AI-based analysis of label-free live cell imaging of T-cell mediated tumor killing assay enables competitive and robust hit calling.

PD-1 inhibitors have reshaped the treatment landscape of classical Hodgkin lymphoma, yet a substantial proportion of patients exhibit primary or acquired resistance driven by tumor-intrinsic alterations, immunosuppressive microenvironmental signals, metabolic constraints, and EBV-mediated modulation. This review summarizes key mechanisms underlying PD-1 resistance and highlights emerging biomarkers-including early 18F-FDG PET response, circulating tumor DNA kinetics, molecular subtyping, and spatial immune profiling-that enable early identification of nonresponders and support precision immunotherapy. Novel therapeutic strategies such as macrophage-targeted agents, metabolic modulators, bispecific antibodies, low-dose PD-1 regimens, and CD30-directed CAR-T cells show promise in overcoming resistance, particularly when integrated into adaptive, biomarker-guided treatment algorithms. Early incorporation of PET and ctDNA monitoring may inform timely treatment adaptation, while combination approaches addressing macrophage-driven suppression or redundant immune checkpoints should be considered in biologically high-risk patients. Overall, a deeper mechanistic understanding coupled with biomarker-driven stratification is essential to optimize PD-1-based therapy and improve long-term outcomes in cHL.

Targeted Radionuclide Therapy (TRT) enables selective delivery of radionuclides for cancer treatment. Alpha particle emitters such as 212Pb are emerging as potential gamechangers, representing highly potent payloads for precision therapy and refractory cancer treatment. While small-molecule carriers are widely explored due to favorable pharmacokinetics, nanoparticle-based TRT remains less studied due to perceived non-ideal pharmacokinetics. We report a unique nanoparticle-TRT pretargeting approach using bispecific antibodies (BsAbs) which "prime" tumor surfaces, enhancing tumor-specific delivery and minimizing off-target deposition of 212Pb. We developed a poly(ethylene glycol) (PEG)-based nanomedicine platform to carry 212Pb, and employed in-house designed and manufactured BsAb (α-epidermal growth factor receptor (EGFR)/α-PEG) to prime tumors to receive the nanocarrier. Sequential administration of each component to EGFR-expressing cells produced enhanced receptor-mediated internalization of the BsAb upon nanomedicine binding, resulting in improved radionuclide delivery and efficacy in a series of in vitro assays. The pretargeting approach more than tripled tumor retention of the 212Pb-nanomedicine compared to the untargeted nanomaterial in a murine EGFR+ breast cancer xenograft model, evidenced by single-photon emission computed tomography (SPECT) imaging of 212Pb-loaded nanomaterials and gamma analysis of the excised organs. Therapeutic studies demonstrated the 212Pb-nanomedicine to produce well-tolerated and statistically-enhanced therapeutic outcomes for the pretargeting versus conventional 212Pb-nanomedicine, with no observed long term hematological effects. This work establishes a modular strategy for targeted TRT nanomedicine delivery. The platform has potential for broad applicability, including simultaneous delivery of diverse or synergistic payloads. These findings represent an important advance toward precision nanomedicine approaches in radionuclide therapy.

Shaping immunotherapy through the tumor microenvironment: Translational perspectives.

Advances and perspectives in CEA-targeted therapies: From classic biomarker toward actionable therapeutic target.

In this issue of Blood Cancer Discovery, Frenking and colleagues demonstrate the prognostic potential of three risk score models to predict hematotoxicity, infections, response rates, and survival following bispecific T-cell engager (TCE) therapy in multiple myeloma. These risk score models, which the authors validated using retrospective real-world data from 278 patients, can easily be calculated using routine blood testing and offer the potential to improve the efficacy and safety of TCE therapy. See related article by Frenking et al., p. XX .

Bispecific antibodies have revolutionized the treatment of multiple myeloma (MM), however primary treatment failure occurs in 30-40% of patients. In this study, we analyzed correlates of response to teclistamab and strategies to overcome primary resistance. Across two independent cohorts (n=90), we developed Hi-MM, defined by extramedullary disease, plasma cell leukemia, bone marrow plasmacytosis ≥50%, or transfusion within 30 days, as a composite correlate of non-response. Patients without Hi-MM had ORR of 84-96% (vs 20-40% with Hi-MM), and significantly superior progression-free (p<0.001) and overall survival (p<0.001). Debulking chemotherapy was utilized in 19 patients; 79% then responded to teclistamab, including 100% who no longer had Hi-MM. All four patients who were primary refractory to a BCMA bispecific immediately prior to debulking then achieved deep responses to teclistamab. In conclusion, simple clinical parameters correlate with response to teclistamab, while debulking chemotherapy can overcome Hi-MM and successfully bridge patients to teclistamab or salvage non-responders.

Metastatic gastric cancer remains a major global health challenge with poor long-term outcomes. Over the past 5 years, the treatment landscape has rapidly evolved with the integration of biomarker-informed strategies that guide the use of immune checkpoint inhibitors and targeted therapies in molecularly defined subgroups, including microsatellite unstable, PD-L1-expressing, HER2-positive and claudin 18.2-positive disease. Standard first-line treatment continues to rely on fluoropyrimidine-platinum chemotherapy backbones, with biomarker-driven agents selectively layered on for improved efficacy. Despite these advances, most patients continue to have disease progression, and durable responses are uncommon. In addition to identifying and validating new targets such as FGFR2b, ongoing efforts are focusing on novel strategies involving established targets, including HER2 and claudin 18.2, using next-generation treatment modalities such as antibody-drug conjugates, bispecific antibodies and cellular therapies. Complementary platforms including circulating tumour DNA and theranostic agents are also being explored to better guide treatment selection, facilitate non-invasive monitoring and enable early response assessments. In this Review, we summarize the current standard of care for patients with metastatic gastric cancer and also highlight emerging approaches aimed at improving the outcomes in these patients.

T-cell-redirecting bispecific antibodies (bsAb) offer a novel therapeutic approach for relapsed/refractory large B-cell lymphoma (R/R LBCL). However, predictive biomarkers are needed to identify patients most likely to respond. As both bsAbs and chimeric antigen receptor (CAR) T cells represent T-cell-based therapies, we hypothesized that the established CAR-HEMATOTOX (HT) and InflaMix models-reflecting the degree of systemic inflammation-could be of prognostic utility for bsAb therapy. We applied both scores to a multicenter international cohort of 174 patients with R/R LBCL treated with bsAbs across 15 sites. Patients with a high HT score (≥3, 35%) displayed inferior median progression-free (PFS; 1.4 vs 7.4 months; P< .0001) and overall survival (OS; 2.0 vs 21.7 months; P< .0001) compared with patients with a low HT score. When applying the InflaMix score, 49% of the patients were assigned to the inflammatory cluster, translating into a significantly shorter median PFS (1.9 vs 17.8 months; P< .0001) and OS (4.1 vs 21.7 months; P< .0001). In a multivariable Cox regression analysis accounting for various prognostic factors, HT and InflaMix remained independent adverse risk factors for both PFS and OS. Patients presenting with both elevated HT score and the inflammatory signature showed markedly shorter OS and PFS compared with patients deemed low-risk by either one of the scores. In the CAR-pretreated subcohort, the combination of early CAR T-cell relapse (≤3 months) and elevated inflammation led to particularly detrimental outcomes. Overall, these data highlight the prognostic utility of baseline inflammatory markers in identifying patients who may benefit from combinatorial strategies alongside bsAb.

Nasopharyngeal carcinoma (NPC), an Epstein-Barr virus (EBV)-driven malignancy with distinct geographic prevalence, presents as locally advanced or metastatic disease in over 70% of patients. Its unique tumor microenvironment (TME) exhibits dense immune infiltration paradoxically coupled with profound immunosuppression orchestrated by EBV through PD-L1 induction, antigen presentation disruption, immunosuppressive cell recruitment, and extracellular vesicle exploitation. These mechanisms underpin the rationale for immunotherapy, where PD-1/PD-L1 inhibitors have transformed management: phase III trials established PD-1 blockade combined with chemotherapy as first-line standard for recurrent/metastatic NPC, significantly improving survival, while integration into locoregionally advanced disease regimens enhances response rates and outcomes. Novel bispecific antibodies and rational combinations (with radiotherapy, anti-angiogenics, and EBV-targeted therapies) show promise in overcoming resistance. Biomarker advances extend beyond PD-L1 to include radiomics, AI-driven models, liquid biopsy markers (EBV-DNA dynamics, exosomal CA1), and tissue-based features (tertiary lymphoid structures, CD70/CD27 axis). Persistent challenges encompass EBV-mediated resistance, biomarker validation, and therapeutic optimization. This review comprehensively synthesizes the mechanistic basis of NPC immune evasion, clinical progress across diverse immunotherapies, biomarker-driven precision strategies, and emerging approaches to harness the immune microenvironment for improved patient outcomes.

Increased detection of NRG1 fusions in non-squamous non-small cell lung cancer using combined DNA and RNA sequencing in a real-world cohort.

Diffuse large B-cell lymphoma (DLBCL) is an aggressive B-cell malignancy and is the most common subtype of lymphoma. Treatment is administered with curative intent and approximately two thirds of patients are expected to have durable long-term survival. To achieve this, anthracycline-based chemotherapy in combination with rituximab is typically administered as initial therapy. Management is optimized based on the disease stage, prognostic clinical features, and histological or molecular subclassification. In patients with the activated B-cell subtype of DLBCL, polatuzumab vedotin is commonly included in the combination. For those with Myc and BCL-2 rearrangements, a more treatment intense approach is used. Despite this risk-adapted approach, at least one third of patients relapse. Those who relapse within 1 year, or are resistant to initial therapy typically receive chimeric antigen receptor (CAR) T-cell therapy. For those relapsing more than a year post initial treatment, salvage chemotherapy followed by an autologous stem cell transplant is offered. In patients ineligible for cellular therapy, or those who progress after CAR T-cell treatment, management is palliative and includes administration of bispecific antibodies or antibody drug conjugate combinations. To further improve the outcome of DLBCL patients, incorporation of cellular and bispecific therapies into front-line treatment is currently being tested.

A real-world comparison of CD20xCD3 bispecific antibodies versus chemotherapy regimens in 3+ line relapsed/refractory diffuse large B cell lymphoma

Abstract Unlike conventional monoclonal antibodies, complex biologics—such as bispecific antibodies (bsAbs) and fusion proteins—often face challenges including lower expression levels, higher mispairing rates, and greater sensitivity to culture conditions, which can collectively limit both titer and product quality. Leveraging process development case studies, this review systematically explores upstream bioprocessing strategies aimed at mitigating these challenges, with a focus on titer enhancement, lactate metabolism regulation, acidic charge variants control, reduction of aggregates and fragments, and glycosylation optimization. Finally, a perspective towards future upstream development strategies is discussed.

ABSTRACTNon–small cell lung cancer (NSCLC) is characterised by a number of driver mutations, among which epidermal growth factor receptor (EGFR) exon 20 insertion mutations are associated with poor prognosis due to structural alterations that confer resistance to conventional tyrosine kinase inhibitors (TKIs). Recently, amivantamab, a fully human bispecific antibody targeting EGFR and MET, has demonstrated promising efficacy in patients with EGFR exon 20 insertion mutations. However, its therapeutic activity against central nervous system metastases remains uncertain. Here, we report a case of a patient with NSCLC who developed leptomeningeal seeding (LMS) after the failure of first‐line therapy and showed a marked clinical and radiologic response to amivantamab monotherapy.

PurposeThe combination of radiotherapy with immunotherapy holds synergistic potential, yet its role in the neoadjuvant treatment of breast cancer remains underexplored. This single-center, retrospective pilot study aimed to explore the preliminary efficacy and safety of integrating stereotactic body radiotherapy (SBRT) with chemoimmunotherapy as a novel neoadjuvant regimen for a small cohort of patients with high-risk, locally advanced breast cancer.Patients and MethodsBetween June 2023 and August 2025, 20 patients received neoadjuvant SBRT (18Gy/3 fractions for node-positive; 24Gy/3 fractions for node-negative disease) concurrently with chemoimmunotherapy (various anti-PD-1/bispecific antibodies plus chemotherapy), followed by surgery. Key endpoints were pathological complete response (pCR, ypT0/Tis ypN0), near-pCR (Residual Cancer Burden [RCB] class 0 or I), objective response rate (ORR), and safety.ResultsThe cohort included triple-negative breast cancer (TNBC, 35%) and hormone receptor-positive/HER2-negative breast cancer of the Luminal B subtype (Luminal B/HER2-, 50%). Most had cT2 (75%) and node-positive (75%) disease. The median number of chemoimmunotherapy cycles were 8, with 80% receiving an AC-T backbone. The overall pCR rate was 45% (9/20). Efficacy varied by subtype: the pCR rate was 85.7% (6/7) in TNBC and 20% (2/10) in Luminal B/HER2- disease. Notably, 100% of TNBC and 40% of Luminal B/HER2- patients achieved near-pCR (RCB 0/I). ORR was 90%. Grade 3–4 adverse events occurred in 25% of patients, with no treatment-related mortality.ConclusionIn this single-center, retrospective pilot study, early integration of SBRT with chemoimmunotherapy demonstrated promising antitumor activity, particularly high pCR rates in TNBC and induction of deep pathological responses (RCB 0/I) in Luminal B/HER2- disease, with a manageable safety profile. Observed in a heterogeneous cohort, these preliminary findings highlight potential efficacy but require cautious interpretation and warrant validation in larger, prospective trials.

Autoimmune hemolytic anemia: New frontiers in diagnosis and therapy.

The gut microbiota plays a critical role in regulating immune homeostasis and modulating responses to cancer immunotherapies. However, the impact of antibiotic-induced dysbiosis in patients with multiple myeloma (MM) treated with bispecific antibodies (BsAbs) remains unexplored. This multicenter, international study investigated whether antibiotic exposure prior to BsAb initiation alters the gut microbiome and affects clinical outcomes in patients with relapsed or refractory MM. We retrospectively analyzed 237 adult patients with MM treated with CD3-engaging BsAbs across six academic institutions. Antibiotic exposure was defined as the administration of any broad-spectrum, non-prophylactic antibiotic within 30 days before BsAb initiation. Clinical outcomes included overall survival (OS), progression-free survival (PFS), and cumulative incidence of relapse, evaluated using Kaplan-Meier estimates, log-rank tests, and multivariable Cox and competing-risk regression models. Additionally, in a subset of 24 patients, peripheral blood samples were collected prior to BsAb infusion for immunophenotyping, cytokine profiling, and serum short-chain fatty acid (SCFA) quantification, while stool samples for 16S ribosomal RNA (rRNA) sequencing were collected in a subset of 19 patients. Broad-spectrum antibiotic exposure prior to BsAb therapy was associated with significantly inferior 1-year OS (60% (95% CI 44% to 81%) vs 77% (95% CI 71% to 83%), p=0.004) and PFS (26% (95% CI 14% to 47%) vs 53% (95% CI 46% to 61%), p<0.001), and higher relapse incidence (68% (95% CI 48% to 82%) vs 43% (95% CI 36% to 50%), p=0.004). In multivariable analyses, antibiotic exposure remained independently associated with poorer OS, PFS, and higher relapse risk. These associations were also observed within the subgroup of patients treated with CD3/B-cell maturation antibody-targeted BsAbs (n=155). Immunoprofiling revealed lower CD4+ T-cell counts (p=0.017) and reduced circulating cytokine levels among antibiotic-exposed patients. 16S rRNA sequencing demonstrated a marked depletion of SCFA-producing genera, including Roseburia and Eubacterium, accompanied by lower serum SCFA concentrations. Moreover, microbiota composition before BsAb treatment correlated with therapy response and treatment-related toxicity. Antibiotic-induced dysbiosis prior to BsAb therapy is associated with impaired immune reconstitution and inferior clinical outcomes in MM. These findings underscore the importance of antibiotic stewardship and suggest that microbiota-preserving strategies could enhance the efficacy of BsAb therapy in MM.

Bispecific antibodies in solid tumors: An Italian Association of Medical Oncology (AIOM) multidisciplinary perspective on immunology and vaccination.

Pharmacokinetic modeling to optimize development and therapeutic potential of mAb-based therapies.