CHEMICAL ENGINEERING JOURNAL
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2025
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IF 13.2
Zhonghao Liao,Jiang Yu,Fangming Liang,Yubo Liu,Zhenyang Zhao,Yue Wang,Xianbao Shi,Yongjun Wang,Hongzhuo Liu
Adeno-associated virus (AAV) has been extensively employed in gene therapy owing to its outstanding safety profile and low immunogenicity. AAV vector-mediated gene therapy necessitates repeated administrations to achieve prolonged therapeutic efficacy. However, clinical trials have demonstrated that subsequent to the initial administration of AAV, the host mounts an immune response against the vectors, thereby hindering the clinical feasibility of repeated AAV administration. Moreover, when the AAV dosage exceeds 1.0 × 10 13 vg/kg, there is a significant risk of excessive activation of the complement system, which can result in severe adverse reactions. Notably, research indicates that merely depleting IgG antibodies does not enable high-dose re-administration of AAV or alleviate excessive complement activation; IgM antibodies also play a pivotal role in both the immune response against AAV and complement system activation. In this study, we synthesized a series of rapamycin lipid prodrugs utilizing succinic anhydride as a chemical linker and co-assembled them with dexamethasone palmitate to produce co-assembled nanoparticles ranging from 100 to 180 nm in size. These co-assembled nanoparticles are capable of inducing regulatory T cell (Treg) generation while inhibiting T follicular helper (Tfh) cell formation, thus reducing germinal center B (GC B) cell production and consequently suppressing anti-AAV IgG and IgM antibody responses along with complement system activation. The findings suggest that these rapamycin lipid prodrug co-assembled nanoparticles facilitate high-dose re-administration of AAV while enhancing gene expression delivered by the vector.
研究领域:
生物医学工程
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免疫学
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基因治疗
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DOI
高影响因子
ANALYST
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2024
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IF 3.6
Mengmeng Li,Yujian Zhong,Mingyang Zhu,Chunjin Pang,Lu Xiao,Ye Bu,Huapeng Li,Yong Diao,Chaoyong Yang,Dan Liu
To overcome limitations in generalizability and efficiency of current AAV vectors, in this current study, we constructed an AAV variant library by insertion of random heptapeptide sequences in the receptor-binding domain of the AAV9 capsid gene. We then applied a recently developed organ-on-a-chip in vitro model of human blood-brain barrier (BBB) to iteratively enrich for variants that efficiently cross the BBB and transduce astrocyte cells. Through multiple rounds of screening, we obtained two candidate AAV variants, AAV-M6 and AAV-M8, that showed significantly higher BBB penetration efficiency than AAV9 or AAV-PHP.eB. Quantitative PCR (qPCR) assay showed that AAV-M6 could accumulate to 5 times higher titer, while AAV-M8 reached 3 times higher titer, than AAV-PHP.eB in the neural chamber of the model. Transduction assay further verified that the AAV-M6 candidate vector was able to infect HA-1800 cells after crossing the BBB, suggesting it could potentially transduce brain parenchymal cells after crossing the hCMEC/D3 layer at higher efficiency than AAV-PHP.eB. Molecular simulations suggested that the human receptor proteins, LY6D and M6PR, could bind the AAV-M6 heptapeptide insertion with high affinity. This study provides two promising candidate AAV vectors and demonstrates the use of this in vitro BBB model for scalable, high throughput screening of gene therapies. These tools can drive investigations of the mechanisms underlying BBB permeability and the cell type specificity of virus vectors.
研究领域:
神经科学
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基因治疗
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分子生物学
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DOI
PubMed
高影响因子
Nature Communications
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2024
·Vol. 15
·期 1
·1-16
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IF 14.7
Wu Xin,Yu Yuanhuan,Wang Meiyan,Dai Di,Yin Jianli,Liu Wenjing,Kong Deqiang,Tang Shasha,Meng Meiyao,Gao Tian,Zhang Yuanjin,Zhou Yang,Guan Ningzi,Zhao Shangang,Ye Haifeng
Gene therapies provide treatment options for many diseases, but the safe and long-term control of therapeutic transgene expression remains a primary issue for clinical applications. Here, we develop a muscone-induced transgene system packaged into adeno-associated virus (AAV) vectors (AAV MUSE ) based on a G protein-coupled murine olfactory receptor (MOR215-1) and a synthetic cAMP-responsive promoter (P CRE ). Upon exposure to the trigger, muscone binds to MOR215-1 and activates the cAMP signaling pathway to initiate transgene expression. AAV MUSE enables remote, muscone dose- and exposure-time-dependent control of luciferase expression in the livers or lungs of mice for at least 20 weeks. Moreover, we apply this AAV MUSE to treat two chronic inflammatory diseases: nonalcoholic fatty liver disease (NAFLD) and allergic asthma, showing that inhalation of muscone—after only one injection of AAV MUSE —can achieve long-term controllable expression of therapeutic proteins (ΔhFGF21 or ΔmIL-4). Our odorant-molecule-controlled system can advance gene-based precision therapies for human diseases.
研究领域:
基因治疗
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药理学
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分子生物学
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慢性病管理
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DOI
PubMed
高影响因子
Pathogens
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2025
·Vol. 14
·期 2
·131
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IF 3.3
Jingjing Zhang,Mengfei Wang,Dan Wang,Xiangyi Zhang,Yue Ma,Els Pardon,Jan Steyaert,Romany Abskharon,Fei Wang,Jiyan Ma
Prion diseases are fatal neurodegenerative disorders affecting humans and animals, and the central pathogenic event is the conversion of normal prion protein (PrPC) into the pathogenic PrPScisoform. Previous studies have identified nanobodies that specifically recognize PrPCand inhibit the PrPCto PrPScconversion in vitro. In this study, we investigated the potential for in vivo expression of anti-PrPCnanobodies and evaluated their impact on prion disease. The coding sequences of three nanobodies were packaged into recombinant adeno-associated virus (rAAV) and were administered via intracerebroventricular (ICV) injection in newborn mice. We found that the expression of these nanobodies remained robust for over 180 days, with no observed detrimental effects. To assess their therapeutic potential, we performed ICV injections of nanobody-expressing rAAVs in newborn mice, followed by intracerebral prion inoculation at 5–6 weeks of age. One nanobody exhibited a small yet statistically significant therapeutic effect, extending survival time from 176 days to 184 days. Analyses of diseased brains revealed that the nanobodies did not alter the pathological changes. Our findings suggest that high levels of anti-PrPCnanobodies are necessary to delay disease progression. Further optimization of the nanobodies, AAV vectors, or delivery methods is essential to achieve a significant therapeutic effect.
研究领域:
神经科学
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分子生物学
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病毒学
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治疗学
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DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
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2026
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IF 4.9
Li Li,Weijia Huang,Zhenyang Zhao,Yuan Gao,Rui Zhang,Zhonghao Liao,Yuxin Hu,Qing Ma,Yang Yang,Yongjun Wang,Hongzhuo Liu
Antidrug antibodies (ADAs) compromise the pharmacokinetics and efficacy of biologics and can trigger adverse reactions. We engineer a tolerogenic liposomal platform in which rapamycin is covalently conjugated to cholesterol (RAPA-chol) and formulated as nanoliposomes (RA-c@L) to induce antigen-specific immune tolerance to coadministered proteins. Covalent anchoring enables high drug encapsulation (>95%) and improves colloidal stability. In a three-dose weekly tolerization regimen with uricase, followed by a high-dose challenge, RA-c@L markedly suppresses ADA formation: by day 42, antiuricase IgG titers are reduced by 2.15-fold compared to uricase alone and 2.17-fold compared to free rapamycin. Responses to an irrelevant antigen (KLH) remain unchanged, indicating antigen specificity. Importantly, coadministration of RA-c@L with rAAV8-SEAP enables vector readministration, yielding approximately 2-fold higher sustained serum SEAP expression after the second dose compared to rAAV alone. Mechanistically, intravenously delivered RA-c@L preferentially accumulates in the liver and reshapes systemic immunity, with reduced splenic T follicular helper cells and germinal-center B cells and an expansion of CD4+Foxp3+ regulatory T cells. Together, these data show that RA-c@L establishes durable, antigen-specific tolerance to therapeutic proteins and facilitates AAV redosing, offering a practical strategy to mitigate ADA-mediated loss of efficacy in repeated biotherapeutic treatments.
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PubMed
高影响因子
Materials Horizons
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2025
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IF 10.7
Yubo Liu,Fangming Liang,Rui Zhang,Chengcheng Tang,Xin Li,Yongjun Wang,Zhonggui He,Hongzhuo Liu
Recombinant adeno-associated virus (AAV) vectors have become the promising platform to deliver a therapeutic transgene for the treatment of monogenic disorders. However, anti-AAV antibodies that are released by B cells after AAV treatment create a substantial barrier to their long-lasting safe and effective therapy, making AAV gene therapy a “one shot” treatment. Here, we offered that tolerogenic nano-adjuvants (named RICP) composed of rapamycin (RAPA) and itaconate (ITA), to induce specific immune tolerance to overcome the anti-drug antibodies (ADAs). Such strategy blocked the feedback loops of follicular helper T （Tfh） cells and germinal center B cells by metabolic regulation, complementary to induction of regulatory T cells (Tregs) by mTORc1 inhibitor RAPA, consequently abolished the secretion of anti-AAV antibodies. The developed nano-adjuvants can also significantly increase expression of a transgene (such as luciferase, green fluorescent protein and human embryonic alkaline phosphatase reporter transgene) in the liver at re-dosing, compared to mice treated with AAV alone which showed almost no change in transgene expression. The emerging strategy provided AAV re-administration schedule available for not only mitigate toxicities with high vector doses, but re-storing therapeutic benefits in children in significant cell proliferation.
研究领域:
基因治疗
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免疫学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Neural Circuits
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2026
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IF 3
Lingbo Zhou,Gao Tan,Yu Li,Man Yuan,Sen Jin,Wenhui Zhang,Qitian Wang,Yin Shen
Introduction: Sparse and bright labeling of retinal ganglion cell (RGC) is essential for correlating single-cell morphology with brain-wide visual circuitry. This study aimed to develop a cell-type-specific, sparse labeling strategy for parvalbumin-expressing RGCs (PV+ RGCs) in the transgenic mouse retina using recombinant adeno-associated virus (rAAV) and to map the whole-brain projection patterns of single PV+ RGCs via fluorescence micro-optical sectioning tomography (fMOST).Methods: A cell-type-specific dual AAV system was employed, co-packaging a Cre-dependent Flpo plasmid and an Flpo-dependent enhanced yellow fluorescent protein (EYFP) plasmid. Key parameters-including the mixing ratio of core plasmids (ranging from 1/100 to 1/1000), gene copy number of Flpo and EYFP (single versus double), and AAV serotype (AAV2.2 versus engineered AAV2.NN)-were systematically optimized. Transduction efficiency and labeling sparsity under each condition were compared. Whole-retina-to-brain imaging was performed using fMOST on samples injected with the optimal condition (AAV2.2-double-1/1000), enabling the reconstruction of complete axonal trajectories of individual PV+ RGCs from the retina to the brain.Results: The sparsity and signal intensity of labeled RGCs varied significantly with the core plasmid ratio, AAV serotype, and gene copy number. The engineered AAV2.NN serotype increased transduction efficiency and labeling density under equivalent conditions, which facilitated the morphological subclassification of PV+ RGCs into ON, ON-OFF, and OFF types based on their stratification relative to ChAT bands. Axonal projections of single PV+ RGCs were successfully traced to the superior colliculus (SC), dorsal and ventral lateral geniculate nuclei (dLGN/vLGN).Discussion: This viral labeling platform effectively resolves the classical trade-off between sparsity and signal intensity, providing a robust methodology for whole-brain mapping of individual RGC projections. The approach establishes a practical foundation for future mechanistic and therapeutic studies investigating subtype-selective vulnerability in RGCs.
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PubMed
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
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2026
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IF 5.6
Zhidan Shi,Xinyi Cheng,Chu Zhang,Jiayi Hu,Xinqi He,Shuting Chen,Hao Wu,Meng Tian,Chuang Yan,Ling He
Alzheimer's disease (AD) is a neurodegenerative disorder with significant sex-related differences, exhibiting a higher incidence rate in elderly women. Tau pathology has been implicated in sex-associated differences in AD susceptibility and disease progression, contributing to differential pathological burden between males and females. Vorinostat (SAHA), a histone deacetylase inhibitor (HDACi) used in cancer treatment, has shown potential therapeutic effects in various neurological and psychiatric disorders; however, the mechanisms underlying its regulation of tau-associated pathology remain poorly understood, with limited evidence regarding its effects in tau-related neurodegenerative conditions. In this study, we investigated the effects of SAHA treatment on an AAV-P301L-Tau-induced tauopathy model. In vivo, we observed that SAHA administration significantly improved p-Tau levels in the hippocampus and cortex of mice, reduced microglial activation and neuronal damage, and alleviated tauopathy-associated cognitive impairments. Our findings revealed sex-associated differences in Tau-related pathological alterations, with female mice exhibiting greater pathological changes and differential responses to SAHA treatment in several molecular and histological indicators. Further combined in vivo and in vitro experiments revealed that SAHA was associated with modulation of the PI3K/AKT/GSK3β signaling pathway, suppression of NF-κB activation, and attenuation of neuroinflammatory responses, including reductions in experimentally measured inflammatory markers such as interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-α). These molecular alterations may represent important regulatory components contributing to the neuroprotective effects of SAHA, rather than acting as the sole determinants of its therapeutic efficacy. In summary, these results indicate that SAHA can improve Tau-associated pathological alterations in an AAV-P301L-Tau-induced tauopathy model. The potential mechanisms may involve coordinated regulation of multiple pathological processes, including modulation of Tau phosphorylation, neuroinflammatory responses, and aging-associated signaling pathways. The observed sex-dependent responses to SAHA provide additional insights into the potential influence of biological sex on HDACi-based therapeutic strategies, although the molecular mechanisms underlying these differences require further investigation.
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高影响因子
eLife
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2022
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IF 8.713
Weizhao Chen,Qiongping Zheng,Qiaoying Huang,Shanshan Ma,Mingtao Li
Lineage reprogramming of resident glial cells to dopaminergic neurons (DAns) is an attractive prospect of the cell-replacement therapy for Parkinson’s disease (PD). However, it is unclear whether repressing polypyrimidine tract binding protein 1 (PTBP1) could efficiently convert astrocyte to DAns in the substantia nigra and striatum. Although reporter-positive DAns were observed in both groups after delivering the adeno-associated virus (AAV) expressing a reporter with shRNA or CRISPR-CasRx to repress astroglial PTBP1, the possibility of AAV leaking into endogenous DAns could not be excluded without using a reliable lineage-tracing method. By adopting stringent lineage-tracing strategy, two other studies show that either knockdown or genetic deletion of quiescent astroglial PTBP1 fails to obtain induced DAns under physiological condition. However, the role of reactive astrocytes might be underestimated because upon brain injury, reactive astrocyte can acquire certain stem cell hallmarks that may facilitate the lineage conversion process. Therefore, whether reactive astrocytes could be genuinely converted to DAns after PTBP1 repression in a PD model needs further validation. In this study, we used Aldh1l1-CreERT2-mediated specific astrocyte-lineage-tracing method to investigate whether reactive astrocytes could be converted to DAns in a 6-hydroxydopamine (6-OHDA) mouse model of PD. However, we found that no astrocyte-originated DAn was generated after effective and persistent knockdown of astroglial PTBP1 either in the substantia nigra or in striatum, while AAV ‘leakage’ to nearby neurons was easily observed. Our results confirm that repressing PTBP1 does not convert astrocytes to DAns, regardless of physiological or PD-related pathological conditions.
研究领域:
神经科学
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分子生物学
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细胞治疗
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神经退行性疾病
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产品关联
DOI
PubMed
高影响因子
Journal of Inflammation Research
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2022
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IF 4.631
Meng Zhang,Yi Liu,Jing Chen,Lei Chen,Li Zhang,Xianguo Chen,Zongyao Hao,Chaozhao Liang
Background Chronic nonbacterial prostatitis (CNP) has a high incidence, low cure rate, and unclear pathogenesis. Here, we aimed to systematically identify effective diagnostic and therapeutic targets for CNP.Methods Prostate tissues were obtained from established mouse models and negative controls and were used for mRNA array sequencing and immunohistochemistry (IHC) staining. Predominant pathways were identified based on pathway enrichment analysis and pharmaceutical experiments. We also investigated the functional role of CXCL12 on CP, a critical factor belonging to the predominant chemotaxis pathway, and employed IHC staining to explore the influence of the CXCL12/CXCR4 axis on the activation of the NF-κB, AKT, and STAT3 signaling pathways. Serum samples derived from both CNP cases and healthy controls were used to determine the secretion level of CXCL12.Results By employing mRNA array sequencing and immunohistochemistry, we found that CXCR4, CXCL12, CD44, and OFLM4 were highly expressed in the infiltrated inflammatory T cells of the prostate tissues generated from CNP mice, while they were rarely expressed on the epithelial cells. Based on the pathway enrichment results, we applied pathway inhibitors to suppress the activity of these classic pathways. We found that targeting the CXCL12/CXCR4 axis with its specific antagonist AMD3100 remarkably alleviated inflammatory infiltration of the prostate in CNP models. Similar results were obtained when we replaced AMD3100 with adenovirus-associated virus (AAV)-shCxcl12. To clarify the potential mechanisms of how the CXCL12/CXCR4 axis influences the pathogenesis of CNP, we tested the classical downstream pathways. The results suggested that p-Akt, p-STAT3, and p-NF-κB were more highly expressed on the inflammatory cells of the prostate derived from the CNP model and were partly suppressed after applying AMD3100 or delivering AAV-shCxcl12, indicating that the CXCL12/CXCR4 axis potentially functioned through AKT/NF-κB and STAT3 signaling to influence the pathogenesis of CNP.Conclusion Our study provides potential diagnostic biomarkers and therapeutic targets for CNP.
研究领域:
免疫学
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分子生物学
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药理学
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泌尿学
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DOI
PubMed
高影响因子
BIOCHEMICAL PHARMACOLOGY
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2024
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IF 5.3
Xinrui Meng,Jingjing Liu,Jia Kang,Menghan Wang,Zhanghui Guan,Dong Tian,Xinping Chen
Gastric ulcer is a highly prevalent digestive tract disease across the world, which is recurrent and hard to cure, sometimes transforming into gastric cancer if left untreated, posing great threat to human health. To develop new medicines for gastric ulcer, we ran a series of screens with ethanol stress model in GES-1 cells, and we uncovered that lamivudine rescued cells from ethanol toxicity. Then, we confirmed this discovery using the well-established ethanol-induced gastric ulcer model in mice and our findings suggest that lamivudine can directly activate phosphoglycerate kinase 1 (PGK1, EC 2.7.2.3), which binds and stimulates superoxide dismutase 1 (SOD1, EC 1.15.1.1) to inhibit ferroptosis and ultimately improve gastric ulcer. Moreover, AAV-PGK1 exhibited comparable gastroprotective effects to lamivudine. The findings are expected to offer novel therapeutic strategies for gastric ulcer, encompassing both lamivudine and AAV-PGK1.
研究领域:
胃肠病学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
BRITISH JOURNAL OF PHARMACOLOGY
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2024
·
IF 6.8
Yun Zhao,Guobin Zheng,Shu Yang,Shangjing Liu,Yifan Wu,Yaodong Miao,Zhen Liang,Yunqing Hua,Jing Zhang,Jia Shi,Dan Li,Yanfei Cheng,Yunsha Zhang,Yuanli Chen,Guanwei Fan,Chuanrui Ma
Background and purpose Atherosclerosis is the basis of cardiovascular disease. Ferroptosis is a form of programmed cell death characterized by lipid peroxidation, which contributes to atherogenesis. The plant extract PNS ( Panax notoginseng saponins ), containing the main active ingredients of Panax notoginseng , exhibits anti-atherogenic properties. Herein, we determined whether PNS and its major components could attenuate atherosclerosis by suppressing ferroptosis and revealed the underlying mechanism(s). Experimental approach The anti-atherogenic effects of PNS and their association with inhibition of ferroptosis was determined in apoE −/− mice. In vitro, the anti-ferroptotic effect and mechanism(s) of PNS components were demonstrated in the presence of ferroptosis inducers. Expression of ferroptosis markers and the ubiquitination of Keap1 were evaluated in USP2 −/− macrophages. Finally, the anti-atherogenic effect of USP2 knockout was determined by using USP2 −/− mice treated with high-fat diet (HFD) and AAV-PCSK9. Key results PNS inhibited ferroptosis and atherosclerosis in vivo. PNS suppressed ferroptosis and ferroptosis-aggravated foam cell formation and inflammation in vitro. Mechanistically, PNS and its components activated Nrf2 by antagonizing Keap1, which was attributed to the inhibition of USP2 expression. USP2 knockout antagonized ferroptosis and ferroptosis-aggravated foam cell formation and inflammation, thus mitigating atherosclerosis. USP2 knockout abolished inhibitory effects of PNS on foam cell formation and inflammation in vitro. Conclusion and implications PNS reduced USP2-mediated Keap1 de-ubiquitination and promoted Keap1 degradation, thereby activating Nrf2, improving iron metabolism and reducing lipid peroxidation, thus contributing to an anti-atherosclerotic outcome. Our study revealed the mechanism(s) underlying inhibition of ferroptosis and atherosclerosis by PNS. Graphical
研究领域:
心血管疾病
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药理学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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