Chimeric antigen receptor (CAR) T-cell therapy targeting B-cell maturation antigen (BCMA) shows activity in multiple myeloma (MM), yet relapse remains common owing to heterogeneous BCMA expression and soluble BCMA (sBCMA). Improving BCMA-directed CAR-T therapy requires persistence, enhanced antitumor activity, tolerance to low antigen density and resistance to sBCMA-mediated inhibition. We identified anti-BCMA VHHs from a humanized phage display library, constructed monospecific and biparatopic CARs, and evaluated their function and optimal designs in preclinical models, including patient-derived MM cells and xenografts. Biparatopic Nab5822 CAR-T cells exhibited superior cytotoxicity and cytokine secretion (IL-2, IFN-γ, TNF-α) against antigen-heterogeneous MM cells in vitro, maintained activity under a supraphysiological sBCMA challenge at 120 ng/mL, and retained function under repeated exposure to clinically relevant sBCMA levels. Under repeated antigen challenge, Nab5822 preserved lysis, reduced exhaustion with an increased stem-cell memory T-cell compartment, and achieved durable tumor control in xenograft models without detectable toxicity. Mechanistic analyses indicated that the VHHs engage non-overlapping BCMA epitopes and promote improved immunological synapse organization together with coordinated proximal signaling, features that are not fully explained by equilibrium affinity alone and may contribute to sustained long-term T-cell function. Our findings support biparatopic CAR design as a strategy to tolerate low antigen density and resist sBCMA-mediated inhibition, providing a rationale for clinical evaluation of next-generation CAR T-cell therapies.

Background Chronic lymphocytic leukemia (CLL), the most prevalent form of adult leukemia, remains a challenge in terms of optimal therapeutic strategies, particularly for patients with rapid progression or relapsed/refractory disease. While mutations in the splicing factor SF3B1 occur in 10–15% of CLL cases, the pathological role of the more common wild-type SF3B1 remains poorly understood. This is especially significant considering recent studies demonstrating the oncogenic role of wild-type SF3B1 in myeloid and T-cell leukemias. Aberrant RNA splicing has been implicated in cancer-related metabolic reprogramming, yet the mechanistic link between SF3B1-mediated splicing regulation and the metabolic pathways essential for CLL cell survival is underexplored. In this study, we evaluate the therapeutic potential of targeting wild-type SF3B1 using the selective splicing modulator H3B-8800, with a particular focus on its impact on glycolytic metabolism and its combination potential with BTK inhibitors. Methods CLL cell lines (MEC-1, JVM-3) and primary CLL cells from treatment-naïve patients were treated with the SF3B1 modulator H3B-8800 (0–10 μM) and covalent BTKi zanubrutinib or ibrutinib (0–100 μM) for 48 hours. Cell viability was assessed using the CCK-8 assay to determine IC50values. RNA-seq was performed to analyze differential splicing events and gene expression. Key metabolic genes and BCR signaling regulators were further validated by qRT-PCR and Western blot. Metabolic phenotypes were evaluated using glucose and lactate assay kits. The synergistic interaction between H3B-8800 and BTKi was assessed using the zero interaction potency (ZIP) model, and apoptosis was quantified via Annexin V/PI staining. Results Knockdown of wild-type SF3B1 via lentivirus transfection significantly inhibited the growth of MEC-1 and JVM-3 cells, suggesting an essential dependency on SF3B1 in CLL cells. Pharmacological inhibition of SF3B1 by H3B-8800 exhibited potent cytotoxicity across CLL cells, with IC50values of 31.1 nM for primary CLL cells, 32.55 nM for MEC-1, and 68.85 nM for JVM-3. BTKi zanubrutinib showed IC50values of 28.92 μM for primary CLL cells, 56.12 μM for MEC-1, and 36.34 μM for JVM-3; while ibrutinib exhibited IC50 values of 10.92 μM for primary CLL cells, and 25.83 μM and 29.46 μM for MEC-1 and JVM-3, respectively. RNA-seq revealed a SF3B1 splicing-dependent inverse expression pattern of PFKFB1 (a glycolysis suppressor) and PFKFB3 (a glycolysis activator) upon H3B-8800 treatment. These splicing shifts were verified at the protein level: PFKFB1 decreased by 78.6%, while PFKFB3 increased by 186%. Metabolic assays demonstrated that H3B-8800 treatment resulted in a concurrent reduction in glucose uptake (20.7% decrease at 50 nM; 32.7% decrease at 100 nM) and lactate secretion (43.5% decrease at 50 nM; 49.1% decrease at 100 nM). Transcriptomic analysis also revealed that H3B-8800 induced upregulation of key BCR signaling regulators, including CD79A, CD19, and JUN, suggesting that this modulation may increase susceptibility to additional BCR pathway inhibition by BTKi. Given BTKi's role in suppressing glycolysis, we hypothesized that co-targeting SF3B1 and BTK would disrupt leukemia cell metabolic homeostasis. Indeed, combination treatment with H3B-8800 and zanubrutinib yielded a ZIP synergy score of 11 in MEC-1 cells. In primary CLL cells, 20 nM H3B-8800 induced 5.47% apoptosis, while zanubrutinib (20 μM) and ibrutinib (20 μM) induced 16.17% and 46.88% apoptosis, respectively. Combined treatment with H3B-8800 and two BTK inhibitors increased apoptosis to 24.16% and 67.16% respectively, indicating significant synergy. These results suggest that dual targeting of SF3B1 and BTK yields potent anti-leukemia efficacy through mechanisms involving BCR signaling and oncogenic metabolism. Conclusions Our study identifies the wild-type SF3B1 as a critical metabolic regulator in CLL through PFKFB-mediated glycolytic control. The splicing modulator H3B-8800 exerts potent anti-leukemic effects by rewiring the PFKFB1/PFKFB3 balance, leading to metabolic disruption. We also provide a mechanistic link between splicing dysregulation and metabolic vulnerabilities in CLL, highlighting SF3B1 as a viable therapeutic target regardless of mutational status. Furthermore, our data support a dual-targeted strategy combining SF3B1 modulation and BTK inhibition as an effective therapeutic approach for CLL.

BackgroundChronic lymphocytic leukemia (CLL) is the most common adult leukemia. Mutations in splicing factor 3b subunit 1 (SF3B1), especially the K700E variant, are closely associated with aggressive progression and poor prognosis, though the underlying mechanisms remain unclear. SAT1 plays a pivotal role in polyamine metabolism, which is essential for cell growth and survival. This study investigates how the SF3B1 K700E mutation regulates SAT1 expression, leading to immune evasion in CLL. Methods and ResultsWe first conducted RNA profiling on MEC-1 cells expressing SF3B1 K700E and primary CLL samples from SF3B1-mutated patients. Integrated expression and splicing analyses revealed upregulation of SAT1 in SF3B1-mutated samples, accompanied by exon 4 skipping and increased mRNA stability. We then employed targeted metabolomics to assess the impact of the spliced isoform of SAT1 (SAT1sp) on polyamine metabolism. The results showed a significant increase in N1-acetylspermidine levels in SAT1sp-overexpressing MEC-1 cells. Additionally, untargeted metabolomic analysis revealed a concomitant enrichment of glutamine metabolism and oxidative phosphorylation, forming a specific metabolic pattern that supports cell survival and proliferation. Functionally, SAT1sp overexpression enhanced cell proliferation and clonogenic potential in CLL cell lines (MEC-1 and JVM-3) and primary CLL cells, while SAT1 knockdown induced apoptosis in these cells. To assess whether CLL-associated SAT1sp expression disrupts T-cell function ex vivo, activated CD8 T cells from healthy donors were co-cultured with MEC-1 cells for 72 hours, and IFNγ production was measured by ELISA. The data demonstrated that SAT1sp overexpression in MEC-1 cells reduced IFNγ secretion and decreased the percentage of GZMB+ and TNFα+ CD8 T cells following co-culture. In line with this, SAT1sp inhibited T-cell-induced cell lysis, as shown by a reduced proportion of CD19+PI+ CLL cells. To investigate the impact of SAT1 on CLL progression in vivo, MEC-1 cells with overexpressed SAT1sp were injected into NSG mice. Mice injected with SAT1sp-overexpressing MEC-1 cells exhibited more pronounced splenomegaly and increased leukemia infiltration, as indicated by CD19 positive staining. To explore the effect of CLL-derived SAT1sp on T-cell function in vivo, a MEC-1 xenograft mouse model was established, followed by two rounds of CD8 T cell injections in NSG mice. The leukemic burden was significantly reduced, with smaller spleens and reduced infiltration in the spleen and liver, while SAT1sp overexpression largely abrogated these effects. These findings highlight the novel role of SAT1 in dampening T-cell functions and facilitating immune evasion in CLL. Mechanistic studies using RNA-seq revealed differentially expressed genes (DEGs) in MEC-1 cells upon SAT1sp overexpression, which were enriched in gene ontology (GO) terms related to “histone modifying activity” and “histone binding.” Consistently, SAT1sp increased the level of H3 acetylation in MEC-1 cells. ChIP-seq further identified Ac-H3 enrichment at the promoter region of IL18BP, a well-known immune checkpoint. Consequently, SAT1sp induced transcription and secretion of IL18BP in both MEC-1 and primary CLL cells. Notably, SAT1 was recently shown to non-canonically acetylate H3K27 domains in mitosis-regulating genes in ovarian cancer (Zheng et al., Nature Communications 2025), suggesting that SAT1-mediated acetylation could serve as a conserved oncogenic mechanism supporting multiple cancer traits. Furthermore, conditioned medium from SAT1sp-overexpressing MEC-1 cells or primary CLL cells significantly decreased the percentage of activated GZMB+ and TNFα+ T cells. Encouragingly, IL18BP knockdown or blockade rescued this effect, indicating that SAT1 suppresses T-cell activation in an IL18BP-dependent manner. ConclusionThis study provides evidence for a dual mechanism by which SAT1 regulates polyamine metabolism and immune response in SF3B1-mutated CLL. SAT1 enhances polyamine catabolism, supporting leukemia survival through metabolic rewiring. Simultaneously, SAT1 regulates Ac-H3 at the IL18BP promoter, promoting IL18BP expression, inhibiting the IL18/IL18RAP axis, and suppressing CD8 T cell activation, which leads to immune evasion. These findings suggest that targeting SAT1 could be a therapeutic strategy to enhance T-cell-mediated immunity and improve outcomes in SF3B1-mutated CLL.

Background Primary refractory or relapsed disease occurs in 20–40% of classical Hodgkin lymphoma (cHL) patients after first-line therapy. Although PD-(L)1 inhibitors are standard salvage therapy for refractory/relapsed (r/r) cHL, acquired resistance is increasingly prevalent. Post-PD-(L)1 inhibitor failure options remain limited, with brentuximab vedotin (BV) constrained by cost barriers in resource-limited settings. Autologous stem cell transplantation (ASCT) offers curative potential but requires salvage-sensitive regimen (≥partial response, PR) pre-transplantation. Thus, developing an effective, low-toxicity, and cost-efficient re-induction regimen for PD-(L)1 inhibitor-resistant r/r cHL is an urgent unmet need. Aim To assess the efficacy, safety, and survival outcomes of the bendamustine, gemcitabine, and vinorelbine (BeGEV) regimen followed by ASCT in patients with PD-(L)1 inhibitor-resistant r/r cHL. Additionally, the impact of bendamustine on hematopoietic stem cell mobilization was investigated. Methods This retrospective analysis enrolled eight patients with r/r cHL treated at Henan Cancer Hospital between May 2021 and May 2023. All participants had PD-(L)1 inhibitor-refractory/relapsed or intolerant disease, received four 21-day cycles of BeGEV re-induction therapy: bendamustine (90 mg/m² IV on days 2–3), gemcitabine (800 mg/m² IV on days 1 and 4), and vinorelbine (1.5 mg/m² [max 2 mg] IV on day 1). Patients achieving ≥ PR underwent immediate ASCT with G-CSF-only mobilization and BEAM conditioning (BCNU 300 mg/m² day -7, etoposide 200 mg/m² days -6 to -3, cytarabine 200 mg/m² days -6 to -3, melphalan 140 mg/m² day -2). Post-ASCT surveillance included quarterly clinical/radiologic assessments in year 1 and semi-annually thereafter, without maintenance therapy. Results Eight patients were enrolled and median age was 36 years (range: 28-52). All presented with advanced-stage (Ann Arbor III–IV) and a median of 4 prior therapy lines before PD-(L)1 inhibitor. After four cycles of BeGEV chemotherapy, the overall response rate (ORR) was 100% (8/8), with a complete response (CR) rate of 62.5% (5/8). All patients underwent subsequent ASCT, achieving a post-ASCT ORR of 100% (8/8) and an improved CR rate of 87.5% (7/8). The most frequent BeGEV-related adverse event was grade 1–2 neutropenia (occurring in 2/8 patients, 25.0%). No bendamustine-related toxicities such as alopecia or peripheral neuropathy were observed. With a median follow-up of 32 months, neither median progression-free survival (PFS) nor overall survival (OS) had been reached. Six patients (75.0%, 6/8) maintained continuous CR, while one patient (12.5%, 1/8) sustained PR. Only one patient (12.5%, 1/8) experienced disease progression at 17 months post-ASCT; notably, this patient had achieved PR after BeGEV and CR post-ASCT. One patient achieved the longest follow-up duration of 50 months and remained disease-free. All eight patients achieved successful mobilization of peripheral blood stem cells, with a mean CD34+ cell yield of 9.04 × 10⁶/kg. Following ASCT, rapid hematopoietic recovery was observed, with median times to neutrophil engraftment and platelet engraftment of 11 days and 15 days, respectively. Remarkably, total treatment costs were contained at <100,000 CNY (~14,000 USD) per patient, enabled by reimbursement-eligible BeGEV agents under China's healthcare policy and avoidance of post-ASCT maintenance therapy. Conclusion Despite the limited sample size, this long-term study demonstrates that BeGEV-ASCT induces durable remissions in PD-(L)1 inhibitor-resistant cHL with favorable cost profiles, positioning it as a viable therapeutic strategy for resource-limited settings. Prospective multi-center validation is warranted.

Background: First-line treatments such as intensive immunochemotherapy followed by autologous stem cell transplantation (ASCT) provide favorable outcomes in fit younger mantle cell lymphoma (MCL) patients. However, elderly patients and younger high-risk patients (e.g., TP53 mutation, high Ki-67, blastoid morphology) face therapeutic challenges with no consensus frontline regimen. Zanubrutinib, a selective BTK inhibitor, demonstrates improved safety and tolerability. Combining it with anti-CD20 antibodies may enhance efficacy. This study evaluates zanubrutinib-containing regimens in elderly and younger high-risk treatment-naïve MCL patients. Methods: This multicenter, investigator-initiated, prospective study stratified patients into two cohorts based on age and biological risk: an elderly cohort (n=21; age ≥65 years) and a younger high-risk cohort (n=20; age <65 years with at least one high-risk feature, including TP53 mutation, blastoid/pleomorphic variant, high-risk sMIPI score [6–11 points], or Ki-67 >30%). Elderly cohort (n=21): Patients received zanubrutinib 160 mg orally BID combined with rituximab 375 mg/m² intravenously on day 1 of each 21-day cycle for 12 cycles as induction therapy, followed by zanubrutinib maintenance until disease progression or unacceptable toxicity. Younger high-risk cohort (n=20): Patients received zanubrutinib 160 mg BID in combination with the R-BAC regimen (rituximab 375 mg/m² on day 1, bendamustine 70 mg/m² on days 2–3, and cytarabine 500 mg/m² on days 2–4), administered every 28 days for 6 cycles as induction therapy. Patients achieving complete remission (CR) or partial remission (PR) and meeting criteria for ASCT underwent ASCT consolidation, followed by zanubrutinib maintenance therapy. Patients ineligible for transplantation continued zanubrutinib monotherapy until disease progression or unacceptable toxicity. Results: From October 2023 to May 2025, 23 patients with newly diagnosed MCL were enrolled, of whom 22 were evaluable for efficacy. Of these, 9 (39.1%) were allocated to the elderly cohort and 13 (60.9%) to the younger high-risk cohort. The elderly cohort had a median age of 71 years (range, 66–82), comprising 7 males and 2 females. High-risk simplified MIPI (sMIPI ≥6) was observed in 4 patients (44.4%), bone marrow involvement in 4 (44.4%), Ki-67 >30% in 4 (44.4%), and TP53 mutation or deletion in 1 (11.1%). The younger high-risk cohort had a median age of 59 years (range, 43–63), with 11 males and 2 females. High-risk sMIPI was identified in 53.8% (7/13), bone marrow involvement in 84.6% (11/13), Ki-67 >30% in 53.8% (7/13), and blastoid variant in 7.7% (1/13). The elderly cohort had an overall response rate (ORR) of 100.0% (95% CI: 66.4%–100.0%) and a CR rate of 88.9% (8/9; 95% CI: 51.8%–99.7%). Among five patients evaluable for minimal residual disease (MRD), four were MRD-negative at cycle 4, resulting in an MRD negativity rate of 80.0% (4/5), including one patient with a TP53 mutation. At a median follow-up of 11.5 months(as of May 2025), the 1-year progression-free survival (PFS) rate was 85.7% (95% CI: 51.8%–99.7%), with one patient experiencing disease progression at 10 months. In the younger high-risk cohort, ORR was 100.0% (95% CI: 75.3%–100.0%), and the CR rate was 84.6% (11/13; 95% CI: 54.6%–98.1%). Among eight patients assessed for MRD after 2 cycles, 87.5% (7/8) were MRD-negative, including one with blastoid morphology. At a median follow-up of 10.7 months (as of May 2025), no events occurred within 12 months; one patient progressed later at 14.9 months. Median duration of response (DoR) was not reached in either cohort; 81.8% (n=18) maintained response ≥6 months. Grade 3–4 adverse events (AEs) occurred mainly in younger high-risk patients, including neutropenia (38.5%), anemia (15.4%), thrombocytopenia (23.1%), and pneumonia (23.1%), mostly related to R-BAC and all manageable. No grade 3–4 AEs were observed in elderly patients. Common grade 1–2 AEs included arrhythmia and rash, each occurring in 1 of 9 patients (11.1%).No deaths were reported. Conclusions: In treatment-naïve elderly MCL patients, zanubrutinib combined with rituximab demonstrated favorable efficacy and safety, warranting further investigation. For younger high-risk patients, zanubrutinib combined with R-BAC showed encouraging efficacy and high MRD negativity rates in a small cohort, suggesting its potential to improve prognosis and showing promise as a first-line treatment option for this population.

Over the past few years, dual-targeted chimeric antigen receptor (CAR) T-cell therapy has been employed in the management of hematological malignancies to mitigate treatment failure, particularly in cases of antigen escape. The most widely used approaches include CD19/CD20, CD20/CD22, and BCMA/CD19 CAR T-cells. Alternative immune cells, including natural killer T cells and invariant natural killer T cells, exhibit innate anti-tumor activity and reduced toxicity. This review summarizes several recent clinical trial reports and preclinical studies from the 2023 American Society of Hematology (ASH) annual meeting on dual-targeted CAR T-cell immunotherapy for hematological malignancies.

The lymphocyte-specific protein tyrosine kinase (LCK) plays a crucial role in both T-cell development and activation. Dysregulation of LCK signaling has been demonstrated to drive the oncogenesis of T-cell acute lymphoblastic leukemia (T-ALL), thus providing a therapeutic target for leukemia treatment. In this study, we introduced a sophisticated virtual screening strategy combined with biological evaluations to discover potent LCK inhibitors. Our initial approach involved utilizing the PLANET algorithm to assess and contrast various scoring methodologies suitable for LCK inhibitor screening. After effectively evaluating PLANET, we progressed to devise a virtual screening workflow that synergistically combines the strengths of PLANET with the capabilities of Schr & ouml;dinger's suite. This integrative strategy led to the efficient identification of four potential LCK inhibitors. Among them, compound 1232030-35-1 stood out as the most promising candidate with an IC50 of 0.43 nM. Further in vitro bioassays revealed that 1232030-35-1 exhibited robust antiproliferative effects on T-ALL cells, which was attributed to its ability to suppress the phosphorylations of key molecules in the LCK signaling pathway. More importantly, 1232030-35-1 treatment demonstrated profound in vivo antileukemia efficacy in a human T-ALL xenograft model. In addition, complementary molecular dynamics simulations provided deeper insight into the binding kinetics between 1232030-35-1 and LCK, highlighting the formation of a hydrogen bond with Met319. Collectively, our study established a robust and effective screening strategy that integrates AI-driven and conventional methodologies for the identification of LCK inhibitors, positioning 1232030-35-1 as a highly promising and novel drug-like candidate for potential applications in treating T-ALL.

Adhesion molecules play essential roles in the homeostatic regulation and malignant transformation of hematopoietic cells. The dysregulated expression of adhesion molecules in leukemic cells accelerates disease progression and the development of drug resistance. Thus, targeting adhesion molecules represents an attractive anti-leukemic therapeutic strategy. In this study, we investigated the prognostic role and functional significance of cytohesin-1 (CYTH1) in acute myeloid leukemia (AML). Analysis of AML patient data from the GEPIA and BloodSpot databases revealed that CYTH1 was significantly overexpressed in AML and independently correlated with prognosis. Functional assays using AML cell lines and an AML xenograft mouse model confirmed that CYTH1 depletion significantly inhibited the adhesion, migration, homing, and engraftment of leukemic cells, delaying disease progression and prolonging animal survival. The CYTH1 inhibitor SecinH3 exerted in vitro and in vivo anti-leukemic effects by disrupting leukemic adhesion and survival programs. In line with the CYTH1 knockdown results, targeting CYTH1 by SecinH3 suppressed integrin-associated adhesion signaling by reducing ITGB2 expression. SecinH3 treatment efficiently induced the apoptosis and inhibited the growth of a panel of AML cell lines (MOLM-13, MV4-11 and THP-1) with mixed-lineage leukemia gene rearrangement, partly by reducing the expression of the anti-apoptotic protein MCL1. Moreover, we showed that SecinH3 synergized with the BCL2-selective inhibitor ABT-199 (venetoclax) to inhibit the proliferation and promote the apoptosis of ABT-199-resistant leukemic cells. Taken together, our results not only shed light on the role of CYTH1 in cell-adhesion-mediated leukemogenesis but also propose a novel combination treatment strategy for AML.

Introduction: Anti-CD20 monoclonal antibodies (mAbs) have revolutionized the treatment of B-cell lymphomas (BCL). The chemo-free combination with lenalidomide (Len) and rituximab (R2) as a first-line therapy has shown promising activity in indolent lymphomas. However, the efficacy of R2 regimen is yet unsatisfactory in aggressive settings, thereby novel synergistic strategy with next-generation CD20 mAbs and/or newer immunomodulatory drugs (IMiD) are urgently needed. Pomalidomide (Pom) is more powerful than Len in immunomodulatory properties including redirection of tumor-associated macrophages (TAM), which may confer a therapeutic opportunity to potentiate antibody-dependent cellular phagocytosis (ADCP) in elimination of lymphoma cells. This study aimed to evaluate the macrophage-based anti-lymphoma activities of Pom plus CD20 mAbs. Methods: BCL cell lines (SU-DHL-4, Raji and Daudi) were co-cultured with human monocyte-derived macrophages and treated with various combinations with IMiDs (Len or Pom) plus CD20 mAbs (rituximab or obinutuzumab). Phagocytosis and cell viability of lymphoma cells were tested by flow cytometry. Protein mass spectrometry and co-immunoprecipitation (co-IP) were performed to identify Pom-redirected neosubstrates of cereblon (CRBN). IMiD-responsive CrbnI391V mice were bred to Eμ-Myc lymphoma mice to evaluate the in vivo immunomodulatory effects of Pom on macrophages. Results: In vitro co-culture systems showed that Pom plus obinutuzumab elicited the strongest ADCP effects, resulting in robust phagocytosis of tumor cells by macrophages and markedly decreased tumor cell number, when compared to other combinations including Len plus rituximab (R2), Len plus obinutuzumab (GALEN) or Pom plus rituximab. Mechanistically, we identified CD47, a well-known negative regulator of macrophage phagocytosis, as a novel CRBN neosubstrate in BCL cells using mass spectrometry screening and co-IP validation. Interestingly, we found that only Pom rather than Len or thalidomide could efficiently induce degradation of total and surface CD47 in SU-DHL-4 cells. We further confirmed that knockdown of CRBN abrogated Pom-induced CD47 degradation, indicating the on-target effect. More importantly, based on the murine spontaneous lymphoma models (Eμ-Myc CrbnI391V mice), we found administration of Pom elicited a shift from M2 (CD206+) to M1 (CD86+) phenotype of tumor-associated macrophages and significant decrease of SIRPα+ populations in both M1 and M2 subtypes. Conclusion: Our study provides evidence for the superior effects of Pom plus obinutuzumab by harnessing the anti-lymphoma activity of TAMs, and suggests degradation of CD47-SIRPα axis as a novel mechanism underlying the drug synergy. Further preclinical and clinical investigations are needed to determine the in vivo synergistic efficacy and potential toxicity of this combined treatment. The research was funded by: This study was supported by the National Natural Science Foundation of China (No. 81470336 to KZ). Keywords: Aggressive B-cell non-Hodgkin lymphoma, Combination Therapies, Immunotherapy No conflicts of interests pertinent to the abstract.

Objective A SARS-CoV-2 Omicron (BA.5.2) epidemic began in China in December, 2022 following stopping the zero COVID policy. Methods We studied features of the epidemic in 1,121 persons with chronic myeloid leukaemia (CML). Results 1103 (98%) were in chronic, 10 in accelerated and 8 in acute phases. 834 (74%) became infected almost all of whom met criteria for COVID-19. The most common symptoms were fever (91%), cough (90%) and fatigue (82%). 42 infected persons were asymptomatic. Most people quarantined at home and self-medicated. 22 were hospitalized for COVID-19. At admission 5 had mild, 14, moderate and 3, severe/critical disease according to World Health Organization (WHO) criteria. 5 received respiratory assistance, 3 were admitted to the intensive care unit (ICU) and 1 in accelerated phase died from COVID-19. Co-variates associated with a risk of COVID-19 in SARS-CoV-2-infected subjects include age ≥ 65 years, higher education level and imatinib therapy. Conclusion In conclusion, most SARS-CoV-2 Omicron BA.5.2 infections in persons with CML resulted in COVID-19 most of which cases are mild with only 1 death.

DHX15 has been implicated in RNA splicing and ribosome biogenesis, primarily functioning as an RNA helicase. To systematically assess the cellular role of DHX15, we conducted proteomic analysis to investigate the landscape of DHX15 interactome, and identified MYC as a binding partner. DHX15 co-localizes with MYC in cells and directly interacts with MYC in vitro. Importantly, DHX15 contributes to MYC protein stability at the post-translational level and independent of its RNA binding capacity. Mechanistic investigation reveals that DHX15 interferes the interaction between MYC and FBXW7, thereby preventing MYC polyubiquitylation and proteasomal degradation. Consequently, the abrogation of DHX15 drastically inhibits MYC-mediated transcriptional output. While DHX15 depletion blocks T cell development and leukemia cell survival as we recently reported, overexpression of MYC significantly rescues the phenotypic defects. These findings shed light on the essential role of DHX15 in mammalian cells and suggest that maintaining sufficient MYC expression is a significant contributor to DHX15-mediated cellular functions.

RNA-binding proteins (RBP) have emerged as essential regulators that control gene expression and modulate multiple cancer traits. T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy derived from transformation of T-cell progenitors that normally undergo discrete steps of differentiation in the thymus. The implications of essential RBP during T-cell neoplastic transformation remain largely unclear. Systematic evaluation of RBP identifies RNA helicase DHX15, which facilitates the disassembly of the spliceosome and release of lariat introns, as a T-ALL dependency factor. Functional analysis using multiple murine T-ALL models demonstrates the essential importance of DHX15 in tumor cell survival and leukemogenesis. Moreover, single-cell transcriptomics reveals that DHX15 depletion in T-cell progenitors hinders burst proliferation during the transition from doublenegative to double-positive cells (CD4-CD8- to CD4+CD8+). Mechanistically, abrogation of DHX15 perturbs RNA splicing and leads to diminished levels of SLC7A6 and SLC38A5 transcripts due to intron retention, thereby suppressing glutamine import and mTORC1 activity. We further propose a DHX15 signature modulator drug ciclopirox and demonstrate that it has prominent anti-T-ALL efficacy. Collectively, our data highlight the functional contribution of DHX15 to leukemogenesis through regulation of established oncogenic pathways. These findings also suggest a promising therapeutic approach, i.e., splicing perturbation by targeting spliceosome disassembly, may achieve considerable anti-tumor efficacy.

Inflammatory bowel disease (IBD) is closely associated with dysregulation of genetic factors and microbial environment. Here, we report a susceptible role of ubiquitin-specific protease 2 (USP2) in experimental colitis and bacterial infections. USP2 is upregulated in the inflamed mucosa of IBD patients and in the colon of mice treated with dextran sulfate sodium salt (DSS). Knockout or pharmacologic inhibition of USP2 promotes the proliferation of myeloid cells to activate IL-22 and IFNγ production of T cells. In addition, knockout of USP2 in myeloid cells inhibits the production of pro-inflammatory cytokines to relieve the dysregulation of extracellular matrix (ECM) network and promote the gut epithelial integrity after DSS treatment. Consistently, Lyz2-Cre;Usp2fl/fl mice exhibit hyper-resistance to DSS-induced colitis and Citrobacter rodentium infections compared to Usp2fl/fl mice. These findings highlight an indispensable role of USP2 in myeloid cells to modulate T cell activation and epithelial ECM network and repair, indicating USP2 as a potential target for therapeutic intervention of IBD and bacterial infections in the gastrointestinal system.

To prolong service life of M50 steel for the main shaft bearing, ultrasonic surface rolling process (USRP) was utilized to improve surface performances. The effects of USRP on surface integrity and tribological behavior were investigated. The results show the effect of USRP parameters on surface performances follows the order: feed rate > load > rolling times. During the USRP treatment, plastic deformation layer with 24 mu m thickness is formed, and fine grain strengthening increases the microhardness (15.56%) and residual stress (79.61%) and decreases the surface roughness (-68.09%), respectively. Friction coefficient and wear rate of USRP sample are lower than those of untreated sample, because USRP helps form a smooth tribolayer and changes the main wear mechanism to slight adhesive wear.

Targeting B-cell receptor signalling using Bruton tyrosine kinase (BTK) inhibitors (BTKis) has become a highly successful treatment modality for B-cell malignancies, especially for chronic lymphocytic leukaemia. However, long-term administration of BTKis can be complicated by adverse on- and/or off-target effects in particular cell types. BTK is widely expressed in cells of haematopoietic origin, which are pivotal components of the tumour microenvironment. BTKis, thus, show broad immunomodulatory effects on various non-B immune cell subsets by inhibiting specific immune receptors, including T-cell receptor and Toll-like receptors. Furthermore, due to the off-target inhibition of other kinases, such as IL-2-inducible T-cell kinase, epidermal growth factor receptor, and the TEC and SRC family kinases, BTKis have additional distinct effects on T cells, natural killer cells, platelets, cardiomyocytes, and other cell types. Such mechanisms of action might contribute to the exceptionally high clinical efficacy as well as the unique profiles of adverse effects, including infections, bleeding, and atrial fibrillation, observed during BTKi administration. However, the immune defects and related infections caused by BTKis have not received sufficient attention in clinical studies till date. The broad involvement of BTK in immunological pathways provides a rationale to combine BTKis with specific immunotherapies, such as immune checkpoint inhibitor or chimeric antigen receptor-T-cell therapy, for the treatment of relapsed or refractory diseases. This review discusses and summarises the above-mentioned issues as a reference for clinicians and researchers.

Immunomodulatory drugs (IMiDs) such as thalidomide, lenalidomide and pomalidomide are antitumor compounds that have direct tumoricidal activity and indirect effects mediated by multiple types of immune cells in the tumor microenvironment (TME). IMiDs have shown remarkable therapeutic efficacy in a set of B-cell neoplasms including multiple myeloma, B-cell lymphomas and chronic lymphocytic leukemia. More recently, the advent of immunotherapy has revolutionized the treatment of these B-cell neoplasms. However, the success of immunotherapy is restrained by immunosuppressive signals and dysfunctional immune cells in the TME. Due to the pleiotropic immunobiological properties, IMiDs have shown to generate synergetic effects in preclinical models when combined with monoclonal antibodies, immune checkpoint inhibitors or CAR-T cell therapy, some of which were successfully translated to the clinic and lead to improved responses for both first-line and relapsed/refractory settings. Mechanistically, despite cereblon (CRBN), an E3 ubiquitin ligase, is considered as considered as the major molecular target responsible for the antineoplastic activities of IMiDs, the exact mechanisms of action for IMiDs-based TME re-education remain largely unknown. This review presents an overview of IMiDs in regulation of immune cell function and their utilization in potentiating efficacy of immunotherapies across multiple types of B-cell neoplasms.

Recombination activating genes 1 ( Rag1 ) and Rag2 are expressed in immature lymphocytes and essential for generating the vast repertoire of antigen receptors. Yet, the mechanisms governing the transcription of Rag1 and Rag2 remain to be fully determined, particularly in thymocytes. Combining cDNA microarray and ChIP-seq analysis, we identify Rag1 and Rag2 as novel Notch1 transcriptional targets in acute T-cell lymphoblastic leukemia (T-ALL) cells. We further demonstrate that Notch1 transcriptional complexes directly bind the Rag1 and Rag2 locus in not only T-ALL but also primary double negative (DN) T-cell progenitors. Specifically, dimeric Notch1 transcriptional complexes activate Rag1 and Rag2 through a novel cis -element bearing a sequence-paired site (SPS). In T-ALL and DN cells, dimerization-defective Notch1 causes compromised Rag1 and Rag2 expression; conversely, dimerization-competent Notch1 achieves optimal upregulation of both. Collectively, these results reveal Notch1 dimerization-mediated transcription as one of the mechanisms for activating Rag1 and Rag2 expression in both primary and transformed thymocytes. Our data suggest a new role of Notch1 dimerization in compelling efficient TCRβ rearrangements in DN progenitors during T-cell development.

The balance between antioxidants and reactive oxygen species (ROS) critically regulates tumor initiation and progression. However, whether and how the tumor-favoring redox status is controlled by cytokine networks remain poorly defined. Here, it is shown that IL-36γ and IL-36Ra reciprocally regulate the progression of non-small cell lung cancer (NSCLC) by modulating glutathione metabolism and ROS resolution. Knockout, inhibition, or neutralization of IL-36γ significantly inhibits NSCLC progression and prolongs survival of the KrasLSL-G12D/+ Tp53fl/fl and KrasLSL-G12D/+ Lkb1fl/fl mice after tumor induction, whereas knockout of IL-36Ra exacerbates tumorigenesis in these NSCLC mouse models and accelerates death of mice. Mechanistically, IL-36γ directly upregulates an array of genes involved in glutathione homeostasis to reduce ROS and prevent oxidative stress-induced cell death, which is mitigated by IL-36Ra or IL-36γ neutralizing antibody. Consistently, IL-36γ staining is positively and negatively correlated with glutathione biosynthesis and ROS in human NSCLC tumor biopsies, respectively. These findings highlight essential roles of cytokine networks in redox for tumorigenesis and provide potential therapeutic strategy for NSCLC.

Clinic therapy of acute myeloid leukemia (AML) remains unsatisfactory that urges for development of novel strategies. Recent studies identified ANP32A as a novel biomarker of unfavorable outcome of leukemia, which promoted leukemogenesis by increasing H3 acetylation and the expression of lipid metabolism genes. It is of great significance to investigate whether targeting ANP32A is a novel strategy for leukemia therapy. To target ANP32A, we identified a peptide that competed with ANP32A to bind to histone 3 (termed as H3-binding peptide, H3BP). Disrupting ANP32A and H3 interaction by the overexpression of H3BP-GFP fusion protein mimicked the effect of ANP32A knockdown, impaired H3 acetylation on multiple locus of target genes, reduced proliferation, and caused apoptosis in leukemia cells. Furthermore, a synthesized membrane-penetrating peptide TAT-H3BP effectively entered into leukemia cells and phenocopied such effect. In vivo, TAT-H3BP showed potent efficacy against leukemia: Intra-tumor injection of TAT-H3BP significantly reduced the volume of subcutaneous tumors in nude mice and recipient mice engrafted with TAT-H3BP-pretreated 6133/MPL W515L cells exhibited ameliorated leukemia burden and prolonged survival. Noticeably, TAT-H3BP efficiently suppressed proliferation and colony-forming unit of human primary AML cells without affecting normal cord blood cells. Our findings demonstrate that intervening the physical interaction of ANP32A with H3 impairs the oncogenicity of ANP32A and may be a promising therapeutic strategy against AML.