BME 695N Lecture 20: GMP and issues of quality control manufacture of nanodelivery systems
15 Nov 2007 | Online Presentations | Contributor(s): James Leary
- What does cGMP mean?
- Why GMP? Controlling processes means more predictable outcomes…
- What can be learned from the semi-conductor industry clean-room and manufacturing?
- What doesn’t fit this paradigm?
- cGMP-level manufacturing
- Predictable methods lead to predictable products
- The CFR (Code of Federal Regulations) sections on GMPs
- What is covered under cGMP?
- So what is special about biomanufacturing?
- Nano-clean water necessary for nano-pharmaceuticals
- Contaminants at the nano-level
- Can you scale up the process?
- Some quality control issues – how to test
- Correctness of size – size matters!
- Composition – atomic level analyses
- Monodispersity versus agglomeration
- Order and correctness of layers
- Correctness of zeta potentials
- Does the nanomedical system contain the correct payload?
- Targeting (and mis-targeting) specificity and sensitivity
BME 695N Lecture 18: Designing nanodelivery systems for in-vivo use
12 Nov 2007 | Online Presentations | Contributor(s): James Leary
- Overview – the in-vitro to ex-vivo to in-vivo paradigm
- In-vitro - importance of choosing suitable cell lines
- Ex-vivo – adding the complexity of in-vivo background while keeping the simplicity of in-vitro
- In-vivo - all the complexity of ex-vivo plus the “active” components of a real animal
- In-vivo systems are open, “active” systems with multiple layers of complexity
- In-vitro and ex-vivo are mostly “closed” systems, but not absolutely
- What is an “open” system?
- Attempts to isolate open systems
- Layers of complexity of in-vivo systems
- Human cells in nude mice – a mixture of in-vitro and in-vivo
- “Model” small animal systems
- Bbetter model larger animal systems
- Examples of the in-vitro to in-vivo experimental pathway
- Kopelman group – multifunctional NPs for MRI and photodynamic therapy
- Langer group – aptamer-targeted NPs for cancer therapy in-vivo
- Leary group – peptide-guided NPs to human tumors in nude mice magnetic nanoparticles as MRI contrast agents in tissue phantoms
- Kopelman, R., Koo, Y-E, Philbert, M., Moffatc, B.A., Reddy,G.R., McConville, P., Hall, D.E.,
Chenevert, T.L., Bhojanie, M.S., Buck, S.M., Rehemtulla, A., Ross, B.D. Multifunctional nanoparticle platforms for in vivo MRI enhancement and photodynamic therapy of a rat brain cancer. Journal of Magnetism and Magnetic Materials 293: 404–410, 2005.
- Farokhzad, O.C., Cheng, J., Teply, B.J., Sherifi, I., Jon, S., Kantoff, P.W., Richie, J.P., Langer, R.
Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. PNAS 103(16), 6315–6320, 2006
BME 695N Lecture 17: Assessing nanotoxicity at the single cell level
06 Nov 2007 | Online Presentations | Contributor(s): James Leary
- Outline – the need for single cell measures of nanotoxicity
- There is more than one way for a cell to die...
- Necrosis" vs. "Apoptosis"
- There are other forms of "toxicity"
- Some other challenges in measuring toxicity of nanomaterials
- Necrosis vs. Apoptosis mechanisms
- Necrosis is unplanned "cell injury"
- Apoptosis is planned "programmed cell death"
- Why it is important to distinguish between necrosis and apoptosis
- Single cell assays for necrosis and apoptosis
- Dye exclusion assays for necrosis
- TUNEL assays for late apoptosis
- Annexin V assays for early apoptosis
- COMET assays for DNA damage and repair
- Light scatter assays
- Nanotoxicity in vivo – some additional challenges
- Single cell nanotoxicity, plus....
- Accumulations of nanoparticles can change toxicity locally to tissues and organs
- Filtration issues of nanoparticles – size matters – toxicity to liver and lung
- Chana, W-H, Nion-Shiao, N-H, Pin-Zhen Lu, P-Z. CdSe quantum dots induce apoptosis in human neuroblastoma cells via mitochondrial-dependent pathways and inhibition of survival signals. Toxicol. Lett. (2006), doi:10.1016/j.toxlet.2006.09.007
- Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F. Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry. 1997 Jan 1;27(1):1-20.
- Kirchner,C. Liedl, T., Kudera, S., Pellegrino,T., Munoz Javier, A., Hermann E. Gaub,H.E., Stolzle,S., N. Fertig, Parak, W.P., Cytotoxicity of Colloidal CdSe and CdSe/ZnS Nanoparticles. Nano Lett., Vol. 5, No. 2, 331-338, 2005.
- Oberdörster,G., Oberdörster, E. Oberdörster, J. Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles. Environmental Health Perspectives 113(7): 2005
- Ryman-Rasmussen, J.P., Riviere, J.E.,Monteiro-Riviere, N.A. Surface Coatings Determine Cytotoxicity and Irritation Potential of Quantum Dot Nanoparticles in Epidermal Keratinocytes. Journal of Investigative Dermatology. 10 August 2006; doi:10.1038/sj.jid.5700508
- Shiohara, A., Hoshino, A., Hanaki, K., Suzuki, K., Yamamoto, K. On the cytotoxicity caused by quantum dots. Microbiol. Immunol. 48(9): 669-675, 2004.
BME 695N Lecture 16: Assessing drug efficacy at the single cell level
02 Nov 2007 | Online Presentations | Contributor(s): James Leary
- Introduction and overview
- Nanomedical treatment at the single cell level requires evaluation at the single cell level
- For evaluation purposes, does structure reveal function?
- The difficulty of anything but simple functional assays
- The need for assays which at least show correlation to functional activity
- Quantitative single cell measurements of one or more proteins per cell by flow and image/confocal cytometry
- Cell surface measures of protein expression on live, single cells
- High-throughput flow cytometric screening of bioactive compounds
- Challenges of measuring protein expression inside fixed, single cells
- When location is important 2D or 3D imaging is required to get spatial location of proteins
- Quantitative multiparameter phospho-specific flow cytometry
- Attempts to measure "functional proteins" by detecting phosphorylation
- Example of phospho-specific, multiparameter flow cytometry
- Example of measuring single cell gene silencing by phospho-specific flow cytometry
- Quantitative measures of gene expression – the promises and the realities
- Is gene expression at the single cell level really possible?
- Is it even useful to measure a single gene's changes?
- Gene arrays of purified cell subpopulations
- RNA amplification techniques to attempt to perform single cell gene arrays
- Steven M. Chan, Janelle A. Olson, and Paul J. Utz. Single-Cell Analysis of siRNA-Mediated Gene Silencing Using Multiparameter Flow Cytometry. Cytometry Part A 69A:59–65 (2005).
- Peter O. Krutzik, Jonathan M. Irish, Garry P. Nolan and Omar D. Perez. Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications. Clinical Immunology 110: 206– 221 (2004).
- Susan M. Young, Mark S. Curry, John T. Ransom, Juan A. Ballesteros, Eric R. Prossnitz, Larry A. Sklar and Bruce S. Edwards. High-Throughput Microfluidic Mixing and Multiparametric Cell Sorting for Bioactive Compound Screening. J Biomol Screening; 9; 103 – 111 (2004).
- Szaniszlo, P., Wang, N., Sinha, M., Reece, L.M., Van Hook, J.W., Luxon, B.A., Leary, J.F. "Getting the Right Cells to the Array: Gene Expression Microarray Analysis of Cell Mixtures and Sorted Cells" Cytometry 59A: 191-202 (2004).
- Szaniszlo, P. Gene Expression Microarray Analysis of Small, Purified Cell Subsets. University of Texas Medical Branch, Galveston, TX April, 2007 (mentor: Dr. Leary)
BME 695N Lecture 15: Nanodelivery of therapeutic genes & molecular biosensor feedback control systems
30 Oct 2007 | Online Presentations | Contributor(s): James Leary
- Introduction and overview
- Some of the advantages of therapeutic genes
- Some of the advantages of molecular biosensor feedback control systems
- Why a nanodelivery approach is appropriate
- The therapeutic gene approach
- What constitutes a "therapeutic gene" ?
- Transient versus stable expression modes
- Molecular feedback control systems
- Drug delivery has traditionally not used feedback controls
- Why feedback control might be a very good idea!
- Positive or negative feedback?
- Molecular Biosensors as a component of a nanomedicine feedback control system
- What is a molecular biosensor?
- How a molecular biosensor functions as a therapeutic gene switch
- Building integrated molecular biosensor/gene delivery systems –some examples
- Required components
- Example of a ribozyme/antivirus system
- Example of an ARE biosensor/DNA repair system
- Prow, T.W., Kotov, N.A., Lvov, Y.M., Rijnbrand, R., Leary, J.F. “Nanoparticles, Molecular Biosensors, and Multispectral Confocal Microscopy” Journal of Molecular Histology, Vol. 35, No.6, pp. 555-564, 2004.
- Prow,T.W., Salazar, J.H., Rose, W.A., Smith, J.N., Reece, L.M., Fontenot, A.A., Wang, N.A., Lloyd, R.S., Leary, J.F. "Nanomedicine – nanoparticles, molecular biosensors and targeted gene/drug delivery for combined single-cell diagnostics and therapeutics" Proc. of SPIE 5318: 1-11, 2004.
- Prow, T.W., Rose, W.A., Wang, N., Reece, L.M., Lvov, Y., Leary, J.F. "Biosensor-Controlled Gene Therapy/Drug Delivery with Nanoparticles for Nanomedicine" Proc. of SPIE 5692: 199 – 208, 2005.
- Prow, T.W., Smith, J.N., Grebe, R., Salazar, J.H., Wang, N., Kotov, N., Lutty, G., Leary, J.F. "Construction, Gene Delivery, and Expression of DNA Tethered Nanoparticles" Molecular Vision 12: 606-615, 2006
- Prow, T.W., Grebe, R., Merges, C., Smith, J.N., McLeod, D.S., Leary, J.F., Gerard A. Lutty, G.A. "Novel therapeutic gene regulation by genetic biosensor tethered to magnetic nanoparticles for the detection and treatment of retinopathy of prematurity" Molecular Vision 12: 616-625, 2006.
- Prow, T.W. “Nanomedicine – Targeted Nanoparticles for the delivery of Biosensors and Therapeutic Genes” PhD Thesis, University of Texas Medical Branch, January 2004.
BME 695N Lecture 14: Challenges of proper drug dosing with nanodelivery systems
29 Oct 2007 | Online Presentations | Contributor(s): James Leary
- Overview of drug dosing problem
- Problems of scaling up doses from animal systems
- Basing dosing on size, area, weight of recipient
- Vast differences between adults in terms of genetics, metabolism
- Dosing in children – children are NOT smaller adults!
- Pharmacokinetics – drug distribution, metabolism, excretion, breakdown
- Conventional dosing assumes drug goes everywhere in the body
- Targeted therapies – a model for future nanomedical systems?
- From the animal dosing to human clinical trials
- Importance of picking an appropriate animal model system
- Does drug dosing really scale?
- The human guinea pig in clinical trials and beyond
- Some drug dosing methods
- Attempts to scale up on basis of area
- Attempts to scale up on weight/volume
- Attempts to use control engineering principles
- Genetic responses to drug dosing
- All humans are not genomically equivalent!
- Predicting on basis of family tree responses
- SNPs, chips, and beyond…predicting individual drug response
- After the $ 1000 individual genome scan… more closely tailored individual therapies
- Dosing in the era of directed therapies – a future model for nanomedical systems?
- How directed therapies change the dosing equation
- Current generation directed antibody therapies dosing
- Some typical side effects of directed therapies
- Nanomedical systems are the next generation of directed therapies
- Most directed therapies are nonlinear processes
- Current and pending FDA approved directed therapies
- Some examples of how a few directed therapies work
- Complement directed cytotoxicity
- ADCC-mediated adaptive immunity switch
- Antibody-directed enzyme producing therapy
- Other ways of controlling dose locally
- Magnetic field release of drugs
- Light-triggered release of drugs
- Adams, G.P., Weiner, L.M. Monoclonal Antibody Therapy of Cancer. Nature Biotechnology 23(9): 1147- 1157 (2005)
- Bailey, J.M., Haddad, W.M. Paradigms, Benefits, and Challenges: Drug Dosing Control in Clinical Pharmacology. IEEE Control Systems 35-51, April 2005
- McCoy, C.P., Rooney, C., Edwards, C.R., Jones, D.S., Gorman, S.P. Light-Triggered Molecule-Scale Drug Dosing Devices. J. AM. CHEM. SOC. 2007, 129, 9572-9573
- Miller, A.A Body Surface Area in Dosing Anticancer Agents: Scratch the Surface! Journal of the National Cancer Institute, Vol. 94, No. 24, December 18, 2002
- Nigel Baber & Deborah Pritchard Dose estimation for children. Br J Clin Pharmacol 56 , 489–493 2003
BME 695N Lecture 13: Assessing Zeta Potentials
29 Oct 2007 | Online Presentations | Contributor(s): James Leary
- Introduction – the importance of the zeta potential
- Nanoparticle-nanoparticle interactions
- Nanoparticle-cell interactions
- Part of the initial nanomedical system-cell targeting process
- Low zeta potential leads to low serum protein binding and potentially longer circulation
- Zeta potential basics
- What is the zeta potential?
- How is it measured?
- Some factors affecting the zeta potential
- Ionic strength
- Some zeta potential experiences
- Zeta potential measurement using laser Doppler electrophoresis (LDE).
- Why Measure Zeta Potential?.
- Zeta Potential: An Introduction in 30 minutes (PDF).
- Washington, C., "Zeta Potential in Pharmaceutical Formulation" (PDF).
- Using zeta potential to assess protein adsorption to surfactant coated latex (PDF).
- Prow, T.W., Rose, W.A., Wang, N., Reece, L.M., Lvov, Y., Leary, J.F. "Biosensor-Controlled Gene Therapy/Drug Delivery with Nanoparticles for Nanomedicine" Proc. of SPIE 5692: 199
– 208, 2005.
KIST/PU Multi-Component, Multi-Functional Nanomedical Systems for Drug/Gene Delivery
23 Oct 2007 | Online Presentations | Contributor(s): James Leary
In this brief paper we describe some of our recent efforts to construct multi-component, multi-functional nanomedical systems for delivery of therapeutic genes. We first describe the general philosophy of our approach. Then we describe three specific aspects of the overall construction in simple examples. Finally we show the results of these efforts including the successful delivery and expression of genes as seen through reporter gene expression and fluorescent analysis of single treated cells.
BME 695N Lecture 6: Rare-event targeting of cells in-vitro and in-vivo
26 Sep 2007 | Online Presentations | Contributor(s): James Leary
- Assessing nanomedical system (NMS) targeting at the single cell level
- Fluorescent labeling of NMSs
- First estimates of NMS binding by fluorescence microscopy
- Internal of external binding by confocal microscopy
- Single-cell image/confocal analysis
- Flow cytometric quantitation of NMS binding to specific cell types
- Image confocal analysis of NMS binding to single cells
- Ability to scan/locate cells of interest
- Photobleaching challenges
- Optical sectioning for 3D location of NMSs on/within cells
- A quick overview of flow cytometry
- Basic principles
- Capabilities and limitations
- Use for assessing specificity and sensitivity
- Rare-event analysis of NMS targeting to desired cells
- Basic concepts of rare-event analysis
- Strategies for rare cell detection
- More advanced flow cytometry for ultra-rare cell detection
- Examples of rare cell detection
- Rare cell sampling statistics
- Leary, J.F.: "Strategies for Rare Cell Detection and Isolation" In: Methods in Cell Biology: Flow Cytometry (Edited by Z. Darzynkiewicz, J.P. Robinson, H.A. Crissman), vol. 42: pp. 331-358, 1994.
- Leary, J.F. "Ultra High Speed Cell Sorting" Cytometry Part A 67A:76–85 (2005)
- Rosenblatt, J.A., Hokanson, J.A., McLaughlin, S.R., Leary, J.F.: A Theoretical Basis for Sampling Statistics Appropriate for the Detection and Isolation of Rare Cells Using Flow Cytometry and Cell Sorting Cytometry 26: 1-6; 1997
BME 695N Lecture 10: Nanomaterials for core design
26 Sep 2007 | Online Presentations | Contributor(s): James Leary
- Core building blocks
- Functional cores
- Functionalizing the core surface
- Ferric oxide cores
- Paramagnetic cores
- Superparamagnetic cores
- Ferric nanorods
- Advantages and disadvantages
- C60 and carbon nanotubes
- Size and structure of C60
- Elongation of C60 into carbon nanotubes
- Advantages and disadvantages
- Gold cores
- Gold nanoparticles
- Gold nanorods
- Other shapes (e.g. "stars")
- Gold nanoshells
- Advantages and disadvantages
- Silica cores
- Silica nanoparticles
- Advantages and disadvantages
- Hybrid materials
- Gold-ferrric oxide nanoparticles and nanorods
- Cobalt-Platinum magnetic nanoparticles
- Barnes, A.L., Wassel, R.A., Mondalek, F., Chen, K., Dormer, K.J., Kopke, R.D. Magnetic characterization of superparamagnetic nanoparticles pulled through model membranes. BioMagnetic Research and Technology 5:1-10, 2007.
- Berry, C.C., Curtis, A.S.G. TOPICAL REVIEW: Functionalisation of magnetic nanoparticles for applications in biomedicine. J. Phys. D: Appl. Phys. 36 (2003) R198–R206
- Burda, C., Chen, X., Narayanan, R., El-Sayed, M.A. Chemistry and Properties of Nanocrystals of Different Shapes Chem. Rev. 2005, 105, 1025-1102 (a VERY comprehensive review!)
- Lee, C.S., Lee, H., Westervelta, R.M. Microelectromagnets for the control of magnetic nanoparticles Applied Physics Letters 79(20): 3308-3310, 2001.
- Mornet, S., Vasseur, S., Grasset, F., Duguet, E. Magnetic nanoparticle design for medical diagnosis and therapy. J. Mater. Chem 14: 2161-2175, 2004.
- Osaka, T., Matsunaga, T., Nakanishi, T., Arakaki, A., Niwa, D., Iida, H. Synthesis of magnetic nanoparticles and their application to bioassays. Anal Bioanal Chem (2006) 384: 593–600
- Park, J-I, Cheon, J. Synthesis of “Solid Solution” and “Core-Shell” Type Cobalt-Platinum Magnetic Nanoparticles via Transmetalation Reactions. J. Am. Chem. Soc. 2001, 123, 5743-5746
- Park, S-J, Kim,S., Lee, S., Khim, Z.G., Char, K., Hyeon, T. Synthesis and Magnetic Studies of Uniform Iron Nanorods and Nanospheres. J. Am. Chem. Soc. 2000, 122, 8581-8582
- Wang, L., Wang, K., Santra, S., Zhao, X., Hilliard, L.R., Smith, J.E., Wu, Y., Tan, W. Watching Silica Nanocrystals Glow in the Biological World. Analytical Chemistry 647-654, 2006.
BME 695N Lecture 8: Technologies for measuring nanomedical systems on/within cells
24 Sep 2007 | Online Presentations | Contributor(s): James Leary
- Introduction to measuring technologies for nanomedical system interaction with cells
- The importance of quantitative or at least semi-quantitative single cell measurements
to detect presence and location of nanomedical systems
- Below "optical limit" imaging
- Requirements on the NMS to have X-ray dense, fluorescent, metallic, or magnetic cores
- Can you study living cells?
- Technologies – Advantages and disadvantages
- Flow cytometry – a "zero order" imaging device
- Scanning and Transmission electron microscopy
- Confocal microscopy – one and two-photon
- Surface Plasmon Resonance (SPR) Imaging
- Atomic Force Microscopy
- Magnetic Sorting/MRI contrast agents for in-vivo imaging
- Liu, J. Scanning transmission electron microscopy and its application to the study of nanoparticles and nanoparticle systems. Journal of Electron Microscopy 54(3): 251–278 (2005).
- Alschinger, M., Maniak, M., f. Stietz, F., Vartanyan, T., Trager. F. Application of metal nanoparticles in confocal laser scanning microscopy: improved resolution by optical field enhancement. Appl. Phys. B 76, 771–774, 2003.
- Jain, P.K., El-Sayed, I.H., El-Sayed, M.A. Au nanoparticles target cancer. Nanotoday 2(1): 18-29, 2007.
- Yoon, T-J, Yu, K.N., Kim, E., Kim, J.S., Kim, B.G., Yun, S-H, Sohn, B-Y, Cho, M-H, Lee,J-K, Park, S.B. Specific Targeting, Cell Sorting, and Bioimaging with Smart Magnetic Silica Core–Shell Nanomaterials. Small 2(2): 209 – 215, 2006.
- Rao, A., Schoenenberger, M., Gnecco, E., Glatzel, Th, Meyer, E., Brändlin, D., Scandella, L.
Characterization of nanoparticles using Atomic Force Microscopy. Journal of Physics: Conference Series 61 (2007) 971–976
- Pris, A.D., Porter, M.D. Nanoparticle Coding: Size-Based Assays Using Atomic Force Microscopy. Langmuir 2004, 20, 6969-6973
BME 695N Lecture 7: Normal & facilitated cell entry mechanisms
15 Sep 2007 | Online Presentations | Contributor(s): James Leary
- The general problem of cell entry
- Choosing modes of cell entry
- How does Nature do it? (biomimetics)
- Non-specific uptake mechanisms
- Pinocytosis by all cells
- Phagocytosis by some cells
- Receptor mediated uptake
- Receptor mediated transport of desired molecules
- Example- transferrin receptor transport of iron for metabolism
- Nanoparticle uptake
- Size matters
- Agglomeration reduces uptake
- Drug delivery by "shedding"
- Extracellular drug delivery by shedding
- Intracellular drug release by shedding
- General reference: Alberts et al., Molecular Biology of the Cell. Garland Science 4th Edition, New York, pp. 747-756. 2002.
- Becker, C., Hodenius, M., Blendingera, G., Sechi, A., Hieronymus, T., Müller-Schulte, D., Schmitz-Rode, T., Zenke, M. “Uptake of magnetic nanoparticles into cells for cell tracking.” Journal of Magnetism and Magnetic Materials Volume 311(1): 234-237, 2007.
- Dawson, G.F., Halbert, G.W. “The In Vitro Cell Association of Invasin Coated Polylactide-Co-Glycolide Nanoparticles.” Pharmaceutical Research, Vol. 17, No. 11, 1420-1425, 2000
- Limbach, L., Yuchun, L., Grass, R.N., Brunner, T., Hintermann, M.A., Muller, M., Gunther, D., Stark, W.J. “Oxide Nanoparticle Uptake in Human Lung Fibroblasts: Effects of Particle Size, Agglomeration, and Diffusion at Low Concentrations.” Environ. Sci. Technol. 39: 9370-9376, 2005.
- Medina, C., Santos-Martinez, M.J., Radomski, A., Corrigan, O.I., Radomski1, M.W. “Nanoparticles: pharmacological and toxicological significance.” British Journal of Pharmacology 150, 552–558, 2007
- Mousavi, S.A., Malerod, L., Berg, T., Kjeken, R. “REVIEW ARTICLE: Clathrin-dependent endocytosis.” Biochemical Journal Immediate Publication. Published on 23 Sep 2003 as manuscript BJ20031000
- Nori, A., Kopecek, J. “Intracellular targeting of polymer-bound drugs for cancer chemotherapy.” Advanced Drug Delivery Reviews 57: 609– 636, 2005
- Romberg, B., Hennink, W.E., Storm, G.. “Sheddable Coatings for Long-Circulating Nanoparticles.” Pharmaceutical Research DOI: 10.1007/s11095-007-9348-7, 2007
BME 695N Lecture 5: Cell Targeting
12 Sep 2007 | Online Presentations | Contributor(s): James Leary
- Overview: targeting nanosystems to cells
- Antibody targeting
- Peptide targeting
- Aptamer targeting
- Antibodies – polyclonal and monoclonal
- Where do antibodies come from – in nature?
- How do we make them in the laboratory?
- Monoclonal antibodies
- Therapy problems with mouse monoclonal antibodies
- “Humanizing” monoclonal antibodies to reduce adverse host immune reactions
- Why antibodies may not be a good overall choices for targeting nanosystems to cells
- Peptide targeting
- How does a peptide target?
- Examples of peptide targeting
- Creating new peptides by random peptide phage display libraries
- High-throughput screening of those peptide libraries
- Advantages and disadvantages of peptide targeting
- Aptamer targeting
- What are aptamers and how do they target?
- Some different types of aptamers
- How do you make aptamers?
- How do you screen for useful aptamers?
- Carmen, S. Jermutus, L. Concepts in antibody phage display, BRIEFINGS IN FUNCTIONAL GENOMICS AND PROTEOMICS. VOL 1. NO 2. 189–203. JULY 2002
- Yang, X., Bassett, S.E., Li, X., Luxon, B.A., Herzog, N.K., Shope, R.E., Aronson, J., Prow, T.W., Leary, J.F., Kirby, R., Ellington, A.D., Gorenstein, D.G.: “Construction and selection of bead bound combinatorial oligonucleoside phosphorothioate and phosphorodithioate aptamer libraries designed for rapid PCR-based sequencing” Nucleic Acids Research 30 (23)e132: 1-8, 2003.
- Yang, X., Li, X., Prow, T.W. Reece, L.M., Bassett, S.E., Luxon, B.A., Herzog, N.K., Aronson, J., Shope, R.E., Leary, J.F., Gorenstein, D.G. “Immunofluorescence Assay and Flow Cytometric Selection of Bead Bound Aptamers” Nucleic Acids Research 31 (10): 1-8, 2003
BME 695N Lecture 4: Designing "Theragnostic" Systems
04 Sep 2007 | Online Presentations | Contributor(s): James Leary
- Bridging the gap between diagnostics and therapeutics
- How conventional medicine is practiced in terms of diagnostics and therapeutics
- The consequences of separating diagnostics and therapeutics
- A new approach – "theragnostics" (or "theranostics")
- Examples of current theragnostic systems
- Example: Rituxan ("Rituximab)(an example of not using diagnostics to guide the therapy)
- Example 1: Herceptin ("terastuzumab")
- Example 2: Iressa ("Gefitinib)
- How theragnostics relates to Molecular Imaging
- Conventional imaging is not very specific
- Types of In-vivo Imaging
- X-rays, CAT (Computed Axial Tomography) scans
- MRI (magnetic Resonance Imaging)
- PET (Positron Emission Tomography) scans
- "Molecular Imaging"
- Engineering nanomedical systems for simultaneous molecular imaging
- Using nanomedical cores for MRI contrast agents
- Difficulties in using PET probes for nanomedical devices
- Using cell-specific probes for molecular imaging of nanomedical devices
- Breaking the "diffraction limit" – nano-level imaging
- Theragnostic nanomedical devices
- Using nanomedical devices to guide separate therapeutic device
- When might we want to combine diagnostics and therapeutics?
- BJJ Abdullah, Molecular imaging: spawning a new melting-pot for biomedical imaging
Biomed Imaging Interv J 2006; 2(4):e28 pages 1-7.
- Chul Ahn, Pharmacogenomics in Drug Discovery and Development, Genomics & Informatics Vol. 5(2) 41-45, 2007
- Eric Betzig, George H. Patterson, Rachid Sougrat, O. Wolf Lindwasser, Scott Olenych,
Juan S. Bonifacino, Michael W. Davidson, Jennifer Lippincott-Schwartz, Harald F. Hess
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution
SCIENCE VOL 313: 1642-1645, 2006
BME 695N Lecture 3: Overview of Basic Nanomedical Systems Design
29 Aug 2007 | Online Presentations | Contributor(s): James Leary
- Nanomedical systems – levels of challenges
- Essential elements of a nanomedical system
- Requirements for specific cell targeting
- Consequences of mis-targeting
- Engineering around the consequences of mis-targeting
- Some ways to lower mis-targeting to non-diseased cells
BME 695N: Engineering Nanomedical Systems (Fall 2007)
03 Aug 2007 | Courses | Contributor(s): James Leary
This course will cover the basic concepts of design of integrated nanomedical systems for diagnostics and therapeutics. Topics to be covered include: why nanomedical approaches are needed, cell targeting strategies, choice of core nanomaterials, technologies for testing composition and structure of multilayered nanomedical systems, optimizing zeta potentials, design and testing of cell and intracellular targeting systems, in-vivo issues, drug delivery and proper dosing, assessing efficacy of drug/gene delivery, nanotoxicity, animal testing, and regulatory issues. In addition to attending lectures and participating in classroom discussions, students will write and present an original research nanomedical system design project.
This course will serve as an interdisciplinary training for doctoral students in Biomedical Engineering and other fields for a basic understanding of the principles and challenges of nanomedicine.
BME 695N Lecture 2: Basic Concepts of Nanomedical Systems
28 Aug 2007 | Online Presentations | Contributor(s): James Leary
- Features of Nanomedicine
- Bottoms up rather than top down approach to medicine
- Nano-tools on the scale of molecules
- Cell-by-cell repair approach – regenerative medicine
- Feedback control system to control drug dosing
- Elements of good engineering design
- Whenever possible, use a general design that has already been tested
- Use multiple specific molecules to do multi-step tasks
- Control the order of molecular assembly to control the order of events
- Therefore, perform the molecular assembly in reverse order to the desired order of events
- Building a nanodevice
- Choice of core materials
- Add drug or therapeutic gene
- Add molecular biosensors to control drug/gene delivery
- Add intracellular targeting molecules
- Result is multi-component, multi-functional nanomedical device
- For use, design to de-layer, one layer at a time
- The multi-step drug/gene delivery process in nanomedical systems
- The challenge of drug/gene dosing to single cells
- Precise targeting of drug delivery system while protecting non-targeted cells from exposure to the drug
- How to minimize mis-targeting
- How to deliver the right dose per cell
- One possible solution – in situ manufacture of therapeutic genes
- Prow, T.W., Rose, W.A., Wang, N., Reece, L.M., Lvov, Y., Leary, J.F. "Biosensor-Controlled Gene
Therapy/Drug Delivery with Nanoparticles for Nanomedicine" Proc. of SPIE 5692: 199 – 208, 2005.
BME 695N Lecture 1: Need for New Perspectives on Medicine
03 Aug 2007 | Online Presentations | Contributor(s): James Leary
- The Progression of Medicine
- Conventional "modern" medicine
- "Personalized" or "molecular" medicine
- Nanomedicine "single-cell" medicine
- How Conventional Medicine Works for Diagnosis of Disease
- Identification of the "diseased state"
- Simple measurements of body structure and function
- Follow-up clinical tests
- Internal examinations by non-invasive in-vivo imaging
- Molecular tests for specific gene properties
- Comparison of individual results with "normal ranges"
- How Conventional Medicine Works for Treatment of Disease
- Stabilization of patient – "heal thyself"
- Surgical repair of injuries
- Treatment with drugs locally
- Treatment with drugs systemically
- Treatment with targeted therapies
- Factors Limiting the Progress of Medicine
- Some Specific Problems with Conventional Medicine
- Consequences of waiting for patient symptoms
- Trained people and modern drugs are expensive
- Diagnostic technologies, if available, are still relatively primitive and/or expensive
- Crude targeting of drugs
- Personalized Medicine
- Based on genetic characteristics of the individual patient
- Specific gene rearrangments or mutations
- Specific SNPs (Single Nucleotide Polymorphisms)
- Medicine performed at single cell level
- Possibility of "regenerative medicine"
- Blurring of distinction between prevention and treatment
- Prow,T.W., Salazar, J.H., Rose, W.A., Smith, J.N., Reece, L.M., Fontenot, A.A., Wang, N.A.,
Lloyd, R.S., Leary, J.F. "Nanomedicine – nanoparticles, molecular biosensors and targeted
gene/drug delivery for combined single-cell diagnostics and therapeutics" Proc. of SPIE 5318: 1-11, 2004.
- Moein Moghimi,SM, Hunter,AC, Murray, JC: Nanomedicine: current status and future
prospects. FASEB J. 19: 311–330, 2005.
- Liu, H. Webster, TJ Nanomedicine for implants: A review of studies and necessary
experimental tools. Biomaterials 28: 354–369, 2007.
- Ulrich Pison, U., Tobias Welte, T., Michael Giersig, M., David A. Groneberg, D.A.
Nanomedicine for respiratory diseases. European Journal of Pharmacology 533: 341–350, 2006.
- Logothetidis S: Nanotechnology in Medicine: The Medicine of Tomorrow and Nanomedicine.
HIPPOKRATIA 10(1): 7-21, 2006.
Nanotechnologies, Science and Society: Promises and Challenges
10 May 2007 | Online Presentations | Contributor(s): James Leary
James Leary (2007), "Nanotechnologies, Science and Society: Promises and Challenges," Purdue Bioethics Seminar Series, http://nanohub.org/resources/2691.
Bioethics at Purdue
The Graduate School of Purdue University - Puskas Fellowship
Engineering Nanomedical Systems
06 Mar 2006 | Online Presentations | Contributor(s): James Leary
This tutorial discusses general problems and approaches to the design of engineered nanomedical systems. One example given is the engineering design of programmable multilayered nanoparticles (PMNP) to control a multi-sequence process of targeting to rare cells in-vivo, re-targeting to intracellular sites, and controlling of final gene/drug delivery. Therapeutic genes can be manufactured inside living cells as a "nanofactory" under the control of these molecular biosensors providing feedback- controlled single cell medicine.
Nanofactories - In - Situ Production of Therapeutic Genes...
27 Jul 2005 | Online Presentations | Contributor(s): James Leary