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Polymer-locking fusogenic liposomes for glioblastoma-targeted siRNA delivery and CRISPR–Cas gene editing

Nature Nanotechnology volume 19, pages 1869–1879 (2024) Cite this article

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Abstract

In patients with glioblastoma (GBM), upregulated midkine (MDK) limits the survival benefits conferred by temozolomide (TMZ). RNA interference (RNAi) and CRISPR–Cas9 gene editing technology are attractive approaches for regulating MDK expression. However, delivering these biologics to GBM tissue is challenging. Here we demonstrate a polymer-locking fusogenic liposome (Plofsome) that can be transported across the blood–brain barrier (BBB) and deliver short interfering RNA or CRISPR–Cas9 ribonucleoprotein complexes into the cytoplasm of GBM cells. Plofsome is designed by integrating a ‘lock’ into the fusogenic liposome using a traceless reactive oxygen species (ROS)-cleavable linker so that fusion occurs only after crossing the BBB and entering the GBM tissue with high ROS levels. Our results showed that MDK suppression by Plofsomes significantly reduced TMZ resistance and inhibited GBM growth in orthotopic brain tumour models. Importantly, Plofsomes are effective only at tumour sites and not in normal tissues, which improves the safety of combined RNAi and CRISPR–Cas9 therapeutics.

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Fig. 1: Plofsomes for GBM-targeted siRNA delivery and CRISPR–Cas gene editing.
The alternative text for this image may have been generated using AI.
Fig. 2: MDK facilitates TMZ resistance and malignant progression of GBM cells.
The alternative text for this image may have been generated using AI.
Fig. 3: Characterization of Plofsomes.
The alternative text for this image may have been generated using AI.
Fig. 4: Plofsomes fuse with the plasma membrane when receiving H2O2 stimulation and deliver siRNA into the cytoplasm.
The alternative text for this image may have been generated using AI.
Fig. 5: Antitumour efficacy of Plofsome@siRNA in an orthotopic LN229R-Luc tumour-bearing mouse model.
The alternative text for this image may have been generated using AI.
Fig. 6: Antitumour efficacy of Plofsome@RNP in an orthotopic LN229R-Luc tumour-bearing mouse model.
The alternative text for this image may have been generated using AI.

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Data availability

Human genome 38 (hg38, https://hgdownload.soe.ucsc.edu/goldenPath/hg38/bigZips/) was used in this study. Transcriptome expression profiling and clinical information data from patients diagnosed with glioma were obtained from TCGA website (https://www.cancer.gov/ccg/research/genome-sequencing/tcga) and cBioPortal (https://www.cbioportal.org/). The gene expression profiling of parental and TMZ-resistant glioma cells was obtained from the GSE113510 dataset. The raw data of single RNA sequencing can be found on the Gene Expression Omnibus at accession number (GSE271379). Source data are provided with this paper.

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Acknowledgements

This study was supported by the (1) National Natural Science Foundation of China (numbers 82372901 and 82073298 for J.C., numbers 82003022 and 82272988 for X.M., number 82230086 for C.J. and number 22007051 for Y.Z.), (2) Heilongjiang Provincial Key R&D Project (number GA21C002 for J.C. and number 2022ZX06C18 for C.J.), (3) Heilongjiang Provincial Key Project of The Educational Science 14th Five-Year Plan (number GJB1422780 for J.C.), (4) Heilongjiang Provincial Natural Science Foundation (number LH2022H022 for X.M.), (5) China Postdoctoral Science Foundation (numbers 2019M660074 and 2022T150173 for X.M.), (6) Heilongjiang Postdoctoral Science Foundation (numbers LBH-Z19029 and LBH-TZ2218 for X.M.) and (7) Harbin Medical University Marshal Initiative Funding (number HMUMIF-22009 for X.M.).

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  1. These authors contributed equally: Yu Zhao, Jie Qin.

Authors and Affiliations

  1. Department of Neurosurgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, China

    Yu Zhao, Jie Qin, Daohan Yu, Yuxiang Liu, Dan Song, Kaifu Tian, Hao Chen, Qile Ye, Xinyu Wang, Tianye Xu, Hanwen Xuan, Nan Sun, Wenbin Ma, Junzhe Zhong, Penggang Sun, Yu Song, Jingze Hu, Yunlei Zhao, Xintong Hou, Xiangqi Meng, Chuanlu Jiang & Jinquan Cai

  2. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York, USA

    Yu Zhao

  3. The Sixth Affiliated Hospital of Harbin Medical University, Harbin, China

    Chuanlu Jiang

  4. Key Laboratory of Functional Polymer Materials of Ministry of Education, State Key Laboratory of Medicinal Chemical Biology, College of Chemistry, Nankai University, Tianjin, China

    Yu Zhao

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Contributions

Yu Zhao, J.Q., X.M., C.J. and J.C. conceived the idea, designed the study, analysed and interpreted the data, and wrote the paper. D.Y., Y.L. and D.S. helped perform the in vivo experiment. K.T., H.C., Q.Y., X.W., T.X., H.X., N.S., W.M., J.Z., P.S., Y.S., J.H., Yunlei Zhao and X.H. helped collect tumour tissues. All authors discussed the results and commented on the paper.

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Correspondence to Yu Zhao, Xiangqi Meng, Chuanlu Jiang or Jinquan Cai.

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Zhao, Y., Qin, J., Yu, D. et al. Polymer-locking fusogenic liposomes for glioblastoma-targeted siRNA delivery and CRISPR–Cas gene editing. Nat. Nanotechnol. 19, 1869–1879 (2024). https://doi.org/10.1038/s41565-024-01769-0

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