Journal of
Materials Chemistry B
Trehalose-polyamine/DNA nanocomplexes:
impact of vector architecture on cell and organ
transfection selectivity†
A novel family of precision-engineered gene vectors with well-defined structures built on trehalose and
trehalose-based macrocycles (cyclotrehalans) comprising linear or cyclic polyamine heads have been
synthesized through procedures that exploit click chemistry reactions. The strategy was conceived to
enable systematic structural variations and, at the same time, ensuring that enantiomerically pure
vectors are obtained. Notably, changes in the molecular architecture translated into topological
differences at the nanoscale upon co-assembly with plasmid DNA, especially regarding the presence of
regions with short- or long-range internal order as observed by TEM. In vitro and in vivo experiments
further evidenced a significant impact on cell and organ transfection selectivity. Altogether, the results
highlight the potential of trehalose-polyamine/pDNA nanocomplex monoformulations to achieve
targeting transfection without the need for any additional cell- or organ-sorting component.
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BSV163/DOPE-mediated TRAIL gene transfection acts
synergistically with chemotherapy against cisplatinresistant ovarian cancer
Abstract
Ovarian cancer is the seventh most frequently diagnosed cancer among women
worldwide. Most patients experience recurrence and succumb eventually to resistant
disease, underscoring the need for an alternative treatment option. In
the presented manuscript, we investigated the effect of the TRAIL-gene,
transfected
by an innovative bioinspired lipid vector BSV163/DOPE in the presence
or absence of cisplatin, to fight against sensitive and resistant ovarian cancer.
We showed that BSV163/DOPE can transfect ovarian cancer cell lines (Caov3,
OVCAR3, and our new cisplatin-resistant,
CR-Caov3)
safely and efficiently. In
addition, TRAIL-gene
transfection in association with cisplatin inhibited cellular
growth more efficiently (nearly 50% in Caov3 cells after the combined treatment,
and 15% or 25% by each treatment alone, respectively) owing to an increase in
apoptosis rate, caspases activity and TRAIL’s death receptors expression. Most
importantly, such synergistic effect was also observed in CR-Caov3
cells demonstrated
by an apoptosis rate of 35% following the combined treatment in comparison
with 17% after TRAIL-gene
transfection or 6% after cisplatin exposition.
These results suggest this combination may have potential application for sensitive
as well as refractory ovarian cancer patients.
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KEYWORDS
BSV163/DOPE, cisplatin-resistance,
lipid vector, ovarian cancer, TRAIL, transfection
An efficient low cost means
of biophysical gene transfection
in primary cells
Efficient, facile gene modification of cells has become an indispensable part of modern molecular
biology. For the majority of cell lines and several primary populations, such modifications can be
readily performed through a variety of methods. However, many primary cell lines such as stem
cells frequently suffer from poor transfection efficiency. Though several physical approaches have
been introduced to circumvent these issues, they often require expensive/specialized equipment
and/or consumables, utilize substantial cell numbers and often still suffer from poor efficiency. Viral
methods are capable of transducing difficult cellular populations, however such methods can be time
consuming for large arrays of gene targets, present biohazard concerns, and result in expression
of viral proteins; issues of concern for certain experimental approaches. We report here a widely
applicable, low-cost (< $100 CAD) method of electroporation, applicable to small (1–10 μl) cell
volumes and composed of equipment readily available to the average investigator. Using this system
we observe a sixfold increase in transfection efficiency in embryonic stem cell lines compared to
commercial devices. Due to efficiency gains and reductions in volume and applied voltage, this process
improves the survival of sensitive stem cell populations while reducing reagent requirements for
protocols such as Cas9/gRNAs transfections.
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