinases with various degrees of cross reactivity among its family members. We therefore screened a number of compounds that inhibit NLK as an off-target and found that these NLK inhibitors improved erythroid expansion in DBA models. Of these inhibitors, OTS-167 performed optimally, improving erythropoiesis by 2-fold at 300nM, with an EC50 of 146nM.

Previous studies of OTS-167 in xenograft models of neuroblastoma for one month did not result in neutropenia, suggesting very little to no toxicity to myeloid cells. The goal in treating DBA patients with NLK inhibitors is to sufficiently raise the hemoglobin to prevent the need for chronic red cell transfusions or treatment with steroids. Our results suggest that pharmacologic inhibition of NLK is a potential approach to treat patients with DBA. We are currently investigating other NLK inhibitors in preclinical models for future clinical application.

Disclosures

Glader: Agios: Consultancy.
Difference in Therapeutic Effects between Roxadustat and Daprodustat, HIF-Ph Inhibitors, Depending on the Blood Type in Hemodialysis (HD) Patients
Authors: Funakoshi Satoshi
Published: November 2021

Human blood group antigens are glycoproteins and glycolipids expressed on the surface of red blood cells and a variety of human tissues. This study aimed to determine if there is an association between ABO blood type and the efficacy of HIF-PH inhibitors. Roxadustat and daprodustat are potent inhibitors of HIF-PH and capable of stimulating erythropoiesis in patients on patients with impaired renal function. These two compounds are reported to act mechanistically similar but display differences in their effects on cells, and the differences may affect their efficacy in the treatment of renal anemia in HD patients. In this study we compared the response rate by blood type between roxadustat and daprodustat, respectively.

Sixty-eight HD patients treated with roxadustat (20-100mg, 3/week) and ninety-five treated with daprodustat (1-12mg, daily) were recruited in our observational study. We defined >1.5g/dL increase in hemoglobin as effective, and <1.5g/dL decrease as ineffective.

As shown in the figure, type A had a significantly high response rate at 47% in HD patients treated with roxadustat. On the other hand, type O had a significantly high response rate at 55% in those who were treated with daprodustat.

We found the association in the effectiveness of roxadustat on the treatment for anemia in HD patients in type A, while the effectiveness was higher in type O treated with daprodustat. The results suggest that the therapeutic effect of HIF-PH inhibitors may differ depending on the blood type.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Targeting Metabolic Dysregulation for Ruxolitinib Failure in MPN
Authors: Xu Na , Luo Jie , Luo Dongmei , Liang Hanying , Tan Yaxian , Liu Qifa , Liu Xiaoli
Published: November 2021

Background:Metabolic alterations are increasingly recognized as an important pathogenic process that underlies fibrosis and cancer cells. MPN models were demonstrated to exhibit metabolic vulnerabilities due to a high dependence on enhanced levels of glycolysis and oxidative phosphorylation. MPN patients often gain weight during ruxolitinib treatment effects on leptin signaling. Ruxolitinib provides symptomatic relief and can improve survival but generally fails to resolve the malignant clone,nealy 50% patients resistance to ruxolitinib.Here,we will study transcriptomic and metabolomic program in JAK2-mutant MPN who resistance to ruxolitinib.

Methods:The collection of blood samples and clinical data from MPN patients, Mice used in this study were kept in accordance with Swiss federal regulations. Multi-omics detection by transcriptome sequencing and liquid chromatography .

Results: We found that active lipid metabolism obviously in patients resistance to ruxolitinib in comparison with patients sensitive to ruxolitinib.Transcriptomic and metabolomic analyses identified numerous metabolic nodes in JAK2-mutant hematopoietic stem and progenitor cells that were altered in comparison with wild-type controls. We studied the consequences of key regulatory enzyme of Fatty acid metabolism gene(SREBP,FASN, ACC, SCD1) and found that pharmacological inhibition of SREBP(Betulin) reduced hematopoietic manifestations of MPNs.

Conclusion: Our findings reveal the contribution of metabolic alterations to JAK2 inhibitor failure and suggest that abnormally active lipid metabolism of mutant cells represent vulnerabilities that can be targeted for treating MPNs.

Keywords: metabolomic;resistance;resistance;Myeloproliferative

Disclosures

No relevant conflicts of interest to declare.
Thyrointegrin αVβ3 Antagonist: Implications in Acute Myeloid Leukemia
Authors: Darwish Noureldien , Glinsky Gennadi V. , Mousa Shaker A
Published: November 2021

Leukemic cells are able to receive and send several signals within bone marrow niche that play an important role in their survival. One of the important crosstalk is the interaction between the bone marrow microenvironment proteins (vitronectin, fibronectin, fibrinogen, and ostepontin) and thyrointegrin αVβ3 on leukemic cells, generating ligand-specific outside-in signals that are relevant to a variety of cell functions, including gene transcription, cell division, cell attachment, and motility

Our previous experiment using in vivo AML animal models with primary AML cells and cell lines have shown significant reduction of leukemic cell burden 74% and >95% (P<0.0001), respectively, after daily subcutaneous treatment with thyrointegrin αvβ3 antagonist fb-PMT (Ki 0.23 nM) at 3 and 10 mg/kg, for 3-4 weeks. In this study we focused on evaluations of the molecular effects of fb-PMT in leukemic cells. Acute myeloid leukemia cell lines (K562-Luc and KG1a cells) were cultured in 50 cm² cell culture flasks with 10 mL phenol red free RPMI media containing 10% fetal bovine albumin. The leukemic cells were treated (at 50% confluence) with 30 µM fb-PMT for 48 hours. Total RNA was immediately isolated from harvested cells using Triazole and used for microarray analysis. Overall, there were 370 significantly down-regulated gene expression records and 273 significantly up-regulated gene expression records, expression of which were changed at least 1.5-fold in fb-PMT-treated human leukemic cells. Significant examples of the fb-PMT-induced gene expression signatures (GES) of pathway's interference include SNAI, MYC, HIF1A, TWIST1, and TFAP2C (P<0.05). Notably, inference of potential contribution to the fb-PMT anticancer activity of the interference with these pathways seems highly congruent with their known biological functions such as cell cycle control (MYC), survival and maintenance of stem cells (HIF1A, TFAP2C), and essential features of the malignant phenotype (TWIST1, SNAI) (Figure 1).

Consistently, examples of the fb-PMT-induced GES of transcriptional pathway's activation include RB1, IRF9, MAML1, RAP1A, and GATA4 pathways (P<0.05), known biological functions of which appear highly consistent with the hypothesis that activation of these pathways might contribute to fb-PMT anticancer activity. Finally, we found that fb-PMT interfered with estrogen signaling in human AML cells. The fb-PMT was associated with decreased phosphorylation and nuclear enrichment of Erα (Figure 1).

Collectively, our in vivo study and genomic data have shown the key role thyrointegrin αvβ3 in leukemogenesis. The thyrointegrin αvβ3 antagonist fb-PMT demonstrated potent anticancer actions on human AML through the molecular interference mechanism with multiple signaling pathways supporting growth and survival of leukemic cells

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Early Transfusion with Mesenchymal Stem Cell Derived Extracellular Vesicles: A New Transfusion Strategy for Life-Threatening Hemorrhage and Traumatic Brain Injury
Authors: Dekker Simone E , Bambakidis Ted , Williams Aaron M , Biesterveld Ben , Bhatti Umar , Li Yongqing , Pickell Zachary , Buller Benjamin , Alam Hasan B
Published: November 2021

Background: Life-threatening hemorrhage and traumatic brain injury (TBI) have a significantly increasing global burden and remain leading causes of preventable deaths. Effective interventions may protect the brain against ongoing damage and improve the long-term outcomes. A growing area of interest is transfusion of cell-based therapies, particularly with bone marrow-derived mesenchymal stem cells (MSC). Transfusion using MSC derived extracellular vesicles (EVs) have shown to improve neurologic outcomes in animal models of life-threatening hemorrhage, stroke, and TBI. However, the precise mechanisms remain poorly characterized. In the present study, we aimed to elucidate some of the key cerebral genes, pathways, and networks that were modulated after transfusion of EVs in a porcine model of hemorrhagic shock (HS) and TBI.

Methods: Swine were subjected to HS (40% blood volume) and severe TBI (8-mm cortical impact). After 1 hour of shock, animals were randomized (n=4/group) to treatment with either lactated Ringer's (LR) or LR+EV. Both groups received fluid resuscitation after 2 hours of shock, and autologous packed red blood cells 5 hours later. After 7-days, brains were harvested and RNA-sequencing was performed. The transcriptomic data was imported into the iPathway pipeline for bioinformatics analyses.

Results: 5,273 genes were differentially expressed in the LR+EV group vs. LR alone (total 9,588 measured genes, Figure 1). Table 1 lists the top 10 genes exhibiting the greatest up- and down-expression based on fold change. Genes with the greatest up-regulation were involved in synaptic transmission and neuronal development and differentiation, while down-regulated genes were involved in inflammation. Gene Ontology terms experiencing the greatest modulation were involved in inflammation, brain development, and cell adhesion. Pathway analysis revealed significant modulation in the glutamatergic and GABAergic systems. Network analysis revealed down-regulation of inflammation (Figure 2), and up-regulation of neurogenesis, and neuron survival and differentiation.

Conclusions: In a porcine model of HS+TBI, EV transfusion was associated with an attenuation of cerebral inflammatory networks and a promotion of neurogenesis and neuroplasticity. These transcriptomic changes could explain the observed neuroprotective and neurorestorative properties associated with EV transfusion. EV transfusion reduces the hyper-inflammatory response and may have great promise in improving outcomes in concurrent life-threatening hemorrhage and severe TBI. Further testing of this novel strategy and its implications in transfusion medicine are warranted.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Analysis of Polymorphism in the Genes TLR3, TLR4 and TLR9 in Patients with Acute Myeloid Leukemia
Authors: Rybka Justyna , Bogunia-Kubik Katarzyna , Kuszczak Bartłomiej , Kalicińska Elżbieta , Łacina Piotr , Dratwa Marta , Wróbel Tomasz
Published: November 2021

Acute myeloid leukemia (AML) is a bone marrow malignancy characterized by uncontrolled proliferation of transformed myeloid precursors. It is the most common acute leukemia and is a highly heterogeneous disease. AML leads to impaired hematopoiesis, which may result in increased risk of infection. Toll-like receptors (TLRs) are major elements of the innate immune system which recognize pathogen-associated molecular patterns. They also have a role in autoimmune diseases and cancer through their recognition of endogenous danger-associated molecular patterns. Various TLRs were found to be expressed in AML cells, and their expression was found to affect growth, differentiation, and immunostimulatory capacity in AML.

We retrospectively analysed 90 patients (48 females and 42 males) with diagnosis of AML, hospitalized in Department and Clinic of Hematology, Blood Neoplasms and Bone Marrow Transplantation of Wroclaw Medical University. The median age of patients was 62 (range:18-81). Infectious complications after induction chemotherapy occurred in 62 patients. In the present study, we detected the effect of seven single nucleotide polymorphisms (SNPs) located in the genes coding for TLR3 (rs5743305, rs3775296, 3775291), TLR4 (rs4986791, rs4986790) and TLR9 (rs187084, rs5743836). DNA was isolated from samples of peripheral blood taken on EDTA. Melting curve genotyping with the use of LightSNiP asays(TIB MOLBIOL, Germany) was employed to determining TLR3, TLR4 and TLR9 genotypes, and the reactions were performed LightCycler 480 device (Roche Applied Science, Germany).Statistical analyses were performer using Real Statistics Resource Pack for Microsoft Excel.

We found that TLR3 rs5743305 AA homozygosity was more common in AML patients with infectious complications than in patients unaffected by infections (p=0.015). TLR4 rs4986791 C and TLR9 rs5743836 C were associated with lower levels of the inflammation marker CRP (p=0.015 and p=0.048, for rs4986791 and rs5743836, respectively), TLR4 rs4986791 C and was less frequently detected among patients with CRP>5 (p=0.028). We also found TLR9 rs187084 C to be more common in patients with adverse cytogenetic risk according to ELN criteria (0.012). Additionally, a trend for higher prevalence of extramedullary disease was observed for carriers of TLR3 rs3775296 T (p=0.054), TLR3 3775291 GG (p=0.068), TLR4 rs4986790 G (p=0.078), as well as TLR9 rs187084 TT (p=0.053).

In conclusion, our results suggest that SNPs in the genes coding for TLR3, TLR4 and TLR9 may be implicated in clinical outcome of AML and is related with increased risk of infectious complications.

Disclosures

Wróbel: BMS: Honoraria; Roche: Honoraria, Research Funding, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau; Takeda: Honoraria, Speakers Bureau; BeiGene: Honoraria.
Obesity and Oral Contraceptives Synergistically Increase Thrombotic Risk By Downregulating Protein S
Authors: Thangada Lekha , Kumar Manoj , Pilli Satish Sekhar , Majumder Rinku
Published: November 2021

Background:

The increased thrombotic effects of estrogen-based oral contraceptives and obesity have been documented independently. However, obesity and oral contraceptives combined are associated with a far greater thrombotic risk, but we have a poor understanding of the mechanism of this greater effect. Increasingly, women are using oral contraceptives, and the national obesity rate has been skyrocketing. Thus, it is imperative to explain how obesity and oral contraceptives work together to significantly elevate thrombotic risk. We know that hypoxia-inducible factor 1-ɑ (HIF-1ɑ) and the estrogen receptor (Erɑ) bind to the promoter of the Protein S gene; binding occurs at sites within ~450 nucleotides of each other, and the two transcription factors downregulate Protein S expression independently. We hypothesize that the two factors, by binding the promoter simultaneously, synergistically downregulate Protein S to a degree much greater than the downregulation mediated by each factor separately.

Aims:

The goal of this project is determining whether estrogen and obesity-induced hypoxia work synergistically to downregulate Protein S transcription and increase thrombotic risk.

Methods:

We measured the effects of obesity and oral contraceptives on hepatocarcinoma (HEP G2) cells because the liver is the major producer of Protein S. Estrogen of varying concentrations (25-150 μM) was used to mimic the effects of oral contraceptives, and cobalt chloride of varying concentrations (25-150 μM) was used to stimulate hypoxia and HIF-1ɑ expression. Cells were exposed to estrogen only, cobalt chloride only, and estrogen and cobalt chloride together for 24 hours, after which the cells were harvested and subjected to q-PCR and immunoblot blot analyses to measure Protein S transcription and protein expression. Although cobalt chloride is a reliable inducer of hypoxia and HIF-1ɑ expression, we also performed hypoxia experiments by incubating cells in a chamber with varying O 2 concentrations (20%, 15%, 10%, 5%, 1%).

Results and Conclusions:

Immunoblot analysis of cells treated with either CoCl 2 and estrogen supplementation revealed a ~20% reduction in Protein S levels compared to control conditions and a more significant reduction in (~60%) Protein S expression in cells treated with estrogen and CoCl 2 together. These results supported our hypothesis that obesity and estrogen-based contraceptives increase thrombotic risk by downregulating anticoagulant Protein S transcription and subsequently decreasing Protein S level.

Disclosures

No relevant conflicts of interest to declare.
Telemedicine in Hematology: Tool to Improve Access and Patient Satisfaction
Authors: Master Samip R , Ramadas Poornima , Mansour Richard Preston
Published: November 2021

Introduction: Telemedicine can transform the future of medicine, especially in rural settings by improving access to medical care. Here were present the patient satisfaction data from both telemedicine and regular visits in our Hematology & Oncology clinics. In Hematology, we have used tele medicine for sickle cell disease, benign hematology cases like iron deficiency, thrombocytopenia, leukocytosis etc., and in malignant hematology for new patient consults, toxicity checks, follow up of plasma cell disorders, chronic leukemias and low intensity chemotherapies.

Methods: We retrospectively collected patient satisfaction survey data from March 2020 to April 2021 for both telemedicine and regular visits. The patients were allowed to give a score from 1 to 99 regarding their experience at visit. We collected data on the following: likelihood of recommending, care provider, access and overall assessment.

Results: A total of 53 patient satisfaction survey were available for telemedicine and 462 for regular visits. Over 60 % of tele visits were non cancer related and 40% were cancer related. The average score for likelihood of recommending, care provider, access, and overall assessment for tele visit were 73.5, 41.5, 73.5 and 62 respectively. The average score for likelihood of recommending, care provider, access, and overall assessment for regular visit were 13.75, 7.5, 26.5, and 11 respectively.

Conclusion: The patient satisfaction scores for likelihood of recommending, care provider, access and overall assessment were significantly better for tele visits compared to regular visits.

Disclosures

Master: Blue Bird Bio: Current holder of individual stocks in a privately-held company.
PI3K/AKT Activation Mediates B-Cell Transformation By the "T" Splice-Variant of Fyn Kinase
Authors: Kumar Preet , Li Lingxiao , Schatz Jonathan H.
Published: November 2021

The SRC-family kinase FYN has key roles downstream of growth-factor receptor activation and is implicated in various cancers. Activating FYNutations were reported in angioimmunoblastic T-cell lymphoma (AITL), but its role in transforming lymphocytes is poorly characterized. FYN also may play a role in signaling downstream of the B cell Receptor (BCR) receptor in Diffuse Large B-Cell Lymphoma (DLBCL), and laboratory studies show inhibition of FYN and other SRC family kinases (SFKs) can block BCR-mediated lymphomagenesis. To investigate FYN's role in oncogenic singling, we introduced FYN alleles to IL3-dependent FL5.12 murine pro-B cells and tested for selective advantage upon cytokine withdrawal and ability to transform the cells to cytokine independence. FYN B and T variants result from alternate splicing of exon 7, with B originally detected in brain and T in hematopoietic cells, but both are expressed and may be deregulated associated with disease states in diffuse tissues. We employed wildtype (WT) alleles for both spliceoforms along with mutations detected recurrently in AITL (L174R and R176C), kinase active (Y528H), and kinase dead (K299A). No FYN B alleles showed any selective advantage in the prolymphocyte cells subjected to cytokine withdrawal. In contrast, all FYN T alleles other than kinase dead were strongly enriched through multiple rounds of IL3 withdrawal and rescue. Indeed, complete withdrawal of cytokine resulted in transformation to IL3-independent growth, generating stable cell lines addicted to FYN kinase activity for survival. The SFK inhibitor PP2 was toxic to all FYN-T transformed cells but not to baseline FL5.12 cells growing in cytokine. WT- and L147R-transformed cells were similarly sensitive to PP2, while the other AITL-derived mutant R176C demonstrated reduced sensitivity. Immunoblotting demonstrated PP2-sensitive phosphorylation of AKT and the MTORC1 target 4EBP1, suggesting PI3K/AKT activation as a key survival pathway downstream from transformation by FYN T alleles. Consistently, the AKT inhibitor ipatasertib and PDK1 inhibitor GSK2334470 were highly active against all FYN-transformed lines, and AKT phosphorylation was sensitive to PDK1 inhibition, showing convergence on AKT activation in all transformants. Pan-PI3K inhibition was active against FYN transformants, while the delta isoform-specific inhibitor idelalisib showed only partial activity, suggesting activation of multiple PI3K isoforms downstream of FYN activity. These results suggest that AKT, PI3K, and PDK1 are important targets downstream of FYN, and that the PI3K/AKT pathway is the key survival mechanism in lymphomagenesis mediated by FYN. We show that WT and activating mutants of the FYN T spliceoform specifically, but not FYN B, induce drug-sensitive B lymphocyte malignant transformation by activating PI3K/AKT/MTORC1 signaling.

Disclosures

No relevant conflicts of interest to declare.
Caplacizumab Is Effective in Treating Immune-Mediated Thrombotic Thrombocytopenic Purpura in Real-World Scenarios and May Lower Mortality: An Aggregative Analysis
Authors: Völker Linus A. , Balduin Gesa , Kühne Lucas , Kaufeld Jessica , Schönermarck Ulf , Menne Jan , Knöbl Paul , Brinkkoetter Paul
Published: November 2021

Introduction: The treatment of acquired thrombotic thrombocytopenic purpura has been a subject to change lately due to the introduction of caplacizumab as a novel, and potentially life-saving agent. Here, we compare patient-centered outcomes of a historic cohort of 119 aTTP-patients and an extended real-world caplacizumab-treated cohort of 113 aTTP-patients from Germany and Austria in the context of multiple recently published international cohorts of aTTP patients.

Methods: From October 2018 until May 2021, data from German and Austrian patients presenting with an acute episode of aTTP as defined by an ADAMTS13 activity below 10 % treated with caplacizumab were gathered retrospectively. We aggregated data from two published randomized controlled trials (TITAN, HERCULES), one single-arm prospective trial and two retrospective cohorts with a historical control cohort. In total, we report 846 aTTP cases, 396 of those treated with caplacizumab and 450 without. For data analysis, placebo groups from randomized trials and historical controls from retrospective analyses were combined.

Results: All cohorts combined reported 30 aTTP-related deaths, predominantly in the non-caplacizumab cohort. Mortality in the caplacizumab-cohort (8 cases total) was mainly driven by 5 reported deaths in the UK-cohort, which have been ascribed to a delayed use of caplacizumab by the authors. The number of daily PEX was significantly less in the caplacizumab-treated cohort (7.2 vs 10.2 days). In line, the number of hospital (14.0 vs 18.1) and ICU (4.7 vs 8.9) days differed significantly. Exacerbations (14% vs 39%) were significantly reduced in the caplacizumab-group. In view of all treatment-related outcomes, the data presented herein argues strongly in favor of an early and consistent use of caplacizumab in all instances of aTTP.

Disclosures

Völker: Sanofi-Genzyme: Honoraria, Other: counselling fees.
Effect of Hydroxyurea Therapy on the Incidence of Infections in Ugandan Children with Sickle Cell Anaemia
Authors: Namazzi Ruth , Conroy Andrea L. , Bond Caitlin , Goings Micheal J. , Datta Dibyadyuti , Ware Russell E. , Jang Jeong Hoon , John Chandy C. , Opoka Robert O.
Published: November 2021

Hydroxyurea is efficacious against sickle cell anaemia (SCA)-related complications in African children. Prior studies demonstrated conflicting results on the effect of hydroxyurea on risk of infection, the most common cause of morbidity and mortality in African children with SCA.

We evaluated the incidence of infections before and after starting hydroxyurea in 117 children aged 1-5 years with SCA enrolled in the Zinc for Infection Prevention in Sickle cell anaemia (ZIPS) clinical trial that received zinc or placebo treatment for one year (Clinicaltrials.gov, NCT03528434). Children were enrolled between March 2018 and November 2019 and initiated on hydroxyurea (20 mg/kg/day) if they met Uganda SCA guideline criteria for hydroxyurea treatment at any time during the study. We compared the incidence of infections before and after hydroxyurea therapy, adjusting for zinc treatment.

Overall, the mean duration on hydroxyurea was 223.8 (85.2) person days. The mean(SD) incidence of any severe/invasive infections (infections meeting strict clinical and laboratory or radiological diagnostic criteria) was 6.2(9.0) vs. 1.9(2.3) infections per child per year before and after hydroxyurea (incidence rate ratio [IRR]: 0.40, 95%CI: 0.29-0.54, p<0.001), including a decrease in the incidence of bacteremia [IRR: 0.05] malaria, [IRR 0.28], cellulitis [IRR: 0.24], gastroenteritis [IRR: 0.41], pharyngitis [IRR: 0.59] and sinusitis [IRR: 0.38] (all p<0.05). Similarly, the incidence of clinically defined infections (infections meeting clinical but not laboratory/radiological criteria) decreased after hydroxyurea, (IRR: 0.49, 0.41-0.59), influenced primarily by large reductions in the incidence of lower and upper respiratory tract infections (p<0.001 for both). As expected, hydroxyurea treatment was also associated with a decrease in complications of SCA, including vaso-occlusive crises, hospitalizations and transfusions (all p<0.001). There was no interaction between zinc and hydroxyurea therapy on risk of infection and SCA-related complications.

In Ugandan children with SCA, hydroxyurea therapy not only decreases the incidence of SCA-related complications, but also substantially reduces the incidence of infections. Research to understand the underlying mechanisms of protection from hydroxyurea against infection and exploration of its potential use for infection prevention is warranted.

Disclosures

Ware: Bristol Myers Squibb: Research Funding; Addmedica: Research Funding; Hemex Health: Research Funding; Nova Laboratories: Research Funding; Novartis: Other: DSMB Chair; Editas: Other: DSMB Chair.
Cigarette Smoking Exacerbates Flu-Induced Thrombo-Inflammation
Authors: Kaminski Tomasz W. , Brzoska Tomasz , Vats Ravi , Tutuncuoglu Egemen , Nickolich Kara , Robinson Keven M. , Nyunoya Toru , Sundd Prithu
Published: November 2021

Rationale: Epidemiological evidence suggests that prior exposure to cigarette smoke (CS) or habitual smoking increases the risk of influenza A virus (IAV)-triggered respiratory failure (severe flu). Although emerging evidence supports the role of thrombo-inflammation in the development of CS and IAV-triggered lung injury, the innate immune mechanism that contributes to this morbidity remains poorly understood.

Materials and methods: We have developed a two-hit model of CS-induced severe flu in mice. Mice were exposed to four weeks of room air (air) or CS followed by intra-nasal administration of A/PR/8/34 (H1N1) IAV. The body weight was measured every day for two weeks after IAV administration followed by assessment of lung injury at day 7 and 14. Lungs were harvested for histological assessment of lung injury and estimation of viral titer by RT-PCR. Quantitative fluorescence intravital lung microscopy (qFILM) was conducted at 2-, 3- and 4-days post IAV-infection to visualize dynamics of neutrophil and platelet recruitment in the lung of mice IV administered with fluorescent dextran, anti-Ly6G Ab and anti-CD49Ab.

Results: Mice exposed to CS+IAV manifested significantly more weight loss, lung injury, lung congestion and alveolar hemorrhage compared to mice administered room-air+IAV. QFILM revealed that severity of lung injury was associated with significantly more entrapment of neutrophil-platelet aggregates within the pulmonary microcirculation and infiltration into the air spaces of CS+IAV than room-air+IAV administered mice.

Conclusion: These initial results suggest that CS primes innate immune signaling in neutrophils and platelets to promote their recruitment in the lung following flu. Currently, studies are underway to identify innate immune pathways in neutrophils and platelets that drive this severe thrombo-inflammatory response.

Disclosures

Sundd: CSL Behring Inc: Research Funding; Bayer: Research Funding; Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding.
PD-L1 and Tumor Mutational Burden Are Not Predictive of Thrombosis in a Cohort of 1,221 Patients with Solid Organ Malignancies
Authors: Thawani Rajat , Kartika Thomas , Elstrott Benjamin , Batiuk Elizabeth , Chen Lilian , Tun Nattapron , Taflin Nicholas , Tao Derrick Law , West Malinda T , Shatzel Joseph J
Published: November 2021

Introduction: Malignancy is a well-known risk factor for thrombosis. While many clinical risk factors for cancer-associated thrombosis have been described, it remains unknown how certain tumor specific observations (such as programmed death ligand 1 expression (PD-L1) and tumor mutational burden (TMB)) correspond to thrombotic risk. To determine the relationship between PD-L1 and TMB quantification and thrombosis in patients with solid tumors, we evaluated a large cohort of clinical samples from an NCI designated cancer center, correlating tissue sample pathology with the development of venous or arterial thromboembolism.

Methods: We performed a retrospective cohort study of patients between the ages of 18-89 who underwent histopathologic examination and next generation sequencing for a diagnosis of a solid organ malignancy at the Knight Cancer Institute between June 2019 and Feb 2021. Medical records were reviewed to document clinical and demographic information as well as for the development of thrombosis after cancer diagnosis. Multivariable logistic regression was performed to assess if PD-L1 expression and TMB are independent predictors of thrombosis. Subgroup analysis was then performed to evaluate whether this relationship differed by the organ of the primary malignancy. All analyses were conducted in R (R Core Team 2019).

Results: We identified 1,221 patients with solid organ malignancies (mean age 62, 53.6% male). The most common malignancies in the cohort were lung cancer (13.8%), pancreatic cancer (12.3%) and colon cancer (12.3%). Thrombotic events occurred in 206 patients (16.8%) after diagnosis of their malignancy (100 deep vein thrombosis, 42 pulmonary embolism, 46 visceral vein thromboses, 6 superficial vein thromboses, and 11 arterial events). The mean time from initial biopsy to thrombosis was 224 days. TMB (mean 8.5 mutations/Mb) was evaluated in all patients in the cohort, while PD-L1 expression testing (mean 23.0%) was available for 255 patients. On multivariable logistic regression adjusting for age and patient sex, neither TMB (ORadj: 0.97, CI: 0.92-1.03) nor PD-L1 expression (ORadj 1.00, CI: 1.00-1.02) were significant predictors of thrombotic events. Subgroup analysis by primary malignancy type did not demonstrate any specific primary sites for which TMB or PD-L1 were predictive of thrombosis.

Discussion: Our analysis found no predictive relationship between TMB and PD-L1 expression with thrombotic events in patients with solid organ malignancy. Further analysis is needed to determine if specific treatment protocols, such as the use of immunotherapy in patients with varying levels of PD-L1 and TMB, alter thrombosis risk.

Disclosures

Shatzel: Aronora Inc,: Consultancy.