Linear-branched poly(β-amino esters)/DNA nano-polyplexes for effective gene transfection and neural stem cell differentiation

Abstract

Controllable regulation of stem cell differentiation is a critical concern in stem cell-based regenerative medicine. In particular, there are still great challenges in controlling the directional differentiation of neural stem cells (NSCs) into neurons. Herein, we developed a novel linear-branched poly(β-amino esters) (S4-TMPTA-BDA-DT, STBD) through a two-step reaction. The synthesized STBD linear branched polymers possess multiple positively charged amine terminus and degradable intermolecular ester bonds, thus endowing them with excellent properties such as high gene load, efficient gene delivery, and effective gene release and transcription in cells. In the mCherry transfection test, a high transfection efficiency of approximately 70% was achieved in primary NSCs after a single transfection. Moreover, STBD also showed high biocompatibility to NSCs without disturbing their viability and neural differentiation. With the high gene delivery property, STBD is capable of delivering siRNA (shSOX9) expression plasmid into NSCs to significantly interfere with the expression of SOX9, thus enhancing the neuronal differentiation and maturation of NSCs. The STBD/DNA nano-polyplex represents a powerful non-viral approach of gene delivery for manipulating the differentiation of stem cells, showing broad application prospects in NSC-based regenerative therapy for treating neurodegenerative diseases.

A special issue focusing on CAS key laboratory of nano-bio interface at Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO)

“…In this special issue, Cao et al developed a novel linear-branched poly (β-amino esters) (called STBD) nanocarrier with multiple positively charged amine terminus and degradable intermolecular ester bonds for efficient gene loading and delivery, and effective gene release and transcription in neural stem cells (NSCs). Thereby, the levels of neuronal differentiation and maturation of NSCs were significantly enhanced after delivering siRNA (shSOX9) expression plasmid into NSCs for silencing the SOX9 gene in NSCs by the STBD nanocarrier [9]. The STBD/deoxyribonucleic acid nano-polyplex may serve as a powerful nonviral approach for gene delivery in NSCs, showing broad application prospects in NSC-based regenerative medicine.…”

Nontoxic, Biodegradable Hyperbranched Poly(β-amino ester)s for Efficient siRNA Delivery and Gene Silencing

“…A hyperbranched polymer formulation suitable for DNA delivery might not be effective for siRNA delivery, possibly due to differences in their chemical/physical properties and mechanisms of action . Slight structure modifications within pBAEs can significantly affect their efficiency in delivering different nucleic acids. , Therefore, identifying the optimal hyperbranched pBAE polymer formulation for siRNA delivery is crucial. Incorporating optimal monomer combinations and hyperbranching architecture in the pBAE polymer shall achieve efficient gene silencing at a lower polymer/siRNA weight ratio or reduced siRNA dose.…”

“…Numerous studies have demonstrated that tuning the polymer structure of pBAEs from linear to a hyperbranched architecture can significantly enhance gene transfection efficiency when delivering plasmid DNA to various cell types, including cancer cells, 37−39 skin cells, 38,40,41 retinal pigment epithelial cells, 42 and stem cells. 43,44 This enhancement may be attributed to hyperbranched pBAEs providing a higher number of tertiary amine groups and terminal end groups, thus promoting better DNA condensation due to their increased cationic charge density and enhanced molecular flexibility. 45 We hypothesize that a hyperbranched architecture could address the limitation of the current pBAE polymer in siRNA delivery by reducing the required amount of polymer carrier or siRNA dose for efficient gene silencing.…”