Department of Pharmaceutics, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA.
Department of Pharmaceutics, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA.
Biomaterials. 2018 Jan;152:1-14. doi: 10.1016/j.biomaterials.2017.10.028. Epub 2017 Oct 20.
Vectors used for stem cell transfection must be non-genotoxic, in addition to possessing high efficiency, because they could potentially transform normal stem cells into cancer-initiating cells. The objective of this research was to bioengineer an efficient vector that can be used for genetic modification of stem cells without any negative somatic or genetic impact. Two types of multifunctional vectors, namely targeted and non-targeted were genetically engineered and purified from E. coli. The targeted vectors were designed to enter stem cells via overexpressed receptors. The non-targeted vectors were equipped with MPG and Pep1 cell penetrating peptides. A series of commercial synthetic non-viral vectors and an adenoviral vector were used as controls. All vectors were evaluated for their efficiency and impact on metabolic activity, cell membrane integrity, chromosomal aberrations (micronuclei formation), gene dysregulation, and differentiation ability of stem cells. The results of this study showed that the bioengineered vector utilizing VEGFR-1 receptors for cellular entry could transfect mesenchymal stem cells with high efficiency without inducing genotoxicity, negative impact on gene function, or ability to differentiate. Overall, the vectors that utilized receptors as ports for cellular entry (viral and non-viral) showed considerably better somato- and genosafety profiles in comparison to those that entered through electrostatic interaction with cellular membrane. The genetically engineered vector in this study demonstrated that it can be safely and efficiently used to genetically modify stem cells with potential applications in tissue engineering and cancer therapy.
用于干细胞转染的载体必须是非遗传毒性的，除了具有高效性，因为它们可能潜在地将正常干细胞转化为起始癌症的细胞。本研究的目的是生物工程一种有效的载体，可以用于干细胞的遗传修饰而不会对体细胞或遗传造成任何负面影响。两种类型的多功能载体，即靶向和非靶向，从大肠杆菌中通过基因工程和纯化得到。靶向载体被设计为通过过表达的受体进入干细胞。非靶向载体配备了 MPG 和 Pep1 细胞穿透肽。一系列商业合成的非病毒载体和腺病毒载体被用作对照。所有载体都针对其效率和对代谢活性、细胞膜完整性、染色体畸变（微核形成）、基因调控和干细胞分化能力的影响进行了评估。本研究的结果表明，利用 VEGFR-1 受体进行细胞进入的生物工程载体可以高效转染间充质干细胞，而不会诱导遗传毒性、对基因功能或分化能力产生负面影响。总的来说，利用受体作为细胞进入端口的载体（病毒和非病毒）在体和遗传安全性方面的表现明显优于通过与细胞膜静电相互作用进入的载体。本研究中的基因工程载体表明，它可以安全有效地用于基因修饰干细胞，具有在组织工程和癌症治疗中的潜在应用。
