The DNA barcode system enables high-throughput in vivo screening of mitochondrial-targeted gold nanoparticles. Thirty types of nanoparticles with different shapes, sizes, and ligands were individually barcoded and combined. The combined library was applied to tumor-bearing mice, and the barcodes from mitochondrial extracts were amplified and sequenced to quantify the relative enrichment of each nanoparticle.
This study was published in Advanced Materials under the title "High-Throughput In Vivo Subcellular Analysis of Gold Nanoparticles for Tumor Mitochondrial Targeting".
This study developed a DNA barcode screening system to quickly identify which gold nanoparticles are best at targeting tumor mitochondria. The researchers labeled 30 different types of gold nanoparticles with different DNA barcodes (balls, rods, triangles, cubes, double cones), mixed them, and injected them into tumor-bearing mice. After 24 hours, the barcodes in tumor mitochondria were extracted and sequenced, allowing for a one-time assessment of the mitochondrial targeting efficiency of all nanoparticles. It was found that the 80 nm cube (CL-FA) and 80 nm sphere (PL-FA) performed the best, and they took different routes - the cube mainly entered the cell through endocytosis mediated by clathrin and specific membrane curvature sensing proteins, while the sphere extended the circulation time by adsorbing specific protein caps, indirectly increasing tumor accumulation. By attaching CL-FA with siRNA targeting the mitochondrial ATP6 gene and combining it with mild photothermal therapy (42-48°C), a single treatment reduced the tumor by 99%, and some mice had their tumors completely disappear. Surprisingly, CL-FA was also extensively engulfed by tumor-associated macrophages, shifting the immunosuppressive M2-type macrophages to the pro-inflammatory M1-type, effectively killing the tumor while modulating the immune response. This high-throughput screening method saved 30 times less mice than traditional methods, and the results for subcutaneous and orthotopic tumors were highly consistent, with no differences in the left and right breast tumors, providing a very practical platform for the precise screening of subcellular-targeting nanomedicines. References:
DOI: 10.1002/adma.202517706