Abstract LB067: A modular Multi-Antigen T Cell Hybridizers (MATCH) platform enables tunable and potent immunotherapy across hematologic and solid tumors

Cancer Research · 2026-04-17

Abstract T cell-engaging immunotherapies have demonstrated remarkable efficacy in cancer treatment but remain constrained by antigen heterogeneity, immune escape, and dose-limiting toxicities. Here, we present Multi-Antigen T Cell Hybridizers (MATCH), a modular immune-engager platform that decouples tumor recognition from immune activation through programmable molecular hybridization, enabling tunable multi-antigen targeting without permanent multispecific antibody engineering. For multiple myeloma (MM) and acute myeloid leukemia (AML), we developed BCMA- and CD33-directed MATCH systems consisting of tumor-targeting antibody fragments conjugated to a 25-base morpholino oligonucleotide MORF1 (Fab’BCMA-MORF1 or Fab’CD33-MORF1) and a complementary CD3-engaging T cell module conjugated to complementary MORF2 (Fab’CD3-MORF2). Hybridization between MORF1 and MORF2 brings tumor cells and T cells into close proximity, facilitating efficient immune synapse formation. Both BCMA and CD33 MATCH induced potent, antigen-dependent cytotoxicity against multiple cancer cell lines and primary patient samples at nanomolar concentrations. Importantly, because the ratio of tumor-targeting and immune-engaging components can be independently adjusted, cytokine release was precisely controlled through sequential versus premixed administration and dose modulation, demonstrating improved tunability compared with conventional bispecific T cell engagers. Cytokine secretion followed a regulated temporal sequence, including IL-2, TNF-α, IFN-γ, and IL-6. In a humanized NRG MM model, T cell dosing was optimized to control graft-versus-host effects, and MATCH treatment significantly prolonged survival compared with untreated controls. Efficacy was also demonstrated in solid tumors, including breast and lung cancers, with PD-L1-targeted MATCH treatment. Beyond CD3-mediated T cell redirection, MATCH enables modular incorporation of costimulatory signaling via Fab’CD28-MORF2. The combined PD-L1/CD3/CD28 trispecific MATCH configuration significantly enhanced T cell activation and tumor cell killing compared with CD3 engagement alone, achieving up to a 20-fold reduction of IC50 values, particularly at low antigen density and immunosuppressive conditions characteristic of solid tumors. Activation of tumor cell apoptotic pathways and T cell activation signaling pathways was confirmed. Collectively, these findings establish MATCH as a versatile immune-engager platform capable of addressing both hematologic and solid tumors through programmable antigen targeting and modular immune activation. Citation Format: Shannuo Li, Yingduo Yang, Joseph Shami, Douglas Sborov, Paul Shami, Jiyuan Yang, Jindřich Kopeček. A modular Multi-Antigen T Cell Hybridizers (MATCH) platform enables tunable and potent immunotherapy across hematologic and solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(8_Suppl):Abstract nr LB067.

Self‐Assembling Multi‐Antigen T Cell Hybridizers for Precision Immunotherapy of Multiple Myeloma

Advanced Healthcare Materials · 2025-08-01 · 3 citations

Abstract Bispecific T‐cell engagers show promise in treating multiple myeloma (MM), but challenges remain in adaptability and targeting flexibility. This paper presents a novel T‐cell based immunotherapy, M ulti‐ A ntigen TC ell H ybridizers (MATCH), a modular, self‐assembling T‐cell engager designed for versatile and patient‐specific cancer targeting. MATCH consists of two components: a B‐cell‐targeting Fab’ fragment conjugated to a 25‐base morpholino oligonucleotide (Fab’ B cell antigen ‐MORF1) and a T‐cell engaging anti‐CD3 Fab’ fragment conjugated to the complementary morpholino oligonucleotide (Fab’ CD3 ‐MORF2). Upon hybridization of MORF1 and MORF2, MATCH enables pre‐targeting of malignant cells followed by in situ post‐assembly of the bispecific complex, facilitating targeted T‐cell recruitment. To enhance antigen specificity based on MM patient expression profile, a panel of Fab’‐MORF1 conjugates targeting key MM surface markers (Fab’ BCMA ‐MORF1, Fab’ SLAMF7 ‐MORF1, Fab’ CD38 ‐MORF1) is developed, which pairs interchangeably with Fab’ CD3 ‐MORF2 for T‐cell engagement. MATCH effectively induces immune synapse formation and exhibits potent, antigen‐specific cytotoxicity across MM cells. Ex vivo validation in patient‐derived bone marrow samples confirms significant tumor cell depletion. Preliminary in vivo studies in humanized mouse model demonstrated effective cancer inhibition along with favorable pharmacokinetics and distribution profiles. These findings support MATCH as a flexible and customizable immunotherapy platform with strong translational potential for the treatment of MM.

Immune cell tumor engagers (SITE) via morpholino-guided self-assembly for precision immunotherapy of multiple myeloma

Blood · 2025-11-03

Abstract Introduction: Bispecific T cell engagers have shown significant clinical efficacy and are currently FDA approved for the treatment of relapsed and refractory multiple myeloma (RRMM). However, challenges remain, including targeting flexibility, immune-related toxicity, and durability of response. To overcome these limitations, we developed Self-Assembled Immune Cell-Tumor Engagers (SITE), a two-component system that targets both immune cell and MM cell surface antigens; each component is tagged with a complementary morpholino oligonucleotide (M1 or M2). Upon administration, these components hybridize in situ via M1-M2 pairing to form bispecific complexes to redirect immune cells to MM cells. Utilization of this pre-targeting approach enables the sequential engagement of immune cells, including T cells, NK cells, and macrophages. Additionally, by simultaneously targeting of multiple antigens (i.e. BCMA, GPRC5D and CD38), this strategy minimizes the risk of antigen loss and immune escape. Herein, we report the efficacy of SITE across multiple models, including in vitro cell lines, ex vivo patient-derived samples, and in vivo mouse models, demonstrating its potential as an innovative, cost-effective platform for orchestrating a coordinated and robust anti-tumor immune response. Methods: Anti-MM antibodies were digested enzymatically to F(ab')2 fragments, further reduced with tris(2-carboxyethyl)phosphine to generate Fab'MM-thiols. A pair of 25 bp M1/M2 was customized from Genetools. Following 3'-end maleimido modification, a panel of Fab'MM-M1 targeting BCMA, GPRC5D and CD38 was obtained. Similarly, α-hCD3 and α-hCD314 were used to generate immune cell targeting Fab'immune-M2. Stability of M1-M2 hybrids was assessed using circular dichroism (CD) spectroscopy in PBS, and size-exclusion chromatography (SEC) following incubation in mouse serum. Primary human T cells and NK cells were isolated from healthy donor blood. In vitro cytotoxicity and immune activation of SITE were assessed by flow cytometry on MM.1S and RPMI-8226 cells. To assess therapeutic efficacy, both patient-derived bone marrow mononuclear cells and a preclinical NRG mouse model were utilized. Mice were sub-lethally irradiated and i.v. injected with 3×106 MM.1S-Luc cells. Human T cells (9×106,3×106, or1×106) were administered to study the influence of T cells. Different dosing regimens (premix vs. consecutive) of T cell-specific SITE constructs (Fab'BCMA-M1/Fab'CD3-M2) were evaluated and compared with conventional teclistamab treatment. Cytokine production (IL-2, IFN-γ, TNF-α) was analyzed, and immune cell subsets were profiled by flow cytometry. Results: Conjugates Fab'MM-M1 and Fab'immune-M2 were successfully synthesized; hybridization of Fab'MM-M1/Fab'immune-M2 upon 1:1 mixing was confirmed by a shift in elution profiles in SEC. Complexes remained stable in 10% mouse serum for over 24 h. CD spectra showed a positive peak at 260 nm and a negative minimum at 210 nm, indicating formation of A-form double helices under physiological conditions. In vitro, SITE constructs successfully engaged T cells and NK cells, triggering potent cytotoxicity towards MM.1S and RPMI-8226 cells. Flow cytometry demonstrated increased expression of activation markers and induction of apoptosis in target cells. Ex vivo studies using patient-derived samples confirmed significant clearance of MM cells by patient's own T cells. In vivo, NRG mice bearing MM.1S-Luc and treated with SITE-T cell therapy extended survival and inhibited tumor growth compared to teclistamab-treated groups. Flow cytometric analysis of bone marrow confirmed a marked reduction in MM.1S cell populations in SITE-treated mice. T cell-related toxicity was found to be associated with injected T-cell numbers: reducing the dose from 9×106 to 3×106 or lower, no body weight loss occurred, highlighting the advantageous flexibility of two-component SITE. Overall, SITE demonstrated a superior anti-tumor effect compared to teclistamab, highlighting its potent efficacy in controlling MM progression in vivo. Conclusion: The SITE platform represents a promising advance in immunotherapy for MM, offering a versatile, potent, and modular approach to address the disease's complex antigenic landscape. By enabling multi-antigen targeting and the orchestration of diverse immune effectors, the platform effectively overcomes challenges such as antigenic heterogeneity, immune escape, and cytokine release syndrome.

PD-L1 targeted antibody-polymer-Epirubicin conjugate prolongs survival in a preclinical murine model of advanced ovarian cancer

Journal of Controlled Release · 2025-04-04 · 7 citations

T cell activation contributed to a more effective antitumor response. Repeated dosing amplified immunomodulatory effects, leading to durable immunity. These results highlight U6244-051 as a next-generation pADC with high translational potential, offering enhanced efficacy and reduced on-target, off-tumor toxicity.

CD38-targeted antibody-polymer drug conjugates for enhanced treatment of multiple myeloma

Biomaterials · 2025-06-04 · 8 citations

Multiple myeloma (MM) remains a formidable disease, especially in relapsed or refractory cases when there are limited treatment options. In this study, we introduce two polymer-antibody drug conjugates (pADCs), ISA-P-EPI (U6244-021) and DARA-P-EPI (U6244-031), which contain semitelechelic N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-epirubicin (EPI) conjugate attached to CD38-targeting antibodies Isatuximab (ISA) and Daratumumab (DARA). These pADCs enhance therapeutic efficacy by combining the specificity of ISA and DARA with the cytotoxic potency of EPI while preserving antibody function. The EPI is linked to the HPMA polymer backbone via a tetrapeptide spacer cleavable by lysosomal enzymes, enabling drug release upon endocytosis within tumor cells. This design achieves a higher drug-to-antibody ratio than conventional ADCs for safer delivery of drug payload. In vitro studies demonstrate efficient binding, internalization, and cytotoxic efficacy of these pADCs in MM cell lines. Mechanistic investigations revealed significant therapeutic effects, including cell cycle arrest, immunogenic cell death, and preserved antibody-dependent cellular cytotoxicity (ADCC). In addition, pADCs were effective in 5 out of 8 primary samples, with their efficacy closely correlating with CD38 surface expression levels. To enhance therapeutic outcomes, we employed panobinostat to upregulate CD38 expression, which further improved pADC efficacy. In a preclinical NRG mouse model inoculated with MM.1S-luc cells, pADC treatment significantly delayed tumor progression and prolonged survival, with all treated mice remaining alive at the 100-day endpoint. These findings underscore the potential of CD38-targeted pADCs as a novel approach to combining chemotherapy with immunotherapy for MM treatment, warranting further investigation into their optimization and clinical application.

Comparative analysis of the uptake of the H+/OC antiporter substrate oxycodone across and into brain endothelial and parenchymal cells with in vitro–in vivo extrapolation

Fluids and Barriers of the CNS · 2025-10-30 · 1 citations

In vitro evaluation of substances utilizing the putative proton‑coupled organic cation (H+/OC) antiporter for active uptake across the blood-brain barrier (BBB) and brain cell membranes requires a thorough understanding of cellular pharmacokinetics supported by reliable translational readouts. This study assessed the rate and extent of uptake of the antiporter substrate oxycodone in brain endothelial and parenchymal cells at clinically relevant concentrations, exploring the suitability of various cell models for investigating active drug transport. Transcellular transport studies were performed using primary brain endothelial cells (BECs) from pig, rat, and mouse, alongside uptake assays in immortalized human cerebral microvascular endothelial cells (hCMEC/D3) and rat brain slices. Drug uptake was estimated by combining transport data with non-specific binding data via equilibrium dialysis. The effect of interleukin-6 (IL-6) on oxycodone uptake was tested in hCMEC/D3 cells. The unbound intracellular-to-extracellular concentration ratio (Kp,uu,cell) and efflux ratio were used to compare the extent of net uptake across models and evaluate the presence of active uptake. Based on cellular pharmacokinetic parameters, both primary BECs and hCMEC/D3 demonstrated active uptake of oxycodone. Mean permeability across primary BECs ranged between 0.9 × 10⁻⁵ (pig) and 1.8 × 10⁻⁵ (rat) cm/s. Transport extent, reflected by 1/Efflux Ratio values of 1.5 (pig) and 2.4 (rat), aligned with in vivo unbound brain-to-plasma concentration ratios, Kp,uu,brain, indicating predominant active uptake. In hCMEC/D3 cells, the uptake rate was time- and concentration-dependent within 60 min and 10–5000 nM concentration range, while the uptake extent, assessed by Kp,uu,cell, was 1.7 and independent of both. IL-6 increased the extent of uptake to 148% of control, without affecting the rate. The extent of uptake into parenchymal cells was also concentration-independent, with Kp,uu,cell values around 1.8, similar to endothelial cells. These findings provide insight into oxycodone distribution across the BBB and into brain parenchymal cells, particularly emphasizing the contribution of probable active transport mechanisms at multiple barriers. The results highlight the importance of assessing both the rate and extent of transport and support the utility of Kp,uu,cell as a key metric for comparing drug transport across models. Both primary BECs and hCMEC/D3 cells are reliable tools for assessing substrate uptake in drug development, particularly for H+/OC antiporter substrates, and hold translational potential for mechanistic studies.

Spotlighting rising researcher Shannuo Li: MATCH: a flexible and safer approach to T cell–based immunotherapy for multiple myeloma

AAPS Open · 2025-08-18

Multiantigen T-Cell Hybridizers: A Two-Component T-Cell-Activating Therapy

ACS Nano · 2024 · 4 citations

• Cancer research
• Biology
• Medicine

model, insights regarding optimal T-cell-to-target cell ratio were gleaned when a ratio of 5:1 T-cell-to-target cell MATCH-treated mice significantly delayed the onset of disease compared to higher and lower ratios.

Region-independent active CNS net uptake of marketed H+/OC antiporter system substrates

Frontiers in Cellular Neuroscience · 2024 · 1 citations

• Pharmacology
• Chemistry
• Neuroscience

, values consistently exceeding unity in all assessed regions. Despite minor regional differences, no significant distinctions were found when comparing the whole brain to investigated regions of interest, indicating region-independent active transport. Furthermore, we observed intracellular accumulation via lysosomal trapping for all studied drugs. These results provide new insights into the CNS regional neuropharmacokinetics of these drugs, suggesting that while the brain uptake is region-independent, the active transport mechanism enables high extracellular and intracellular drug concentrations, potentially contributing to neurotoxicity. This finding emphasizes the necessity of thorough neuropharmacokinetic evaluation and neurotoxicity profiling in the development of drugs that utilize this transport pathway.

Antioxidant Activity and Antitumor Potential of <i>Hedyotis chrysotricha</i>Extract in Human Colorectal Cancer and Breast Cancer Cell Lines

The Natural Products Journal · 2024-01-16

Background: Cancer is a serious disease severely endangering human health and life in the world in the world, and conventional anticancer drugs have frequently suffered from severe side effects and resistance. Therefore, developing novel and effective therapeutic strategies is urgently needed. As a Chinese herbal medicine with important medical values, Hedyotis chrysotricha has been suggested to be useful for the treatment of many diseases, especially cancer. Objective: Our aim was to investigate the antioxidant and antitumor activities of the phytochemical extracts of H. chrysotricha. Methods: 2,2-diphenyl-1-picrylhydrazyl free radical scavenging assay was applied to determine antioxidant property. 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay, wound healing assay, transwell matrix assay, flow cytometry and cell morphology were established to observe anti-proliferative, anti-migration, anti-invasion and apoptosis induction effects. The intracellular reactive oxygen species generation was detected by reactive oxygen species detection assay. Results: The ethyl acetate fraction showed promising antioxidant activity with IC50 of 98.67±0.49 μg/ml, and petroleum ether fraction exhibited the most potent antiproliferative effect on various human cancer cell lines, especially MDA-MB-231 (IC50 = 8.90±1.23 μg/ml) and HCT-116 (IC50 = 9.69±3.69 μg/ml) cancer cells. Further investigation revealed that the petroleum ether fraction suppressed the proliferation, migration and invasion of the cancer cells significantly. Additionally, it promoted cell apoptosis by increasing reactive oxygen species levels. Conclusion: H. chrysotricha extract possessed excellent antioxidant and antitumor activities. Therefore, it could be useful as a source of antioxidants and compounds for cancer therapy.