Chen S, Peng Y, Yan D
… +3 more, Lyu L, Scherngell T, Hu Y
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42211693
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Small interfering RNA (siRNA) represents a transformative modality in next-generation therapeutics, yet challenges in delivery efficiency still limit its clinical translation. Systematic evaluation tools are needed to re...Small interfering RNA (siRNA) represents a transformative modality in next-generation therapeutics, yet challenges in delivery efficiency still limit its clinical translation. Systematic evaluation tools are needed to reveal technological progress in this area. Given the innovation momentum inherent in patents, this study develops a patent-based evaluation model for siRNA delivery technologies from a technology management perspective. By integrating the Hierarchical Decision Model (HDM) with patent landscape analysis, the model incorporates 15 multidimensional criteria across technological, commercial, and legal perspectives, with the commercial perspective assigned the greatest weight (41%), followed by technical (32%) and legal (27%) perspectives. A large-scale dataset comprising 20,319 siRNA delivery-related patent documents was quantitatively assessed using this model. The results reveal that high-value patents are primarily concentrated in lipid-based carriers and ligand-siRNA conjugates, with firms such as Alnylam and Arbutus emerging as dominant innovation leaders. Furthermore, the analysis highlights that therapeutic indication, assignee's technological accumulation, technological impact, and drug delivery method are key drivers of patent value. Overall, the proposed model supports strategic decision-making in patent portfolio management and technology forecasting by identifying high-value innovations.
Sasset L, Cameron AD, Sussman C
… +8 more, Rubinelli L, Maji D, Miller R, Thompson A, Campbell D, Walker MR, Drozdz MM, Liberatore RA
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42211692
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Incretin receptor agonists (IRAs) have recently come to prominence as powerful weight and glucose control drugs and are approved for the treatment of type 2 diabetes (T2D) and obesity. Semaglutide and tirzepatide, curren...Incretin receptor agonists (IRAs) have recently come to prominence as powerful weight and glucose control drugs and are approved for the treatment of type 2 diabetes (T2D) and obesity. Semaglutide and tirzepatide, currently the most widely prescribed within this class, are both potent molecules but have a short half-life, requiring weekly dosing. This requirement can negatively affect quality of life and the adherence to therapy, as well as create a significant financial burden for patients who may have a life-long need for treatment. The MYO Technology platform was developed to overcome these barriers. It consists of therapeutic-encoding plasmid DNA, and a proprietary medical device for intramuscular injection and electroporation of muscle cells. This leads to the uptake of injected DNA, expression of the therapeutic, and its secretion into peripheral circulation. Here, we demonstrate that MYO Technology-delivered IRAs are efficacious in promoting long-lasting weight and glucose control in mouse models of diet-induced obesity. Moreover, engineering the IRAs to facilitate blood-brain barrier penetration further enhances treatment efficacy, with benefits persisting beyond 1 year following a single administration. Together, these findings highlight MYO Technology's potential to transform standard of care by enabling long-lasting therapeutic effects with minimal dosing.
Dolgin V, Eremenko E, Narkis G
… +3 more, Mazor-Oring M, Gradstein L, Birk OS
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42211691
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The development of antisense oligonucleotide (AON) therapy for monogenic diseases such as retinitis pigmentosa (RP) is effective but labor-intensive. Rapid, efficient technologies to identify optimal AONs are needed. We...The development of antisense oligonucleotide (AON) therapy for monogenic diseases such as retinitis pigmentosa (RP) is effective but labor-intensive. Rapid, efficient technologies to identify optimal AONs are needed. We present a novel approach focusing on intronic splicing mutations. We identified a novel heterozygous intronic non-canonical splice variant in causing dominant RP in a four-generation pedigree. Using a Dual Fluorescence Reporter Assay, we transfected HEK293T cells with a reporter plasmid containing either the wildtype intron (resulting in GFP fluorescence due to normal splicing) or the mutant intron (showing no fluorescence due to splicing defects). As an alternative to standard AONs, we used uridine-rich 7 (U7) small nuclear RNA (snRNA) derivatives with optimized Sm protein-binding site and a modified antisense sequence (U7 Sm OPT snRNA) as modulators of pre-mRNA splicing. Eight AONs were designed to target the aberrant transcript, to restore normal splicing and gene function. To evaluate treatment efficacy, HEK293T cells with the reporter GFP plasmid were co-transfected with U7snRNA cassettes, each containing an incorporated AON sequence. FACS analysis quantified splicing modulation and gene expression rescue, identifying the most effective AON. Our study broadens the genetic understanding of RP, highlighting the significance of personalized medicine in genetic disorder treatment.
Yang Y, Wang J, Yu H
… +17 more, Quan Z, Zhang Z, Guo J, Wu H, Liu S, Li X, Hu X, Zou H, Zhu M, Wang T, Zhou Z, Liu J, Wang J, Wang K, Yang L, Han Z, Sun X
The costimulatory receptor CD28 is indispensable for regulating T cell effector function, and the absence of co-stimulation is associated with poor immunotherapy response. However, longitudinal molecular imaging studies...The costimulatory receptor CD28 is indispensable for regulating T cell effector function, and the absence of co-stimulation is associated with poor immunotherapy response. However, longitudinal molecular imaging studies targeting CD28 as a costimulatory biomarker using positron emission tomography (PET) have not been reported. Here, we developed the co-stimulation-associated radiotracer [Zr]Zr-DFO-anti-CD28 to enable early prediction of therapeutic response by identifying recovery of T cell responsiveness following programmed cell death protein 1 (PD-1) blockade in lung cancer. [Zr]Zr-DFO-anti-CD28 demonstrated high specificity for CD28 tumor-infiltrating lymphocytes (TILs) in vitro and in vivo. By monitoring CD28 expression and its loss within the tumor microenvironment, [Zr]Zr-DFO-anti-CD28 PET identified responses to anti-PD-1 therapy earlier than changes in tumor volume observed at day 5, and increased radiotracer uptake correlated with more favorable treatment responses. Repeated PET imaging was performed to monitor dynamic changes in CD28 TILs induced by different immunotherapies at day 14, and [Zr]Zr-DFO-anti-CD28 PET captured the CD28-dependent restoration of T cell effector function, further supporting its potential for early prediction. Taken together, costimulatory biomarker-specific CD28 PET molecular imaging may enable noninvasive early prediction of responses to multiple immunotherapies and facilitate the application of precision medicine in lung cancer.
Bentayebi K, Louati S, Suwan K
… +2 more, Eljaoudi R, Hajitou A
Mol Ther Oncol
· 2026 Jun · PMID 42199604
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Medulloblastoma accounts for approximately 20% of all pediatric brain tumors. Standard treatment typically involves a multimodal approach including surgery, radiation therapy, and chemotherapy. Despite advances in molecu...Medulloblastoma accounts for approximately 20% of all pediatric brain tumors. Standard treatment typically involves a multimodal approach including surgery, radiation therapy, and chemotherapy. Despite advances in molecular classification and the development of targeted therapies, long-term survival and quality of life remain suboptimal, particularly for patients with high-risk subgroups. Although targeted therapies hold significant promise, their effective delivery to the brain remains a major challenge. The unique cerebellar microenvironment, including the blood-brain barrier, the blood-brain-tumor barrier, and the blood-cerebrospinal fluid barrier, restricts drug penetration and therapeutic efficacy. To overcome these limitations, innovative strategies are being explored to bypass or transiently modulate these barriers in a subgroup- and compartment-specific manner. These approaches include nanoparticle-based delivery systems, focused ultrasound, intrathecal administration, and rational combination therapies. Emerging platforms such as transmorphic phage/adeno-associated virus vectors and lysosomal targeting strategies are under active investigation and may further enhance therapeutic precision. This review provides a comprehensive overview of current brain-targeted drug delivery systems for medulloblastoma, highlighting barrier heterogeneity, recurrence patterns, and emerging technologies with the potential to improve therapeutic efficacy and long-term outcomes for children affected by this aggressive pediatric brain tumor.
The clinical potential of bispecific T cell engagers (BTEs) is limited by their short serum half-life and the complexity and cost of recombinant protein manufacturing. Currently, BTEs rely on external production and repe...The clinical potential of bispecific T cell engagers (BTEs) is limited by their short serum half-life and the complexity and cost of recombinant protein manufacturing. Currently, BTEs rely on external production and repeat dosing, limiting scalability and patient access. Here, we present a synDNA platform that enables in vivo assembly and long-term secretion of bispecific antibodies directly from host muscle. The system includes complementary "knob" and "hole" Fc chains on separate plasmids that pair only when co-expressed, ensuring controlled heterodimer formation and reducing homodimer assembly. Using ovarian cancer (OC) as a model of antigen heterogeneity, we generated dKBTEs targeting follicle stimulating hormone receptor (dK_FSHRxCD3) and Her2 (dK_Her2xCD3). Artificial intelligence simulations supported successful knob-hole pairing and presented significant challenges for homodimer formation. Co-transfected Expi293F cells secreted fully assembled bispecifics that bound antigen and CD3, activated primary human T cells, and induced potent antigen-specific cytotoxicity. OC patient peripheral blood mononuclear cells responded strongly to dKBTEs in vitro. In mice, dKBTEs showed superior pharmacokinetics and tumor control over first-generation BTEs. Combination therapy with both dKBTEs further improved tumor control. synDNA delivery allows simultaneous in vivo production of two dKBTEs without compromising expression. These findings establish synDNA delivery as a programmable approach for sustained delivery of complex biologics.
Zonari E, Naldini MM, Barcella M
… +21 more, Volpin M, Vincenti F, Desantis G, Hadadi L, Caserta C, Galasso I, Martini B, Midena F, Farina G, Vacca R, Tucci F, Ormoli L, Visigalli I, Vezzoli M, Lazarevic D, Merelli I, Xie SZ, Dick JE, Di Micco R, Montini E, Gentner B
Ex vivo expansion of mobilized peripheral blood (mPB) hematopoietic stem cells (HSCs) represents a promising approach to advance cell and gene therapy strategies yet is hampered by loss of stem cell function when applyin...Ex vivo expansion of mobilized peripheral blood (mPB) hematopoietic stem cells (HSCs) represents a promising approach to advance cell and gene therapy strategies yet is hampered by loss of stem cell function when applying commonly used culture protocols. We performed in-depth characterization of mPB expansion cultures by single-cell RNA sequencing, which highlighted differentiation trajectories with preservation of lineage fidelity in committed progenitors. Defining a putative HSC cluster allowed an estimation of transduction efficiency in ex vivo cultures, which correlated with long-term gene marking in xenografts and patients enrolled in a gene-therapy study. We then developed a clinically translatable, good manufacturing practice-compliant process to expand lentiviral (LV)-transduced HSCs from mPB of pediatric patients and adult donors by biologically informed protocol improvements of cytokine supplementation, media choice, timing of LV transduction, and combinations of small molecules preventing the activation of differentiation programs. Our optimized process outperforms validated state-of-the-art cord-blood expansion protocols when applied to mPB. LV integration site analysis and genomic barcode-based clonal tracking provided definitive proof for symmetric HSC self-renewal divisions occurring during ex vivo culture. These results warrant clinical testing of this HSC transduction/expansion process in an upcoming clinical gene-therapy trial for autosomal recessive osteopetrosis (EU CT 2024-518972-30).
Boespflug-Tanguy O, Valent A, Rambaud J
… +23 more, Léger PL, Plu I, Seilhean D, Frémeaux-Bacchi V, Levade T, Marinello M, Denard J, Derome M, Brun PL, Guerchet N, Veron P, Blatzer M, Fragnoud R, Prieto L, Corbel T, Knuchel-Legendre N, Beaurain B, Lelait M, Sanz M, Abergel K, Rivière A, Buj-Bello A, Perret G
Spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME) is an ultra-rare, fatal autosomal-recessive disorder caused by ASAH1 mutations, with no curative treatment. We report the first-in-human intravenous a...Spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME) is an ultra-rare, fatal autosomal-recessive disorder caused by ASAH1 mutations, with no curative treatment. We report the first-in-human intravenous administration of an AAV9 vector carrying the human ASAH1 coding sequence in a 15-year-old female with advanced SMA-PME (heterozygous ASAH1 c.456A>C and c.918-2A>G mutations). The patient presented with severe neuromuscular impairment, deafness, and recurrent myoclonic status epilepticus, without detectable anti-AAV9 neutralizing antibodies. A marked clinical deterioration warranted compassionate use (GNT-017-ASAH-CU). A dose of 2.2 × 10 vector genomes per kilogram of body weight was administered under prophylactic prednisolone and sirolimus. The patient experienced rapid onset of complement activation leading to cytokine-mediated capillary leak syndrome, culminating in refractory shock, multiorgan failure, and death on day 8. A postmortem examination revealed acute circulatory failure as the cause of death without myocarditis or thrombotic microangiopathy. Some endothelial injury was suggested by a rise in von Willebrand factor from days 4 to 8, paralleled by increased hyaluronic acid on days 7 and 8. These findings underscore the potential for life-threatening innate immune activation in patients with advanced SMA-PME receiving high-dose systemic AAV9 gene therapy, highlighting the need to identify high-risk patients and proactively monitor biomarkers of endothelial injury.
Base editing enables the precise, permanent modulation of gene expression, offering a promising therapeutic avenue for targeting lipid-regulating genes. Recently, asialoglycoprotein receptor 1 (ASGR1), a liver-specific r...Base editing enables the precise, permanent modulation of gene expression, offering a promising therapeutic avenue for targeting lipid-regulating genes. Recently, asialoglycoprotein receptor 1 (ASGR1), a liver-specific receptor, has emerged as a potential therapeutic target for controlling circulating lipid levels. Here we employed adeno-associated virus serotype 8 (AAV8) to deliver an all-in-one adenine base editor (ABE) to target Asgr1 in mice. Systemic administration of an Asgr1-targeting ABE achieved 54.7% ± 2.2% editing efficiency, leading to a marked depletion of hepatic ASGR1 and significant reductions in serum and hepatic total cholesterol (TC) and triglyceride (TG). These effects were mediated, in part, by the modulation of biliary cholesterol excretion pathways. No overt liver injury was observed following in vivo base editing of Asgr1. Moreover, combined treatment with Asgr1 base editing and the lipid-lowering drug ezetimibe produced an additive reduction in TC levels. Finally, lipid nanoparticle (LNP)-delivered base editing of ASGR1 in human HepG2 cells achieved >91% editing and near-complete protein knockout, which significantly lowered cellular cholesterol. Collectively, our results demonstrate that ASGR1 base editing represents a promising strategy for blood lipid control.
In recent years, plant-derived microRNAs (miRNAs), as a new type of cross-kingdom regulatory molecules, have shown great potential in anti-tumor therapy. These non-coding RNAs, about 20-24 nucleotides (nt) long, can stab...In recent years, plant-derived microRNAs (miRNAs), as a new type of cross-kingdom regulatory molecules, have shown great potential in anti-tumor therapy. These non-coding RNAs, about 20-24 nucleotides (nt) long, can stably exist in the human body through dietary intake and resist nuclease degradation through unique protective mechanisms, thereby achieving cross-kingdom gene regulation. Studies show that they can directly target tumor-related signaling pathways to inhibit cancer cell proliferation and induce apoptosis and indirectly exert anti-tumor effects by regulating long non-coding RNAs and immune cell functions in the tumor microenvironment. However, oral delivery faces multiple biological barriers. Researchers have developed innovative delivery strategies to improve miRNA stability, targeting, and bioavailability. Despite its promising application, clinical translation is limited. Future efforts should combine multi-omics technologies, optimize delivery systems, and adopt synergistic therapy to promote it as an oral anti-tumor biologic.
Old blood can induce senescence in young mice, yet the underlying mechanisms remain unclear. Here, we demonstrate that non-protein components (NPs) of aged human serum (ONPs) induce skeletal aging in young mice, with met...Old blood can induce senescence in young mice, yet the underlying mechanisms remain unclear. Here, we demonstrate that non-protein components (NPs) of aged human serum (ONPs) induce skeletal aging in young mice, with methylmalonic acid (MMA) identified as a key driver capable of independently recapitulating the skeletal aging phenotypes observed in ONP-treated mice. Photoaffinity labeling confirms Kindlin-2 as a direct intracellular MMA receptor. MMA binds directly to Kindlin-2 via a positively charged motif in its UBL domain, promoting Kindlin-2 ubiquitination and degradation, which accelerates skeletal aging. Notably, the endogenous MMA scavenger coenzyme A (CoA), a clinically approved molecule, effectively reverses MMA- and ONP-induced skeletal aging. Furthermore, CoA treatment effectively protects against skeletal aging and bone mass loss in aged or estrogen-deficient mouse models. Our study establishes MMA as a central aging factor and Kindlin-2 as its intracellular receptor, offering a potential therapeutic strategy for age-related diseases such as osteoporosis.
Facioscapulohumeral muscular dystrophy (FSHD) is a progressive muscle-wasting disease caused by aberrant activation of the DUX4 gene in skeletal muscle. Antisense oligonucleotides (ASOs) targeting DUX4 have shown therape...Facioscapulohumeral muscular dystrophy (FSHD) is a progressive muscle-wasting disease caused by aberrant activation of the DUX4 gene in skeletal muscle. Antisense oligonucleotides (ASOs) targeting DUX4 have shown therapeutic potential, but challenges related to systemic delivery and efficacy limit their clinical utility. In this study, we explored lipid nanoparticle (LNP)-mediated delivery of optimized locked nucleic acid (LNA) and 2'-O-methoxyethyl (MOE) gapmers to enhance their therapeutic potential. In FSHD patient-derived muscle cells, gapmers reduced DUX4 expression and improved myogenic differentiation indices relative to non-treated controls. In vivo studies demonstrated that LNP encapsulation increased gapmer bioavailability in skeletal muscle tissues compared to bare gapmers while maintaining a favorable safety profile. A single systemic injection in iDUX4pA mice moderately reduced DUX4 levels without significant functional improvement. However, repeated weekly injections resulted in up to ∼60% knockdown of DUX4 in skeletal muscles, repression of its downstream targets, improved grip strength and treadmill performance, and amelioration of muscle pathology. This study indicates that LNP-mediated gapmer delivery could be a candidate strategy to improve skeletal muscle biodistribution in iDUX4pA mice. This work provides preliminary proof of concept for further preclinical optimization and additional safety and efficacy studies.
Engineered cell therapies have revolutionized the treatment of immune disorders; most notably, chimeric antigen receptors (CARs) have been used to generate antigen-specific T cells capable of targeting and eliminating tu...Engineered cell therapies have revolutionized the treatment of immune disorders; most notably, chimeric antigen receptors (CARs) have been used to generate antigen-specific T cells capable of targeting and eliminating tumors. Ongoing research extends similar principles to induce immune tolerance in autoimmune diseases and transplantation by leveraging the immunosuppressive properties of regulatory T cells (Tregs) and co-opting conventional T cells for tolerogenic applications. In this review, we highlight the diverse uses of engineered antigen receptors to generate human T cell-based therapies, spanning a variety of disease contexts and focusing primarily on CAR Tregs. We further summarize work that aims to improve therapeutic potency and safety, including approaches to enhance suppressive pathways and optimize antigen receptor design and regulation. Finally, as CAR Tregs and similar therapies move to the clinic, we discuss the practical implications of translation and methods that utilize off-the-shelf products and in vivo gene delivery.
Cancer patients receiving chemotherapy and targeted therapy often experience severe side effects due to systemic damage of healthy cells throughout the body. Here, inspired by the programmable design of RNA molecules, we...Cancer patients receiving chemotherapy and targeted therapy often experience severe side effects due to systemic damage of healthy cells throughout the body. Here, inspired by the programmable design of RNA molecules, we present microRNA (miRNA)-guided RNA-toxin constructs (miR-RTCs), a novel therapeutic platform that achieves superior tumor selectivity through miRNA-controlled toxin expression. miR-RTCs operate as miRNA-responsive molecular switches that specifically respond to dysregulated intracellular miRNA expression profiles in tumor cells to conditionally express toxic proteins, enabling precise cancer cell elimination while sparing healthy tissues. In vivo, systemic delivery of miR-RTCs effectively restricts protein expression to tumor regions in hepatocellular carcinoma models, resulting in significant tumor regression with minimal off-target effects. This study establishes miR-RTCs as a programmable RNA therapeutic platform that overcomes the selectivity limitations of traditional chemotherapy while maintaining potent antitumor efficacy, with potential for further enhancement through immunotherapy combinations.
Phosphatidylserine is a representative eat-me signal on the outer membrane of apoptotic cells promoting phagocytosis/efferocytosis. It is recognized by various molecules, such as MFG-E8, which produce an immune-tolerizin...Phosphatidylserine is a representative eat-me signal on the outer membrane of apoptotic cells promoting phagocytosis/efferocytosis. It is recognized by various molecules, such as MFG-E8, which produce an immune-tolerizing environment. In order to develop immunological treatments targeting this mechanism, we designed a novel chimeric protein that consists of a phosphatidylserine-recognition domain from MFG-E8 and receptor-associated protein (RAP), which is a ligand of CD91 known as an immunostimulatory phagocytic receptor. This chimeric protein, PStRAP, can theoretically link phosphatidylserine-exposed apoptotic cells to phagocytes not through authentic immunosuppression but through an immunostimulative route via CD91. The inoculation of dying tumor cells expressing PStRAP, which served as an immunogen, induced potent adaptive immune responses against parental tumor cells in murine tumor models. The presence of PStRAP increased the phagocytosis of apoptotic cells in vitro and cross-priming to cytotoxic T-lymphocytes in vivo compared with the absence of it. Furthermore, PStRAP protein was supplied systemically by intravenous injection of PStRAP mRNA contained in lipid nanoparticles and along with cytotoxic drug to treat tumor-bearing mice. PStRAP significantly enhanced antitumor effects of cytotoxic drug alone. These results suggest that PStRAP can induce CD91-mediated phagocytosis of the apoptotic tumor cells and initiate meaningful adaptive immune responses against the tumor cells.
Most neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 target the receptor-binding domain (RBD). However, due to high immune pressure, the RBD accumulates mutations, thus significantly reduc...Most neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 target the receptor-binding domain (RBD). However, due to high immune pressure, the RBD accumulates mutations, thus significantly reducing antibody efficacy against emerging variants/subvariants. Although some RBD-targeting antibodies bind conserved epitopes, they usually exhibit weak-to-moderate neutralization. Similarly, antibodies against the N-terminal domain (NTD) or S2 subunit often retain broad binding but generally lack potent neutralization. To address this, we initially identified the broadly reactive nanobody, N103, which exhibited weak neutralizing potency. Structural and functional analyses revealed that N103 targets a conserved NTD epitope and triggers S1 subunit shedding, thereby destabilizing the spike trimer through an allosteric mechanism. Leveraging this insight, we engineered a trispecific antibody combining N103 with antibodies targeting the conserved RBD and S2 epitopes. This design synergistically integrated their distinct binding profiles and mechanisms, achieving potent and broad neutralization against both pseudoviruses and authentic viruses, including the immune-evasive BA.2.86 subvariant. Furthermore, challenge studies in human angiotensin-converting enzyme 2 knockin mice demonstrated robust in vivo protection. Our findings highlight a cooperative multi-target strategy in which antibodies with limited individual potency can collectively achieve broad and potent neutralization through rational design. This approach provides a promising framework for next-generation antibody therapeutics.
Diaz JA, Chilcott EM, Barbanoj AA
… +11 more, Keegan A, Gurung S, Pauzuolyte V, Waddington Z, Kyriacou M, McTague A, Cross JH, Schorge S, Lignani G, Waddington SN, Karda R
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42181696
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Dravet syndrome (DS) is a severe childhood developmental and epileptic encephalopathy. Symptoms usually manifest in the first year of life and include prolonged severe seizures, developmental delay, severe intellectual d...Dravet syndrome (DS) is a severe childhood developmental and epileptic encephalopathy. Symptoms usually manifest in the first year of life and include prolonged severe seizures, developmental delay, severe intellectual disability, and increased mortality. Approximately, 90% of patients carry a heterozygous loss-of-function mutation in , encoding a voltage-gated sodium ion channel, Na1.1. Na1.1 is expressed in the brain and at a lower level, in the heart. Previous studies have identified a long non-coding RNA (lncRNA), which specifically downregulates expression. This natural antisense transcript (NAT) can be modulated by AntagoNATs, small synthetic oligonucleotides. AntagoNATs have shown to improve seizure frequency in DS mice after repeated administration. Here, we have developed new AntagoNATs and incorporated these into an adeno-associated virus serotype 9 (AAV9) gene therapy vector. We administered two new AAV9-AntagoNAT-H and AntagoNAT-K vectors to newborn mice via intracerebroventricular (i.c.v.) and intravenous (i.v.) injection to deliver vector to the brain and heart. AAV9-AntagoNAT-H significantly increased survival, decreased the frequency of febrile and spontaneous seizures. In this proof-of-concept study, we have demonstrated for the first time the delivery of AntagoNAT via an AAV9 vector. Thus, offering the possibility of a one-time treatment for DS patients.
Knop O, Ben Haim N, Kalter N
… +9 more, Rosenberg M, Breier D, Baider K, Lee YN, Adam E, Somekh I, Nagler A, Somech R, Hendel A
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42181695
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X-linked severe combined immunodeficiency (SCID-X1) is a severe primary immunodeficiency caused by mutations in the gene, a shared subunit of cytokine receptors critical for the development and function of T and natural...X-linked severe combined immunodeficiency (SCID-X1) is a severe primary immunodeficiency caused by mutations in the gene, a shared subunit of cytokine receptors critical for the development and function of T and natural killer (NK) cells. The standard treatment, allogeneic hematopoietic stem cell transplantation (HSCT), requires a compatible donor and is often associated with significant transplant-related complications. We aimed to develop a more robust and universal gene therapy by modification of patient hematopoietic stem and progenitor cells (HSPCs) using the CRISPR-Cas9/rAAV6 gene-editing platform. To evaluate efficiency, we utilized a feeder-free, platform enabling T and NK cell differentiation from modified HSPCs. We investigated two approaches: the cut-site method (inserting a corrective cassette downstream of the start codon within the locus) and a replacement method (replacing the entire gene under endogenous regulation). We demonstrated that the cut-site insertion strategy is preferable for SCID-X1 correction, achieving superior homology-directed repair rates, lower toxicity, and significantly improved T and NK cell differentiation, based on phenotypic marker expression. Importantly, corrected patient-derived HSPCs from two SCID-X1 patients, modified using the cut-site approach, successfully demonstrated phenotypic evidence of T cell differentiation. Therefore, our findings firmly establish the feasibility of the cut-site strategy as a universal, high-efficiency, therapeutic solution for SCID-X1.