利用层层包被策略开发靶向EGFR阳性膀胱癌细胞的弹性蛋白样多肽核酸递送系统

Development of an Elastin-like Polypeptide-Based Nucleic Acid Delivery System Targeted to EGFR+ Bladder Cancer Cells Using a Layer-by-Layer Approach

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BIOMACROMOLECULES

影响因子:5.4

分区:化学1区 Top / 有机化学1区高分子科学1区生化与分子生物学2区

发表日期:2024 Sep 09

作者: Aayush Aayush, Saloni Darji, Kiera M Estes, Emily Yeh, David H Thompson

DOI: 10.1021/acs.biomac.4c00165

摘要

核酸（NA）类疗法因其能在基因水平上调控细胞功能，正在革新生物医学研究。本研究展示了一种多功能弹性蛋白样多肽（ELP）载体系统，采用层层包被（LbL）制剂策略，可递送从siRNA到质粒的不同大小的核酸货物。该系统的组成可以重新配置，以调节载体的生物化学和生物物理特性，从而匹配目标的独特生物特征。我们对Murine和人膀胱肿瘤细胞系中的LbL ELP核酸纳米粒子（LENNs）进行了物理表征和生物性能评估。由于膀胱持续的尿液流入，药物在膀胱中的接触时间通常不足2小时，给局部灌注递送带来挑战。研究发现，LENNs在30分钟内即可结合膀胱肿瘤细胞，并在60分钟内被快速内吞，释放核酸货物。结果表明，已建立了一种具有高度适应性和灵活性的核酸递送系统，能调控目标能力、货物大小及解离动力学。此方法为替代脂质纳米粒或病毒载体提供了新途径，后者常因效率低、物理化学稳定性差或制造及免疫排斥而受限。这种弹性蛋白样纳米载体具有生物制造、可降解、生物相容和高度可调的优点，能通过与过表达的细胞表面受体结合，实现靶向递送。

Abstract

Nucleic acid (NA)-based therapies are revolutionizing biomedical research through their ability to control cellular functions at the genetic level. This work demonstrates a versatile elastin-like polypeptide (ELP) carrier system using a layer-by-layer (LbL) formulation approach that delivers NA cargos ranging in size from siRNA to plasmids. The components of the system can be reconfigured to modulate the biochemical and biophysical characteristics of the carrier for engaging the unique features of the biological target. We show the physical characterization and biological performance of LbL ELP nucleic acid nanoparticles (LENNs) in murine and human bladder tumor cell lines. Targeting bladder tumors is difficult owing to the constant influx of urine into the bladder, leading to low contact times (typically <2 h) for therapeutic agents delivered via intravesical instillation. LENN complexes bind to bladder tumor cells within 30 min and become rapidly internalized to release their NA cargo within 60 min. Our data show that a readily adaptable NA-delivery system has been created that is flexible in its targeting ability, cargo size, and disassembly kinetics. This approach provides an alternative path to either lipid nanoparticle formulations that suffer from inefficiency and physicochemical instability or viral vectors that are plagued by manufacturing and immune rejection challenges. This agile ELP-based nanocarrier provides an alternative route for nucleic acid delivery using a biomanufacturable, biodegradable, biocompatible, and highly tunable vehicle capable of targeting cells via engagement with overexpressed cell surface receptors.