Thursday, May 15, 2025
5/15/2025
ONI NanoImager - PROJECT ABSTRACT Children’s Hospital Los Angeles (CHLA), an affiliate of the University of Southern California (USC), requests funds to purchase a Nanoimager (ONI, Inc.) for the Extracellular Vesicle (EV) Core Facility of The Saban Research Institute (TSRI). EVs and their artificial counterpart, lipid nanoparticles (LNPs), represent a breakthrough in several therapeutic applications including, but not limited to, regenerative medicine, vaccine development, and biomarker discovery. The successful therapeutic application of EVs and LNPs will require overcoming several technical and quality control challenges inherent to their very nature. Because of their small size, their purification requires advanced techniques that do not guarantee their structural integrity and reproducible purity. Visualization of their structure and molecular makeup has historically been limited to costly, low throughput, and technically challenging electron microscopy techniques. The ONI Nanoimager utilizes dSTORM (direct Stochastic Optical Reconstruction Microscopy), a Single-Molecule Localization Microscopy technique (SMLM), to resolve EVs and nanostructures at the single-molecule level with a resolution of 20nm. This allows the revealing of the spatial structure of single particles, including their molecular determinants. The Nanoimager will enable the researchers to develop quality control and reproducibility assays by simultaneously visualizing the structural integrity of the particles and the presence of markers that support the mode of action of novel therapeutics. There is a strong rationale for the request. First, the current instrumentation housed at the EV Core does not allow for an in-depth analysis of the particles’ quality and heterogeneity, limiting our analytical capabilities to the sizing and counting by Nanoparticle Tracking Analysis (Nanosight NS300) and to the detection of the cargo by low-resolution immunophenotyping of single EVs captured on a chip (ExoView R100, limited to EVs). Second, by instituting the first core lab in the country dedicated to EV research, CHLA has shown its commitment to supporting researchers focused on the investigation and development of EV and LNP-based therapeutics. After detailed testing of three dSTORM imaging instruments (ONI Nanoimager, AbbeLight, Bruker Vutara), we determined that the Nanoimager is optimal for the inclusion in a core lab at CHLA because of its integrated specimen preparation and analysis capabilities, and the demonstrated commitment of ONI to the development of automated workflows for EV and nanoparticle research. Lastly, the EV Core has built a solid user base of NIH-funded and other CHLA and USC researchers that will immediately benefit from this instrument. The ONI Nanoimager will be fully supported financially by CHLA and managed by Ph.D.-level personnel, with the direct assistance of the Cellular Imaging Core. This instrument will significantly benefit users at CHLA and beyond by allowing us to characterize the composition of EVs and meet the long- term objective of deciphering the mechanisms of pediatric disease.
HHS’ Tracking Accountability in Government Grants System (TAGGS) website provides access to detailed descriptions of grants, loans, aggregated direct payments and other types of financial assistance awarded by the HHS Operating Divisions (OPDIVs) and the Office of the Secretary Staff Divisions (STAFFDIVs).