Discoveries at the Nano/Bio interface as the impetus to innovation in Nano Safety and Nano Therapeutics André Nel M.B.,Ch.B; Ph.D Professor of Medicine and Chief of the Division of NanoMedicine at UCLA Director of the NSF and EPA funded Center for the Environmental Implications of Nanotechnology (UC CEIN) Director of the NIEHS funded Center for NanoBiology and Predictive Toxicology Co Director UCLA NanoMacchine Center NanoMachine Center Copyright 2010 The Regents of the University of California. All Rights Reserved. Contact cein@cnsi.ucla.edu to obtain permission to use copyrighted material.
The Nano/Bio interface is an Evolutionary conserved Toolbox ready to be exploited for rapid advances in NanoMedicine Nano-bio interface Evolutionary conserved Much of life conducted at Nanoscale level Intrinsic material ENM properties Biopoesis on nanoscale platform Cell membrane Nel et Nature Mat. 2009 A protein interacting with ENM surface architectures A protein corona as the initial Nano/bio interface Multiple Nano/bio interfaces at cellular level
Examples of Nanomedicine Platform Development Nanotechnology Long-term Impacts and Research Directions: 2000 2020 http://www.wtec.org/nano2/ Nanomaterial Synthesis and Device Design Diagnostic and Imaging Tools Targeted Therapy and Drug Delivery Systems Tissue Engineering Regenerative Medicine High sensitivity Fast processing Multiplexing Portable/Lab on a chip Non-Invasive structure and function imaging Hydrophobic drugs Targeted Delivery Controlled delivery Theranostics Nucleic acid delivery Drug/siRNA delivery Architectures ECM Mimics Pro-Morphogens Cell sheet engineering and repair Stem cell differentiation Point of Medical Care: Improved diagnosis, treatment and prevention
Vision for Nanomedicine in the Next Decade Screening Diagnosis & Staging Treatment & Monitoring Followup Current Relatively non-specific biomarkers Limited number Macroscale imaging Established disease Macroscale site imaging Diagnostic surgery/biopsy Intervention Batch testing Insensitive biomarkers Surgery Catheter lab Radiation Debilitating chemo Total body effects Macroscale site imaging Relatively non-specific markers Relatively late Intervals Nano Impact Specific markers Large number lab-on- a-chip Molecular whole body imaging Disease inception Image structure & function Whole body Non-invasive Sensitivity (femto/attomol) Tissue targeting Targeted drug delivery On-demand drug delivery Imaging while delivering Non-invasive quantitative imaging Specific molecular markers Early Continuous
How do we Speed up Discovery that allows the Nano Toolbox to deliver more, better and safer ENM? Response: In part through our ability to translate gazillions of possibilities at the Nano/bio interface.. e - Nel et al Nature Mat. 2009 Cell membrane Proteins DNA and nucleus Cell interior (organelles etc) h Electronic states dissolution Material composition Hydrophobic/ philic Charge & Surface functions crystallinity Size curvature angle into structure-activity relationships defining the unique ENM properties advantageous biological outcomes unique ENM properties biological hazard and safe design options
Key Challenge to Nanotechnology: Human and Environmental Safety where do we stand? No known human disease or serious environmental impact Currently, 6 base materials constitute >90% of nano products Presently > 10 3 consumer nano products >10 4 by 2020 > 10 5 ultimately? Definitive experimental evidence of ENM hazard Incomplete knowledge about ENM properties that can lead to hazard and risk Need for a scientific platform that can generate knowledge more rapidly to be commensurate with the growth of nanotechnology Ti/TiO 2 Silica Zn/ZnO Au 6% 6% Carbon Ag
Platform for Knowledge Generation at the nano/bio interface In Vivo (Whole Animal) Screening 10 2 animals per experiment (weeks to months) Validity of predictions Material physicochemical properties Structure/ Activity relationships Structure/ Activity relationships mechanisms of injury bio-pathways/networks Cellular or Bio-molecular Injury Endpoints Up to 10 5 measurements per day Nel et al Science 2006. Huan Meng et al ACS Nano. 2009
Pharma s Approach to the use of In Vitro Discovery to Develop new Therapies in Humans Cellular Systems for Predictions Butcher et al. Nature Biotech. 2004 Pathway analysis and clustering by drug function Genomic analysis and clustering by Gene function Network architecture and connectivity Uses: Target ID and validation Network/pathway organization Structure-activity relationships Uses: Target and compound validation Lead optimization
The In Vitro/In Vivo Platform being used by the UC Center for the Environmental of Nanotechnology to speed up knowledge generation for ENM hazard.. 100 nm 350 1000 100 600 300 80 800 80 500 Number of Particles 250 200 150 100 50 Number of Particles 60 40 20 Number of Particles 600 400 200 Number of Particles 60 40 20 Y Axis Title 400 300 200 100 0 20 25 30 35 40 45 Size (nm) 0 6 8 10 12 14 16 18 20 22 24 Size (nm) 0 0 5 10 15 20 25 Size (nm) 0 8 10 12 14 16 18 Size (nm) 0 8 10 12 14 16 18 20 22 24 26 X Axis Title Producing material libraries to study property-activity relationships Using biological Pathways & networks Using high throughput approaches Using computational power to enhance and speed up decision-making Use of this exercise for hazard ranking, risk analysis and building a sustainable technology
2 1 Injury Paradigms for High Throughput Analysis Cationic Lysosome Cationic toxicity Organellar damage S S 4 3 SH SH O 2 h O e 2 Redox activity Oxidant injury Inflammazone assembly 5 MeO, QDs Dissolution and Metal Ion Tox Immunogenicity Inflammation Membrane disruption Nel et al. Science. 2006 Nel et al. Nature Materials. 2009
Compositional and Property-based Nanomaterial Libraries to make high throughput discoveries at the nano-bio interface Compositions Me Oxides TiO 2, CeO 2, ZnO Sizes d=10 nm Cube Shapes Combinatorial Library: Me & MeO Property variation by doping Metals d=30 nm Rod solubility Au, Ag, Pt, Co Carbon Nanotubes SWCNT, MWCNT Silica Amorphous, Crystalline, Mesoporous d=130 nm Wire Band gap metal doping Redox activity photoactivation 50 nm FSP TiO 2, ZnO
High Throughput Screening using Injury Pathways, Cellular circuits, Networks Robotic automation Cell culture, yeast, Simple organisms Endocytosis Proton sponge Membrane damage O 2.- Lysosome [Ca 2 ] i Mitochondria Toxic ions Cytotoxicity O 2.- Caspase 9 Energy failure Control Trt Fluoresence and luciferase Imaging
Use of in silico Approaches for Data Transformation, Clustering, Display, Predictions (Nano-informatics) MeO, QDs Heatmap Zebrafish embryo Metal Ions Fe-doped Dissolution ZnO HTS Plate Map Cell 1 Profiling Cell 2 Material Cluster Mapping George et al ACS Nano
Predictive Platform linking cellular to whole organism outcome NSF: EF 0830117 ENM libraries compositions properties Hazard profling Iterative Add new properties Change hazardous properties In vivo/animal testing disease pathogenesis pathology involving related mechanisms, pathways etc In vitro/cellular screening mechanisms pathways clusters property-activity relationships
Can the Pharma Approach for in vitro in vivo Translation be used to improve Nano-therapeutics in vivo Butcher et al. Nature Biotech. 2004
Mesoporous Silica Nanoparticles: A multi-functional platform using design features based on events taking place at the intracellular nano/bio interface Multifunctional platform The basic particle fitted with a valve hν NanoMachine Center wrapped in a polymer and serving dual functions
Fitted with a ph Responsive Cyclodextrin Valve that opens intracellularly and releases Doxirubicin 7 6.5 6 5.5 5 Valve design based on pka values of valve components and ph of intracellular compartments 5.2 6.2 4.5 Huan Meng, Min Xue et al. JACS 2010 4 Normal cells NH 4 Cl NH 4 Cl NH 4 Cl Green particles Red Dox
Dual Delivery of a Cancer Drug as well as a sirna to knock out the gene responsible for resistance to that cancer drug Pgp-siRNA Dox Pgp-siRNA Dox Dox Pgp transporter N Xia et al. ACS Nano. 2009
Design strategy tuning ENM properties and using the nano-bio interface in vitro to improve in vivo delivery Size tuning-smaller particle Pore interior adapted for cargo Functionalize surface Steric hindrance Valves Monodisperse Liganding In vitro/cell testing uptake targeting controlled delivery outcome imaging safety Iterative testing Optimize Add/tune functions Improve safety In vivo/animal testing bio-distribution limit RES uptake optimize EPR effect targeting imaging efficacy safety NanoMachine Center
Thank You Center for NanoBiology and Predictive Toxicology NanoMachine Center Jeff Zink Fuyu Tamanoi Fraser Stoddart
The Power of the Nano/bio interface at Global Level Phytoplankton NP attached to Phytoplankton Iron oxide NP on bacterium Mount Manitoba Electron transfer Phytoplankton Bloom