bluuebunny/crossref_metadata_2025 huggingface dataset:
Maintenance Therapy with Chidamide Decreases the Relapse in Fusion Gene-Positive AML Patients with MRD-Positive or ELN-High Risk
Authors: Yu Guopan , Yin Zhao , Huang Jixian , He Huiqing , Jiang Xuejie , Zhou Hongsheng , Huang Pengcheng , Zhang Yu
Published: December 2023

Purpose: Acute myeloid leukemia (AML) patients with fusion gene-positive including core binding factor (CBF), mixed-lineage leukemia (MLL) gene rearrangement have a high incidence of relapse if they are continious measurable residual disease (MRD)-positive or in ELN-high risk and could not be bridged to allogenetic heamatopoitic stem cell transplantation (allo-HSCT). As known histone deacetylase (HDAC) is abnormally recruited in these fusion gene-positive AML, also RUNX1 mutated AML, and could be a therapy target. Aim of this study is to assess the effect and safety of HDAC inhibitor chidamide as maintemance therapy in these AML patients.

Methods: A multi-center retrospective study was performed. Patients with MRD-positive CBF-AML, MLL rearrangement or RUNX1 mutation AML, without being bridged to allo-HSCT were included. Maintemance with chidamide was started one to three months after consolidation for at least six months, at a dose of 10mg/day or 30mg twice a week.

Results: Twelve patients were enrolled, included 8 with AML1-ETO-positive, 3 with RUNX1 mutation, 1 with MLL rearrangement. The median age was 33(16-62) years old. The ratio of male to female was 6 to 5. They previously experienced median 6(4-8) cycles of chemotherapy, included one with allo-HSCT, one with autologous-HSCT. They were all maintemanced with chidamide, except one with chidamide plus sorafenib. Eleven patients were available for outcome assessment. With a median follow-up of 14(2-67) months, among the 9 patients with MRD-positive, 6(66.7%) achieved MRD-negative, 1 stable disease, 2 relapsed and died. The median time to achieved MRD-negative was 2.5(1-12) months. The other 2 patients was contenious MRD-negative. The cumulative incidence of relapse was 2/11(18.2%). The rate of MRD-nagetive was 8/11(72.7%). The mortality rate was 2/11(18.2%). During the maintemance, 4 patients underwent bone marrow suppression, including 2 with grade 3, 2 with grade 1/2. One patients had an anorexia of grade 2.

Conclusion: Maintenance therapy with chidamide might decrease the relapse in fusion gene-positive AML patients with MRD-positive or ELN-high risk, who could not take allo-HSCT, and prolong their survival, with a well toleration.

Key words: chidamide, maintenance, fusion gene, acute myeloid leukemia
Prospective Cohort Study on Paediatric and Adult Haemophilia a Patients Correlating Emicizumab Levels with Clinical Experience
Authors: P'ng Stephanie , Carter Tina , Kaminskis Lisa , Modica Natasha , Michalopoulos Nick , Goruppi Marina , Pepperell Dominic
Published: December 2023

Aim:

The efficacy of Emicizumab in preventing bleeds in patients with Haemophilia A has been demonstrated in the HAVEN studies. Emicizumab is not universally available at the recommended dosages and there is a push towards reducing frequency and dose to increase availability. This study aims to provide data on Emicizumab levels and corresponding clinical response in the real world.

Method:

Moderate-severe Haemophilia A patients who transitioned to Emicizumab were identified from 2 Centres in Western Australia between November,2017 to December,2022. Bleeds requiring factor replacement were recorded 12 months prior to transition and 12 months following. These were obtained from the Australian Bleeding Disorders Registry and patient hospital notes. Available steady state Emicizumab levels were also recorded.

Results:

31/32 paediatric patients had the same or improved bleeding rates since transition to Emicizumab. No patient had a spontaneous bleed since transition. One patient continued to have high bleeding rates due to adherence issues, with an Emicizumab level of 10µg/ml. Of the 16/32 patients had Emicizumab levels measured with a mean 55.54µg/ml (range 38.8 - 81).

21/24 adult patients experienced the same or improved bleeding rates post transition. 7 patients did not have completed bleed data. 3 patients had worse bleeding rates since transition with 1 bleed each. Their Emicizumab levels ranged 58.54-79.15µg/ml. Emicizumab levels were recorded in 16, with a mean of 53.08µg/ml (range 18.47 - 85.22).

6 adult patients had surgical procedures with 2 bleeds; 1 bleed occurred 10 days after colonoscopy with emicizumab level 18.47ug/mL and 2nd bleed occurred 1 day after pharyngeal level 42.42ug/mL.

Conclusion:

In both paediatric and adult patients, Emicizumab was an effective treatment in preventing bleeds. The mean and range of Emicizumab levels in both groups who had an effective response were similar. The wide range of therapeutic levels may be of important for dosing changes, particularly with the paediatric cohort
Histiocytic Necrotizing Lymphadenitis with Hemophagocytic Lymphohistiocytosis: A Single-Center Analysis of Five Cases
Authors: Miao Yi , Chen Qingqing , Shi Wenyu , Zhang Jing , Li Jianyong
Published: December 2023

Introduction: Histiocytic necrotizing lymphadenitis (HNL) is a benign, self-limiting disease that presents with few nonspecific symptoms such as fever and swollen lymph nodes in the neck. Histiocytic necrotizing lymphadenitis with hemophagocytic lymphohistiocytosis (HNL-HLH) is rare and case series have been reported in children. HNL-HLH in adults has not been systematically analyzed.

Methods: We aimed at exploring the clinical, laboratory, and radiological features and outcomes of adult patients with HNL-HLH. We collected the clinical data of patients with HNL-HLH admitted to the First Affiliated Hospital of Nanjing Medical University from October 2010 to June 2015. All the patients underwent lymph node biopsy and have a pathological diagnosis of HNL. The age, gender, clinical presentation, lymph node signs, laboratory findings and imaging data, and pathological findings of the patients were collected.

Results: In this study, we reported five adult patients with HNL-HLH, including 3 females and 2 males. The median age was 37 years (range: 20-57). All five patients showed enlarged lymph nodes and prolonged fever. Laboratory findings were consistent with the diagnosis of HLH. All five patients had hyperferritinemia and the levels of ferritin were all higher than 1000 ug/L. Serum lactate dehydrogenase (LDH) and aminotransferase were significantly elevated in all five patients. 18F-FDG PET/CT showed enlarged lymph nodes with increased FDG uptake and splenic hypermetabolism could be present. Pathological findings were consistent with HNL (Fig A) and hemophagocytosis in the lymph node was present in two cases (Fig B). All the patients responded well to corticosteroids and had a good prognosis. Two of the five patients were diagnosed with systemic lupus erythematosus (SLE) during the follow-up.

Conclusion: Our study demonstrated that adult patients with HNL-HLH showed distinct clinical, laboratory, and radiological features. And the prognosis is good and patients could be managed with steroids and supportive care.
Dynamic Changes in Lysine Succinylation As Important Regulators of Erythropoiesis
Authors: Hu Bin , Gong Han , Yang Chaoying , Nie Ling , Zhang Ji , Liang Long , Narla Mohandas , Sheng Yue , Liu Jing
Published: December 2023

Lysine succinylation has emerged as a recently discovered protein modification that significantly impacts the chemical environment and exhibits diverse functions in various biological processes. However, the specific role of lysine succinylation in erythropoiesis has not been fully elucidated. In this study, we investigated the levels of six common acylations (acetylation, crotonylation, succinylation, propionylation, butyrylation, and malonylation) in human erythroid cells. Interestingly, we observed a prominent accumulation of lysine succinylation during human erythroid differentiation, suggesting its potential importance in this process. To explore the functional significance of succinylation, we inhibited succinylation in human erythroid progenitor cell line by disrupting the expression of the key succinyltransferases and desuccinylases. The results revealed that succinylation inhibition led to suppressed cell proliferation, increased apoptosis, and disrupted differentiation, indicating the essential role of succinylation in erythropoiesis. Furthermore, integrative proteome and succinylome analysis identifies 939 quantifiable proteins with 2,871 Ksu sites. Notably, we observed inconsistencies between alterations in protein levels and succinylation levels, suggesting that the role of succinylation in proteins' function regulation. These succinylated proteins are widely distributed in various cellular compartments and involved in multiple cell processes, indicating that succinylation is a prevalent modification in erythropoiesis. Mechanically, we identified CYCS as a key target of succinylation during erythropoiesis, emphasizing its essential role in this process. Specially, we implicated KAT2A-mediated histone succinylation in chromatin remodeling, further highlighting the regulatory significance of lysine succinylation in erythropoiesis at the epigenetic level. Collectively, our comprehensive investigation of the succinylation landscape during erythropoiesis provides valuable insights into its regulatory role and offer potential implications for erythroid-related diseases and therapeutic strategies.
One-Pot Era-CRISPR/Cas12a System for Rapid and Sensitive Detection of Multiple NPM1 Mutations
Authors: Liu Yin , Liu Xinyi , Huang Xingxu , Li Kui , Wang Xinjie , Zhou Fuling
Published: December 2023

Nucleophosmin 1 (NPM1) gene c.863_864 4-bp insertion mutations occur in 30% of adult acute myeloid leukemia (AML) patients, which is associated with a unique pathological phenotype and good prognostic significance. The detection of NPM1 mutations is important for risk stratification, treatment guidance, and minimal residual disease monitoring in AML. However, the current detection methods have their limitations in terms of equipment requirements, sensitivity, turnaround time, price, etc. Moreover, simultaneous detection of multiple NPM1 mutation types is challenging. Here, we developed a one-pot ERA-CRISPR/Cas12a-based assay in which only one guide crRNA is needed to detect hundreds of NPM1 gene variants, covering over 97% of NPM1-mutated AML cases. By optimizing multiple parameters, this assay finally achieved a sensitivity of as low as 0.01% within 30 min detection under constant 39℃, without wild-type related cross signal. In clinical detection, 21 NPM1 mutation positive cases from 70 AML patient were successfully screened out using our method, including two cases not detected by Sanger sequencing. This method is one-step, rapid, sensitive, specific, inexpensive, and without the need of Sequencer or temperature cycler. It holds the potential to assist clinicians in point of care testing.
Zanubrutinib Combined Regimen As Bridging Treatment on Promising Clinical Outcome of CD19 Chimeric Antigen Receptor T-Cell Therapies in Relapsed or Refractory Large B-Cell Lymphoma
Authors: Lu Yan , Ye Shiguang , Zhou Lili , Li Ping , Li Shaoguang , Liang Aibin
Published: December 2023

Background

Relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL) shows poor clinical outcomes. Anti-CD19 chimeric antigen receptor-T cells (CAR-T) therapy is emerging as the second-line therapy of choice, but in real-world studies its efficacy and long term follow is not promising. Effective bridging therapy can reduce tumor burden and improve CAR-T efficacy. Zanubrutinib, a selective Bruton tyrosine kinase (BTK) inhibitors, combined regimens have established therapeutic activity by targeting B-cell receptor signaling and have individually shown efficacy in patients with R/R DLBCL. Thus, the present study evaluated the efficacy and safety of zanubrutinib-combined regimen as bridging treatment in CD19 CAR-T therapy.

Methods

This retrospective study includes all patients participant in the clinical trail (NCT 02537977) between January 2020 and December 2022. All 21 heavily treated patients with r/r DLBCL who received zanubrutinib-combined regimens as bridging therapy are included. Patient clinical characteristics were collected upon patient enrollment. Efficacy outcomes included overall response rate (ORR), progression-free survival (PFS), and overall survival (OS), whereas safety outcomes included incidence of adverse events. The zanubrutinib combined regimens are zanubrutinib with IMIDs, chemotherapy and/or rituximab.

Results: Of the 21 evaluable patients, median age of patients is 57 year old and median treatment line is 3. 19 patients (90%) are non-GCB subtype and 18 patients(86%) are IPI score >3. 14 patients(67%) are double expression and 7 patients (33%) have TP53 mutation. Objective response rate(ORR) is 81%; of whom, 10 patients(48%) got complete remission(CR) and 7 patients(33%) got partial remission(PR). With a median follow-up of 25 months, the median PFS was 12.8 months and median OS was not yet reached. 8 CR patients and 3 PR patients maintain long-term remission and are still under followup. 18 patients (86%) has grade 1-2 cytokines release syndrome(CRS) and 3 patients(14%) has grade 1 immune effector cell-associated neurotoxicity syndrome (ICANS). No severe CRS and >1 grade ICANS were observed. The most common hematologic toxicities included grade ≥3 neutropenia (90%) and grade 3/4 thrombocytopenia (71%). No patients had grade ≥3 non hematologic toxicities.

Conclusion:

Zanubrutinib-combined regimens are effective and safe as bridging therapy in CD19 CAR-T therapy. More prospective studies are further command.
A Real-World Efficacy of Letermovir Primary Prophylaxis for Cytomegalovirus Infection after Allogeneic Hematopoietic Stem Cell Transplantation in China: A Single-Center Retrospective Analysis
Authors: Zhang Xiaoyu , Yang Donglin , Pang Aiming , He Yi , Wei Jialin , Zhang Rongli , Zhai Weihua , MA Qiaoling , Chen Xin , Han Mingzhe , Feng Sizhou , Jiang Erlie
Published: December 2023

Introduction: Cytomegalovirus (CMV) infection is a common complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT) and is associated with increased mortality. Letermovir (LET) is a novel antiviral drug used to prevent CMV infection.

Methods: We retrospectively analyzed 167 consecutive patients who underwent allo-HSCT between August 2022 and May 2023, who received LET 240 mg/day, to evaluate the efficacy and safety of LET in primary prophylaxis for clinically significant CMV infection (csCMVi), defined as CMV disease or CMV viremia reactivation, in real-world situations in China.

Results: Among the enrolled 167 patients, 95.21% (159/167) of patients underwent a haploidentical donor (HID) transplant and 4.79% (8/167) a matched unrelated donor (MUD) transplant. 96.41% of the recipients were CMV seropositive, and only 6 recipients were CMV seronegative but donor seropositive. The median interval from allo-HSCT to the administration of LET was 7 (2-55) days. A total of 121 patients received LET within 14 days after HSCT. With the median follow-up 162(74-397) days post-HSCT, the incidence of CMV viremia was 11.98% (20/167). 3 patients developed CMV viremia after discontinuation of LET medication. Among the 20 patients with CMV viremia, the median duration of preemptive therapy was 9 (3-28) days, and the peak level of CMV-DNA was 1867 (1228-22383) copies/ml. Only 1 patient developed refractory CMV viremia with prolonged preemptive anti-CMV treatment. No patients were diagnosed with clinically significant CMV disease. Patients who developed CMV viremia or not were similar with respect to the CMV IgG status of the recipient at HSCT, donor CMV IgG, and GVHD incidence. Notably, the CMV viremia incidences of patients who received LET within 14 days and after 14 days post-HSCT were 7.44% and 23.91%, respectively (p=0.005). The incidence of EBV viremia was 17.16% (29/169), and 9 developed PTLD. The six-month overall survival was 93.4±2.1% in all patients who received LET prophylaxis. Meanwhile, LET prophylaxis was well tolerated. No drug-related death or permanent discontinuation was reported due to toxicity.

Conclusion: Letermovir was effective and safty as primary prophylaxis of csCMVi, particularly initiating early post-HSCT.
Homoharringtonine Overcomes the Negative Impact of Genetic Patterns on Venetoclax Plus Azacitidine Regimen in Relapsed/Refractory Acute Myeloid Leukemia: A Multi-Center, Cohort Study
Authors: Yu Guopan , Zhang Yu , Jin Hua , Yin Zhao , Weng Guangyang , Yu Sijian , Xu Na , Du Xin , Lin Dongjun , Xiao Jie , Sun Zhiqiang , Zhang Hongyu , Liang Xinquan , Guo Ziwen , Zhao Weihua , Dai Min , Jiang Xuejie , Xuan Li , Shi Pengcheng
Published: December 2023

Objective: Whether other agents added to venetoclax (VEN) plus hypomethylating agents (HMA) regimen could overcome the negative impact of genetic patterns on response in patients with relapsed/refractory acute myeloid leukemia (RR-AML) remains unclear. Aim of this study was to evaluate whether homoharringtonine (HHT) added to VEN plus azacitidine (AZA) regimen could improve the response and overcome the negative impact of genetic patterns in patients with RR-AML .

Methods: A multi-center, cohort study of the response and the genetic patterns of response of VEN plus AZA and HHT (VAH) versus VEN plus AZA (VA) regimens as salvage treatment in the patients with RR-AML was performed. Patients were enrolled from four studies from October 2018 to December 2022 at nine medical centers in china.

Results: A total of 321 patients were analyzed, including 150 females and 171 males, with a median age at 46 (IQR, 35-61) years. There were 172 patients in the VAH and 149 in the VA group. VAH significantly improved CRc rate (66.3% vs. 44.3%, P<0.001) and prolonged OS (median OS, not reach vs. 14.3 months, P=0.004), to compared with VA. Multivariate analysis showed that VAH was the independently protective factor for CRc and survival. In addition, VAH significantly overcame the negative impact of FLT3-ITD/TKD, N/KRAS, TET2, DNMT3A mutations, and t(8;21)/AML1-ETO, as well as non-adverse ELN risk, also apparently in adverse ELN risk or complex karyotype, on the response of VA regimen.

Conclusion: The impact of genetic patterns on the response presented diversely in different VEN-based regimens. HHT added to VA regimen might improve the response and overcome the negative impact of part genetic patterns in RR-AML.

Key words: homoharringtonine, venetoclax, relapsed/refractory, acute myeloid leukemia, genetic pattern
Targeting RBM39-MEK5 Axis Synergizes with Bortezomib to Inhibit Multiple Myeloma Malignancy
Authors: Liu Jia , Zhang Zilu , Xu Wenbin , Jia Ming-Yuan , Wu Chengyu , Hua Yan
Published: December 2023

Multiple myeloma (MM) is a type of haematological malignancy characterised by the accumulation and expansion of malignant plasma cells in the bone marrow (BM). Despite recent advances in chemotherapy, MM remains an incurable disease for most patients and therefore requires novel therapies. Abnormal alternative splicing is a hallmark of cancer and is associated with poor survival and an ultra-high-risk population in MM (Bauer et. al. Haematologica 2021). Targeting splicing is a promising therapeutic strategy in MM. Our previous study showed that RBM39 is highly expressed in primary MM cells and cell lines and correlates with poor overall survival (Tong et. al. Haematologica 2020). To investigate the underlying role of RBM39 in MM, we knocked down RBM39 in MM cell lines. We found that knockdown of RBM39 inhibited MM cell proliferation, induced apoptosis and cell cycle arrest at the G2/M phase, and inhibited subcutaneous tumour growth. Indisulam has been reported to induce RBM39 degradation via the proteasomal pathway by acting as a molecular glue to link RBM39 to the E3 ubiquitin ligase DCAF15 (Han et. al. Science 2017). In our study, Indisulam treatment in MM cells was shown to degrade RBM39 via DCAF15. Indisulam could induce apoptosis and G2/M phase arrest in MM cell lines and inhibite tumour growth in vivo. It also significantly decreased MM cell viability in a co-culture system with MM BM-derived mesenchymal stromal cells. Indisulam was also able to inhibit primary CD138+ cells isolated from newly diagnosed MM patients.

To explore the mechanistic role of RBM39 in promoting MM malignancy, we performed RNA immunoprecipitation (RIP)-Seq and RNA sequencing in RBM39 knockdown MM cells to investigate the alternative splicing targets of RBM39. KEGG pathway analysis of differentially expressed and alternatively spliced genes revealed the enrichment of the MAPK pathway. We then identified MEK5, a non-redundant member of the MAPK family, as a potential target of RBM39, which is highly expressed in MM. Full-length MEK5 contributes to increased proliferation and survival of MM cells. We further demonstrated that MEK5 is bound and alternatively spliced by RBM39 and that splicing of full-length MEK5 is dependent on RBM39. Given the importance of the RBM39-MEK5 axis in MM, we also found that inhibition of RBM39 with Indisulam or MEK5 with the inhibitor BIX02189 increased the cytotoxicity of bortezomib in vivo and in vitro.

In conclusion, we have demonstrated that the RBM39-MEK5 axis, in which RBM39 maintains splicing of full-length MEK5, is essential for MM cell survival and proliferation. Mechanistically, shRNA-mediated knockdown or Indisulam-mediated degradation of RBM39 caused extensive altered splicing, including mis-splicing of MEK5. Targeting RBM39 or MEK5 could not only inhibit MM malignancy, but also enhance the cytotoxicity of bortezomib for MM.
Targeted, Safe, and Efficient Gene Delivery to Human Hematopoietic Stem and Progenitor Cells In Vivo Using Engineered Avid Adenovirus Vector Platform
Authors: Yao Jia , Atasheva Svetlana , Wagner Nicole , Di Paolo Nelson C , Stewart Phoebe L , Shayakhmetov Dmitry M.
Published: December 2023

Precise editing of human genome using CRISPR-Cas nucleases, base editors, prime editors, or other technologies represents a promising approach for ameliorating or correcting many human genetic diseases. Although numerous pre-clinical and clinical proof-of-concept studies demonstrated feasibility of gene correction in the desired regions of the human genome ex vivo, targeted delivery to and cell-type-specific expression of gene editing proteins in various cell types in vivo represent major challenges for all viral and non-viral delivery platforms developed to date. Here, we describe the development and analysis of AVIDs, a novel adenovirus-based gene delivery platform, that allows for a highly targeted, safe, and efficient gene delivery to human hematopoietic stem and progenitor cells (HSPCs) in vivo after intravenous vector administration. The AVID vectors attach to human HSPCs via engineered fibers that bind to either CD46 or DSG2 receptors, which are highly expressed on human HSPCs. The efficient entry of AVIDs into these cells is mediated via a mutated penton that was engineered to interact with CD49f, α6 integrin class, whose expression is restricted to cells with long-term repopulating and stem cell capacity. To improve virus safety after intravenous administration, as well as to increase the efficacy of virus production for a cost-effective high-yield vector manufacturing, AVIDs further comprise mutations in hypervariable loops of the major capsid protein hexon. While in vitro infection with AVIDs leads to about 30% transduction of a pool of human CD34 + cells, the efficacy of transduction of CD34 +CD38 -CD45RA -CD90 + subsets, highly enriched for hematopoietic stem cells (HSCs), ranged from 80% to 100% for individual donors. Single intravenous administration of AVIDs to humanized mice, grafted with human CD34 + cells, after mobilization led to up to 20% transduction of CD34 +CD38 -CD45RA- HSPC subsets in the bone marrow. Importantly, targeted in vivo transduction of CD34 +CD38 -CD45RA -CD90 -CD49f + subsets, highly enriched for human HSCs, reached up to 19%, which represented a 1900-fold selectivity in gene delivery to HSC-enriched over lineage committed CD34-negative cell populations in the bone marrow. Because the AVID platform allows for regulated, cell type-specific expression of gene editing technologies as well as for expression of immunomodulatory proteins to ensure persistence of corrected HSCs in vivo, the HSC-targeted AVID platform may enable development of novel curative therapies through safe and effective in vivo correction of disease-causing mutations in human HSCs after a single intravenous administration.
Selinexor in Combination with Salvage Regimen for Refractory Peripheral T-Cell Lymphoma (PTCL) Patients
Authors: Wang Li , CAI Mingci , Huang Yaohui , Xu Pengpeng , Chen Shu , Zhao Weili
Published: December 2023

Background

Peripheral T-cell lymphomas, not otherwise specified (PTCL-NOS) are a diverse and aggressive type of non-Hodgkin lymphomas with a poor prognosis when treated with conventional chemotherapy. The standard CHOP chemotherapy regimen typically yields an overall response rate (ORR) of 60-80%, a complete response (CR) rate of 30-40%, and long-term survival, measured by 5-yr OS，in the range of 20-30% in non-ALCL (anaplastic large cell lymphoma) subtypes. Azacitidine plus CHOP has achievied a 75% CR in newly diagnosed PTCL, but patients with DNMT3A mutation showed adverse progression-free survival (PFS), indicating that genetic mutations may influence resistance to chemotherapy. Selinexor has shown promising results in refractory lymphoma patients by inhibiting oncogenes, therefore, the efficacy of selinexor combined salvage regimen in refractory PTCL remains great interest.

Method

Refractory PTCL patients were enrolled to be treated with selinexor plus GDP/ICE/Gemox since December 2021 at our center. Selinexor was administered orally (40 mg day 1, 8, 15) plus GDP (gemcitabine 1g/m2*d1, cisplatin 25mg/m2 d1-d3, dexamethasone 40mg d1-d4), ICE (ifosfamide 1.5g/m2*3d, carboplatin (AUC 5) and etoposide 100mg/m2*3d), Gemox (gemcitabine 1g/m2*d1, oxaliplatin 100mg/m2*d1) every 3 weeks as per physician's choice. Objective response rate (ORR) was evaluated every 3 cycles based on Lugano 2014 criteria and Adverse events (AE) were rated according to the NCI CTCAE 5.0.

Results

10 patients were treated with selinexor plus salvage regimen in this study and received more than two cycles of the treatment, making them eligible for response evaluation in this poster. The median age of the 10 patients was 65 years (range from 42 to 79), 8 (80%) were male, 9 (90%) had disease stage III-IV at screening, and 60% of all patients had an IPI intermediate to high risk. All of the patients were refractory to the last line of treatment. Among 10 patients, 2 were angioimmunoblastic T-cell lymphoma (AITL), 3 with PTCL-NOS, 3 with ALK-ALCL, 2 with monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL). As shown in Figure 1, 3 patients had TP53 alterations, 2 patients with DNMT3A mutation. Patients received a median of three cycles (range, 2-6) of selinexor 40mg once a week combined with salvage regimen（4 patients received Selinexor-GDP, 3 with selinexor-ICE, 3 with selinexor-Gemox）.

The best overall and complete response rates were 70% and 50%, respectively. In terms of MEITL refractory to conventional CHOP treatment, 1 achieved PR but progressed and died after stop treatment for three months due to Covid19 and 1 in CR with selinexor maintenance.

Conclusions

There are limited options for the treatment of refractory PTCL. We identified a dosing schedule of selinexor (40mg qw) with GDP/ICE/Gemox that is efficiency inrefractory PTCL and is worthy of further study on its mechanism of action.
Targeting NUP214 Eradicates Leukemia Stem Cells By Inducing Ferroptosis
Authors: Chen Zhe , Wu Feng , Li Lei , Hou Yu
Published: December 2023

Acute myeloid leukemia (AML) arises from and are maintained by leukemia stem cell (LSCs), which are also critical causes for drug resistance and relapse of AML. The critical genes involved in LSC maintenance remain largely unknown. Here, we performed a pooled negative-selection screen with a customized small hairpin RNA (shRNA) library against 1191 known pan-essential genes of AML blast cells. We found that several Nucleoporins (NUPs) genes were essential for LSC survival, and NUP214 was highly expressed in LSCs. Knock-out of NUP214 completely eradicated LSCs in MLL-AF9 and HOXA9-MEIS1 AML mouse models, but also comprised HSC maintenance and function. Haploinsufficiency of NUP214 resulted in the increased death of LSCs (~4% to ~33%), reduced LSC numbers, and extended survival of AML mice. In contrast, the haploinsufficiency of NUP214 had a negligible effect on normal hematopoietic stem cells (HSCs) and hematopoiesis. Haploinsufficiency of NUP214 substantially induced lipid peroxidation and resulted in ferroptosis of LSCs, attributed to the remarkably increased transcription of lipoxygenase-15 (ALOX15) and heme oxygenase-1 (HMOX1), evidenced by the results of RNA-seq, proteome, and functional experiments. Immunofluorescence revealed that NUP214 was a mobile nucleoporin and largely located inside the nucleoplasm in LSCs, while NUP214 was mainly located in the nuclear periphery. Co-immunoprecipitation and Cleavage Under Targets and Tagmentation (CUT&Tag) showed that NUP214 directly interacted with transcriptional coactivator Sub1 in LSCs, binding the gene locus of HMOX1 and ALOX15, eventually inhibiting their transcription. Collectively, our findings reveal the essential roles of NUP214 in LSC maintenance and provide a valuable target for AML therapy.
Clot-Based Assay for Factor XI (FXI) Activity Is Highly Sensitive to Lupus Anticoagulant (LA) and Heparin with Potential Misdiagnosis of FXI Deficiency: The Importance of Recognizing Non-Parallelism
Authors: Tushek Michael , Mitsios John V. , Francischetti Ivo
Published: December 2023

Introduction: Clot-based assays are employed to estimate coagulation factor activity in clinical pathology laboratories. These assays are PT- or aPTT-based and utilize dilutions of the patient's plasma in factor-deficient plasma. The curves of activity for each dilution follow a parallelism or non-parallelism pattern and the latter is typically seen in the presence of Lupus Anticoagulant (LA). Identification of this pattern is critical to avoid misdiagnosis of factor deficiency in patients who actually have an inhibitor. Materials and Methods: We observed low Factor XI (FXI) levels in patients with LA, while other factors were not affected. In order to determine the reason(s) why FXI levels are sensitive to LA, we collected 34 plasma samples from patients with dRVVT confirm ratio > 1.2, and estimated FVIII and FXI levels. FXI activity was also estimated with Dade ® Actin ® FS, a reagent that is not sensitive to the LA. The coefficient of variance (CV) for both FXI and FVIII levels were compared. To estimate the effects of heparin, plasma was spiked (0.1-1.0 IU/ML) and tested in the PTT, FXI, and FVIII assays. We tested with Dade ® Actin ® FS 4 known normal patients, 3 known FXI deficient patients, 20 known plasma positive for LA, and 10 with heparin. All assays were performed on a Siemens CS-5100 analyzer. Results: The results show that FXI is highly sensitive to LA, with non-parallelism and high CV, not observed with FVIII activity. As expected, FXI levels were not affected when LA-insensitive Dade ® Actin ® FS was used. Heparin (0.3-0.7 IU/ML) also showed non-parallelism in FXI activity, but not for FVIII. Only the highest dose of heparin (1.0 IU/ML) affected FVIII levels. Notably, the sensitivity of FXI activity to LA and heparin was explained by a few dilutions of the patient's plasma (5X) in the FXI assay, compared to other coagulation factors (20X). Of note, less diluted plasma is required for FXI activity because relatively higher levels of FXI is required to sustain and produce clot in the FXI-deficient plasma. Conclusion: This simple, yet not recognized explanation based on a literature search, is helpful to increase awareness of this analytical variable. It is also important to avoid misdiagnosis of FXI deficiency in patients with LA.
Preliminary Study on the Role of Kynurenic Acid in Inhibiting the Physiological Changes of Astrocyte Induced By HIV-1 Envelope Protein gp120
Authors: Chen Xiaoliang , Tang Yi , Wang Zixuan , Long Min , Cao Hong
Published: December 2023

Background

Since the introduction of combined antiretroviral therapy, acquired immunodeficiency syndrome (AIDS) caused by human immunodeficiency virus (HIV) has transformed from an incurable fatal disease to a chronic treatable disease. HIV-associated neurocognitive disorder (HAND) is a common long-term complication of AIDS. Astrocytes are believed to play an essential role in the development of HAND. Therefore this study aims to find out how astrocytes change and how to resist this change in the process of hand.

Methods

Cortical astrocytes were dissected from Sprague-Dawley (SD) neonatal rats (1-3 days) with the aseptic operation and digested with 0.25% trypsin 37℃ for 20 min. The cell suspension was dispersed through the cell strainer and seeded into the poly-D-lysine coated T-75 flasks supplemented with 10% FBS and 1% penicillin/streptomycin. After ten days of incubation, the cells were purified in a shaking incubator (250 r/min) at 37℃ for 24 h, and astrocytes were cultured at desired densities in 6-well plates. The cells were identified as >98% pure astrocytes by Cy3-conjugated GFAP immunostaining. Cultured astrocytes were treated with 150 pM HIV-1 gp120 CM for 12 h before applying 5 or 25 μM kynurenic acid(KYNA) for 12 h. Afterward, western blotting technology was used to detect the expression levels of different proteins in each group, and Fuji Image J software was used to analyze the bands.

Results

Compared with the control group, the expression levels of AKT, PI3K and P62 proteins in astrocyte of mice added with gp120 were significantly reduced, which were 38%, 41% and 40%, respectively, while KYNA alone had no significant change; Compared with the addition of gp120 group, the trend of decreased AKT (P<0.05) and PI3K protein expression was inhibited when both gp120 and KYNA were added, with an increase of 43% and 52%, respectively. We observed that the expression level of p-AKT had a similar trend but the expression level of p-PI3K had an opposite trend. This suggests that the effect of gp120 on the PI3K/AKT pathway may have begun with an upregulation of PI3K activation levels.

Conclusion

HIV-1 gp120 can cause changes in the physiological status of astrocytes, and KYNA can counteract this change, which indicates that KYNA may have a potential therapeutic effect on HAND. ( Acknowledgements:National Natural Science Foundation of China, No. 82172259 to H.C.; Undergraduate Training Program for Innovation and Entrepreneurship of SMU, China, No. 202212121247; Corresponding author: Hong Cao, gzhcao@smu.edu.cn )

Keywords：HIV-1 gp120, Kynurenic acid, Astrocyte, AKT, PI3K disorder
Adoptive Transfer of CMV-Specific TCR-T Cells for the Treatment of CMV Infection after Haploidentical Hematopoietic Stem Cell Transplantation
Authors: Chao Ma , Daihong Liu , Dou Liping
Published: December 2023

Background: CMV reactivation after unmanipulated haploidentical stem cell transplantation (SCT) frequently occurs, causing life-threatening morbidities and transplantation failure. Preemptive therapy upon the detection of CMV viremia using antiviral agents is currently the standard of care but its efficacy is quite limited and associated with significant toxicity. The CMV antigen-specific cytotoxic T lymphocytes (CTLs) therapy was limited by the time-consuming manufacture process and relatively low success rate. More effective and safer approaches for the treatment of CMV reactivation after haploidentical SCT are in urgent need.

Methods: A single-arm, open-label Phase I clinical trial evaluating the safety and efficacy of CMV-targeting TCR-T (CMV-TCR-T) cell therapy as the first-line preemptive therapy for patients with CMV reactivation post haploidentical PBSCT was conducted in the Chinese PLA General Hospital (ClinicalTrials.gov Identifier: NCT05140187). Six patients with CMV reactivation after haploidentical SCT were adoptively transferred by 1 to 3 doses of SCT donors-derived CMV-TCR-T cells. This trial was a dose-escalation study with doses ranging from 1×10 3 CMV-TCR-T cells/kg body weight per dose to 5×10 5 CMV-TCR-T cells/kg per dose

Results: Except for the Grade 1 CRS observed in one patient and mild fever in 2 patients, no other adverse events were observed. Four patients had complete response within a month after CMV-TCR-T cells infusion without the administration of any antiviral agents. The other two patients who initially didn't completely respond to CMV-TCR-T cell therapy had salvage ganciclovir and foscarnet administration and then had rapid CMV clearance. The CMV-TCR-T cells displayed overall robust expansion and persistence in the peripheral blood after infusion. The CMV-TCR-T cells were firstly detected in the peripheral blood of these patients 3 to 7 days after the 1 st dose of CMV-TCR-T infusion, rapidly expanded and persisted for at least 1~4 months, providing long-term protections against CMV reactivation. In one patient, the CMV-TCR-T cells started to expand even when the anti-GvHD reagents were still being used , further indicating the proliferation potential of CMV-TCR-T cells.

Conclusions: Our study firstly showed CMV-targeting TCR-T cell as a highly feasible, safe and effective first-line preemptive treatment for CMV reactivation after haploidentical PBSCT.
The Mechanism of Setd2 in Mesenchymal Stromal Cell in MDS-Associated Ineffective Hematopoiety
Authors: Wang Roujia , Wang Lan , Chang Chunkang
Published: December 2023

Myelodysplastic syndrome (MDS) is a heterogeneous, clonal, hematopoietic stem progenitor cell malignant disease. Bone marrow microenvironment participates in the occurrence and development of MDS and plays a key role.

Histone methyltransferase Setd2 is the only enzyme that can catalyze the methylation of mammalian histone H3K36Me3. Previous studies have confirmed that Setd2 is involved in the the transformation of MDS to leukemia, and the expression of S100A9 decreases after Setd2 knockout. This study is based on NHD13 mice with mesenchymal stem cell (MSC) targeted knockout of Setd2. Through experimental methods such as flow cytometry, peripheral blood routine, Wright staining, HE staining, and Gomori staining, it finds that Setd2 targeted knockout mice undergo faster leukemia transformation compared to NHD13 mice; Peripheral blood white blood cells increase, while hemoglobin and platelets decrease; The differentiation of hematopoietic stem progenitor cells in bone marrow is abnormal, with abnormal hematopoiesis and increased blast cells; Spleen fibrosis aggravates and megakaryocyte infiltrates; and Extramedullary hematopoiesis in liver. Through flow cytometry,β- Galactosidase staining and other experimental methods, we finds that compared with NHD13 mice, the MSC morphology of knockout mice is irregular, and the support function for hematopoietic cells is weakened. This study demonstrates that MSC targeted knockout of Setd2 has a promoting effect on the progression of MDS.

We used mass spectrometry to identify proteins that are significantly upregulated in the targeted knockout mouse bone marrow supernatant compared to the NHD13 mouse bone marrow supernatant, and and select the protein Uba5 related to hematopoiesis from them. The expression of Uba5 in the bone marrow supernatant of knockout mice is significantly higher than that in the NHD13 group through WB validation. It is speculated that the hematopoietic changes in Setd2 targeted knockout mice may be related to the downstream target protein Uba5 of Setd2.

This study takes the change of bone marrow microenvironment as an entry point to explore the possible mechanism of MDS ineffective hematopoiesis, hoping to provide microenvironment regulation therapy for some MDS patients with genes that hematopoietic stem cell transplantation that cannot cure(such as TP53, NRAS, KRAS), and provide new ideas for MDS treatment. There are still many limitations in this study, further clinical validation and vivo and vitro rescue experiments on downstream target genes and proteins are still needed in the future.
Comparison of Long-Term Outcome between Imatinib and Dasatinib Prophylaxis after Allogeneic Stem Cell Transplantation in Patients with Philadelphia-Positive Acute Lymphoblastic Leukemia-a Multi-Center Retrospective Study
Authors: Guan Fangshu , Yang Luxin , Zhu Panpan , Lai Xiaoyu , Liu Lizhen , Shi Jimin , Zhao Yanmin , Yu Jian , Fu Huarui , Zhao Yi , Lan Jianping , Chen Yi , Lu Ying , Ouyang Guifang , Xu Yang , Qiu Xi , Cai Zhen , Huang He , Luo Yi
Published: December 2023

Background: Though the prognosis of Philadelphia-positive (Ph-positive) acute lymphoblastic leukemia (ALL) has been improved since the introduction of tyrosine kinase inhibitors (TKIs) and combined with allogeneic stem cell transplantation (allo-HSCT), relapse after HSCT remains a concern. Prophylactic TKIs after allo-HSCT seemed a promising strategy to prevent relapse. But the optimal choice of the agent is still obscure.Methods: We retrospectively analyzed the clinical data of Ph-positive ALL patients who underwent allo-HSCT in CR1 from 6 medical centers in China between September 2010 and October 2021. Patients were divided into two cohorts according to different choice of TKI after allo-HSCT, namely Ima cohort with imatinib prophylaxis and Das cohort with dasatinib prophylaxis. Survival and safety outcomes of two cohorts were compared, and prognostic factors were determined by univariate and multivariate analyses.Results: One hundred and forty-one patients, with median age of 37 years (range, 14-61 years), were included, of whom 91 were in Ima cohort and 50 were in Das cohort. After median follow-up of 50.6 months, 5-year cumulative relapse rate (CIR), non-relapse mortality rate (NMR), leukemia-free time (LFS) and overall survival (OS) in Ima and Das cohort were 16.1% and 12.5%; 5.2% and 9.8%; 78.8% and 77.6%; 86.5% and 77.6%, respectively, with no significant difference. Mild cGVHD was higher in Das cohort compared to Ima cohort, while the total, moderate or severe cGVHD were similar. The most common adverse event was neutropenia (66.4%). Das cohort owned higher incidence of gastrointestinal bleeding and gastrointestinal reaction than Ima cohort. The proportion of patients treated on schedule was 30% in Das cohort, lower than that of in Ima cohort (p<0.001). Drug intolerance was the main reason for not completed treatment as intended.Conclusions: For patients with Ph-positive ALL receiving allo-HSCT in CR1, imatinib prophylaxis after transplantation could obtain the same long-term outcome compared to dasatinib, showing better tolerance.
Granulocyte Colony-Stimulating Factor and Decitabine-Containing Conditioning Improves Survival in Myelodysplastic Syndrome Patients with Iron Overload Undergoing Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Zhao Wenshu , Pan Danqi , Zeng Xiangzong , Xuan Li , Fan Zhiping , Huang Fen , Xu Na , Sun Jing , Liu Qifa , Dai Min
Published: December 2023

Background: Iron overload is considered as an unfavorable prognosis in myelodysplastic syndrome (MDS) even in those undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although iron chelation therapy has improved the prognosis of these patients to some extent, the effect has not yet been satisfactory. This study aimed to investigate whether granulocyte colony-stimulating factor and decitabine (G-DAC)-containing conditioning improve the prognosis of iron-overloaded MDS patients undergoing allo-HSCT.

Study Design: One hundred and ninety-seven patients were enrolled in this retrospective study. Based on the level of serum ferritin (SF) and conditioning regimen, all patients enrolled were divided into 4 groups: SF﹤1000µg/L with G-DAC conditioning (cohort 1), SF﹤1000µg/L with non-G-DAC conditioning (cohort 2), SF ≥ 1000µg/L with G-DAC conditioning (cohort 3) and SF ≥ 1000µg/L with non-G-DAC conditioning (cohort 4). The clinical features and prognosis between cohorts were analyzed.

Results: The 2-year overall survival (OS) was 77.0%, 72.1%, 73.8% and 38.7% ( P = 0.001), and disease-free survival (DFS) was 75.7%, 62.8%, 71.7% and 35.5% ( P = 0.001), and the cumulative incidence of non-relapse mortality (NRM) was 17.3%, 25.0%, 15.2% and 53.1% ( P = 0.001), and the incidence of relapse was 8%, 13.6%, 13.1% and 12.5% ( P = 0.592), respectively, in the four groups. To be specific, cohort4 had worse OS and DFS and higher NRM than the other three groups (all P﹤0.05). Multivariate analysis revealed that SF ≥ 1000µg/L was a risk factor for OS, DFS and NRM ( P = 0.022, P = 0.025, P = 0.036), while G-DAC-containin g conditioning was a protective factor ( P = 0.009, P = 0.003, P = 0.004). Intriguingly, when cohort 1 to cohort4 were included in the multivariate analysis, only cohort4 was a risk factor for OS, DFS and NRM (all P﹤0.05), but cohort3, namely iron-overloaded patients who received G-DAC-containing conditioning had no difference in prognosis compared with patients with SF﹤1000µg/L.

Conclusions: The poor prognosis of patients with iron overload could be overcome by G-DAC-containing conditioning partly.
Corelationship of Lymphocyte Subsets and Prognosis in Juvenile Myelomonocytic Leukemia
Authors: Chen Yunlong , Zhang Jingliao , Liu Fang , Zhang Yingchi , Zhu Xiaofan , Yang Wenyu
Published: December 2023

Objective: Juvenile Myelomonocytic Leukemia (JMML) is a rare hematopoietic stem cell malignant clonal disease in children and has the characteristics of MDS/MPN. Due to the poor response of JMML to chemotherapy, hematopoietic stem cell transplantation (HSCT) is currently the only cure method for JMML. Currently, there are some reports on the distribution of lymphocyte subsets in MDS. A significant lymphocytopenia was observed in MDS patients compared with similarly aged healthy people. Another study form Polish found patients with MDS have more CD3+T cells than healthy donors. But there are few related reports in JMML. The objective of this study is to analyze relationship between the distribution of lymphocyte subsets and prognosis in JMML patients, and explore new risk stratification indicatorsfor JMML personalized therapy.

Method: This study retrospectively collected JMML patients clinical data from Institute of Hematology & Blood Diseases Hospital of CAMS during Jan 2008 and Nov 2022. All completed complete blood count and peripheral blood classification, fetal hemoglobin (HbF), granulocyte colony-stimulating factor (GM-CSF) high sensitivity, bone marrow morphology, immune typing, molecular biology, cytogenetics and other tests. We analyzed the relationship between distribution of lymphocyte and prognosis in these patients.

Result: 96 newly diagnosed JMML patients were admitted to this study. The median age at diagnosis was 25 (2-82) months, and male to female was 3.8:1. 66 cases underwent lymphocyte subsets by flow cytometry. Median percentage of lymphocytes is 34.3%(1.8%-90.4%, normal range: 20%-40%), median percentage of CD3+T cells is 54.85%(10.3%-91%, normal range: 57%-88%), median percentage of CD3+CD4+ T cells is 30.95%(3.54%-53%, normal range: 27%-58%), median percentage of CD3+CD8+ T cells is 18.4%(2%-49.1%, normal range: 16%-45%), median percentage of NK cells is 13.95%(2.3%-50.3%, normal range: 3%-32%), and median percentage of B cells is 23.25%(4%-69.6%, normal range: 4%-18%). These cases were followed-up to Jan 2023, with a median follow-up time of 12 months (1-104 months). The overall survival (OS) rate was 52.67%, with a median OS time of 12 months (0-103 months). There were significant differences in the percentage of CD3+T cells (30.89% ± 9.46% vs 59.05% ± 10%, P<0.01) and CD3+CD4+T cells (14.87% ± 5.82% vs 35% ± 5.41%, P<0.01) between children with single NRAS mutation and compound mutations (≥2 classical five Ras pathway mutation ). In PTPN11 mutation patients, there is a significant positive correlation between OS and the percentage of lymphocytes (P=0.018).

Conclusion: Our study shows differences in percentage of CD3+ T cells and CD3+CD4+ T cells between patients with NRAS mutation and compound mutations. Patients with PTPN11 mutation and higher percentage of lymphocytes will have better prognosis.