DNA flowerstructure co-localizes with human pathogens in infected macrophages

Area of Science:

• Biotechnology
• Nanomedicine
• Cellular Biology

Background:

• DNA structures offer novel therapeutic possibilities.
• Understanding cellular uptake mechanisms is crucial for nanomedicine development.
• Macrophage-targeted delivery is key for treating intracellular pathogens.

Purpose of the Study:

• To characterize the cellular uptake and intracellular fate of DNA nanostructures.
• To evaluate the potential of DNA nanostructures for treating infectious diseases.
• To investigate the interaction of DNA nanostructures with pathogenic bacteria and parasites.

Main Methods:

• Rolling circle DNA amplification to create nanoflowers.
• Fluorescent labeling (ATTO488-dUTP) for intracellular tracking.
• Mammalian cell culture, including macrophages, for uptake studies.
• Confocal microscopy to determine localization and co-localization.
• Viability assays to assess nanostructure toxicity.

Main Results:

• DNA nanostructures (nanoflowers) of ~300 nm diameter were synthesized.
• Nanoflowers exhibited non-toxicity across tested mammalian cell lines.
• Specific uptake by macrophages via phagocytosis was observed within hours.
• Nanoflowers compartmentalized in phagolysosomes with a half-life of 12 hours.
• Co-localization with Mycobacterium tuberculosis and Leishmania infantum was confirmed.

Conclusions:

• DNA nanostructures demonstrate efficient and specific uptake by macrophages.
• Nanoflowers remain stable within phagolysosomes, interacting with intracellular pathogens.
• These findings highlight a potential application of DNA nanostructures in targeted therapies against tuberculosis and leishmaniasis.