Sopida Pimcharoen

All Publications

• CRISPR-Cas13d-Mediated Targeting of a Context-Specific Essential Gene Enables Selective Elimination of Uveal Melanoma. bioRxiv : the preprint server for biology Stauber, D., Sosnick, L., Ma, Y., Pimcharoen, S., Lawanprasert, A., Murthy, N., Myung, D., Qi, L. S. 2025

  Abstract

  Uveal melanoma, the most common eye cancer in adults, remains limited to surgical intervention and chemotherapy, with a dismal survival rate that has not improved in over 50 years. To address this therapeutic impasse, we systematically analyzed public gene expression, RNAi, and CRISPR knockout datasets and identified RASGRP3 as an essential gene specifically for uveal melanoma. RasGRP3 is uniquely overexpressed and essential for survival in uveal melanoma cells, but dispensable in healthy cells. RasGRP3 remains "undruggable" due to its intracellular localization and lack of targetable binding pockets. To overcome this, we developed a CRISPR-Cas13d RNA-targeting therapeutic that specifically knocks down RasGRP3 mRNA. This Cas13d-based therapeutic mediates selective uveal melanoma killing through two synergistic mechanisms: (i) direct silencing of the essential RasGRP3 transcript, and (ii) collateral RNA degradation triggered by the cleavage of overexpressed RasGRP3. When delivered via optimized lipid nanoparticles encoding Cas13d mRNA and guide RNA, this strategy eliminated >97% of uveal melanoma cells while sparing healthy cells, including retinal pigment epithelial cells. This approach outperformed conventional Cas9 and siRNA methods in potency without inducing permanent genomic alterations. Our findings establish a RNA-targeting therapeutic for uveal melanoma and a framework for Cas13d-based interventions against broad "undruggable" cancers.

  View details for DOI 10.1101/2025.08.21.671629

  View details for PubMedID 40894775

  View details for PubMedCentralID PMC12393512

• Acid-degradable lipid nanoparticles enhance the delivery of mRNA. Nature nanotechnology Zhao, S., Gao, K., Han, H., Stenzel, M., Yin, B., Song, H., Lawanprasert, A., Nielsen, J. E., Sharma, R., Arogundade, O. H., Pimcharoen, S., Chen, Y. J., Paul, A., Tuma, J., Collins, M. G., Wyle, Y., Cranick, M. G., Burgstone, B. W., Perez, B. S., Barron, A. E., Smith, A. M., Lee, H. Y., Wang, A., Murthy, N. 2024

  Abstract

  Lipid nanoparticle (LNP)-mRNA complexes are transforming medicine. However, the medical applications of LNPs are limited by their low endosomal disruption rates, high toxicity and long tissue persistence times. LNPs that rapidly hydrolyse in endosomes (RD-LNPs) could solve the problems limiting LNP-based therapeutics and dramatically expand their applications but have been challenging to synthesize. Here we present an acid-degradable linker termed 'azido-acetal' that hydrolyses in endosomes within minutes and enables the production of RD-LNPs. Acid-degradable lipids composed of polyethylene glycol lipids, anionic lipids and cationic lipids were synthesized with the azido-acetal linker and used to generate RD-LNPs, which significantly improved the performance of LNP-mRNA complexes in vitro and in vivo. Collectively, RD-LNPs delivered mRNA more efficiently to the liver, lung, spleen and brains of mice and to haematopoietic stem and progenitor cells in vitro than conventional LNPs. These experiments demonstrate that engineering LNP hydrolysis rates in vivo has great potential for expanding the medical applications of LNPs.

  View details for DOI 10.1038/s41565-024-01765-4

  View details for PubMedID 39179796

  View details for PubMedCentralID 10190134