The development of effective cellular immunotherapies for solid tumors requires the presence of robust infiltration, persistence, proliferation, and antigen-specific cytotoxicity. Here, we engineered induced pluripotent...The development of effective cellular immunotherapies for solid tumors requires the presence of robust infiltration, persistence, proliferation, and antigen-specific cytotoxicity. Here, we engineered induced pluripotent stem cell (iPSC)-derived cytotoxic T cells transduced with a chimeric antigen receptor (iCAR-T cells) and identified an optimal cytokine armoring strategy. Co-expression of interleukin-15 (IL-15) and IL-21 synergistically enhanced STAT1 phosphorylation, leading to increased transcriptional activation of the chemokine receptor CXCR3 and thereby improving tumor homing capacity. Furthermore, the engineered iCAR-T cells maintained a CD45RACD45ROCCR7CD62L memory T cell-like phenotype in tumors, contributing to the prolonged survival of the animal model. These findings demonstrate that cytokine synergy can be engineered into iPSC-derived T cells to overcome significant barriers in solid tumor immunotherapy, offering a scalable approach to developing next-generation off-the-shelf CAR-T therapies.
Daquinag AC, Ghosh SC, AghaAmiri S
… +7 more, Farmer SM, Zhang S, Vargas SH, Ramesh AK, An Z, Azhdarinia A, Kolonin MG
Mol Ther Oncol
· 2026 Jun · PMID 42124952
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There is a lack of approaches to detect and kill metastatic cells. As an agent for metastasis targeting, a cyclic peptide BLMP6 has been previously characterized. As a step toward translation, we designed AZDye555-labele...There is a lack of approaches to detect and kill metastatic cells. As an agent for metastasis targeting, a cyclic peptide BLMP6 has been previously characterized. As a step toward translation, we designed AZDye555-labeled BLMP6 and demonstrated its homing to metastases of human MDA-MB-231 cells in mice. We show that Ga-radiolabeled BLMP6 can be used for the detection of MDA-MB-231 metastases. We designed a peptide-drug conjugate consisting of monomethyl auristatin E (MMAE) and BLMP6. We show that MMAE-BLMP6 kills MDA-MB-231 cells in cell culture and . In mouse models of lung metastases, treatment with MMAE-BLMP6 suppressed metastasis growth and improved survival. Based on BLMP6 similarity to latent transforming growth factor β binding protein 4 (LTBP4), we identified fibulin-4 as a BLMP6 target. We show that BLMP6 mimics the LTBP4 domain binding to fibulin-4 and selectively binds to fibulin-4 . Fibulin-4 knockout in cancer cells abrogated BLMP6 homing to lung metastases in mice. Fibulin-4 expression was found to be increased in invasive and metastatic human breast cancer and correlated with the binding of AZDye555-BLMP6 in human tissue sections. Our results suggest that fibulin-4 and BLMP6 may be further developed for the detection and targeting of metastatic human cancers.
Keshta A, Hashimoto R, Kitai Y
… +5 more, Nakata Y, Sakamoto A, Gotoh S, Takeda M, Takayama K
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42112103
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Respiratory syncytial virus (RSV) is one of the major causes of lower respiratory tract infections, particularly in infants and older adults. However, the host factors mediating infection remain poorly defined. It has be...Respiratory syncytial virus (RSV) is one of the major causes of lower respiratory tract infections, particularly in infants and older adults. However, the host factors mediating infection remain poorly defined. It has been suggested that four host surface proteins, namely intercellular adhesion molecule-1 (ICAM-1), epidermal growth factor receptor (EGFR), nucleolin (NCL), and insulin-like growth factor 1 receptor (IGF1R), may interact with the RSV fusion (F) protein. To investigate these roles under physiologically relevant conditions, we employed human induced pluripotent stem cell (iPSC)-derived respiratory organoids as a model for RSV infection. In this model, ICAM-1 and EGFR were genetically depleted using the CRISPR-Cas9 genome editing technique, while NCL and IGF1R were inhibited with neutralizing antibodies. Suppression of ICAM-1 or NCL significantly reduced nucleoprotein gene expression, whereas inhibition of EGFR or IGF1R had no observable effect on viral gene expression. Notably, simultaneous suppression of ICAM-1 and NCL resulted in a more substantial reduction in infectious viral titers and RSV F protein expression than inhibition of either protein alone. Our results suggest that both ICAM-1 and NCL may play important roles during RSV infection in human iPSC-derived respiratory organoids.
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42112102
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N-acetylgalactosamine-conjugated small interfering RNA (GalNAc-siRNA) therapeutics have emerged as a groundbreaking modality with unparalleled efficacy for battling previously "undruggable" diseases. The unique pharmacok...N-acetylgalactosamine-conjugated small interfering RNA (GalNAc-siRNA) therapeutics have emerged as a groundbreaking modality with unparalleled efficacy for battling previously "undruggable" diseases. The unique pharmacokinetic (PK) and pharmacodynamic (PD) characteristics of GalNAc-siRNA therapeutics provide an opportunity to leverage PK/PD modeling strategies for drug development. By utilizing the wealth of literature data, we developed and validated a mechanistic computational model-driven platform to guide the development of new GalNAc-siRNA therapeutics, optimizing their clinical translation. This platform integrates preclinical and clinical data from all seven FDA-approved GalNAc-siRNA drugs-fitusiran, givosiran, inclisiran, lumasiran, vutrisiran, nedosiran, and plozasiran-spanning multiple species (mouse, rat, monkey, and human). To enhance user accessibility, we further implemented a web-based Shiny application. The platform was used to inform the development of an investigational new angiotensinogen-silencing GalNAc-siRNA (SAL0132). Multiple PK/PD datasets from rats and monkeys were satisfactorily fitted, and extrapolated to humans. The platform successfully predicted the PK and simulated the PD profiles of SAL0132 in humans, which demonstrated model-informed strategies to support efficient drug development of this modality. In conclusion, this platform enables users to predict GalNAc-siRNA PK/PD profiles across species by inputting specific model parameters, providing a powerful resource to guide the development of next-generation GalNAc-siRNA therapeutics.
Chen S, Liu H, Cong D
… +11 more, Dong A, Wang Y, Bi J, Guo S, Yang J, Wang X, Ren G, Zhang K, Wang H, Lai F, Dang Y
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42112101
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Gapmer antisense oligonucleotides (ASOs) enable sequence-specific degradation of target mRNAs, offering therapeutic access to previously "undruggable" genes and holding great promise for treating chronic and genetic dise...Gapmer antisense oligonucleotides (ASOs) enable sequence-specific degradation of target mRNAs, offering therapeutic access to previously "undruggable" genes and holding great promise for treating chronic and genetic diseases. However, the rapid development of ASO therapeutics remains limited by challenges in rational sequence design and translational validation. Here, we present ClinASO, an integrated computational-experimental platform that unifies key determinants of ASO efficacy-including RNase H1 cleavage preference, SNP avoidance, off-target filtering, and cross-species conservation-into a single, data-driven workflow. This system enables rapid identification of potent ASO leads and direct validation in wild-type animal disease models. Using ClinASO, we efficiently identified potent ASOs against and , both exhibiting superior silencing activity compared to their clinical counterparts. Furthermore, ClinASO generated a potent ASO targeting genes implicated in metabolic dysfunction-associated liver disease (MASLD). In multiple human cells, the ASO achieved robust silencing effect. Notably, by conjugating GalNAc, this ASO demonstrated durable, liver-specific knockdown, significantly ameliorating hepatic steatosis and normalizing systemic lipid profiles in MASLD mouse model. Together, these findings establish ClinASO as an efficient, experimentally validated online tool for the rational design and rapid development of translatable ASO therapeutics.
Shcherbinina E, Fong M, Biscans A
… +1 more, Sarshad AA
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42112100
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RNA molecules are dynamic regulators of gene expression, with duplex RNAs such as siRNAs and saRNAs functioning as versatile effectors that can both silence and activate gene expression through Argonaute (AGO) proteins....RNA molecules are dynamic regulators of gene expression, with duplex RNAs such as siRNAs and saRNAs functioning as versatile effectors that can both silence and activate gene expression through Argonaute (AGO) proteins. While cytoplasmic RNA interference (RNAi) is well established, the mechanisms and outcomes of nuclear RNAi and RNA activation (RNAa) are only emerging. This review integrates recent discoveries on nuclear RNAi and RNAa, outlining how siRNAs can target nuclear long noncoding RNAs, promoter-associated transcripts, and pre-mRNAs to mediate silencing, transcriptional repression, or alternative splicing. Conversely, saRNAs can recruit AGO2 and transcriptional cofactors to activate gene expression through chromatin remodeling and RNA polymerase II engagement. Beyond their mechanistic roles, we highlight the growing therapeutic potential of duplex RNAs, which can be harnessed to selectively silence or activate gene expression, offering new strategies for RNA-based precision medicine.
Gondaliya P, Luo Y, Sayyed AA
… +8 more, Zinn DA, Qie Y, Yan IK, Driscoll J, Patel PB, Li S, Qin H, Patel T
Mol Ther Oncol
· 2026 Jun · PMID 42110477
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An immunosuppressive microenvironment driven by tumor and stromal cells expressing programmed death-ligand 1 (PD-L1) contributes to immune evasion and poor prognosis in cholangiocarcinoma. Although antibodies to PD-L1 ar...An immunosuppressive microenvironment driven by tumor and stromal cells expressing programmed death-ligand 1 (PD-L1) contributes to immune evasion and poor prognosis in cholangiocarcinoma. Although antibodies to PD-L1 are used clinically, their benefit is limited by immune exclusion within the local microenvironment. To overcome this, we evaluated engineered T cells directed toward PD-L1 that simultaneously target tumor cells and the immunosuppressive microenvironment. Human donor T cells were transduced with a lentiviral vector encoding a chimeric antigen receptor (CAR) consisting of an anti-PD-L1 scFv, CD4 transmembrane domain, and 4-1BB/CD3ζ signaling domain. The antitumor efficacy of these CAR-T cells was assessed in a murine orthotopic tumor model, and their specificity and effect were validated in human malignant cholangiocytes with varying PD-L1 expression. PD-L1 CAR-T cells retained T cell identity, demonstrated antigen-specific cytotoxicity, and effectively reduced tumor burden . Cytotoxicity was abrogated in PD-L1 knockout cells, confirming target specificity. PD-L1 CAR-T cells significantly reduced tumor cell viability within multicellular spheroids. Gemcitabine pretreatment upregulated PD-L1 expression and enhanced CAR-T-mediated cytotoxicity. These findings demonstrate the feasibility of second-generation PD-L1 CAR-T cells, demonstrating preclinical efficacy and specificity, and validating a therapeutic strategy that targets the tumor microenvironment for these challenging cancers.
Bouillon M, Lapoujade C, Basset L
… +4 more, Fontaine A, Hollenstein M, Garcion E, Rousseau A
Mol Ther Oncol
· 2026 Jun · PMID 42110475
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Glioblastoma, the most common primary central nervous system tumor, is the leading cause of death in neuro-oncology. Gene fusions, caused by chromosomal rearrangements, may act as drivers of tumorigenesis in glioblastoma...Glioblastoma, the most common primary central nervous system tumor, is the leading cause of death in neuro-oncology. Gene fusions, caused by chromosomal rearrangements, may act as drivers of tumorigenesis in glioblastoma. These fusions result from the juxtaposition of two genes, leading to the production of a chimeric protein and, in most cases, constitutive activation of a tyrosine kinase receptor. Despite the use of tyrosine kinase inhibitors to block the oncogenic activity of gene fusions, clinical responses in glioblastoma remain poor compared to those in other cancers, highlighting the need for innovative therapeutic strategies. RNA interference, using small interfering RNAs or micro-RNAs, offers a promising approach to target these oncogenic fusions. Through specific silencing, small interfering RNAs spare healthy cells, avoiding the adverse effects associated with tyrosine kinase inhibitors. Recent advances in biotechnology (e.g., antisense oligonucleotides and aptamers) and delivery systems have improved small interfering RNA stability, specificity, and ability to cross the blood-brain barrier. This review discusses these advances and their potential applications to target oncogenic gene fusions in glioblastoma. RNA interference-based therapy represents a critical area of research that could improve the survival of patients with glioblastoma.
Wu A, Dumo CJ, Cook WH
… +8 more, Richards I, Macapagal Foliaki J, Cooper E, Fong DM, Mouravlev A, Park TI, Dragunow M, Young D
Mol Ther Oncol
· 2026 Jun · PMID 42110474
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Glioblastoma (GBM) is a highly aggressive type of brain cancer associated with poor prognosis due to limited effective treatment options. Adeno-associated viral (AAV) vector-based gene therapy strategies have demonstrate...Glioblastoma (GBM) is a highly aggressive type of brain cancer associated with poor prognosis due to limited effective treatment options. Adeno-associated viral (AAV) vector-based gene therapy strategies have demonstrated promise in preclinical models of GBM, but inter- and intratumor cellular heterogeneity poses a challenge for tumor-specific targeting. Here, we screened a panel of 15 AAV serotypes against the widely used human GBM cell line U-87 MG, and five primary patient-derived GBM cell lines to characterize their transduction efficiencies. AAV1, AAV2, AAV1/2, AAV6, and AAV6.2 were the most efficient and consistent serotypes across cell lines, capable of differentially transducing CD44-expressing cells in culture, albeit non-selectively. Interestingly, we observed a reduction in cell number and proliferation in certain cell lines, which correlated with AAV transduction efficiency. This phenomenon was shown to be a cytostatic rather than cytotoxic effect on GBM cells, occurring independently of promoter sequence and transgene expression. The anti-proliferative effect was particularly pronounced in two patient-derived cell lines, where cell proliferation decreased by nearly 80%. Altogether, our results provide a foundation for future studies optimizing AAV-mediated transduction in the diverse GBM cell population and demonstrate the potential to harness the natural properties of AAV in treatment development for brain cancers.
Wei P, Zhang K, Hu Y
… +19 more, Gao X, Hou Y, Riaz F, Zhang Z, He J, Zhang M, Li Y, Li F, Wang A, Zhao T, Zhang B, Li X, Jiang A, Wu L, Chen YH, Fan Z, Huang J, Wang Q, Pan F
Autoimmune and inflammatory diseases are characterized by dysregulated T cell-mediated immune responses leading to tissue damage. Despite therapeutic advancements, patients remain resistant to treatment, highlighting the...Autoimmune and inflammatory diseases are characterized by dysregulated T cell-mediated immune responses leading to tissue damage. Despite therapeutic advancements, patients remain resistant to treatment, highlighting the urgent need for alternative therapeutic strategies. Here, we identified neuritin (NRN), an immunosuppressive molecule, capable of restraining effector CD4+ T cell responses and mitigating autoimmune and inflammatory diseases. NRN is downregulated in CD4+ T cells of rheumatoid arthritis (RA) patients, driving T cell-mediated autoimmune pathology. NrnCD4 mice exacerbate both experimental autoimmune encephalomyelitis (EAE) and dextran sulfate sodium (DSS)-induced colitis, characterized by regulatory T cell (Treg) depletion and expansion of interferon (IFN)-γ+ and interleukin (IL)-17+ pro-inflammatory cells. Mechanistically, NRN selectively binds to cannabinoid receptor 2 (CB2), but not to CB1, specifically through its threonine residues at positions T78 and T81. Meanwhile, mice lacking CB2 (CB2 or CB2CD4) exhibit worsened colitis, with an increased IFN-γ+ and IL-17+ cells, mirroring the NrnCD4 phenotype. T cell-specific Nrn knockin (NrnCD4) or exogenous NRN administration ameliorated disease severity in multiple autoimmune models, including DSS-induced colitis, IMQ-induced psoriasis, and collagen-induced arthritis, by promoting Treg expansion and suppressing IFN-γ+ and IL-17+ pro-inflammatory cells. However, these protective effects of NRN were abolished in CB2-deficient mice. Overall, NRN is an immunosuppressive molecule with therapeutic potential in autoimmune and inflammatory diseases.
Mol Ther Nucleic Acids
· 2026 Jun · PMID 42100560
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Long interspersed nuclear element-1 (LINE-1), a prominent class of retrotransposons, is abundantly distributed throughout mammalian genomes and plays critical roles in development, aging, and disease. In this review, we...Long interspersed nuclear element-1 (LINE-1), a prominent class of retrotransposons, is abundantly distributed throughout mammalian genomes and plays critical roles in development, aging, and disease. In this review, we provide a comprehensive overview of the multifaceted functions of LINE-1, with a focus on its regulatory impact on gene transcription and cellular responses across the DNA, RNA, and protein levels. We highlight recent advances in understanding the transcriptional and epigenetic mechanisms controlling LINE-1 transcription. Furthermore, we explore the current challenges and future opportunities in LINE-1 research, emphasizing its potential as a novel therapeutic target. Continued investigation into LINE-1 biology holds promise for deepening our understanding of human health and disease.