Poly(amino acid)-based Delivery Systems for Genome Editing and Therapeutic Efficacy Evaluation in Circulating Malignant Cells

Li-jin Qi; Di Han; Xiao-he Ren; Xiao-yan He; Tao Guo; Xian-zheng Zhang; Si-xue Cheng

doi:10.11777/j.issn1000-3304.2024.24196

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Poly(amino acid)-based Delivery Systems for Genome Editing and Therapeutic Efficacy Evaluation in Circulating Malignant Cells

Research Article | 更新时间：2025-01-24

- Poly(amino acid)-based Delivery Systems for Genome Editing and Therapeutic Efficacy Evaluation in Circulating Malignant Cells
- Acta Polymerica Sinica Vol. 56, Issue 2, Pages: 253-265(2025)
- 作者机构：
  
  1.生物医用高分子材料教育部重点实验室武汉大学化学与分子科学学院武汉 430072
  2.安徽医科大学生命科学学院合肥 230011
  3.安徽医科大学第一附属医院合肥 230011
- 作者简介：
  
  Si-xue Cheng, E-mail: chengsixue@whu.edu.cn
- 基金信息：
- DOI：10.11777/j.issn1000-3304.2024.24196
  CLC：
- CSTR：32057.14.GFZXB.2024.7292
- Published：20 February 2025，
  
  Published Online：22 November 2024，
  
  Received：14 July 2024，
  
  Accepted：2024-09-06
- 稿件说明：
移动端阅览
LI-JIN QI, DI HAN, XIAO-HE REN, XIAO-YAN HE, TAO GUO, XIAN-ZHENG ZHANG, SI-XUE CHENG. Poly(amino acid)-based Delivery Systems for Genome Editing and Therapeutic Efficacy Evaluation in Circulating Malignant Cells. [J]. Acta polymerica sinica, 2025, 56(2): 253-265.
DOI：

LI-JIN QI, DI HAN, XIAO-HE REN, XIAO-YAN HE, TAO GUO, XIAN-ZHENG ZHANG, SI-XUE CHENG. Poly(amino acid)-based Delivery Systems for Genome Editing and Therapeutic Efficacy Evaluation in Circulating Malignant Cells. [J]. Acta polymerica sinica, 2025, 56(2): 253-265. DOI： 10.11777/j.issn1000-3304.2024.24196. CSTR： 32057.14.GFZXB.2024.7292.

摘要

肿瘤异质化和每位患者的特异性是肿瘤研究和治疗中的重要挑战，而个性化精准诊疗是解决这一挑战的关键途径之一. 本研究中，设计制备了基于聚氨基酸的肿瘤靶向传递体系，利用

-聚-

-赖氨酸负载敲除促进肿瘤发展的黏蛋白1 (MUC1)的CRISPR-Cas9质粒，外层修

饰键接了AS1411适配子的透明质酸，通过靶向肿瘤细胞表面过表达的核仁素(nucleolin)和CD44实现对肿瘤细胞的特异性高效传递和基因编辑. 进一步，利用这一靶向传递载体负载检测MUC1、肺转移标志物组织蛋白酶C (CTSC)、骨转移标志物锯齿状典型Notch配体1 (JAG1)的mRNA的3种分子信标，对基因编辑效果进行检测. 针对肿瘤细胞系BT549及乳腺癌患者外周血中的循环恶性细胞开展了基因编辑及疗效评估研究，结果表明，基因编辑传递体系可以有效靶向恶性细胞，敲除MUC1基因，抑制MUC1的表达、显著下调肺转移和骨转移标志物. 本研究利用少量全血进行个性化体外研究，可便捷高效地评估特定治疗措施的疗效，为肿瘤个性化精准治疗提供依据和参考.

Abstract

Tumor heterogeneity and the individual specificity of each patient pose a significant challenge in tumor research and treatment. Personalized precision therapy is one of the key approaches to address this challenge. In this study

tumor targeting delivery systems based on

-poly-L-lysine (

-PL) for tumor therapy and diagnosis were designed and prepared. In the therapeutic system

-PL was used to complex with the CRISPR-Cas9 plasmid for knocking out mucin 1 (MUC1)

a protein promotes tumor development

and then the complexes were decorated with AS1411 conjugated hyaluronic acid. The presence of AS1411 and hyaluronic acid chain facilitated the tumor targeting delivery of the genome editing plasmid into nucleolin and/or CD44 overexpressed tumor cells. Moreover

this target

ed delivery vector was loaded with three types of molecular beacons to detect mRNAs of MUC1

cathepsin C (CTSC

a lung metastasis biomarker)

and Notch ligand Jagged1 (JAG1

a bone metastasis biomarker)

thereby evaluating the therapeutic effects of gene editing.

In vitro

and

ex vivo

studies were conducted in the BT549 tumor cell line and circulating malignant cells from breast cancer patients. The results demonstrate that the gene editing delivery system effectively targets malignant cells

leading to knockout of MUC1 gene and downregulation of MUC1 expression. Furthermore

the genome edited malignant cells show markedly reduced mRNA biomarkers associated with lung and bone metastasis

suggesting MUC1 knockout is a promising strategy for inhibiting tumor progression and metastasis. By employing a small volume of whole blood for personalized

ex vivo

studies

this research offers a safe and convenient method for effectively evaluating the therapeutic efficacy of a specific intervention. This approach is also applicable for evaluating other treatment modalities as well as detecting various nucleic acid biomarkers

thereby offering precise information for personalized therapy.

关键词

聚氨基酸肿瘤靶向传递载体基因治疗mRNA检测

Keywords

Poly(amino acid)Tumor targetingDelivery vectorGene therapymRNA detection

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