Self-Assembled DNA–Protein Hybrid Nanospheres: Biocompatible Nano-Drug-Carriers for Targeted Cancer Therapy

Abstract

We developed hybrid nanospheres comprised of two of the most important biomolecules in nature, DNA and proteins, which have excellent biocompatibility, high drug payload capacity, in vivo imaging ability, and in vitro/in vivo cancer targeting capability. The synthesis can be done in a facile one-pot assembly system that includes three steps: step-growth polymerization of two DNA oligomers, addition of streptavidin to assemble spherical hybrid nanostructures, and functionalization of hybrid nanospheres with biotinylated aptamers. To test the feasibility of cancer targeting and drug-loading capacity of the hybrid nanospheres, MUC1-specific aptamers (MA3) were conjugated to nanosphere surfaces (apt-nanospheres), and doxorubicin (Dox) was loaded into nanospheres by DNA intercalation. The successful construction of nanospheres and apt-nanospheres was confirmed by agarose gel electrophoresis and dynamic light scattering (DLS). Their uniform spherical morphology was confirmed by transmission electron microscopy (TEM). Fluorescence spectra of nanospheres demonstrated high Dox-loading capability and slow-release characteristics. In vitro MUC1-specific binding of the apt-nanospheres was confirmed by flow cytometry and confocal microscopy. Dox-loaded apt-nanospheres significantly increased cytotoxicity of the MUC1-positive cancer cells due to aptamer-mediated selective internalization, as shown via cell viability assays. Apt-nanospheres could also be imaged in vivo through the synthesis of hybrid nanospheres using fluorescent dye-conjugated DNA strands. Finally, in vivo specific targeting ability of apt-nanospheres was confirmed in a MUC1-positive 4T1 tumor-bearing mouse model, whereas apt-nanospheres did not cause any sign of systemic toxicity in normal mice. Taken together, our self-assembled DNA–streptavidin hybrid nanospheres show promise as a biocompatible cancer targeting material for contemporary nanomedical technology.

Nucleolin-Targeting AS1411 Aptamer-Conjugated Nanospheres for Targeted Treatment of Glioblastoma

“…After the formation of a linear DNA chain, STV and an equivalent amount of DNA monomer were mixed, stored at 4 °C for 3 h, and then used. The complementary strand of sequences and synthesis conditions were performed as previously reported [ 35 , 64 ]. FAM-labeled nanospheres were used as follows: 5′-FAM-ACGGCTGCGCGACGTAGGTACGGCAACTCGCGGCTATGCA-3′.…”

“…Here, we apply previous research techniques to synthesize nanocarriers using only biopolymers that induce biotin–streptavidin molecular binding based on DNA structures. The objective of this innovation is to fabricate hybrid nanocarriers by employing the probabilistic pairing of streptavidin (STV) and double-stranded DNA (dsDNA) long chains [ 35 ]. STV, existing as homotetramers, exhibits the capacity to bind with up to four biotin units [ 36 ].…”

“…An effective nanocarrier design encompasses considerations of size and ζ potential. Building upon our previous work, we utilized a hybrid nanosphere composed of DNA and streptavidin in this study, anticipating enhanced suspension stability with a near-zero ζ potential [ 35 ]. Consistent with our prior findings, we expected the incorporation of streptavidin into dsDNA to align the nanodimer′s ζ potential with streptavidin, and the inclusion of aptamer and doxorubicin (Dox) to further decrease the ζ potential, facilitating tumor targeting.…”

Dual-Oxygenation/Dual-Fenton Synergistic Photothermal/Chemodynamic/Starvation Therapy for Tumor Treatment

“…Lack of O 2 in the TME limits the efficacy of PDT, SDT, and ST, and researchers have investigated various strategies to increase O 2 , such as applying hemoglobin 33,34 and O 2 -saturated perfluoropentane 35,36 as O 2 donors to increase the O 2 concentration in the TME, thus improving the efficacy of PDT, SDT, and ST. Good tumor therapies/drug delivery systems all require the ability to target the treatment site. 37,38 CD44 receptor is highly expressed on a variety of tumor cells, and the specific binding between hyaluronic acid (HA) and CD44 receptor is commonly used to design drug delivery systems or nanotherapeutic platforms that provide the ability to target tumor cells. 39 Furthermore, HA plays an important role in biomedical, clinical, and pharmaceutical applications because of its favorable properties, including water solubility, biocompatibility, nonimmunogenicity, biodegradability, and structural similarity to tissue components.…”

“…Good tumor therapies/drug delivery systems all require the ability to target the treatment site. , CD44 receptor is highly expressed on a variety of tumor cells, and the specific binding between hyaluronic acid (HA) and CD44 receptor is commonly used to design drug delivery systems or nanotherapeutic platforms that provide the ability to target tumor cells . Furthermore, HA plays an important role in biomedical, clinical, and pharmaceutical applications because of its favorable properties, including water solubility, biocompatibility, nonimmunogenicity, biodegradability, and structural similarity to tissue components.…”