bluuebunny/crossref_metadata_2025 huggingface dataset:
Transcriptomic Analysis of the Functions of CSF1R-Dependent Macrophages in Postnatal Development in the Rat
Authors: Carter-Cusack Dylan , Huang Stephen , Keshvari Sahar , Patkar Omkar , Sehgal Anuj , Allavena Rachel , Byrne Robert , Morgan Paul , Bush Stephen , Summers Kim Z , Irvine Katharine M , Hume David A
Published: December 2024

Adaptation to existence outside the womb is a key event in the life of a mammal. The absence of macrophages in rats with a homozygous mutation in the Csf1r gene (Csf1rko) severely compromises pre-weaning somatic growth and maturation of organ function. Transfer of wild-type bone marrow cells (BMT) at weaning rescues tissue macrophage populations permitting normal development and long-term survival. To dissect the phenotype and function of macrophages in postnatal development, we generated transcriptomic profiles of all major organs of wild-type and Csf1rko rats at weaning and selected organs following rescue by BMT. The transcriptomic profiles revealed subtle effects of macrophage deficiency on development of all major organs. Network analysis revealed a common signature of CSF1R-dependent resident tissue macrophages that includes the components of complement C1Q (C1qa/b/c genes). Circulating C1Q was almost undetectable in Csf1rko rats and rapidly restored to normal levels following BMT. Tissue-specific macrophage signatures were also identified, notably including sinus macrophage populations in the lymph nodes that have not previously been identified. Their loss in Csf1rko rats was confirmed by immunohistochemical localisation of CD209B (SIGNR1). By 6-12 weeks, Csf1rko rats succumb to emphysema-like pathology associated with the selective loss of interstitial macrophages and granulocytosis. This pathology was prevented by BMT. Along with physiological rescue, BMT precisely regenerated the abundance and expression profiles of resident macrophages. The exception was the brain, where BM-derived microglia-like cells had a distinct expression profile compared to resident microglia. In addition, the transferred BM failed to restore blood monocyte or CSF1R-positive bone marrow progenitors. Considering the integrated data we provide insight into the inter-related systemic consequences of developmental delay in bone, liver and pituitary and potential contributions to somatic growth deficiency in Csf1rko rats.
A Multicenter Clinical Study of Cccg - APL-2017 in the Treatment of Children with Acute Promyelocytic Leukemia
Authors: Chang Lixian , GAO Ju , Lei Xiaoying , He Yingyi , Zhuang Shuquan , Li Chunhuai , Weng Kaizhi , Wang Lingzhen , Guo Xia , Liu Qihui , Wang Pengfei , Zhuang Yong , Yan Mei , Liu Wei , Chen Hui , Zhang Min , Shen Shuhong , Zhu Xiaofan , Ju Xiuli , Zhang Li , Wang Zhuo
Published: December 2024

The standard therapy for childhood acute promyelocytic leukemia (APL) involves the combination of all-trans retinoic acid (ATRA) with arsenic trioxide (ATO) or Compound Huangdai tablet (Realgar-Indigo Naturalis formula, RIF). However, the effectiveness and safety of combining RIF with ATRA in a larger population of pediatric APL patients remains undocumented. Additionally, there is variability in the oral timing and dosage of RIF in different studies, which should be standardized.

We conducted a multicenter clinical trial (ChiCTR-OIC-16010014) in China. Individuals newly diagnosed with APL were categorized into two risk groups based on their initial white blood cell (WBC) count. Those with a WBC count exceeding 5×10^9/L received hydroxycarbamide at a dosage ranging from 20 to 50 mg/kg per day until their WBC levels dropped to 10×10^9/L or lower. The primary outcomes assessed were event-free survival (EFS) and overall survival (OS) over six years.

We recruited 200 patients diagnosed with APL. After a median follow-up time of 43.2 months, it was observed that the six-year OS rate was 100% in the low-risk cohort and 97.6% in the high-risk cohort. The six-year EFS rate was found to be 98.3% in the low-risk group and 97.6% in the high-risk group. It was observed that plasma levels of arsenic remained stable after the administration of RIF at a dosage of 60 mg/kg/d for seven days and returned to baseline levels within fourteen days after discontinuation of RIF administration. Furthermore, treatment strategies aimed at controlling WBC counts to maintain levels at or below 30×10^9/L were found to reduce the incidence of induced differentiation syndrome (DS) or alleviate its associated symptoms.

In conclusion, The CCCG-APL-2017 regimen, which combines RIF with ATRA, demonstrates both efficacy and safety in treating children with APL. A dosage of 60mg/kg/d of RIF is considered sufficient for the management of pediatric APL.
Hematopoietic Stem Cell Transplantation (HCST) Is Underutilized in Black Patients with Myeloid Neoplasms. Results from a Retrospective Study at Chicago's Leading Minority-Serving Hospital
Authors: Turk Ekrem , Elsebaie Maha , Deenadayalan Vaishali , Obeidat Khaldun , Nanaa Ahmad , Zia Maryam , Birch Noah
Published: December 2024

Background: Access to hematopoietic stem cell transplantation (HSCT) is crucial for patients with myeloid neoplasms, yet disparities in its utilization are evident, particularly among ethnically diverse, minority-serving hospitals. These disparities can exacerbate treatment inequities. This study aims to understand how race and ethnicity impact access to and decisions regarding HSCT. Methods: In this retrospective study, we examined patients over 18 with myeloid neoplasms or premalignant conditions, who had undergone genetic profiling. Transplant eligibility was decided through a comprehensive evaluation of patient's risk classification, comorbidities, and donor availability by the leukemia service and transplant referral center. In eligible patients who had an indication for HSCT but did not undergo transplant, we collected information on reasons for non-transplant. Clinical data were presented according to race and ethnicity and compared using Pearson's χ2 or Fisher exact tests.

Results: 94 patients, including 45 females (47.9%) were included. 38 patients (40.4%) identified as Black, 28 (29.8%) as White, and 28 (29.8%) as Other (10.6% Asian, 9.6% Mexican/South American, and 9.6% undetermined). 30 patients (32%) identified as Hispanic and 64 (68%) as non-Hispanic. Diagnoses included Acute myeloid leukemia (AML, 32%), Myelodysplastic syndromes (MDS, 13%), Myeloproliferative neoplasms (MPN, 40%), MDS/MPN overlap syndrome (7%), and Clonal Hematopoiesis of Indeterminate Potential (7%). 27 patients (29%) were deemed eligible for HSCT. 38% of eligible white patients received HSCT compared to none of the black or other race eligible patients (p=0.06). Lack of necessary social support (28.6%) and loss of follow-up (14.3%) were the most common barriers to transplant in eligible Black patients. In contrast, lack of insurance (25%) and patient refusal (25%) were common reasons for non-transplant despite eligibility in white patients. An equal number of Hispanic (18%) and non-Hispanic (18.7%) transplant-eligible patients ended up receiving transplant. Lack of insurance/financial support was common reason for non-transplant despite eligibility (44.4%) in Hispanic compared to non-Hispanic patients (7.6%,P= 0.021).

Conclusions:The study highlights significant racial and ethnic disparities in HSCT use among myeloid neoplasm patients, suggesting systemic healthcare access barriers. Key obstacles for Black patients were insufficient social support and loss of follow-up, whereas Hispanics faced predominantly insurance-related barriers. These findings highlight the urgency of targeted strategies to overcome these disparities and ensure equitable HSCT access for all racial and ethnic groups.
Osteoblast-Derived Erythroferrone Regulates Stress Erythropoiesis
Authors: Na Phatthalung Pinanong , Ginzburg Yelena , van Caloen Gabrielle , Planoutene Marina , Gumerova Anisa A. , Witztum Ronit , Ingber Eva , Sultana Farhath , Korkmaz Funda , Levy Maayan , Yuen Tony , Zaidi Mone
Published: December 2024

Erythroferrone (ERFE) secreted by erythroblasts is known as an erythroid regulator of iron metabolism through hepcidin suppression. Our earlier studies indicated that ERFE highly expresses in osteoblasts and Erfe-/- mice displayed decreased bone mineral density, indicating the osteoprotective effect of ERFE in bone homeostasis. However, the contribution of ERFE in osteoblasts to bone formation, iron metabolism, and erythropoiesis is unknown. To explore the role of osteoblast-derived ERFE as a physiological regulator of bone remodeling and as an erythroid regulator of iron homeostasis, we generated osteoblast-specific Erfe knockout mice (Col2.3-Cre;Erfefl/fl) by crossing Erfefl/fl (floxed) with osteoblast-selective Cre (Col2.3-Cre). Unlike Erfe-/- mice, skeletal phenotyping revealed that Col2.3-Cre;Erfefl/fl mice had a WT-like phenotype with no differences in spleen weight, hemoglobin, reticulocyte counts, and hepcidin expression. Meanwhile, under in vitro studies, Col2.3-Cre;Erfefl/fl osteoblasts exhibit high levels of mineralization and induction of Col1a1 and Alp expression, consistent with Erfe-/- osteoblasts. A previous study has shown that ERFE is highly induced in erythroblasts in response to phlebotomy and erythropoietin (EPO) treatment. Our study is the first to demonstrate that Erfe expression in osteoblasts is stimulated by hypoxia and EPO treatment. In the normal condition, Epor in Col2.3-Cre;Erfefl/fl osteoblasts was found higher than wild type osteoblasts. EPO and its receptor, EPOR are crucially involved in the regulation of erythropoiesis. To confirm that ERFE in osteoblasts might participate in stress erythropoiesis, Col2.3-Cre;Erfefl/fl mice were phlebotomized to induce anemia. We found that Col2.3-Cre;Erfefl/fl mice do not suppress hepcidin and had a decrease in serum ERFE after 16 hours phlebotomy. Interestingly, after 6 day phlebotomy, Col2.3-Cre;Erfefl/fl mice had higher levels of red blood cells than control mice (Erfefl/fl) with no change in hemoglobin and mean corpuscular hemoglobin (MCR) concentrations. Taken together, our data demonstrate that ERFE in osteoblasts might regulate red blood cell regeneration in response to stress erythropoiesis.
TRIM28 Serves As a Key Regulator of Protein Homeostasis and Mediates Proteasome Inhibitor Resistance in Multiple Myeloma
Authors: Fang Teng , Liu Lanting , Sun Hao , Sun Xiyue , Yu Zhen , Qiu Lugui , Hao Mu
Published: December 2024

Introduction:

The maintenance of intracellular protein homeostasis is a critical biological process necessary for the survival of multiple myeloma (MM) cells. Tripartite motif (TRIM) proteins, a subfamily of the RING-type E3 ubiquitin ligase family, are integral to protein quality control. In this study, we examined the role of TRIM28, a member of the TRIM family, as a regulator of protein homeostasis in MM.

Methods:

Bioinformatic analysis was utilized to elucidate the association between TRIM28 and genes involved in the proteasome and autophagy mediated protein degradation pathway. ShRNA knockdown (KD) and overexpression (OE) experiments were conducted to investigate the role of TRIM28 in MM pathogenesis. ChIP-seq and IP-MS analyses were performed to identify potential downstream targets of TRIM28.

Results:

Firstly, our survival analysis revealed that MM patients exhibited poor outcomes with high levels of TRIM28 in both the MMRF-CoMMpass and in-house datasets, particularly in the context of bortezomib treatment. Overexpression of TRIM28 promoted cell proliferation and induced drug resistance in MM cell lines, while knock-down of TRIM28 had the opposite effect. Bioinformatic analysis demonstrated a strong correlation between TRIM28 expression and the expression of proteasome subunits and autophagy-related genes in MM datasets. RNA-seq data showed down-regulation of both the proteasome pathway and autophagy pathway after TRIM28 knockdown, leading to decreased proteasome activities and accumulation of ubiquitinated proteins. Immunofluorescence assay confirmed nuclear localization of TRIM28, while ChIP-seq identified its binding to proteasome gene promoters such as PSMB1, PSMD2, and PSMD4. Knock-down of TRIM28 reduced the expression of proteasome subunits, indicating its critical role in transcriptional activation of these genes in MM cells.

Additionally, a decrease in autophagosome formation was observed in TRIM28 KD cells, indicating the involvement of TRIM28 in autophagy activity. Further IP-MS and Co-IP assays demonstrated the interaction between TRIM28 and 14-3-3ζ. A significant reduction in the polyubiquitination level of 14-3-3ζ was noted in TRIM28 KD cells. The 14-3-3ζprotein serves as an adaptor that modulates cellular signaling by binding to a wide array of proteins. Therefore, acting as an E3 ligase, TRIM28 promotes ubiquitin-dependent degradation of 14-3-3ζ, thereby enhancing autophagy activity in MM. These findings underscore the crucial role of TRIM28 in maintaining protein homeostasis by regulating both proteasome and autophagy activities involved in myeloma pathogenesis.

Conclusion:

Our research has provided new and important insights into the crucial role of TRIM28 in maintaining protein homeostasis and its involvement in myeloma pathogenesis. TRIM28 activates the proteasome and autophagy pathways, making it a potential promising therapeutic target in MM.
The C553-C581 Disulfide of Human Factor XI Is Crucial for Its Activation and Is Targeted By Thiol Isomerase Pdir
Authors: Zhang Yuxin , Liu Shuo , Zhao Zhenzhen , Lv Keyu , Fang Chao , Han Yue , Wu Juhong , Wu Depei , Zhang Jingyu , Yang Aizhen , Wu Yi
Published: December 2024

The activation of the coagulation system is a key step in the process of thrombosis. Using genetically-modified mouse models and inhibitors, we and others have reported that thiol isomerases PDI, ERp57, ERp72, ERp5 and ERp46 promotes coagulation while TMX1 has an inhibitory effect. However, the mechanisms by which these enzymes regulate coagulation factors remain largely unknown. More efforts are needed to understand whether the thiol-disulfide exchange of coagulation factors contribute to their activation. There are 17 pairs of disulfide bonds on the factor XI (FXI). In this study we performed a comprehensive evaluation of the functions of all disulfide bonds on FXI. When FXI carrying single cysteine with the disruption of disulfide bonds were expressed in 293T cells, we found that the C122-C128 disulfide bond was critical for FXI synthesis process; the C2-C85, C28-C58 and C32-C38 of the A1 domain, the C92-C175 and C118-C147 of the A2 domain, the C182-C265, C208-C237 and C212-C218 of the A3 domain, the C273-C356, C299-C328 and C303-C309 of the A4 domain, as well as the C398-C414, C496-C563 and C527-C542 of the catalytic domain were necessary for the secretion of FXI. Although the ablation of C362-C482 and C553-C581 did not cause the defect of synthesis and secretion, disrupted C362-C482 and C553-C581 completely lost the activity in FXI-mediated thrombin generation and aPTT, suggesting that C362-C482 and C553-C581 are crucial functional disulfides of FXI. C362-C482 links heavy chain and light chain, which is necessary for calalytic domain getting access to the heavy chain containing the binding site for FIX. As shown by comparative molecular dynamics simulations, the C553-C581 breakage induced the significant conformational change of the loop 553CKGDS557, which probably interrupts the hydrogen bond of catalytic residues H413-S557 and the substrate binding affinity . To investigate whether FXI is targeted by thiol isomerases, FXI was incubated with PDI, ERp57, ERp72, ERp5, ERp46 and TMX1 as well as other thiol isomerases, followed by chromogenic substrate assay, we found that FXI activity was selectively inhibited by PDIr. As shown by 3-(N-maleimido-propionyl) biocytin (MPB) labeling, PDIr marked reduced the FXI disulfides. Furthermore, using thiol differential labeling and mass spectrometry, PDIr was found to reduce C553-C581 of FXI. In a laser-induced cremaster arteriole injury model, PDIr deficiency enhanced fibrin formation at the site of injury suggesting that PDIr inhibits coagulation in vivo, consistent with its inhibition and reduction of FXI. Taken together, this study identified the role of every single FXI disulfide in protein synthesis, secretion and enzymatic activity, and provides the new evidence showing that the C553-C581 of human factor XI is a novel crucial disulfide for its activation and is targeted by the thiol isomerase PDIr for coagulation.
Donor Neutrophil Subsets in G-CSF Mobilized Peripheral Blood Stem Cell to Predict the Risk of Agvhd
Authors: Zhang Haiyan , Wu Qiaoyuan , Dai Huajuan , Liu Xinyu , Fan Zhiping , Lin Ren , Xu Na , Liu Qianwei , Liu Qifa , Jin Hua
Published: December 2024

Purpose:

Recently, a few studies began to focus on the role of neutrophil subsets in the immune tolerance; however, the role of these subgroups in acute graft versus host disease (aGVHD) is unclear. In our study, we explore the effect of neutrophil subsets in G-CSF mobilized peripheral blood stem cell transplants on the occurrence of aGVHD and establish a prediction model for the occurrence of II-IV aGVHD within 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Patients and Methods:

A prospective cohort trial was performed in our hospital from December 2023 to March 2024. The proportion of neutrophil subsets in the donor-derived G-CSF-mobilized peripheral blood stem cell grafts, including: neutrophil progenitor 1 (proNeu1), neutrophil progenitor 2 (proNeu2), neutrophil precursor cells (preNeu), immature neutrophils (immatureNeu), and mature neutrophils (matureNeu), were detected by flow cytometry. Univariate logistic regression and restricted cubic spline (RCS) were used to screen variables related to the onset of aGVHD. A multivariate logistic regression equation was constructed with the screened variables to establish a prediction model, and the prediction performance of the model was evaluated. This trial is registered with ClinicalTrials.gov (NCT 06394895) and is completed.

Results:

A total of 139 patients were enrolled in the study, of which 98 patients were randomly selected as the training cohort and 41 patients as the validation cohort. We found that the matureNeu ratio, proNeu2 ratio, donor type and donor age in the graft were independent influencing factors for II-IV aGVHD. Based on these four variables, we established a prediction model for the risk of II-IV aGVHD.In the prediction model,the area under the receiver operating characteristic (ROC) curve (AUC) of the model was 0.789 and 0.777 in the training set and validation set, respectively. The optimal cutoff value of this prediction scoring system was 0.4. A prediction score greater than 0.4 was defined as a high-risk population for II-IV aGVHD. The 100-day cumulative incidence of grade II-IV aGVHD was 19.1% (95% CI 9.2-27.9%) and 60.6% (95% CI 47.4-70.4%) in the high- and low-risk groups of this cohort, respectively (P < 0.0001).

Conclusion:

The proportion of matureNeu subgroup, proNeu2 subgroup, donor type and donor age in peripheral blood stem cell transplants are independent influencing factors for II-IV aGVHD after allo-HSCT. The prediction model based on these variables has good predictive ability and can intuitively and quickly identify the high-risk population for II-IV aGVHD.
Sintilimab, Rituximab and Lenalidomide (sR2) As First-Line Treatment in Elderly Patients with Diffuse Large B-Cell Lymphoma: A Single-Arm, Open-Label, Phase 2 Trial
Authors: Wei Xiaolei , Guo Xutao , Wei Qi , Wei Yongqiang , Feng Ru
Published: December 2024

The chemo-free combination of rituximab and lenalidomide (R2) has been showed activity in a significant proportion of older patients with diffuse large B-cell lymphoma (DLBCL). Lenalidomide could enhance the cytotoxicity induced by checkpoint blockade in vitro. Herein, we did a phase 2 trial of sintilimab, rituximab, and lenalidomide (sR2) to evaluate the efficacy and safety in older patients with de novo DLBCL.

Eligible patients were aged older than70 years or older than 60 years with ECOG PS ≥ 2, newly diagnosed DLBCL. Patients were administered a maximum of 8 28-day cycles of 10 mg oral lenalidomide from days 1 to 21, IV rituximab 375 mg/m2 and sintilimab 200mg on day 1, with response assessment after cycles 4 and 8. Patients with partial response or complete response (CR) at cycle 8 were administered lenalidomide 10 mg/d from days 1 to 21 for every 28 cycles for a total of 2 years or until progression or unacceptable toxicity. The primary endpoint in this study was complete response rate (CRR) after 8 cycles or at the end of the induction treatment. The secondary end point was 2-year overall survival, progression-free survival, overall response rate (ORR) and treatment-emergent adverse events.

Until April 9, 2024, A total of 18 older DLBCL patients were enrolled and 14 of them was evaluated. The median age of the patients in this study was 75 years old (range 68-70) and 2 (11.1%) patients had a poor performance status with an ECOG score of 2 or greater. 12 (85.7%) patients achieved an overall response at the end of the induction treatment, with 11 (78.6%) achieving a complete response. With a median follow up of 8.9 months, the 1-year progression-free survival and overall survival were 80.8% and 92.9%, respectively. Common grade 3-4 adverse events were hematological toxicities (neutropenia in 4 patients (28.6%), thrombocytopenia in 3 patients (21.4%), anemia in 3 (21.4%). No treatment-related deaths were reported.

A chemotherapy-free sR2 regimen is clinically effective and safe and warrants further investigation in older patients with DLBCL.
Vincristine Overcomes Cytarabine Resistance in Acute Myeloid Leukemia By Suppression of ARHGAP18-Mediated Cellular Senescence
Authors: Ren Xiyang , Yan Haotian , Zhou Mei , Wu Fan , Zhang Qing , Ding Yangyang , Li Qian , Wang Huiping , Xu Caixian , Wang Jiyu , Li Cong , Zhang Wanqiu , Liu Linlin , Chen Ruotong , Dong Yi , Dai Wenjie , Zhai Zhimin , Tao Qianshan
Published: December 2024

Background : Cytarabine is the cornerstone of chemotherapy in patients with acute myeloid leukemia (AML), yet the outcomes remain unsatisfactory because of resistance. Cellular senescence is a novel mode of cytarabine resistance in AML, but its mechanism of action and solution strategy remain unclear.

Methods : We observed the proportion of cellular senescence and the expression level of ARHGAP18 in relapsed and refractory AML patients after treatment with cytarabine. In vitro, we established stable AML cell lines with knockdown and overexpression of ARHGAP18 gene using lentiviral transduction. In vivo, we developed an mouse xenograft model to explore the role of ARHGAP18 gene in the process of cytarabine resistance. Additionally, we used the microtubule inhibitor targeting ARHGAP18 protein to evaluate the potential applicability of overcoming cytarabine resistance in AML.

Results: In this study, we demonstrated that cellular senescence primarily occurred in leukemia cell subsets in AML patients resistant to cytarabine, accompanied by high expression of ARHGAP18gene and ARHGAP18 protein. Cellular senescence (including cellular senescence specific staining, cell cycle arrest, and cellular senescence related secretory phenotypes) was rare in ARHGAP18 gene wild-type and knockdown leukemia cells but frequent in ARHGAP18 gene overexpressed leukemia cells. Under the stress of cytarabine, both in vitro and in vivo, ARHGAP18 gene knockdown significantly decreased the incidence of cellular senescence and increased chemotherapy sensitivity compared to the wild-type ARHGAP18gene. Conversely, ARHGAP18 gene overexpression significantly increased the incidence of cellular senescence, chemoresistance, the ability to resume proliferation, and extramedullary invasion, and resulted in shorter survival times. Mechanistically, the p16/Rb and p53/p21 signaling pathways were involved in ARHGAP18-mediated cellular senescence. Importantly, given previous research indicates that the ARHGAP18 is located on microtubules, we found that the microtubule protein inhibitor vincristine significantly disrupted the ARHGAP18 protein, reduced cellular senescence, and increased chemotherapy sensitivity.

Conclusions: ARHGAP18 induces cellular senescence via the p16/Rb and p53/p21 signaling pathways, mediating cytarabine resistance in AML. However, vincristine overcomes cytarabine resistance by suppression of ARHGAP18-mediated cellular senescence.
Early Responders (Time to Best Response ≤3 months) Predicts Functional High-Risk Multiple Myeloma
Authors: Xu Yang , Fan Yili , Wang Luyao , Zhang Jiawei , Chen Boxiao , Yang Luyu , Zhang Xuzhao
Published: December 2024

Introduction: Despite advancements in risk evaluation and management of multiple myeloma (MM), functional high-risk (FHR) MM often shows a suboptimal response to induction therapy or relapses early after initial treatment. Early recognition of FHRMM may help to tailor treatment approaches and improve survival outcomes. However, the characteristics of FHRMM is not fully understood, and the impact of current treatment on FHRMM remains unclear. Our objective is to identify baseline risk factors and treatment response kinetics associated with FHRMM.

Method: The patients with newly diagnosed multiple myeloma (NDMM) between January 2017 and December 2022 were included. The patients' characteristics, treatment protocols, best responses, and survival data were collected and analyzed. High-risk factors were defined based on the NCCN MM guideline 2024v4, which includes various cytogenetic abnormalities and disease characteristics (R-ISS III, extramedullary disease, circulating plasma cells, and cytogenetic abnormalities including Del(1p32), t(4;14), t(14;16), t(14;20), Del17p, 1q21 gain/1q21 amp, and MYC translocation). Among MM without baseline risk factors, those progressed or relapsed ≤18 months after treatment initiation were considered FHR, while those progressed or relapsed >18 months were classified as standard risk (SR). Cox regression analysis was used to assess the impact of multiple variables on prognosis, and log-rank test was performed to explore risk factors for FHRMM.

Results: Among 211 NDMM patients, there were 30 patients (14.2%) in the FHR group, 119 (56.4%) in the SR group, and 62 (29.4%) in the high-risk (HR) group. With a median follow-up of 44.0 months, the progression-free survival (PFS) and overall survival (OS) for FHR group were 14.0 months and 39.0 months, respectively. Notably, the FHR group had a significantly higher proportion of patients with time to best response (TBR) ≤3 months compared to the SR group, P=0.039. Furthermore, among non-HR groups stratified by TBR, there were significant differences in PFS and OS between TBR ≤3 months and TBR >3 months, with median PFS of 30.0 months and 56.0 months, and median OS of 58.0 months and 94.0 months, respectively, both P<0.001. Autologous stem cell transplantation (ASCT) significantly improved the PFS of FHRMM patients, P=0.033.

Conclusions: At present, it is not easy to differentiate FHR from SR patients based on the baseline characteristics of NDMM. Nevertheless, the analysis of response kinetics suggests that TBR ≤3 months could be a feasible indicator for early identification of FHRMM. ASCT has been found to improve the prognosis of FHRMM.
Single Cell Profiling of Hematopoietic Stem and Progenitor Cells in Pediatric Acquired Aplastic Anemia
Authors: Chan Derek , Omur Meltem , Lin Diana , Wasserman Wyeth , Karsan Aly , Vercauteren Suzanne
Published: December 2024

Acquired severe aplastic anemia (AA) is a bone marrow failure disorder featuring a paucity of blood cells that significantly predisposes affected individuals to life threatening infections and/or bleeding, evolution to myelodysplastic syndrome/leukemia, risk of treatment relapse and/or death. The latest successful addition of Eltrombopag, a unique thrombopoietin receptor agonist, to frontline immunosuppressive therapy among adult patients with acquired AA has provided proof-of-principle that there may be additional ways to recover hematopoiesis in this setting. To examine the causes of and to identify novel strategies to rescue haematopoiesis in pediatric AA, we employed single cell profiling using cellular indexing of transcriptomes and epitopes (CITE-seq) on lineage-negative and treatment-naïve diagnostic severe paediatric AA (n = 5) and age-approximated healthy donor (n = 4) primary bone marrow samples and obtained transcriptomes of 2,009 and 13,861 single cells respectively. This profiling uncovered a widespread loss of primitive hematopoietic stem and progenitor cells (HSPCs) in pediatric AA with significant enrichment for IFN-γ, IL6/JAK/STAT, unfolded protein response and PI3K/AKT/MTOR pathways. Residual HSCs in pediatric AA showed enrichment of a recently described HSC inflammatory memory transcriptional signature. Granzymes were distinctly upregulated in residual HSPCs within pediatric AA. Further profiling of additional samples is required to capture additional events among rare HSPC populations. Altogether, these results demonstrate significant underlying inflammation and rationalize testing of Ruxolitinib, Sirolimus and granzyme blockade in pediatric AA. Future work will involve establishing a bone marrow organoid model of pediatric AA for therapeutic screening and drug discovery.
A Novel Approach Combining Type II JAK Inhibitor Chz-868 with Nutrient Restriction to Augment Cell Death in Philadelphia like Acute Lymphoblastic Leukemia
Authors: Alhaddad Muneera , Buono Roberta , Huynh Van Thu , Tasian Sarah K. , Fruman David A
Published: December 2024

Background: There is an urgent clinical need to improve therapeutic outcomes in Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL). Approximately 60% of Ph-like cases harbor rearrangements in the CRLF2 locus, often with accompanying mutations in JAK kinases. Current clinical trials are assessing whether ruxolitinib, a type I JAK inhibitor, can enhance responses to chemotherapy in CRLF2-r acute leukemias. Notably, preclinical studies have shown that type II JAK inhibitors provide greater anti-leukemic effects than ruxolitinib. Other work has demonstrated promising ability of nutrient limitation to enhance the efficacy of tyrosine kinase inhibitors (TKIs) in solid tumors and chemotherapy in hematologic malignancies. Indeed, there is growing excitement for short-term application of fasting-mimicking diet (FMD), since clinical studies have shown promising therapeutic efficacy with higher compliance rates than other dietary interventions. The objectives of this study were to compare the cytotoxicity of type I and II JAK inhibitors in CRLF2-r Ph-like ALL, and to determine whether nutrient restriction via FMD enhances JAK inhibitor efficacy.

Methods: We conducted both in vitro and in vivo experiments. For in vitro experiments, we used the human B-precursor ALL cell lines MUTZ5 and MHH-CALL-4 as well as patient derived Ph-like ALL cells that carry CRLF2 translocation. We used flow cytometry techniques to measure cell viability. Western blotting analysis was used to quantify pro-survival and pro-apoptotic proteins in the treated cell lines. For in vivo experiments, we used mouse patient-derived xenograft (PDX) model in which NSG mice were injected with patient derived Ph-like ALL cells.

Results: In vitro experiments confirmed that the type II JAK inhibitor CHZ-868 had more potent cytotoxic effects than ruxolitinib in CRLF2-r cell lines and cells derived from PDX models. Short-term starvation with low glucose and low serum strongly sensitized CRLF2-r cell lines to CHZ-868 cytotoxicity. Mechanistically, the combination of CHZ-868 and starvation was associated with decreased expression of the pro-survival protein MCL-1 and increased expression of the pro-apoptotic protein BIM. In vivo experiments using a CRLF2-r PDX model showed that FMD reduced leukemic burden and enhanced the efficacy of CHZ-868.

Conclusion: Efficacy of the type II JAK inhibitor CHZ-868 in CRLF2-r cells is enhanced by nutrient restriction. These results provide proof-of-concept for testing FMD as a diet-based approach to enhance TKI efficacy in Ph-like B-ALL patients.
Anti-CD3/B-Cell Maturation Antigen Bispecific Antibody CM336 in Relapsed/Refractory Multiple Myeloma: An Open-Label, Phase I/II Study
Authors: An Gang , Huang Zhongxia , Liang Aibin , Fang Baijun , Jing Hongmei , Qiu Lu-Gui
Published: December 2024

Introduction: Multiple myeloma is an incurable malignancy characterized with high relapse rates. Therapeutic options to improve clinical outcomes remain limited for patients with relapsed/refractory multiple myeloma (RRMM). CM336 is a novel bispecific antibody targeting both CD3 and B-cell maturation antigen (BCMA), redirecting T cells to BCMA-expressing myeloma cells. Here, we report the dose-escalation results of CM336 from a first-in-human, open-label, phase I/II study (NCT05299424).

Methods: This study included a dose-escalation and a dose-expansion part. For dose escalation, eligible subjects (18 years or older) were diagnosed with RRMM (per International Myeloma Working Group criteria), had a measurable disease, and had exposed to at least one proteasome inhibitor, immunomodulatory drug, and anti-CD38 antibody. Subjects received CM336 at doses ranging from 0.04 to 160 mg using an accelerated titration design for the first 3 cohorts, followed by a “3+3” design. CM336 was administrated subcutaneously weekly, with a step-up dosing scheme for 40 mg and higher doses.

Results: As of June 30, 2024, 25 subjects received CM336 (median age, 63 years [range 41-76]; 72% female) in the dose-escalation part. Subjects had received a median of 4 (range 2-15) prior lines of therapy, with all being triple-class exposed and 20% with extramedullary plasmacytoma. The median CM336 exposure duration was 4.8 months (range 0.03-12). Maximum tolerated dose was not reached. The most common treatment-emergent adverse events (TEAEs) included cytokine release syndrome (CRS) (any grade: 68%; grade ≥3: only one subject), anaemia (68%; 32%), lymphocyte count decreased (64%; 56%), neutrophil count decreased (56%; 44%), pneumonia (44%; 32%), and pyrexia (44%; 0). All CRS events were resolved completely, and no immune effector cell associated neurotoxicity syndrome (ICANS) events were reported. At a median follow-up of 8.0 months, the confirmed overall response rate (ORR) was 60.9% (14/23; 95% CI, 38.5%-80.3%). Notably, for subjects in the 80 mg and 160mg cohorts who received at least one target dose (n=8), the ORR was 100%. Of the 10 subjects (43.5%) with a confirmed stringent complete response (sCR), 9 were evaluable for minimal residual disease (MRD), and the MRD negativity rate (at a threshold of 10−5 cells) was 88.9% (8/9). The median time to response was 1.4 months (range 0.7-3.8). The median duration of response and median progression-free survival were not reached. All subjects in remission have not experienced disease progression yet.

Conclusions: Treatment with CM336 was well tolerated with preliminary promising anti-myeloma efficacy. CM336 is expected to improve the prognosis of subjects with RRMM. With longer follow-up, additional efficacy/safety data will be presented.
Impact of Hospital-Acquired Venous Thromboembolism on Surviving Medical Admission: Findings from the Medical Inpatient Thrombosis and Hemostasis Study
Authors: Martin Karlyn A. , Sparks Andrew , Wilkinson Katherine , Packer Ryan , Terrell Deirdra R , Gergi Mansour , Ferraris Augusto , Wood William A. , Repp Allen B , Roetker Nicholas S , Zakai Neil
Published: December 2024

Introduction

Approximately 200,000 venous thromboembolism (VTE) events occur each year during or within 3 months after medical (non-surgical) hospitalizations in the U.S. The consequences of hospital-acquired VTE (HA-VTE, VTE occurring while hospitalized), such as likelihood of dying during admission, are unknown. The objective of this study was to examine the association between HA-VTE and risk of in-hospital mortality.

Methods

Adults admitted to medical services for at least 1 midnight at one of 5 hospital systems from 2016-2022 were included. Admissions for primary diagnosis of VTE were excluded. Each admission with HA-VTE was matched with up to 4 admissions without HA-VTE based on day of hospitalization, age (± 5 years), sex, race and ethnicity, hospital system, level of care (medical services versus intensive care unit [ICU]), calendar year, and COVID-19 status at admission. The primary outcome was in-hospital mortality. We used conditional logistic regression to estimate the association of HA-VTE with in-hospital mortality, adjusting for the matching factors, anticoagulation level at admission (therapeutic, prophylactic, or none), and the Elixhauser Comorbidity Index. We tested whether the association varied by COVID-19 status.

Results

The matched cohort included 2,061 medical admissions with HA-VTE and 7,908 medical admissions without HA-VTE. In the matched admissions, the mean age was 58 years (SD 16), 57.1% identified as male, 54.5% as White race, 26.6% as Black race, 24.0% were admitted to an ICU, and 10.8% were admitted with COVID-19. The mean Elixhauser Comorbidity Index score was 14.3 (SD 18.4) and 12.2 (SD 17.3) in admissions with and without HA-VTE, respectively. In-hospital mortality occurred in 12.4% of HA-VTE admissions and 5.1% of non-HA-VTE admissions. Compared to non-HA admissions, the adjusted odds ratio (OR) for in-hospital mortality with HA-VTE was 2.71 (95% confidence interval [CI] 2.23, 3.29). The association of HA-VTE with in-hospital mortality was similar for admissions with COVID-19 (OR 2.93; 95% CI 2.33, 3.70) and admissions without COVID-19 (OR 2.18; 95% CI 1.49, 3.18; pinteraction = 0.12).

Conclusions

HA-VTE was associated with a nearly 3-fold increased odds of death during hospitalization in a diverse patient cohort from 5 hospital systems. While increased illness severity may account for some of the increased mortality risk, HA-VTE itself or interventions to treat HA-VTE (anticoagulation) likely contribute. Efforts should be made to understand whether reducing HA-VTE can translate to decreased mortality.
First-in-Human Trial of BCMA-CD19 Compound CAR (cCAR) in Desensitization of Donor-Specific Anti-HLA Antibodies (DSAs) in Haploidentical Stem Cell Transplant Recipients
Authors: Liu Fang , MA Yu , LI Yecheng , Pinz Kevin G. , Wang Xiao , Wada Masayuki , Ma Yupo
Published: December 2024

Background: DSAs are recipient antibodies targeted against donor class I/II human leukocyte antigens (HLAs). The existence of DSAs before HLA haploidentical hematopoietic stem cell transplantation (haplo-HCT) is known to cause primary graft failure. Currently there is no established method of DSA desensitization. cCAR targets the “root cause” of DSA production, B cells and plasma/long-lived plasma cells, and thus may achieve desensitization through a dual reset of humoral and B cell immune system.

Aims:We present the safety and efficacy of cCAR, which targets antibody- producing “root cause”, B cells and plasma/long-lived plasma cells in our first-in-human clinical trial.

Methods:cCAR is comprised of a complete BCMA CAR fused to a complete CD19 CAR, separated by a self-cleaving P2A peptide. CAR T cells were manufactured in a cGMP facility.

Beginning May 2019 through October 2020, 4 haplo-HCT candidates with high titer of DSA (MFI>5000） were enrolled in this study. All patients received conditioning with fludarabine (30 mg/m2/d) and cyclophosphamide (300 mg/m2/d). CAR T cells were given by dose escalation at 2～4.5×106/kg with a single dose.

The primary endpoint is to evaluate toxicity of cCAR and the second endpoint is to observe the reduction of DSA after cCAR therapy and engraftment post-haplo-HCT.

Results:The median patient age was 35 years (range, 21～48). All were female; 2 with acute myeloid leukemia (AML), 1 with acute lymphocytic leukemia (ALL) and 1 aplastic anemia/paroxysmal nocturnal hemoglobinuria (AA/PNH). All donors were from haploidentical family members.

Cytokine release syndrome (CRS) occurred in 3 patients, all grade 1, and resolved within 3～8 days after supportive care or administration of low dose steroids. No neurotoxicity occurred. One patient suffered grade Ⅳ cytopenia which resolved in 2 weeks. No other toxicity occurred.

All patients demonstrated B cell and plasma cell depletion at 4 weeks post-cCAR infusion. The average median fluorescence intensity (MFI) value was 82187 (range, 7820～249840) and 2995 (range, 0～4343) before and after cCAR therapy, respectively.

All patients received a haplo-HCT within 49～135 days and achieved successful engraftment with full chimerism. The median time for neutrophil and platelet engraftment was12 and 13 days, respectively.

Conclusion:Our first-in-human clinical trial of cCAR demonstrated profound efficacy in reducing DSA levels in haplo-HCT candidates. Remarkable depletion of antibody-producing “roots cause”, B cells and plasma/long-lived plasma cells was observed in all patients. Our results suggest cCAR has the potential to benefit patients receiving solid organ transplants or those with other antibody-mediated diseases.
Activity of Orally Available CBP/p300 Degraders in Pre-Clinical Models of Multiple Myeloma
Authors: Tiwari Praveen Kumar , Harrison Drew A , Ojeda Samuel , Kawale Ajinkya S , Doda Sai Reddy , Vannam Raghu , Karakyriakou Barbara , Koglin Ann-Sophie , Rizvi Sarah , Ott Christopher J
Published: December 2024

The enhancer lysine acetyltransferases CBP and p300 are compelling targets for therapy in multiple myeloma. Chemical inhibition of these factors, either through the bromodomain or the catalytic acetyltransferase domain, show promising activity in pre-clinical models of myeloma and other cancers. Some compounds, including inobrodib, have progressed to early-stage clinical trials. Yet CBP/p300 inhibitors may only have partial effects on protein activity. Using heterobifunctional chemical approaches to induce targeted protein degradation we have developed dCBP-1, a potent tool compound that induces complete degradation of both CBP and p300 via hijacking the activity of the E3 ligase CRBN. This compound shows promising effects in myeloma cell lines with induction of apoptosis occurring in some lines within 48-72 hours of treatment. Tested across >300 cancer cell lines, myeloma cells are among the most sensitive to dCBP-1 treatment, and this activity is more pronounced when compared to CBP/p300 inhibitors. These antimyeloma effects are at least in part a consequence of rapid disruption of oncogenic transcriptional signaling networks mediated by essential transcription factors IRF4 and MYC. Yet dCBP-1 fails to achieve adequate exposures in mice to use as an in vivo tool. We have thus embarked on a chemical optimization strategy to incorporate newer heterobifunctional design strategies to create a compound with better pharmacokinetic properties for use in animal models. Our strategy included derivatization of the CRBN-binding moiety and progressive linker optimization. Through these efforts we have defined several advanced analogs of dCBP-1 that have dramatically improved potency in vitro, along with improved in vivo properties including oral bioavailability. With intermittent dosing strategies, advanced compounds lead to potent and sustained loss of CBP and p300 in myeloma cell lines and tumor xenografts and achieve tumor reduction as single agents when delivered orally. With these optimized compounds, we can now address systematically (in vitro and in vivo) the relative consequences of CBP/p300 inhibition and degradation both in tumor cells and normal tissue/hematopoietic cells.
Chromatin Remodeling By GATA1s Alters Megakaryocytic Gene Expression
Authors: de Castro Carini Picardi Morais , Martínez-Cebrián Gerard , Quijada-álamo Miguel , Freed Grace , Lorenzi Lucía , Malinge Sébastien , Wagenblast Elvin , Cuartero Sergi
Published: December 2024

The GATA1 transcription factor is crucial for the regulation of gene expression during the differentiation of hematopoietic progenitors to erythrocytic and megakaryocytic lineages. Mutations in GATA1 are pervasively found in myeloid leukemia of Down syndrome (ML-DS), a subtype of acute myeloid leukemia that is common in children with Down syndrome. These mutations typically occur in the N-terminal region of the GATA1 gene and lead to the production of a truncated isoform known as GATA1s, which retains the DNA-binding capacity. Expression of the truncated form GATA1s disrupts normal hematopoietic differentiation and leads to the over-production of megakaryocytic progenitors in the fetal liver. In Down syndrome children, this results in a non-lethal blood disorder known as transient abnormal myelopoiesis (TAM), which in many cases progresses to myeloid leukemia.

Our study investigates the impact of the GATA1s isoform on megakaryopoiesis and its underlying molecular mechanisms. We use mouse hematopoietic progenitors and CRISPR-edited human HSPCs and perform transcriptomic and epigenomic analyses to understand how GATA1s differentially controls megakaryocytic gene expression. Our findings indicate that the expression of GATA1s promotes a premature increase of megakaryocyte-specific transcription already in hematopoietic stem cells. The upregulation of megakaryocyte markers is observed in HSCs from both fetal liver and adult bone marrow, though with varying effects across subpopulations. Moreover, the response to megakaryocyte-inducing acute stressors, such as interferon, differs between cells expressing GATA1 and those expressing GATA1s. Despite this early bias, GATA1s-expressing cells show incomplete differentiation into megakaryocytes in culture. ChIP-seq of normal and mutant GATA1 shows that despite both isoforms bind to a majority of shared binding sites, a fraction of these sites is isoform-specific. This is linked to changes in chromatin accessibility, which present different dynamics of gain and loss along megakaryocyte differentiation and which are associated to the number of GATA1 motifs on DNA. Furthermore, Hi-C analysis of global chromatin interactions shows changes in long-range genomic interactions associated to the deregulation of Mk-specific transcripts.

In summary, our work illustrates how GATA1s disrupts normal megakaryopoiesis by inducing early lineage bias and impairing differentiation through remodeling of the chromatin landscape and gene expression. These findings enhance our understanding of GATA1s-related disorders, such as myeloid leukemia of Down syndrome, and may guide future therapeutic strategies.