Molecular Therapy最新文献

Multiple lines of genetic and neuropathological evidence strongly implicate α-synuclein (αSyn) as a causal factor in synucleinopathies like Parkinson disease (PD), with numerous studies supporting the therapeutic concept of αSyn reduction as a disease-modifying strategy. Artificial miRNAs can be durably expressed from adeno-associated viral (AAV) vectors and have shown increasing promise as a highly specific modality to reduce levels of target mRNA. This modality is therefore well suited for the long-term treatment of PD and related diseases. Here, we assessed the therapeutic potential of the partial reduction of αSyn by AAV-mediated artificial microRNA (amiRNA) expression by using the pre-formed fibril model of α-synucleinopathy to induce the seeding and spreading of αSyn pathology in wild-type animals. We demonstrate that a 50% decrease in endogenous SNCA mRNA levels is sufficient to fully block the spread of pathogenic αSyn protein and prevent the loss of dopaminergic neurons in the substantia nigra. To support clinical translation, human-specific amiRNAs were designed and evaluated in vitro and in vivo. Concatenation of amiRNAs was evaluated to optimize the potency of individual guide sequences. Several candidates had excellent expression, processing, and SNCA reduction, supporting the continued development of αSyn-lowering therapeutics as a safe and efficacious approach for treating patients with synucleinopathies.

Non-invasive imaging of gene expression remains a critical challenge in gene therapy. Here, we report the first in vivo demonstration of a genetically encoded chemical exchange saturation transfer (CEST) MRI reporter expressed from a recombinant adeno-associated virus (rAAV). The genetically encoded peptide-based reporter superCESTide enables detection of transgene expression without the need for exogenous contrast agents. Mice were systemically administered rAAV encoding superCESTide and fluorescence reporter tdTomato, and they were evaluated using CEST MRI, fluorescence imaging, and reverse transcription-PCR. CEST MRI revealed dose-dependent signal enhancement in the liver, with a strong correlation between CEST contrast and superCESTide expression (r = 0.75, p < 0.05). Mice receiving the highest dose (1.4 × 10¹³ vector genomes/kg) exhibited the strongest MTR_asym signal (mean = 0.0225), and histogram analyses showed distinct expression patterns. These results establish CEST MRI as a powerful, non-invasive method for monitoring rAAV-mediated transgene expression in vivo, opening new possibilities for imaging-guided gene therapy.

Engineered bacterial systems have been widely utilized as versatile biotherapeutic platforms in disease intervention, yet their therapeutic efficacy remains limited. Recent advancements in synthetic biology have enabled the systematic deconstruction of bacteria through rational computational modeling and modular-genetic circuit design, thereby overcoming intrinsic functional limitations. These engineered bacteria represent a promising therapeutic strategy characterized by enhanced spatiotemporal precision, biosafety profiles, and scalable production. Moreover, the advent of sophisticated genetic engineering technologies has facilitated the expansion of bacterial therapeutic applications into new areas, including vaccine development and tumor-microenvironment modulation. Here, we systematically examine recent advances in the development and testing of engineered bacteria across different organs on the basis of tissue specificity. We further summarize the therapeutic roles of engineered bacteria in tumor treatment and metabolic-disorder management. Additionally, we outline advanced technologies for engineered bacterial development, emphasizing their critical roles in optimizing the overall clinical efficacy of therapeutic agents after system-level optimization.