Mol Ther Nucleic Acids
· 2026 Jun · PMID 42239496
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Decades of research have cemented antisense oligonucleotides (ASOs) as a cornerstone of molecular medicine. Advancements in synthesis and chemical modification, together with an improved understanding of the human genome...Decades of research have cemented antisense oligonucleotides (ASOs) as a cornerstone of molecular medicine. Advancements in synthesis and chemical modification, together with an improved understanding of the human genome and transcriptome, have enabled their emergence as highly specific and tailorable therapeutics across a wide variety of conditions. Although the greatest strength of ASOs lies in their nucleic acid composition that confers the theoretical ability to target any known genetic sequence, effective ASO design requires a holistic approach considering the molecular mechanism underlying the desired therapeutic outcome. Initially considered straightforward inhibitors of gene expression, ASOs have now evolved into versatile modulators capable of exploiting an increasingly diverse array of molecular processes. Despite this progress, their full therapeutic potential remains far from realized. Emerging research, driven by a deepening understanding of RNA biology, continues to expand the repertoire of mechanisms through which ASOs can modulate gene expression. Collectively, these studies demonstrate that ASOs can be designed to modulate numerous pre-mRNA processing events, including splicing, polyadenylation, microRNA activity, and translation initiation and termination, thereby broadening the range of conditions and patients that may benefit from ASO-based therapeutics.
Chen B, Tian Y, Liang L
… +6 more, Ding P, Yin R, Xu R, Wang T, Chen H, Lu D
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42239495
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Canonical base editors (BEs) enable programmable nucleotide conversions without inducing DNA double-strand breaks (DSBs), yet the significant off-target (OT) effects restrict their applications. The transformer BE (tBE),...Canonical base editors (BEs) enable programmable nucleotide conversions without inducing DNA double-strand breaks (DSBs), yet the significant off-target (OT) effects restrict their applications. The transformer BE (tBE), a BE derivative, minimizes DNA and RNA OT mutations, but its dual single-guide RNA (sgRNAs) requirement limits its targeting scope. To address this, we integrated SpRY, a protospacer-adjacent motif (PAM)-relaxed Cas9 variant, into tBE. The resulting tBE-SpRY achieved robust C-to-T editing across various cell lines in a PAM-unlimited manner, with minimal OT effects at the tested genomic loci and negligible transcriptome-wide RNA editing. We further demonstrated its versatility and . Microinjection of tBE-SpRY components into zygotes efficiently generated albino C57BL/6J mice, while dual adeno-associated virus (AAV)-mediated delivery in mice resulted in modest C-to-T modifications (up to 2.96%) in ocular tissues. Thus, tBE-SpRY provides a safe and PAM-flexible platform for generating genetic mouse models and exploring potential therapeutic applications.
Ahn S, Oh TS, Suh S
… +22 more, Jeon JY, Lah S, Ryu K, Choi S, Kim H, Lee EG, Lee H, Lee J, Kim Y, Kim S, Park SR, Yoo H, Choi SY, Lee JE, Kim DK, Ku BM, Jung W, Ahn MJ, Jung JU, Kim YS, Oh BH, Jeong BS
Antibodies that recognize peptide-loaded class I major histocompatibility complex (MHC) molecules could enable therapeutic targeting of intracellular oncogenic proteins, yet their discovery has been hampered by the small...Antibodies that recognize peptide-loaded class I major histocompatibility complex (MHC) molecules could enable therapeutic targeting of intracellular oncogenic proteins, yet their discovery has been hampered by the small size of peptide antigens and the need for allele-specific recognition. Here, we describe an integrated in silico-in vitro workflow for generating high-affinity, selective antibodies to Kirsten rat sarcoma viral oncogene homolog (KRAS) G12D presented by human leukocyte antigen (HLA)-C∗08:02, a clinically validated cancer neoantigen. In the in silico stage, multiple human antibody-derived variable fragments optimally docked to the target were generated, followed by limited sequence design of complementarity-determining regions (CDRs). In the in vitro stage, limited CDR diversity was introduced to construct a yeast surface-displayed library, which was subjected to iterative selection. This workflow yielded KRAS G12D/HLA-C∗08:02-specific antibodies with high affinity. Comprehensive specificity profiling using a phage display library confirmed the absence of human off-target reactivity, and in silico analysis predicted low immunogenicity. When reformatted as chimeric antigen receptors or bispecific T cell engagers, these antibodies mediated selective cytotoxicity against target-positive cells. Together, these findings establish a practical design-to-function pipeline for T cell receptor (TCR)-like antibody discovery and demonstrate the feasibility of therapeutically targeting KRAS G12D-driven malignancies.
Pathogenic variants in splicing factors are the second most common cause of autosomal dominant retinitis pigmentosa, with mutations in PRPF31 being the most prevalent. Here, we characterize a novel intronic variant in PR...Pathogenic variants in splicing factors are the second most common cause of autosomal dominant retinitis pigmentosa, with mutations in PRPF31 being the most prevalent. Here, we characterize a novel intronic variant in PRPF31 (c.1074-11C>G) that creates a cryptic 3' splice site, resulting in an aberrantly spliced transcript predicted to encode a protein with an altered C terminus. However, the pathogenic protein is unstable and undetectable in patient-derived induced pluripotent stem cells (iPSCs). In addition, expression of the full-length PRPF31 protein was reduced in patient-derived retinal pigment epithelium (RPE). To correct the splicing defect, we designed a panel of antisense oligonucleotides (ASOs) targeting putative RNA-binding sites in exon 10 and intron 10 and identified a candidate that corrects PRPF31 splicing in a minigene reporter system as well as in patient-derived iPSCs and RPE. We further showed that ASO treatment enhances PRPF31 protein expression in patient-derived iPSC and RPE carrying the intronic mutation, supporting the potential of the ASO-based approach to restore PRPF31 expression in patients with the same or similar splicing defects.
Metastasis causes 90% of cancer-related deaths, yet preventive strategies remain limited. Here, we report that a single dose of therapeutic Salmonella, a prominent anti-tumor bacterial therapy, provides long-lasting prot...Metastasis causes 90% of cancer-related deaths, yet preventive strategies remain limited. Here, we report that a single dose of therapeutic Salmonella, a prominent anti-tumor bacterial therapy, provides long-lasting protection against metastasis in mice by inducing "trained" natural killer (NK) cells. Integrated functional and multi-omics analyses revealed that Salmonella-trained NK (stNK) cells establish an enduring reprogrammed epigenome characterized by enhanced pro-survival signaling and immune effector functions, resulting in more potent IFN-γ release and cytotoxicity upon secondary stimulation. We further showed that this training requires a transient pulse of IL-12 combined with sustained IL-18 signaling. Crucially, stNK cells significantly outperform conventional immune checkpoint therapies, including PD-1 and TIGIT blockade, in preventing metastasis, underscoring the unique immunological mechanisms in combating metastasis. These findings highlight the potential of bacterial cancer vaccines or trained NK cells as effective strategies for metastasis prevention.
Sarko LE, Givand D, Rattin B
… +14 more, Shepley C, Tommasi A, Attar A, Taylor R, Kutler B, Traynor RM, Upadhyaya A, Mnuk M, Gehrke C, Murren N, Ulland TK, Capitini CM, Kotanchek T, Saha K
Cell therapy manufacturing of primary T cells often results in heterogeneous cell populations in the final product, with many cells lacking desired receptor expression or exhausted and other dysfunctional phenotypes. Her...Cell therapy manufacturing of primary T cells often results in heterogeneous cell populations in the final product, with many cells lacking desired receptor expression or exhausted and other dysfunctional phenotypes. Here, we design a novel cell-intrinsic strategy to genetically reprogram primary human T cells to autonomously detect and eliminate dysfunctional cells. This integrated detection and elimination process, known as directed fratricide, is programmed via non-viral CRISPR genome editing to eliminate the T cell receptor (TCR) alpha chain (TRAC gene knockout) and integrate a chimeric antigen receptor (CAR) against the urokinase-type plasminogen activator receptor (uPAR), also known as CD87. In these cell products, strong T cell stimulation or activation during manufacturing causes a small subset of cells to express uPAR, which triggers CAR-mediated killing within the product. This fratricide induces proliferation in the desired cells and destroys undesired cells, a process that could be modeled computationally and controlled robustly via supplements to the culture media. The strategy enabled enrichment of anti-uPAR and anti-disialoganglioside (GD2) CAR-T cell products up to ≥99% CAR+/TCR-, favoring a memory-like phenotype. Understanding growth dynamics among T cell subsets and reprogramming them via CRISPR could accelerate the biomanufacturing of potent cell products without extensive selection methods.
Chen Q, Sedloev D, Yao H
… +12 more, Mao L, Depke D, Wang L, Scheller M, He B, Liu Y, Unglaub JM, Schmitt A, Jayavelu AK, Müller-Tidow C, Schmitt M, Sauer T
Chimeric antigen receptor T (CART) cell therapy has demonstrated clinical efficacy in hematologic malignancies; however, primary or secondary treatment failure remains a major obstacle to durable responses. Defining the...Chimeric antigen receptor T (CART) cell therapy has demonstrated clinical efficacy in hematologic malignancies; however, primary or secondary treatment failure remains a major obstacle to durable responses. Defining the optimal cellular composition of CART products therefore represents a critical unmet need. Here, we show that CARTs targeting acute leukemias achieve maximal efficacy when composed exclusively of CD4 T cells. Pure CD4 CARTs exhibit superior anti-tumor activity and proliferation compared with CD8-containing CART products. To elucidate the molecular basis of this functional divergence, we applied a combinatorial exploratory approach integrating bulk RNA sequencing and quantitative proteomics. Transcriptomic analyses revealed that pure CD4 CARTs adopt a highly proliferative state characterized by a cytotoxic effector-like polarization. On the protein level, we are demonstrating coordinated loss of cell-cycle machinery and induction of apoptotic pathways in CD4 CART cells co-cultured together with CD8 CART cells, while pure CD4 CART cultures maintained a proliferative, cytotoxic phenotype. Using mechanistically discriminative co-culture systems, we demonstrate that CD8 CART-mediated impairment of CD4 CART functionality is primarily driven by competitive access to shared antigen. Collectively, these findings identify antigen competition between CART subsets as a previously unrecognized mechanism limiting CD4 CART efficacy and provide a framework for optimizing CART product composition to enhance therapeutic persistence and durability.
Targeted protein degradation (TPD) has reshaped modern pharmacology by shifting therapeutic strategies from occupancy-driven inhibition toward event-driven elimination of disease-causing proteins, including targets long...Targeted protein degradation (TPD) has reshaped modern pharmacology by shifting therapeutic strategies from occupancy-driven inhibition toward event-driven elimination of disease-causing proteins, including targets long regarded as undruggable. Beyond canonical modalities such as proteolysis-targeting chimeras and molecular glues, the TPD landscape has rapidly expanded to encompass diverse molecular scaffolds-including small molecules, antibodies, peptides, and nucleic acids-as well as multiple degradation pathways spanning the ubiquitin-proteasome system, lysosomal routes, and autophagy. This growing mechanistic and structural diversity has created an urgent need for an integrative framework to rationalize platform selection and translational potential. Here, we introduce a two-dimensional classification matrix that organizes TPD strategies according to degrader scaffold and degradation mechanism, revealing mechanistic "white spaces" that may represent underexplored opportunities for future TPD development. We further propose the TPDS analytical framework-encompassing target class broadening, platform refinement, degradation mechanism reprogramming, and scaffold expansion-to systematically map over 30 emerging TPD platforms. Finally, we discuss areas of clinical maturity in TPD, with representative examples from hematologic, hormone-driven, and kinase-driven malignancies, as well as key translational bottlenecks and emerging solutions, ranging from delivery engineering to AI-assisted degrader design. Together, these insights provide a strategic roadmap for the rational development of next-generation TPD therapeutics in precision oncology.
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42233109
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Messenger RNA (mRNA) vaccines using lipid nanoparticles (LNPs) are globally approved with acceptable safety profiles for preventing respiratory diseases. Here, we describe tissue distribution and kinetics of mRNA-LNP vac...Messenger RNA (mRNA) vaccines using lipid nanoparticles (LNPs) are globally approved with acceptable safety profiles for preventing respiratory diseases. Here, we describe tissue distribution and kinetics of mRNA-LNP vaccines after intramuscular dosing using three products formulated with same LNP matrix: mRNA-1273 (Spikevax), mRNA-1647 (a candidate cytomegalovirus [CMV] vaccine), and a reporter mRNA (nascent peptide-luciferase) drug product. Consistent biodistribution patterns were observed: tissues with highest exposures were the injection site, draining lymph nodes, and spleen, with minimal distribution to other organs. Levels of vaccine components declined rapidly in circulation and tissues, with model-based projections indicating concentrations at low or below quantifiable levels by ∼2 weeks. Unbound (free) spike protein was transient after mRNA-1273 vaccination (modeled decline < 5 days) and did not accumulate with repeated dosing. The ionizable lipid in the LNP, lipid H, underwent biotransformation and was excreted renally and hepatically, with no human-specific metabolites. Collectively, these results indicate similar biodistribution patterns across products within this dataset using a shared LNP composition, despite different mRNA cargo. Furthermore, in a SARS-CoV-2 infection-free model, no sustained spike protein was observed. Overall, the data establish a framework that can be used to scientifically justify leveraging data across a shared platform.
Splendiani E, Maiorano G, Spinello Z
… +23 more, Vecchiotti D, Gugliuzza G, Moretti M, Asquino A, Izzo M, Po A, Feverati N, Raia T, Besharat ZM, Verzella D, Mardente S, Zicari A, Ciccarella G, Citarella A, Zazzeroni F, De Smaele E, Venneri MA, Mastronuzzi A, Gigli G, Angelucci A, Catanzaro G, Palamà IE, Ferretti E
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42233108
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MicroRNAs play a crucial role in tumorigenesis, functioning as oncogenes or tumor-suppressors while orchestrating the differentiation states of cancer cells. In medulloblastoma, the most common malignant pediatric brain...MicroRNAs play a crucial role in tumorigenesis, functioning as oncogenes or tumor-suppressors while orchestrating the differentiation states of cancer cells. In medulloblastoma, the most common malignant pediatric brain tumor, the downregulation of miR-326 is associated with a loss of tumor suppression that leads to uncontrolled proliferation and survival or de-differentiation. In this study, we explore an innovative approach for microRNA delivery using polycaprolactone/chitosan-based nanoparticles. We evaluate the biocompatibility of the nanocarrier in medulloblastoma cell lines and report results on nanoparticles administration, demonstrating minimal toxicity in the most exposed organs. findings reveal that nanoparticles-mediated miR-326 delivery significantly reduces stemness markers (Nanog and Sox2) and promotes neuronal differentiation, as evidenced by increased β3-tubulin expression. These results highlight polycaprolactone-based nanoparticles as a promising platform for miR-326 delivery, establishing a foundation for future investigations on therapeutic strategies in medulloblastoma.
de Klerk JA, Slieker RC, Beulens JWJ
… +7 more, Peerlings JHD, Mei H, Elders PJM, van Zonneveld AJ, van Raalte DH, Bijkerk R, 't Hart LM
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42233107
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Chronic kidney disease (CKD) is a common complication of type 2 diabetes, characterized by reduced kidney function and/or albuminuria, yet its progression varies widely among individuals. While sodium-glucose cotransport...Chronic kidney disease (CKD) is a common complication of type 2 diabetes, characterized by reduced kidney function and/or albuminuria, yet its progression varies widely among individuals. While sodium-glucose cotransporter-2 (SGLT2) inhibitors are known to protect against kidney decline, the molecular mechanisms underlying their renoprotective effects remain incompletely understood. Circulating small non-coding RNAs (sncRNAs), particularly microRNAs, have been linked to CKD but the contribution of other sncRNA classes is less explored. We profiled plasma sncRNAs in 263 participants with type 2 diabetes from the Hoorn DCS cohort without CKD at baseline, followed for ∼9 years ( = 122, = 141). sncRNA profiling was also performed before and after treatment of SGLT2 inhibitors in three trials ( = 65, total) to assess drug-induced molecular changes in the circulation. Eleven sncRNAs were nominally associated with incident CKD, most strongly and . In an independent SGLT2 inhibitor trial, exploratory analyses identified 34 sncRNAs that changed following treatment, including . Preliminary analyses linked these snoRNAs to co-regulated proteins, suggesting potential functional relevance. Our results identify snoRNAs, particularly , as potential novel markers of CKD risk and SGLT2 inhibitor response in type 2 diabetes, revealing an unexplored sncRNA axis and providing a foundation for future functional studies into their mechanistic role in CKD progression.
Zhang Y, Xiong N, Song Y
… +3 more, Lu X, Ni K, Tang Z
Mol Ther Oncol
· 2026 Jun · PMID 42232064
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R-loops, three-stranded nucleic acid structures comprising an RNA-DNA hybrid and a displaced single-stranded DNA strand, play context-dependent roles in cancer-serving essential physiological functions while also driving...R-loops, three-stranded nucleic acid structures comprising an RNA-DNA hybrid and a displaced single-stranded DNA strand, play context-dependent roles in cancer-serving essential physiological functions while also driving tumorigenesis when dysregulated. Their pathological effects are mediated through replication stress, genomic instability, transcriptional disruption, and defective RNA processing. This review highlights the emerging potential of targeting R-loops as a therapeutic strategy in oncology. We survey advanced methodologies for R-loop mapping, addressing technical limitations of current approaches, and advocate for multi-omics integration to elucidate R-loop dynamics and functional networks in cancers such as lung, bladder, and prostate malignancies. Critically, we explore two promising therapeutic avenues: (1) direct inhibition of R-loop resolvers to trigger excessive R-loop accumulation and replication catastrophe, and (2) synthetic lethality strategies that capitalize on cancer-specific R-loop handling defects. Clinical evidence supporting these approaches is discussed, along with challenges including tumor heterogeneity, detection limitations, and adaptive resistance. We argue that a multi-omics-driven understanding of R-loop biology will accelerate the translation of R-loop-directed therapies into precision oncology.
Brunialti E, Panzeri A, Rizzi N
… +17 more, Villa A, Meda C, Sfogliarini C, Persano S, Arlati F, Guevara Lopez ML, Martelli C, Cannavale G, Rebecchi M, Di Vito C, Venturini L, Ottobrini L, Mavilio D, Fagerholm S, Minucci S, Vegeto E, Ciana P
Immunization engages a highly coordinated series of innate and adaptive immune responses across multiple anatomical sites, a complexity that has contributed to the predominantly empirical nature of vaccine development. T...Immunization engages a highly coordinated series of innate and adaptive immune responses across multiple anatomical sites, a complexity that has contributed to the predominantly empirical nature of vaccine development. To address this limitation, we developed and validated a STAT6 reporter mouse that enables dynamic whole-body imaging and ex vivo characterization of STAT6-mediated anti-inflammatory signaling. We then integrated this model with an established NF-κB reporter system to dissect the immune activation triggered by two lipid nanoparticle (LNP)-formulated mRNA vaccines differing in RNA chemistry (unmodified versus N-methylpseudouridine-modified). The dual-reporter approach enabled the construction of a spatiotemporal atlas of vaccine-induced signaling, revealing distinct immune dynamics driven by RNA chemistry and identifying the liver as an early hub for both NF-κB and STAT6 activity following systemic administration. Correlation with serological data showed that early STAT6 activation followed by rapid signal resolution was associated with favorable humoral responses. These findings establish NF-κB and STAT6 reporter mice as rapid in vivo screening tools for early evaluation of vaccine immune potential and highlight their ability to inform more mechanistic and rational design of mRNA vaccine platforms, while emphasizing the role of the liver both as a primary LNP target and as an immunologically relevant organ.
Jimenez V, Sacristan V, Garcia M
… +15 more, Jambrina C, Casana E, Muñoz S, Vilà L, Grass I, Jaén ML, Roca C, León X, Marcó S, Molas M, Ribera A, Elias I, Rodó J, Ferré T, Bosch F
The decline of organ function during aging limits health span. Despite the potential of lifestyle interventions to improve health, sustained maintenance of health span is challenging, and no gerotherapeutic drugs have be...The decline of organ function during aging limits health span. Despite the potential of lifestyle interventions to improve health, sustained maintenance of health span is challenging, and no gerotherapeutic drugs have been approved. Here, we demonstrated that aged and geriatric male and female mice treated with muscle-directed adeno-associated viral (AAV) vector-mediated fibroblast growth factor 21 (FGF21) gene therapy extended health span and lifespan with sustained organ benefits. This treatment normalized body weight and adiposity, improved insulin sensitivity and glucose homeostasis, preserved hepatic detoxification capacity, counteracted age-related kidney disease, promoted cardiac health and muscular function, and enhanced cognition. Transcriptomic and histopathological analyses indicated improved whole-body energy homeostasis and cellular fitness, which were mediated by tissue-specific adaptations, including enhanced mitochondrial function, restored proteostasis, and reversion of inflammation, fibrosis, and amyloidosis. AAV-FGF21 treatment also activated AMPK signaling. These results highlight FGF21 gene therapy as a potential strategy to promote health span and delay age-related deterioration.
Chimeric antigen receptor (CAR) efficacy depends on the presence of surface antigen. While antigen levels in leukemias can be measured in the blood or bone marrow, expression in solid tumors is often inferred from archiv...Chimeric antigen receptor (CAR) efficacy depends on the presence of surface antigen. While antigen levels in leukemias can be measured in the blood or bone marrow, expression in solid tumors is often inferred from archival tissues or remains undetermined. Neuroblastoma (NB) is an aggressive pediatric solid tumor for which we have co-developed a CAR targeting glypican 2 (GPC2). Here, we developed a radiotheranostic platform comprising an antibody-based positron emission tomography (immunoPET) agent, [Zr]Zr-DFO-mCT3 (Zr-mCT3), for non-invasive detection of GPC2 and [Ac]Ac-macropa-mCT3 (Ac-mCT3) for alpha-particle therapy. In orthotopic NB models with variable GPC2 levels, immunoPET accurately detected tumors with positive versus low/negative GPC2 status, which correlated with in vivo response to CAR. As a therapeutic agent, Ac-mCT3 induced tumor regression in a dose-dependent manner, with the maximal effect at 2.96 kBq (80 nCi). Because Ac-mCT3 does not downregulate GPC2, we assessed GPC2-CAR therapy with Ac-mCT3 (1.48 kBq [40 nCi]) and showed improved survival in mice with GPC2 but not GPC2 tumors. These findings establish Zr-mCT3 as a novel radiotracer for GPC2 monitoring and a predictive tool for CAR responses. Ac-mCT3 enhances GPC2-CAR therapy in tumors with medium antigen levels, offering a rationale for clinical development in NB and other pediatric tumors.
Enhancing the efficacy of adoptively transferred T cells is essential for successful immunotherapy. The cytotoxic activity of these T cells is closely related to their mitochondrial function, which plays a critical role...Enhancing the efficacy of adoptively transferred T cells is essential for successful immunotherapy. The cytotoxic activity of these T cells is closely related to their mitochondrial function, which plays a critical role in T cell differentiation. This study explored the potential of nicotinamide riboside (NR), known to enhance mitochondrial function, to improve the efficacy of adoptively transferred CD8 T cells against hepatocellular carcinoma (HCC). In subcutaneous and metastatic HCC tumor models, NR treatment significantly improved the effectiveness of adoptive T cell therapy (ACT). NR promoted the differentiation of CD44CD62L central memory T cells (T) and upregulated memory-associated genes (Tcf7, Ccr7, and Foxo1). Upon restimulation, NR-treated CD8 T cells exhibited strong recall responses, as evidenced by metabolic reprogramming and increased cytokine production. Furthermore, RNA sequencing revealed an increased expression of Foxo1 and its target genes. Inhibition of Foxo1 partially reversed the beneficial effects of NR on mitochondrial fitness and cytotoxic function, suggesting a role for Foxo1-associated pathways in mediating these effects. In summary, NR improved mitochondrial fitness and promoted T differentiation, in part through a transcriptional program associated with Foxo1, thereby improving the efficacy of ACT. These findings identify NR as a promising and translatable metabolic adjuvant for HCC immunotherapy.
Hereditary tyrosinemia type 1 (HT1) is a life-threatening metabolic disorder caused by the toxic accumulation of tyrosine and its metabolites. While treatment with 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedion...Hereditary tyrosinemia type 1 (HT1) is a life-threatening metabolic disorder caused by the toxic accumulation of tyrosine and its metabolites. While treatment with 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) combined with a strict dietary regimen has improved outcomes, it imposes a significant lifelong burden and is associated with debilitating side effects and incomplete protection. Here, we developed an engineered probiotic with an optimized design, e-EcN-HT, and demonstrated its comprehensive efficacy and safety in HT1 across multiple animal models, including fumarylacetoacetate hydrolase (FAH) mice, FAH rabbits, and Bama minipigs. Our findings indicate that e-EcN-HT not only mitigates multifaceted acute manifestations of FAH mice, including neonatal death and acute liver injury, but also improves chronic liver lesions when combined with NTBC. The therapeutic effect translated successfully to the FAH rabbit model. Moreover, e-EcN-HT administration led to rapid metabolism of orally administered C-tyrosine, confirming robust and active tyrosine consumption in pigs. Comprehensive safety assessments across murine and porcine models showed that e-EcN-HT was well tolerated, with no significant adverse effects, systemic dissemination, or detrimental disruption to the resident gut microbiota. Collectively, our multi-species preclinical data underscore the potential of engineered bacteria as a viable therapeutic strategy for HT1 and possibly other metabolic disorders.
Jiang Q, Yang AY, Mo GL
… +5 more, Mo LG, Deng T, Huang SN, Hou ST, Huang QR
Mol Ther Oncol
· 2026 Jun · PMID 42222348
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Lower-grade glioma (LGG) is a highly heterogeneous disease, making accurate prognosis prediction and the development of precise, personalized treatment plans for patients challenging. The multi-omics data from patients w...Lower-grade glioma (LGG) is a highly heterogeneous disease, making accurate prognosis prediction and the development of precise, personalized treatment plans for patients challenging. The multi-omics data from patients with LGG were analyzed using 10 clustering algorithms to identify subgroups at very high resolution. Ten machine learning algorithms were subsequently integrated to develop a robust artificial intelligence model (AIM). We identified two cancer subtypes (CSs) linked to prognosis through multi-omics clustering. After screening 46 hub genes, we integrated 10 machine learning algorithms into 117 combinations to select the AIM with the highest average C-index. The gradient boosting machine (GBM)-based AIM outperformed previous prognostic signatures in nearly all cohorts. Patients in the low-AIM group had a better prognosis and greater sensitivity to immunotherapy, whereas those in the high-AIM group had a poorer prognosis and lower immunotherapy sensitivity. However, MG132 showed promise as a potential therapeutic agent. studies confirmed the oncogenic role of the hub gene in LGG cells; its knockdown reduced the proliferation, invasion, and migration of these cells. In summary, the AIM we developed is highly valuable for predicting the prognosis of patients with LGG and identifying those who are sensitive to immunotherapy.
Slattery S, Huo W, Arif A
… +2 more, Gordon J, Takeuchi R
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42221546
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Gene editing technologies have opened the possibility of directly targeting viral DNA for therapeutic applications. In chronically infected hepatocytes with hepatitis B virus (HBV), covalently closed circular DNA (cccDNA...Gene editing technologies have opened the possibility of directly targeting viral DNA for therapeutic applications. In chronically infected hepatocytes with hepatitis B virus (HBV), covalently closed circular DNA (cccDNA) serves as the master template for viral transcripts and gene products. In the present study, we evaluated anti-HBV multiplex gene editing with the CRISPR-Cas9 endonuclease from (SaCas9) using primary human hepatocytes (PHHs) and HBV mouse models. Nonviral delivery of SaCas9-encoding mRNA and a pair of HBV-targeting guide RNAs (gRNAs) substantially reduced viral biomarkers and intrahepatic HBV DNA copies and . Hybridization capture sequencing analyses showed that small insertions and deletions (indels) and structural variants including excisions and inversions of the viral sequences were introduced in the residual HBV DNA. These assays also demonstrated that transient expression of the HBV-targeting SaCas9 significantly suppressed random integration of HBV DNA into the host genome, while leading to no detectable increase in chromosomal translocations involving viral copies. Lastly, our gene editing approach blocked viral rebound after stopping treatment with a nucleos(t)ide analogue (NA), entecavir. Our results suggest that anti-HBV multiplex gene editing removes viral DNA from chronically infected hepatocytes, potentially reducing the risk of hepatocarcinogenesis associated with HBV DNA integration.