New England Biolabs (China)

The molecular heterogeneity and distinct features of HER2-low breast cancer are poorly understood, limiting their precise management. To address this issue, longitudinal multiomic profiling of HER2-low breast cancer is performed, including genomics, transcriptomics, proteomics, lactylomics, and phosphoproteomics, using 250 well-characterized samples, and identified three proteomic subtypes: PS1 (estrogen response signaling enriched), PS2 (angiogenesis enriched), and PS3 (proliferation enriched and HER2-high like). These three proteomic subtypes have distinct features and potential therapeutic strategies, namely, endocrine therapy, antiangiogenic therapy, and anti-HER2 therapy, and are validated in external datasets and PDO models. In addition, a detailed description of the genomic characteristics and a map of the lactate modification landscape of HER2-low breast cancer are provided. This research provides complementary information, reveals the molecular characteristics of HER2-low breast cancer, and suggests potential precise therapeutic strategies for patients with this type of cancer.

Abstract Background: Small cell lung cancer (SCLC) is a highly malignant tumor and accounts for approximately 15% of lung cancer cases. The prognosis of invasive SCLC is poor, with a 5-year survival is 7%. SCLC has a high response to chemotherapy but tends to relapse and resist. In recent years, immune checkpoints have brought breakthroughs in first-line treatment and prolonged OS, but options after the second line are still lacking. Delta-like ligand 3 (DLL3) is an inhibitory Notch ligand that is highly expressed in SCLC and other neuroendocrine tumors, and its expression is limited in normal cells. Nowadays, many DLL3-targeting therapies are in development among them Tarlatamab, which is a T cell engager (TCE), has been approved by FDA for the treatment of adult patients with extensive-stage small cell lung cancer (ES-SCLC) with disease progression on or after platinum-based chemotherapy, at the same time several DLL3 targeting ADCs were also showing some promising results. Thus makes DLL3-targeting therapy an attractive strategy to treat SCLC. Methods: Here we report a T cell engager conjugate (TEC), LBL-058, comprised of a DLL3-targeted T cell engager conjugated to a novel topoisomerase I inhibitor (TOP1i) payload. Moreover, LBL-058 was designed with a higher affinity targeting DLL3 and a lower affinity targeting CD3 to avoid potential cytotoxicity to T cells mediated by binding to T cells. The binding affinity of LBL-058 to DLL3 and CD3 was determined with Fortebio, while the T cell based activity was measured using several cell-based assays including reporter gene and TDCC assays. Binding of LBL-058 to DLL3 or CD3 induced Internalization and cytotoxicity were also evaluated by several cell-based assays. The anti-tumor activity of LBL-058 was investigated in xenograft mouse model. Results: The affinity difference between CD3 and DLL3 has been fine-tuned, LBL-058 binds more easily to DLL3-positive cells. In CD3 reporter gene assay, LBL-058 could activate the NFAT reporter element through DLL3 antigen-dependent CD3 cross-linking. LBL-058 not only induce potent TDCC and T cell activation but also selectively deliver cytotoxicity payload to DLL3-positive tumor cells. In animal models, LBL-058 treatment induced durable tumor regression in xenograft tumor model. Conclusion: LBL-058 has the potential to be a First-In-Class DLL3-targeted TCE ADC, which combines the anti-tumor properties of TCE and ADC, and shows a potent and durable anti-tumor activity and is expected to provide clinical benefits for the treatment of patients with SCLC. Citation Format: Sun Jianming, Huang Xiao, Huang Jing, Zhang Peng, Ding Mi, Qin Yurong, Wu Guojin, Mi Ye, Yang Ye, Jordan Zhu, Yue Zhao, Shoupeng Lai, Xiaoqiang Kang, Hong Ling. LBL-058, A novel T cell engager conjugate (TEC) targeting DLL3 with dual tumor suppressive functions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 7322.

Abstract Background: Multiple myeloma (MM) is a clonal plasma cell proliferative disorder characterized by abnormal increase of monoclonal immunoglobulins. Despite newly developed novel therapeutic agents have greatly improved the outcome of MM patients in the past decades, MM is still an incurable hematological malignancy. Several CD3 bispecific immunotherapies that harness T cells against MM are at various stages of preclinical and clinical development, but most of them are hampered by the refractory and resistant mechanisms. Leukocyte immunoglobulin-like receptor B4 (LILRB4) is an immune checkpoint inhibitory receptor, which is highly expressed on monocytic blasts of AML and myeloid-derived suppressive cells. Most recently it is discovered also highly expressed in MM, in particular rrMM(relapsed/refractory MM). It does not express on hematopoietic stem cells and progenitor cells, therefore make it a promising target for hematological malignancies. Herein a novel T cell engager targeting LILRB4, LBL-043, is developed to specifically kill LILRB4 expressing MM cells. Methods: LBL-043 is a bispecific antibody targeting LILRB4 and CD3, designed using proprietary LeadsBodyTM platform, which is a 2:1 format with two VHH arms targeting LILRB4 with high affinity and one scFv arm targeting CD3 with fine-tuned affinity. The binding affinity of LBL-043 was determined with Fortebio and flow cytometry, and the function activity was measured using reporter gene and TDCC assays on MM cell lines. The anti-tumor activity was investigated in B-NDG/hu-PBMC reconstructed mice implanted with NCI-H929. In addition, the potential toxicity profile of LBL-043 was examined in a non-GLP dose range finding (DRF) study in cynomolgus monkeys via IV infusion, once a week for a total of 4 doses. Results: The binding affinity of LBL-043 to LILRB4 and CD3 protein was 0.724 nM and 35.7 nM, respectively. In CD3 reporter gene assays, LBL-043 could activate the NFAT signaling only in the presence of LILRB4+ cells. And LBL-043 was shown a potent TDCC activity in different LILRB4 expression level MM cells. In NCI-H929 CDX mouse model, LBL-043 was shown significant anti-tumor activity. DRF Study in Drug Toxicology Assessments demonstrated that LBL-043 has good safety profile in cynomolgus monkeys. There were no LBL-043 related changes of cardiac parameters, clinical chemistry, hematology, urinalysis, organ weight and gross pathology (except for decreased size of thymus). Conclusion: LBL-043, a novel bispecific antibody targeting CD3 and LILRB4, redirecs T cells to effectively kill MM cells by simultaneously binding MM cells and T cells. It is also shown a great anti-tumor efficacy in NCI-H929 CDX mouse model. DRF of LBL-043 (0.2∼10 mg/kg) in cynomolgus monkeys was well tolerated, and HNSTD was 10 mg/kg. These data for the first time show LBL-043 as a promising therapeutic bispecific antibody for the treatment of MM patients. Citation Format: Yujia Dang, Xiao Huang, Yurong Qin, Xiaoya Liu, Min Chen, Duqing Jiang, Guojin Wu, Mi Ye, Jianming Sun, Hong Ling, Shoupeng Lai, Xiaoqiang Kang, Jordan Zhu. LBL-043, a novel LILRB4xCD3 T cell engager, for the treatment of relapsed/refractory multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 7323.

Chronic, non-healing wounds are a leading cause of prolonged patient morbidity and mortality due to biofilm- associated, polymicrobial infections. Staphylococcus aureus and Pseudomonas aeruginosa are the most frequently co-isolated pathogens from chronic wound infections. Competitive interactions between these pathogens contribute to enhanced virulence, persistence, and antimicrobial tolerance. P. aeruginosa utilizes the extracellular proteases LasB, LasA, and AprA to degrade S. aureus surface structures, disrupt cellular physiology, and induce cell lysis, gaining a competitive advantage during co-infection. S. aureus evades P. aeruginosa by employing aggregation mechanisms to form biofilms. The cell wall protein SasG is implicated in S. aureus biofilm formation by facilitating intercellular aggregation upon cleavage by an extracellular protease. We have previously shown that proteolysis by a host protease can induce aggregation. In this study, we report that P. aeruginosa proteases LasA, LasB, and AprA cleave SasG to induce S. aureus aggregation. We demonstrate that SasG contributes to S. aureus biofilm formation in response to interactions with P. aeruginosa proteases by quantifying aggregation, SasG degradation, and proteolytic kinetics. Additionally, we assess the role of SasG in influencing S. aureus biofilm architecture during co-infection in vivo, chronic wound co-infections. This work provides further knowledge of some of the principal interactions that contribute to S. aureus persistence within chronic wounds co-infected with P. aeruginosa, and their impact on healing and infection outcomes.

BackgroundThe myeloid-derived immune suppression in the tumor microenvironment contributes to tumor immune evasion. LILRB1 (also known as ILT2) and LILRB2 (also known as ILT4) are inhibitory receptors with intracellular tyrosine-based inhibitory motifs (ITIMs) that recognize common MHC-I ligands. Both LILRB1 and LILRB2 are highly expressed on tumor-infiltrating myeloid cells, and LILRB1 is also expressed on tumor-associated T cells, macrophages and NK cells. LILRB1 and LILRB2 receptors inhibit cytotoxic T cells’ function by interacting with human leukocyte antigen-G (HLA-G) and MHC-I. In patients who do not respond to immune checkpoint inhibitor therapies, the upregulation of LILRB1 and LILRB2 expression is observed, suggesting that these receptors may serve as potential mechanisms of resistance. Here we report a novel bispecific antibody LBL-042, targeting PD-1 and LILRB1/2 simultaneously, could block both PD-1 and LILRB1/LILRB2 immune suppression, may improve immune evasion of TME and possibly overcome resistance to IO therapy.MethodsA panel of diverse VHH antibodies against LILRB1/2 were developed using alpaca immunization and phage library screening. The candidate molecules underwent rigorous in vitro evaluation, including binding to LILRB1 and LILRB2, MHC-I and HLA-G blockade, myeloid cell functional assays, etc. The anti-PD-1 and LILRB1/2 bispecific antibodies were developed using the Leads Biolabs’s multi-specific antibody platform. These candidates include different anti-LILRB1/2 locations, valencies and IgG isotypes. The binding and blocking capabilities of these bispecific antibodies were evaluated using ELISA and FACS. The in vitro activities on myeloid cells were studied in macrophage phagocytosis, MDSC reprogramming and DC maturation assays. T cell proliferation and cytokine release were assessed using a mixed lymphocyte reaction assay. Furthermore, in vivo anti-tumor efficacy was investigated in A375-HLA-G PD-1 resistant tumor model.ResultsThe LBL-042 bispecific antibody exhibited high affinities to both PD-1 and LILRB1/2. It could block the binding of LILRB1/2 to MHC-I and HLA-G. Blockade of LILRB1/2 enhances macrophage phagocytosis, MDSC reprogramming and DC maturation. In MLR assay, it enhanced T cells’ activation. LBL-042 also demonstrated potent anti-tumor effects in vivo.ConclusionsThe novel bispecific antibody LBL-042, which targets PD-1 and LILRB1/2, unleash the suppression of T cells through anti-PD-1, while reprogramme or activate macrophages and DCs through LILRB1&2 inhibition. These effects reprogramme the immune-suppressive microenvironment, ultimately leading to a synergistic immune effect with anti-PD-1. It may overcome the resistance to PD-1 monotherapy induced by myeloid cell immune-suppression.

Abstract Protein post-translational modifications (PTMs) are crucial for cancer growth and metastasis. Vitamin B3, a key precursor of NAD + and NADP+, however its epigenetic functions in physiology and disease remain unclear. Herein we report a nicotinic acid (NA), a component of vitamin B3, induces a histone PTM, lysine nicotinylation (Knic), and demonstrate 17 Knic site across core histones in cells. Tandem mass spectrometry and stable isotope tracing revealed that NA-derived nicotinyl-CoA, catalysed by ACSS2, enhances histone Knic in vivo and in vitro. Analysis of chromatin accessibility revealed that histone Knic downregulates chromatin accessibility and therefore inhibits gene expression, for instance, restrain the binding of transcription factor HOXB9 to the promoter of oncogene PPFIA1. PPFIA1 level is correlated with malignancy and poor prognosis of hepatocellular carcinoma. These findings suggest that vitamin B3 supplementation may affects the chromatin accessibility depending on its composition. Collectively, we propose that NA induces histone Knic, a histone mark controlling gene expression.

The Monarch Mag Viral DNA/RNA Extraction Kit provides a rapid and reliable magnetic bead-based process for extracting viral nucleic acids from saliva and respiratory swab samples. The kit combines the efficiency of silica-based nucleic acid purification with the ease of use of magnetic beads. Manual and automated workflows allow samples to be processed in microfuge tubes or 96-well plates. Kit sizes align to 96-well formats (100 preps, 600 preps, and 1800 preps), and the protocol is compatible with high throughput automation on a variety of platforms, including the KingFisher Flex magnetic particle processor and Agilent Bravo and MGISP liquid handler platforms.