Labonia MCI, Martinez de Castilla PE, van der Kraak PH
… +8 more, Brans MAD, Yang Q, Lei Z, de Jager SCA, de Voogt WS, Schiffelers RM, Sluijter JPG, Vader P
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42181694
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The efficiency and safety of modified messenger RNA (modRNA) delivery into the heart using lipid nanoparticles (LNPs) remain undetermined. We previously demonstrated that modRNA encapsulated in C12-200 LNPs outperforms c...The efficiency and safety of modified messenger RNA (modRNA) delivery into the heart using lipid nanoparticles (LNPs) remain undetermined. We previously demonstrated that modRNA encapsulated in C12-200 LNPs outperforms current state-of-the-art intramyocardial modRNA delivery methods. Surprisingly, C12-200 LNPs triggered robust local immune cell activation 5 days post-injection, which was not evident on day 1. To investigate whether this immune response is driven by LNP composition or modRNA transfection efficiency, we systematically compared cardiac transfection efficiency, off-target biodistribution, and immunogenicity of modRNA formulated with clinically validated LNPs from Onpattro, BNT162b2/Comirnaty, and mRNA-1273/Spikevax. All tested formulations outperformed C12-200 in cardiac delivery, with mRNA-1273 and Onpattro showing markedly reduced off-target accumulation in the liver and spleen. Histopathological analysis revealed formulation-dependent immune cell infiltration, most pronounced with C12-200. C12-200 significantly elevated cytokine levels in both serum and heart tissue, whereas Onpattro increased cytokine levels mainly locally. In contrast, cytokine levels in animals treated with BNT162b2 and mRNA-1273 were comparable to those in PBS-treated controls. Importantly, cardiac transfection efficiency did not correlate with cytokine induction or histopathological changes, indicating that LNP-driven immunogenicity is independent of transfection efficacy. These findings provide a foundation for refining LNP formulations to optimize cardiac modRNA delivery while minimizing immune-related adverse effects.
Xu H, Wan MCT, Wong CCC
… +4 more, Qin S, Wen Y, Wang J, Chen Z
Mol Ther Oncol
· 2026 Jun · PMID 42179971
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Nicotinamide mononucleotide (NMN) supplementation has shown clinical benefits by regulating metabolic activities in the energy production process. Its protective effect and underlying immune regulatory mechanisms against...Nicotinamide mononucleotide (NMN) supplementation has shown clinical benefits by regulating metabolic activities in the energy production process. Its protective effect and underlying immune regulatory mechanisms against tumor progression are still poorly understood. Here, we found that the high-dose NMN treatment could alter the level of several key NAD metabolic enzymes in human immune cells. Moreover, high-dose NMN treatment exhibited comparable antitumor efficacy as the PD-1 blockade in the murine mesothelioma challenge model. Subsequent immune profiling in both secondary lymphoid organ and tumor demonstrated that, rather than modulating T cell and NK cell responses, high-dose NMN treatment could reset tumor-associated macrophages toward the inflammatory M1-like phenotype compared with PD-1 blockade or non-treated subjects. These results provided a better understanding of NMN's regulatory effect on immune cells and suggested an alternative strategy of cancer immunotherapy.
Mooney R, Ngai G, Kaur G
… +17 more, Kang EY, Lara J, Abd El-Fattah EE, Huang P, Flores L, McDonald M, Talley M, Hammad M, Laidlaw M, Alamillo A, Nguyen DH, Lesniak MS, Curiel DT, Kortylewski M, Santidrian AF, Dellinger T, Aboody KS
Mol Ther Oncol
· 2026 Jun · PMID 42179969
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Ovarian cancer (OVCA) is an aggressive and often recurrent malignancy with limited long-term responses to standard chemotherapy. To address the urgent need for novel, targeted therapies, we evaluated the efficacy of a tu...Ovarian cancer (OVCA) is an aggressive and often recurrent malignancy with limited long-term responses to standard chemotherapy. To address the urgent need for novel, targeted therapies, we evaluated the efficacy of a tumor-tropic neural stem cell (NSC) platform delivering a conditionally replicative adenovirus (CRAd), NSC.CRAd-S-pk7, in preclinical models of advanced OVCA. A single intraperitoneal dose of NSC.CRAd-S-pk7 delivering 3 × 10 infectious units (IFUs) significantly reduced tumor burden and increased median survival by 33% (from 57 to 76 days) in immunocompetent mice, and 48% (from 54 to 80 days) in immunodeficient mice bearing OVCA peritoneal metastases. No observed adverse effects occurred at or below this dose. Optimization of the NSC transduction protocol enabled at least a 10-fold increase in CRAd-S-pk7 viral payload per cell, reducing the therapeutic cell dose by more than an order of magnitude-from 60 million to just 1 million NSCs. Repeated dosing further decreased tumor burden and increased median survival by 60% (from 57 to 91 days) in immunocompetent mice, suggesting a contribution from innate immune activation. These findings establish NSC.CRAd-S-pk7 as a promising oncolytic viro-immunotherapy treatment for advanced OVCA and support its advancement toward clinical translation.
Safarnezhad Tameshkel F, Sadat Kalaki N, Karimi E
… +6 more, Razizadeh MH, Haidar AEH, Amiri A, Zaki M, Nikkhah M, Karbalaie Niya MH
Mol Ther Oncol
· 2026 Jun · PMID 42179967
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Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality worldwide; molecular biomarkers that reflect hepatitis B virus (HBV)/hepatitis C virus (HCV)-associated tumor biology and predict prognosis or therape...Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality worldwide; molecular biomarkers that reflect hepatitis B virus (HBV)/hepatitis C virus (HCV)-associated tumor biology and predict prognosis or therapeutic vulnerabilities are needed. The research is a secondary analysis of public data (secondary data analysis) based on Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) data to identify differentially expressed genes (DEGs) common to HBV-HCC, HCV-HCC, and non-viral HCC datasets. Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, protein-protein interaction (PPI) network analysis, and hub-gene selection were then performed. Differential expression, prognostic association, immune-infiltration, and drug-sensitivity correlations were also analyzed. Eighty common DEGs (73 upregulated, 7 downregulated) were identified. PPI topological analysis yielded 53 hub genes; five genes and , were prioritized and validated as significantly upregulated in liver HCC and associated with poorer overall survival. Immune deconvolution showed consistent positive correlations between the hub genes and B cells and regulatory T cell subsets, and negative correlations with mucosal-associated invariant T (MAIT) cells, macrophages, and natural killer (NK)/monocyte signatures. Drug-gene correlation analysis revealed positive associations of and with sensitivity to mitogen-activated protein kinase inhibitors and negative correlations with several targeted inhibitors. The identified prognostically unfavorable hub genes in HBV/HCV-associated HCC are associated with an immunosuppressive microenvironment and with patterns of drug sensitivity.
Mol Ther Oncol
· 2026 Jun · PMID 42170678
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Carcinoma of the gallbladder (CAGB) carries a poor prognosis. While alterations in the bile microbiome and lipidome have been linked to CAGB development, the contribution of the fungal microbiome remains unexplored. We i...Carcinoma of the gallbladder (CAGB) carries a poor prognosis. While alterations in the bile microbiome and lipidome have been linked to CAGB development, the contribution of the fungal microbiome remains unexplored. We investigated fungal microbiome alterations and identified key fungal peptides capable of segregating CAGB patients. Bile samples from gallstone (GS) patients ( = 10), CAGB patients ( = 16), and healthy controls ( = 16) underwent fungal peptide-based diversity analysis and metabolomic profiling. Findings were cross-validated in plasma, correlated with clinical parameters and analyzed using machine learning. Six phyla and 24 fungal species were differentially regulated ( < 0.05). Alpha/beta diversity was higher in CAGB compared to GS and controls ( < 0.05). Ninety-three fungal peptides were upregulated and 63 downregulated in CAGB ( < 0.05, fold change [FC] > 1.5). CAGB patients showed significant enrichment of (log2FC > 12.21), (FC > 11.25), Saccharomyces (FC > 10.89), (FC > 10.68), and (FC > 10.38). Fungal-metabolite correlations (r > 0.5, < 0.05) linked these taxa to lysine biosynthesis, taurine and hypotaurine metabolism, fatty acid metabolism in bile, and cysteine/methionine, ascorbate, and purine metabolism in plasma. Fungal peptide panel achieved 96% diagnostic efficiency for mortality prediction with>90% accuracy, sensitivity, and specificity. Bile fungal diversity correlates with CAGB development and identifies fungal peptide panel capable of segregation of CAGB patients.
McClellan BL, Peña Agudelo JA, Mujeeb AA
… +22 more, Dabaja AA, Zhu Z, Raghuram S, Varela ML, Tronrud C, Banerjee K, Wei A, Calatroni C, Zhang LH, Romero LC, Oh P, Alghamri MS, Robbins A, Perricone M, Wang Y, Shay B, Sajjakulnukit P, Lyssiotis CA, Welch JD, Schwendeman A, Lowenstein PR, Castro MG
Mol Ther Oncol
· 2026 Jun · PMID 42170677
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Gliomas account for ∼80% of primary malignant brain tumors. Many CNS WHO grade 2-3 and some grade 4 gliomas harbor mutant isocitrate dehydrogenase 1 (mIDH1), which causes a gain-of-function mutation (IDH1 R132H) leading...Gliomas account for ∼80% of primary malignant brain tumors. Many CNS WHO grade 2-3 and some grade 4 gliomas harbor mutant isocitrate dehydrogenase 1 (mIDH1), which causes a gain-of-function mutation (IDH1 R132H) leading to the production of 2-hydroxyglutarate (2HG). Mutant IDH1-induced 2HG, through epigenetic reprogramming elicits an immune-permissive tumor microenvironment (TME). An immunosuppressive mechanism in the glioma TME involves adenosine production via the ectoenzyme CD73. This study investigates mIDH1's influence on CD73 expression and adenosine levels. We demonstrate that mIDH1 glioma cells exhibit reduced CD73 expression, driven by DNA hypermethylation, leading to reduced adenosine levels. Since wtIDH1 gliomas have high CD73 expression, we evaluated CD73 blockade as an immunotherapy target. We show that CD73 inhibition used as monotherapy did not improve survival in wtIDH1 glioma-bearing mice. However, when combined with immune-stimulatory Ad-TK (adenoviral vectors encoding herpes simplex virus thymidine kinase) and Ad-Flt3L (adenoviral vectors encoding FMS-like tyrosine kinase 3 ligand) gene therapy, CD73 blockade significantly enhanced therapeutic efficacy and increased anti-glioma effector T cell activity. These findings reveal that CD73 inhibition used in combination with immune-stimulatory Ad-TK/Ad-Flt3L gene therapy may be an effective treatment for wtIDH1 gliomas, which could be readily translated to the clinical arena.
Rossi J, Martinello C, Sorrentino R
… +5 more, Guyonnet-Duperat V, Amintas S, Abrey Recalde MJ, Verhoeyen E, Dabernat S
Mol Ther Oncol
· 2026 Jun · PMID 42170676
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Lentiviral vectors (LVs) are emerging as versatile tools for the efficient and stable delivery of therapeutic genes. Although VSV-G-pseudotyped LVs remain the standard for genetic engineering, their broad tropism and se...Lentiviral vectors (LVs) are emerging as versatile tools for the efficient and stable delivery of therapeutic genes. Although VSV-G-pseudotyped LVs remain the standard for genetic engineering, their broad tropism and serum sensitivity limit their applicability for cancer therapy, where precise targeting is essential for efficacy and safety. This review synthesizes two decades of advances in LV pseudotyping, comparing natural viral envelopes, engineered targeting strategies, and multicomponent glycoprotein systems that reshape vector tropism. We discussed donor-derived envelopes such as measles virus, baboon endogenous retrovirus, Nipah virus, and Sindbis virus, highlighting their value for transducing hematopoietic, immune, and tumor cells. We also examined next-generation retargeting innovations, including engineered receptor-binding domains, display of cytokines or antibody fragments, and VSV-G mutants that ablate natural receptor interactions, to enhance specificity for T cells, B cells, hematopoietic stem cells, and tumor-associated antigens. Finally, we review the recent emergence of engineered virus-like particles (VLPs) as precision tools for delivery of CRISPR-Cas9, base-editing, and prime-editing complexes. Collectively, these advances position receptor-targeted lentiviral and VLP systems as promising platforms for the next generation of precision oncology, enabling selective CAR T/NK cell generation, targeted tumor modification, and cell-restricted gene editing.
Massumi M, Li G, Owji H
… +3 more, Yang G, Girda E, Hatefi A
Mol Ther Oncol
· 2026 Jun · PMID 42164422
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Cancer stem-like cells (CSCs) drive ovarian cancer metastasis, therapeutic resistance, and relapse. This study aimed to develop a combination therapeutic strategy capable of eliminating both rapidly proliferating ovarian...Cancer stem-like cells (CSCs) drive ovarian cancer metastasis, therapeutic resistance, and relapse. This study aimed to develop a combination therapeutic strategy capable of eliminating both rapidly proliferating ovarian cancer cells and drug-resistant CSCs, thereby eradicating metastatic disease and preventing relapse. We engineered an adipose-derived mesenchymal stem cell clone (ASC-shCE2:yCD) that homes to tumors and locally converts the prodrugs irinotecan and 5-fluorocytosine (5-FC) into the cytotoxic agents SN38 and 5-fluorouracil (5-FU). This approach was combined with natural killer (NK) cell immunotherapy to eradicate CSCs that survived chemotherapy. Using patient-derived, drug-resistant metastatic ovarian cancer models, we show that exposure to SN38 or 5-FU upregulates NKG2D stress ligands (MICA/B) on CSCs, enhancing their susceptibility to NK-mediated cytotoxicity. Real-time imaging demonstrated rapid homing of engineered adipose-derived stem cells (ASCs) to tumor sites within 3 days. In triple-immunodeficient CIEA NOG mice, ASC-directed enzyme/prodrug therapy followed by NK cell immunotherapy eliminated metastatic ovarian tumors and prevented relapse during the monitoring period. Histopathological and hematological analyses revealed no clinically significant toxicity. Collectively, these findings establish a stem cell-directed chemoimmunotherapy that primes drug-resistant ovarian cancer cells for immune elimination, offering a rationale and effective strategy to safely eradicate metastatic disease and prevent recurrence.
Shea EN, Olafson HR, Muscato DR
… +15 more, Adams LM, Valero MC, McKee KK, Shen X, Dougherty PG, Hicks AN, Liu N, Kheirabadi M, Streeter M, Li X, Mon P, Girgenrath M, Sethuraman N, Qian Z, Wang ET
Steric-blocking antisense oligonucleotides rescue myotonic dystrophy type 1 phenotypes in preclinical models and are under evaluation in clinical trials. However, the rationale for biomarker selection remains a topic of...Steric-blocking antisense oligonucleotides rescue myotonic dystrophy type 1 phenotypes in preclinical models and are under evaluation in clinical trials. However, the rationale for biomarker selection remains a topic of debate. Here, we show that a cyclic cell-penetrating peptide that escapes endosomes enhances muscle delivery of a phosphorodiamidate morpholino oligonucleotide designed to block pathogenic CUG repeat expansions in HSA mice. A single systemic administration rescued mis-splicing and eliminated myotonia 1 week post-injection, with partial splicing rescue evident after 24 h. Interestingly, some exons showed more robust rescue than others, but the relationship between muscleblind-like (MBNL) protein concentration ([MBNL]) and percent spliced in (Ψ) did not fully explain the extent of rescue. We hypothesized that since pre-existing transcripts must be degraded to reveal the full drug effect, rates of transcript replacement might account for these discrepancies. We formulated a mathematical framework and used Bayesian inference to model how apparent Ψ lags behind nascent Ψ as a function of time; faster rates of replacement result in shorter lags. In vivo 5-ethynyl uridine labeling followed by RNA sequencing (RNA-seq) validated these predictions. Overall, we show that transcript turnover influences Ψ during periods of dynamically changing [MBNL] and recommend considering this when selecting splicing biomarkers and interpreting responses to therapeutic interventions.
The local cell-extrinsic and cell-intrinsic roles of endogenous interleukin-33 (IL-33) in tumor progression and metastasis have been controversial, which has delayed scrutiny of the systemic application of IL-33 in tumor...The local cell-extrinsic and cell-intrinsic roles of endogenous interleukin-33 (IL-33) in tumor progression and metastasis have been controversial, which has delayed scrutiny of the systemic application of IL-33 in tumor immunoprevention and immunotherapy. A prominent concern regards its capacity in stabilizing the immunosuppressive phenotype of regulatory T (T) cells. Here, we report that systemic IL-33 treatment potently promotes the effects of prophylactic tumor vaccines and inhibits the progression of mouse solid tumors. Mechanistically, systemic IL-33 treatment reshapes the tumor immune microenvironment, including the increase in suppressor of tumorigenicity 2 (ST2) T cells. Unexpectedly, systemic IL-33 treatment constrains tumor growth in a T ST2-dependent manner, although the endogenous IL-33/ST2 axis in T cells promotes tumor growth. Indeed, IL-33 reprograms T cells toward a cytotoxic and inflammatory phenotype. Lastly, we fail to show that endogenous IL-33 promotes tumor growth. Thus, our findings elucidate a T ST2-dependent mechanism of the anti-tumor effect of systemic IL-33 application and support the use of IL-33 for cancer immunoprevention and immunotherapy.
TILT-123 (Ad5/3-E2F-d24-hTNFα-IRES-hIL2, igrelimogene litadenorepvec) is a chimeric oncolytic adenovirus engineered to selectively replicate in tumor cells and express tumor necrosis factor (TNF) and interleukin-2 (IL-2)...TILT-123 (Ad5/3-E2F-d24-hTNFα-IRES-hIL2, igrelimogene litadenorepvec) is a chimeric oncolytic adenovirus engineered to selectively replicate in tumor cells and express tumor necrosis factor (TNF) and interleukin-2 (IL-2). While oncolytic viruses are commonly administered intratumorally, this approach often restricts practical applicability. Intravenous delivery is less invasive and offers a more practical alternative; however, systemic immune barriers challenge effective tumor targeting. In a cohort of a phase 1 clinical trial, six patients with advanced solid tumors who had exhausted standard treatments received two intravenous doses of TILT-123 (split-dose), per treatment day across seven cycles. Safety was the primary objective, while efficacy outcomes were exploratory and assessed using positron emission tomography/-computed tomography (PET/CT) imaging, along with overall survival. Split-dosing was safe, with chills, fever, fatigue, and lymphocyte reduction being the most common treatment-related events. In imaging, the disease control rate was 83.3% according to PET criteria and 33.3% by RECIST 1.1. The median overall survival was 198 days. Split-dosing increased the area under the curve of TILT-123 and amplified systemic cytokine responses. Proteomic analysis of serum and neutralizing antibody profiling indicated that enhanced humoral immune activity was associated with shorter overall survival. Post-treatment biopsies confirmed virus presence and alterations in immune cell composition. These findings support further evaluation of intravenous TILT-123 administration in clinical trials.
Branched-chain 2-ketoacid dehydrogenase (BCKDH) deficiency (maple syrup urine disease; MSUD) causes lethal encephalopathy by disrupting cerebral metabolism, a process imperfectly reflected by circulating biomarkers. Diet...Branched-chain 2-ketoacid dehydrogenase (BCKDH) deficiency (maple syrup urine disease; MSUD) causes lethal encephalopathy by disrupting cerebral metabolism, a process imperfectly reflected by circulating biomarkers. Diet and liver transplantation stabilize peripheral metabolites but fail to restore brain neurochemistry, demarcating the central nervous system as the decisive therapeutic compartment. To define the pathogenesis of intoxication and its therapeutic response, we performed paired serum-brain metabolomics in Bckdha-/- mice treated with a systemic AAV9 dual-gene vector encoding human BCKDHA and BCKDHB (A-BiP-B). Untreated neonates exhibited a 9-fold elevation of brain 2-ketoisocaproate accompanied by cerebral depletion of glutamate and glutamine, as well as shifts in tricarboxylic acid cycle and ketone body metabolism. These disturbances originated from reversal of branched-chain aminotransferase 2 flux and destabilization of glutamate-2-ketoglutarate mass balance, producing divergent metabolic endophenotypes in blood versus brain. A single intravenous injection of A-BiP-B rescued mice from fatal encephalopathy, partially restored cerebral BCKDHA mRNA expression, and brought core brain neurochemical endpoints within wild-type range despite persistent elevation of serum 2-ketoacids. These findings expose limitations of current MSUD management and establish systemic dual-gene therapy as a means of restoring neurochemical homeostasis while enabling survival on unrestricted protein intake.
Molecules on the cell membrane dictate how cells interact with their environment and other cells and serve as the target for tumor immunotherapy. Enzyme-catalyzed proximity labeling has become a pivotal tool for studying...Molecules on the cell membrane dictate how cells interact with their environment and other cells and serve as the target for tumor immunotherapy. Enzyme-catalyzed proximity labeling has become a pivotal tool for studying intercellular communication and functionalizing cell membranes. To enable more precise and efficient enzymatic labeling for cell membrane engineering, further investigation of novel labeling strategies is crucial. In this study, we developed a precise chemoenzymatic labeling strategy using SrtA-SpA, mediated by antibodies (Ab-SS), as a general method for detecting cell-cell interactions and constructing antibody-cell conjugates (ACCs) for tumor immunotherapy. Firstly, we successfully employed the Ab-SS strategy to label specific cell populations within complex cellular environments. Furthermore, this method can be precisely extended to label cell groups that interact with the target cells. Employing the Ab-SS system, we report a potent ACC platform as an alternative to chimeric antigen receptor technology. This platform precisely equips immune cells with tumor-targeting antibodies to direct their recognition and attack against cancer cells. Collectively, given its flexibility and robust functionality, the Ab-SS enzymatic labeling system provides a generalized strategy for facilitating research on cellular function and advancing targeted immune cell therapies through proximity labeling.
While genome-wide association studies have linked the human PLAC9 gene to body mass index, its physiological function remains largely unexplored. This study identifies PLAC9 as a novel adipokine that is enriched in the s...While genome-wide association studies have linked the human PLAC9 gene to body mass index, its physiological function remains largely unexplored. This study identifies PLAC9 as a novel adipokine that is enriched in the stromal vascular fraction of adipose tissue. Its circulating levels correlate with key metabolic dysregulation markers in humans and mice. We utilized gain- and loss-of-function approaches in diet-induced obesity (DIO) and streptozotocin (STZ)-induced diabetic mouse models to demonstrate that PLAC9 is a critical regulator of systemic metabolism. Notably, knockdown of endogenous PLAC9 exacerbated metabolic impairments, while its overexpression significantly mitigated DIO-associated metabolic dysregulation. Additionally, recombinant PLAC9 protein administration alleviated hyperglycemia in insulin-resistant and insulin-deficient models. Mechanistically, PLAC9 potentiated calcium-dependent insulin secretion in pancreatic beta cells, promoted glucose uptake in the liver and skeletal muscle, and upregulated hepatic Ghr and Igf1 levels to facilitate glucose homeostasis. Based on these hormone-like properties, we propose renaming the protein Placin. Collectively, these findings establish Placin as a promising therapeutic target, offering translational potential for the management of both type 2 and type 1 diabetes.