Former MNIMBS Students

Song Ge, Ph.D. Song Ge obtained his B.S. in physics from Shandong University in China. He then graduated with a Ph.D. in Physics from the University of Michigan under the guidance of Dr. Bradford Orr. Song's research work was developing a SQUID imaging system of targeted magneticnanoparticles for use as contrast agents, which adds in M-NiMBS's effort of early-stage cancer detection. Besides, he invented a hydrothermal synthetic route of iron oxide nanoparticles with controllable size and tunable magnetic properties.

Thesis: Development of a SQUID (Superconducting Quantum Interference Device) Detection System of Magnetic Nanoparticles for Cancer Imaging

Chair: Bradford G. Orr

Committee members: James R. Baker, Jr., Mark M. Banaszak Holl, Luming Duan, Cagliyan Kurdak

In this dissertation, Song Ge presents the development of a SQUID (Superconducting Quantum Interference Device) imaging system of targeted magnetic nanoparticles (NPs) for use as contrast agents. The contrast agents are functionalized for targeting by the conjugation of the magnetic NPs to folic acid (FA) molecules on dendrimer scaffolds. Cellular internalization is accomplished through the high-affinity folic acid receptors (FARs), which are overexpressed in various human carcinomas. SQUID can be applied to detect signals from the magnetic cores of the contrast agents and hence diagnose the tumor. Based on the magnetic properties of the magnetic NPs, two detection methods were developed: remanence and magnetorelaxometry (MRX).

The remanence measurement-based method detects the magnetic NPs according to the magnetic remanence exhibited by particles that are sufficiently large and possess long relaxation mechanism. Samples were vertically oscillated and horizontally translated each in one-dimension. The system was calibrated with g-Fe2O3 NPs (mean diameter 25 nm) and the detection limit was found to be 10 ng at a distance of 1.7 cm and the spatial resolution was ~1 cm. A theoretical model of this system was proposed and applied to image reconstruction of scanned phantoms with two NP injection spots. The developed SQUID system can determine not only the amount and horizontal position of the NPs, but also their depth in the phantoms.

The MRX technique utilizes the NPs superparamagnetic property and records their time course magnetic decay. The system was investigated by using a number of iron oxide NP products with different mean diameters. The results showed that the MRX signal intensity is sensitively dependent on the size of the NPs. The best detection limit of 300 ng of total iron content was found on using a d = 12 nm Fe3O4 NP sample and this result was supported by computer simulations. To produce magnetic NPs for the MRX study, a synthetic approach of size-controllable Fe3O4 NPs was developed. Accordingly, the magnetic property can be tuned from ferromagnetic to superparamagnetic. In vitro experiments were conducted after functionalization of the synthesized NPs, which showed enhancement in cell targeting of the FA-modified NPs.

Awards

Physics Incoming Student Fellowship, University of Michigan   2002-2003

Excellence Graduate Student Award, Peking University   2002

“GuangHua” Scholarship in Condensed Matter Division, Peking University1999-2000

First Prize Scholarship in every semester, Shandong University   1995-1999

“KeNi” Scholarship for excellent students, Shandong University  1998

Publications

S. Ge, X. Shi, K. Sun, C. Li, J. R. Baker, Jr., M. M. Banaszak Holl, and B. G. Orr, A facile hydrothermal synthesis of iron oxide nanoparticles with tunable magnetic properties, Journal of Physical Chemistry, Vol. 113, p. 13593-13599, 2009

S. Ge, X. Shi, J. R. Baker, Jr., M. M. Banaszak Holl, and B. G. Orr, Development of a remanence measurement-based SQUID with in-depth resolution for nanoparticle imaging, Physics in Medicine and Biology, Vol. 54, p. N177-N188, 2009

X. Shi, S. H. Wang, S. D. Swanson, S. Ge, Z. Cao, M. E. Van Antwerp, K. J. Landmark, J. R. Baker, Jr., Dendrimer-functionalized shell-crosslinked iron oxide nanoparticles for in vivo magnetic resonance imaging of tumors, Advanced Materials, Vol. 9999, p.1-8, 2008

S. D. Swanson, J. F. Kukowska-Latallo, A. K. Patri, C. Chen, S. Ge, Z. Cao, A. Kotlyar, A. T. East, J. R. Baker, Jr., Targeted gadolinium-loaded dendrimer nanoparticles for tumor-specific magnetic resonance contrast enhancement, International Journal of Nanomedicine, Vol. 3, p. 201-210, 2008

S. Ge et al., Synthesis and field emission study of large-scaled aligned carbon nanotube films, Acta Scientiarum Naturalium Universitatis Pekinensis, Vol.39 No.3 p.341, 2003

Paul Makidon, DVM, Ph.D. in Biomedical Engineering Nanobiology certificate program Research Investigator Director of Vaccine Development Department of Internal Medicine and Michigan Nanotechnology Institute for Medicine and Biological Sciences Clinical Instructor, Unit for Laboratory Animal Medicine University of Michigan

While earning his DVM degree (1998) at Michigan State University (MSU), Paul Makidon worked on research projects at the Laboratory of Comparative Orthopedic Research and the Endocrinology Section of the Animal Health Diagnostics Laboratory at MSU. He then worked in a clinical practice setting for several years. However, recognizing his deep interest in research, he started the PhD program at the Department of Biomedical Engineering at the University of Michigan (UM) in Fall 2005 and earned candidacy in Fall 2007. He worked under mentorship of Dr. James R. Baker Jr., Director of the Michigan Nanotechnology Institute for Medicine and Biological Sciences (M-NIMBS) and earned his Ph.D. degree in May 2009. He worked concurrently as aresearch fellow with the Unit for Laboratory Animal Medicine (ULAM) at UM where he was funded from a NIH/NCRR T-32 grant.

Paul Makidon, DVM, PhD is now Research Investigator in the Department of Internal Medicine, Division of Allergy & Clinical Immunology and is a Clinical Instructor in the Unit for Laboratory Animal Medicine with a secondary appointment in the Michigan Nanotechnology Institute for Medicine and Biological Sciences. He serves as Director of Vaccine Development at MNIMBS.

Paul is interested in biomedical applications of nanomedicine. He has been actively involved in development of nanoemulsion based, needle-free, nasal spray adjuvant vaccine development for Hepatitis B. His doctoral research project focuses on the development of an antimicrobial nanoemulsion based inhalation therapeutic to prevent infection or treat antibiotic resistant infections in patients with Cystic Fibrosis, a disease that affects more than 53,000 children worldwide. He will continue this research to complete his Ph.D. degree and then hopes to start an academic research career that is similarly clinically oriented, nanomedicine based and further examines treatment options that will improve the health status of Cystic Fibrosis patients.

Paul defended his thesis: Tuesday, December 2, 2008, 2:30 PM

Department of Biomedical Engineering
Chair: James R. Baker, Jr., MD

Other Committe members: Victor DiRita, Ph.D., Joseph Bull, Ph.D.and John LiPuma, MD

OIL-IN-WATER NANOEMULSIONS AS MUCOSAL VACCINE ADJUVANTS:
CHARACTERIZATION, MECHANISM, FORMULATION, AND DEVELOPMENT OF A
NANOEMULSION-BASED BURKHOLDRERIA CENOCEPACIA VACCINE

Surface active oil-in-water nanoscale emulsions have been developed as
mucosal vaccine adjuvants capable of producing robust systemic, mucosal,
and cellular immune responses against diverse microbial and recombinant
antigenic proteins. This dissertation examines the development of
nanoemulsion (NE) as a new generation nasopharyngeal adjuvant. Part of
the thesis is organized to address the characterization of NE-induced
immune response and includes the pre-clinical studies of a novel NE-based
recombinant hepatitis B vaccine (HBsAg-NE). Our results suggest that nasal
immunization with HBsAg-NE may be a safe and effective hepatitis B
vaccine. The adjuvant induces specific IgG, mucosal IgA, and a Th1-biased
cellular immunity. Immunogenicity is comparable to the standard
alum-based vaccine. HBsAg-NE is stable for months at elevated
temperatures because of the physical association of NE and antigen and its
stability was enhanced with buffered salt diluents. We also report that
NE-based vaccines do not require specially engineered delivery devices.
The prolonged stability and ease of delivery are direct advantages for use
of NE-based vaccines in developing populations.

We also evaluate the mechanism of NE adjuvant activity. NE promotes
antigen internalization in nasal epithelium and loading into mucosal DC.
Trafficking of the antigen to the submandibular lymph nodes and thymus
occurs within 24 hours of intranasal vaccination. Administration of NE
was not associated with the typical induction of local inflammation or
histopathological changes. Microarray analysis shows the upregulation of
only 1.6% of genes responsible for the production of acute phase
inflammatory cytokines including IL6. Hallmark inflammatory cytokines
such as IL4, and INF- were not measured in nasal secretions. The role of
IL6 in NE adjuvant activity was examined by evaluating immunogenicity in
IL6 mutant mice.

The final component of the dissertation addresses the development of a
NE-based Burkholderia cenocepacia outer membrane protein (OMP) vaccine.
We demonstrate that NE is as a strong mucosal adjuvant for OMP and OMP-NE
protects against experimental lung infections in mice.

Overall, these findings confirm that NE is an excellent mucosal stimulant
and support the further development of nanoemulsions as nasopharyngeal
adjuvants. We conclude that nanoemulsion exhibits all the major desired
characteristics of an adjuvant.

Honors and Awards

2008      The Maurice Hilleman Young Investigator of the Year Award: 

             National Foundation for Infectious Diseases -First Runner up

1998      The Society of Phi Zeta – Honor society of Veterinary Medicine

1996      Recipient of the Hutton Fund Scholarship

1995      SCAVMA Clinical Competency National Champion

Publications

Makidon, P. E., Nigavekar, S. S., Bielinska, A. U., Mank, N. Shetty, A. M., Suman, J., Knowlton, J., Myc, A., Rook, T. and Baker, Jr.,J. R., Characterization of stability and nasal delivery systems for immunization with nanoemulsion-based vaccines, Journal of Aerosol Medicine and Pulmonary Drug Delivery, Volume 22, Number 4, 2009, Pp. 1-14. DOI: 10.1089=jamp.2009.0766

LiPuma, J.J., Rathinavelu, S., Foster, B.K...Makidon, P.E. (5/7)…Baker, J.R. Jr., In vitro activities of a novel nanoemulsion against Burkholderia and other multidrug-resistant cystic fibrosis-associated bacterial species, Antimicrob Agents Chemother. 2009 Jan;53(1):249-55. Epub 2008 Oct 27. [I participated in the design troubleshooting of the methods and participated in the actual writing of the manuscript]

Makidon, P.E, Bielinska, A.U., Nigavekar, S.S., Janczak, K.W., Knowlton, J., Scott, A.J., Mank, N, Cao, Z., Rathinavelu, S., Beer, M.R., Wilkinson, J.E., Blanco, L.P., Landers, J.J., Baker, J.R. Jr., Pre-clinical evaluation of a novel nanoemulsion-based hepatitis B mucosal vaccine, PLoS ONE. 2008 Aug 13;3(8):e2954.

Bielinska, A. U., Janczak, K. W., Landers, J. L., Makidon, P., Sower, L.E., Peterson, J. W., Baker, J. R. Jr.., Mucosal Immunization with a Novel Nanoemulsion-based Recombinant Anthrax Protective Antigen Vaccine Protects against B. anthracis Spore Challenge, Infect Immun. 2007 Aug;75(8):4020-9.

Makidon, P.E.: “Esophagostomy Tube Placement in the Anorectic Rabbit” A technique article. Lab Animal, 2005 Sept;34:33-36.

Christopher Kelly, Ph.D. Ph.D. Applied Physics Nanobiology certificate program Postdoctoral Associate, Cornell University 2009 - present Observing the IgE and FcεRI signaling pathway via dielectric-filled nanoapertures Advised by Drs. Harold Craighead and Barbara Baird

Christopher graduated with high honors in Physics from Oberlin College in 2003.  After working for a year at NASA Glenn Research Center on thin film solar cells, Christopher started graduate work at University of Michigan with Professors Orr and Banaszak Holl in 2004.  Christopher earned his Masters in Electrical Engineering in 2007 and earned his Ph.D. in 2009 in Applied Physics, focusing on the molecular details of the dendrimer-plasma membrane interaction.  Christopher has been awarded the Biophysics Training Grant and is a graduate fellow of the Graham Institute for Environmental Sustainability.

Christopher V. Kelly’s Thesis: "The Biophysics of nanoparticles interacting with the plasma membrane"

My research aims to guide the design of nanoparticles for both biomedical applications (e.g. targeted chemotherapeutics) and everyday applications (e.g. tennis rackets and sun screen) by understanding the detailed interactions of nanoparticles with the plasma membrane.  In particular, I will discuss the atomistic and molecular-level interactions of poly(amidoamine) (PAMAM) dendrimers with model plasma membranes.  Previous work has demonstrated that larger and more charged dendrimers induce greater membrane disruption, such as nanometer scale pores that allow cytoplasmic release from the cell.  I utilize all-atom molecular dynamics, isothermal titration calorimetry, transmission electron microscopy, dynamic light scattering, differential scanning calorimetry, and atomic force microscopy, to determine nanoparticle moieties and molecular-mechanisms that facilitate membrane degradation.  For example, sixth-generation PAMAM dendrimers have been determined to be a critical size for membrane degradation since larger dendrimers induce a nanoparticle-encased lipid vesicle complex whereas smaller dendrimers primarily flatten on the bilayer.

Awards and Fellowships
Certificate in Nanoscience and Technology with NanoBiology emphasis, University of Michigan 2009
Chair’s Fund Fellowship, Gordon Conference in Polymer Physics 2008
Best Poster in Polymer Synthesis, 31st Annual Symposium of Macromolecular Science and Engineering, University of Michigan 2007
NSF Workshop Fellowship, University of Montana Center for Ethics 2007
Doctoral Candidacy with Highest Distinction 2006
Graham Environmental Sustainability Institute Graduate Fellowship 2006
Biophysics Training Grant, University of Michigan 2005
Sigma Xi, Associate Member 2003
Carl E. Howe Prize in Physics, Oberlin College 2003

Publications

1. Kelly, C. V.; Liroff, M. G.; Triplett, L. D.; Leroueil, P. R.; Mullen, D. G.; Wallace, J. M., Meshinchi, S.; Baker,J. R.; Orr, B. G.; Banaszak Holl, M. M., Stoichiometry and Structure of Poly(amidoamine) Dendrimer-Lipid Complexes. ACS Nano 2009, 3, 7, 1886-1896

2. Kelly, C. V.; Kober, M. M.; Kinnunen, P.; Reis, D. A.; Orr, B. G.; Banaszak Holl, M. M., Pulsed-laser creation and characterization of giant plasma membrane vesicles on living cells. Journal of Biological Physics 2009, 5, 3, 279-295

3. Kelly, C. V.; Leroueil, P. R.; Nett, E. J.; Wereszczynski, J. M.; Baker, J. R.; Orr, B. G.; Banaszak Holl, M. M..; Andricioaei, I., Poly(amidoamine) dendrimers on lipid bilayers I: Free energy and conformation of binding. Journal of Physical Chemistry B 2008, 112, 31, 9337-9345

4. Kelly, C. V.; Leroueil, P. R.; Orr, B. G.; Banaszak Holl, M. M.; Andricioaei, I., Poly(amidoamine) dendrimers on lipid bilayers II: Effects of bilayer phase and dendrimer termination. Journal of Physical Chemistry B 2008, 112 (31), 9346-9353

5. Mullen, D. G.; Desai, A. M.; Waddell, J. N.; Cheng, X.; Kelly, C. V.; McNerny, D. Q.; Majoros, I. J.; Baker, Jr., J. R.; Sander, L. M.; Orr, B. G.; Banaszak Holl, M. M., The Implications of Stochastic Synthesis for the Conjugation of Functional Groups to Nanoparticles. Bioconjugate Chemistry 2008, 19, 9, 1748-1752

6. Landmark, K. J.; DiMaggio, S.; Ward, J.; Kelly, C. V.; Vogt, S.; Hong, S.; Kotlyar, A.; Penner-Hahn, J. E.; Baker, J. R.; Banaszak Holl, M. M.; Orr, B. G., Synthesis, Characterization, and In Vitro Testing of Superparamagnetic Iron Oxide Nanoparticles Targeted Using Folic Acid-Conjugated Dendrimers. ACS Nano 2008, 2 (4), 773-783.

7. Erickson, B.; Kelly, C. V.; Leroueil, P. R.; Berry, S. A.; Baker, J.R.; Orr, B. G.; Banaszak Holl, M. M., Interactions of Poly(amidoamine) Dendrimers with Survanta Lung Surfactant: The Importance of Lipid Domains. Langmuir 2008, 24, 19, 11003-11008

8. Farrell, D. F.; Ijiri, Y.; Kelly, C. V.; Borchers, J. A.; Rhyne, J. J.; Ding, Y.; Majetich, S. A., Small angle neutron scattering study of disordered and crystalline iron nanoparticle assemblies. Journal of Magnetism and Magnetic Materials 2006, 303 (2), 318-322.

9. Kelly, C. V.; Jin, M. H. C.; Banger, K. K.; McNatt, J. S.; Dickman, J. E.; Hepp, A. F., Parametric study on nonvacuum chemical vapor deposition of CuInS2 from a single-source precursor. Materials Science and Engineering B-Solid State Materials for Advanced Technology 2005, 116 (3), 403-408.

10. Ijiri, Y.; Kelly, C. V.; Borchers, J. A.; Rhyne, J. J.; Farrell, D. F.; Majetich, S. A., Detection of spin coupling in iron nanoparticles with small angle neutron scattering. Applied Physics Letters 2005, 86 (24), 243102.

11. Haider, M. B.; Constantin, C.; Al-Brithen, H.; Yang, H.; Trifan, E.; Ingram, D.; Smith, A. R.; Kelly, C. V.; Ijiri, Y., Ga/N flux ratio influence on Mn incorporation, surface morphology, and lattice polarity during radio frequency molecular beam epitaxy of (Ga,Mn)N. Journal of Applied Physics 2003, 93 (9), 5274-5281.

Pascale R. Leroueil, Ph.D. Staff Scientist Michigan Nanotechnology Institute for Medicine and Biological Sciences   B.A., Chemistry, Whitman College, Walla Walla, WA, 2003 M.S., Chemistry, University of Michigan, Ann Arbor, MI 2005 Ph.D., Chemistry, University of Michigan, Ann Arbor, MI, 2008.

Pascale Leroueil’s thesis: Investigation of Nanoparticles with Real and Model Cell Membranes

Co-Chairs: Mark M. Banaszak Holl (Chemistry) and Bradford G. Orr (Physics)

The first component of the thesis examines the binding of a multivalent, folic acid (FA) receptor targeted, generation 5 (G5) PAMAM dendrimer using force pulling spectroscopy.  This targeted nanoparticle, when conjugated to the therapeutic methotrexate and then acetylated to form a neutral complex, has been shown to be effective at reducing FA-receptor expressing KB cell tumors.  Surface plasmon resonance (SPR) showed a 100,000-fold decrease in the dissociation rate of the G5-FAn from a model FA receptor surface, folate binding protein (FBP), as the number of FAs (n) increased from 1-12.  Force pulling spectroscopy was then used to compare the force required to rupture the interaction between the FBP and a single G5-FAn taken from a solution of G5-FAn where n = 0, 4.7, 2.7, and 7.2.  A difference in rupture force was measured but due to the heterogeneity of the number of FAs per G5-FAn within a given solution, it was not possible to assign the measured forces to a specific number of FA-FBP interactions.

The second component of this thesis examines the interaction of a variety of non-targeted, charged nanoparticles with cells and model membranes.  Five polycationic polymers (G5 and G7 PAMAM dendrimer, branched polyethylenimine (PEI), poly-L-lysine (PLL) and diethylaminoethyl-dextran (DEAE-DEX)) were shown to induce nanoscale hole formation in cells as measured by enzyme and dye diffusion assays, as well as in dimyristoylphosphatidylcholine (DMPC) supported lipid bilayers (SLB) as measured by atomic force microscopy (AFM).  In contrast, neutral polymers polyethylene glycol and polyvinyl alcohol did not induce nanoscale hole formation in cells or DMPC SLBs.  This suggests that a possible mechanism for polycationic polymer internalization and/or nanoparticle-induced cytotoxicity of cells is through nanoscale hole formation.  The interaction between SLBs and a variety of other charged nanoparticles (MSI-78, Au-NH2, G3-NH2 PAMAM dendron, and silica-NH2 were also investigated using the AFM-SLB assay.  The general trend taken from the AFM-SLB studies is that surface area of the polycationic nanoparticles is the largest contributing factor to membrane disruption.  In addition, micelles of charged detergents cetyl trimethylammonium bromide bromide (CTAB) and sodium dodecyl sulfate (SDS) were also shown to induce hole formation in SLBs. 

Awards

Fellowship Awardee and Member of the University Education Partnership Program (UEPP), Lawrence Livermore National Lab (2005-2008)

Selected to the Excellence in Graduate Research Symposium, Polymer Chemistry Division of ACS (Spring 2007)

Recipient of Graduate Student Instructor Award, Department of Chemistry (2004)

Recipient of Graduate Assistance in Areas of National Need (GAANN) Fellowship (2003)

Former President of Associated Students of Whitman College (2001-2002)

Publications

Kelly, C., Liroff, M., Triplett, D., Leroueil, P., Mullen, D., Wallace, J., Meshinchi, S., Baker, J.R., Orr, B., Banaszak Holl, M., “Stoichiometry and Structure of Poly(amidoamine) Dendrimer-Lipid Complexes.”  ACS Nano, 2009: 7, 1886-1896.

Erickson B., DiMaggio S., Mullen D., Kelly, C., Leroueil, P., Berry, S., Baker, J.R., Orr, B., Banaszak Holl, M., “Interactions of poly(amidoamine) dendrimers with Survanta lung surfactant: The importance of lipid domains.”  Langmuir, 2008: 24, 11003-11008.

Kelly, C., Leroueil, P., Orr, B., Nett E., Werezcynski J., Baker J.R., Orr, B, Banaszak Holl, M, Andricioaei I., “Poly(amidoamine) dendrimers on lipid bilayers I: Free energy and conformation of binding.” Journal of Physical Chemistry B, 2008: 112, 9337-9345.

Kelly, C., Leroueil, P., Orr, B., Banaszak Holl, M., Andricioaei I.  “Poly(amidoamine) dendrimers on lipid bilayers II: Effects of bilayer phase and dendrimer termination.” Journal of Physical Chemistry B, 2008: 112, 9346-9353.

Leroueil P., Berry S., Duthie K., Han G., Rotello V., McNerny D., Baker J.R., Orr B., Banaszak Holl M.  “Wide varities of cationic nanoparticles induce defects in supported lipid bilayers” Nano Lettedr, 2008: 8, 420-424. 

Leroueil P., Hong S.Y., Mecke A., Baker J.R., Orr BG, Banaszak Holl, M, “Nanoparticle interaction with biological membranes: Does nanotechnology present a janus face?”  Accounts of Chemical Research, 2007: 40, 335-342.

Hong S., Leroueil P, Majoros, I, Orr B., Baker JR, Jr., Banaszak Holl, M., “The Binding Avidity of a Nanoparticle-Based Multivalent Targeted Drug Delivery Platform.” Chemistry and Biology,2007:14, 107–115.

Hong S, Hessler J, Banaszak Holl M, Leroueil P, Mecke A, Orr B: “Physical interactions of nanoparticles with biological membranes: The observation of nanoscale hole formation.” Chemical Health & Safety, 2006:16–20

Hong S, Leroueil P, Janus E, Peters J, Kober M, Islam MT, Orr B, Baker JR, Jr., Banaszak Holl M: “Interaction of Polycationic Polymers with Supported Lipid Bilayers and Cells: Nanoscale Hole Formation and Enhanced Membrane Permeability.”  Bioconjugate Chemistr., 2006:17, 728–734.  (An ACS Most-Cited Article 2006)

Kevin Landmark, Ph.D. Applied Physics   Kevin graduated summa cum laude from Michigan Technological University with a B.S. in Physics in 2000, advised by Dr. Robert Weidman. He then spent two years as a test engineer with Visteon, a Tier-1 automotive components supplier, gaining practical industrial experience before deciding to return to academia.

As a member of the M-NiMBS team, Kevin has successfully synthesized and characterized dendrimer-functionalized magnetic nanoparticles for use as targeted contrast agents. Targeted uptake of the devices was verified using visible fluorescence from dye labels on the dendrimers as well as X-ray fluorescence from iron in the iron oxide cores. The latter experiments were conducted at the Advanced Photon Source, Argonne National Lab. This work has been published in ACS Nano: Landmark, K. J.; DiMaggio, S.; Ward, J.; Kelly, C.; Vogt, S.; Hong, S.; Kotlyar, A.; Myc, A.; Thomas, T. P.; Penner-Hahn, J. E.; Baker, J. R., Jr.; Banaszak Holl, M. M.; Orr, B. G. Synthesis, Characterization, and in Vitro Testing of Superparamagnetic Iron Oxide Nanoparticles Targeted Using Folic Acid-Conjugated Dendrimers. ACS Nano 2008, 2, (4), 773-783.

Kevin Landmark's Thesis:

DENDRIMER-COATED IRON OXIDE NANOPARTICLES AS TARGETED MRI CONTRAST AGENTS

by Kevin J. Landmark

Friday, May 16, 2008, 8:30 am, 335 West Hall

Co-Chairs: Mark M. Banaszak Holl and Bradford G. Orr

Other committee members: James R. Baker, Jr. and Roy Clarke

Targeted MRI contrast agents are anticipated to be critical tools in realizing the dream of predictive and preventative medicine.  Many approaches have been documented regarding superparamagnetic iron oxide nanoparticles (SPIONs) as contrast agents and the use of various ligands to actively target them to specific tissues.  This dissertation explores SPIONs coated and targeted by functionalized dendrimers for specific uptake by cancer cells via the folic acid receptor (FAR).

Monodisperse SPIONs were first prepared in organic solvents (OC-SPIONs).  Amine-terminated generation 5 poly(amidoamine) (G5-PAMAM) dendrimers were conjugated with an average of five folic acid (FA) moieties for targeting and three 6-TAMRA (6T) dye molecules for tracking by optical fluorescence.  To minimize nonspecific interactions, the remaining amino groups were neutralized by capping with acetyl (Ac) groups.  The resulting polymer units, G5-Ac(102)-FA(5)-6T(3), were used to transfer OC-SPIONs from organic to aqueous media, imparting protection, biocompatibility, optical tracking and targeting in a single step.  Following phase transfer, the dendrimer-coated SPIONs (DC-SPIONs) exhibited key properties for effective contrast agents: they retained their size and shape uniformity and exhibited a high saturation magnetization.

The ability of the dendrimer-coated SPIONs (DC-SPIONs) to be specifically internalized by cancer cells overexpressing the FAR was confirmed and quantified in vitro.  Targeted uptake of the dendrimer coatings and SPION cores was independently verified by two distinct but complementary techniques: flow cytometry for the dendrimers (6-TAMRA signal) and X-ray fluorescence (XRF) microscopy for the SPIONs (elemental iron signal).  Using XRF microscopy is unique because it enables quantification of iron uptake at the single-cell level versus analysis on bulk cell samples.  The XRF microscopy data reveal a wide variation in iron uptake that correlates well with the uptake distribution from flow cytometry and is consistent with the variability in uptake observed for neat G5-Ac(102)-FA(5)-6T(3).

Publications

Synthesis, Characterization, and In Vitro Testing of Superparamagnetic Iron Oxide Nanoparticles Targeted Using Folic Acid-Conjugated Dendrimers

Eric Tkaczyk University of Michigan MD/PhD program May 2010 Nanobiology certificate program Eric Tkaczyk graduated magna cum laude with degrees in mathematics and Electrical Engineering from Purdue University and minors in German and French. Now, he is a fellow in the University of Michigan MD/PhD program.  His research interests include flow cytometry, pulse-shaping, coherent control, ultrafast optics, multiphoton microscopy, and their medical applications.  Previously, he pursued quantum chemistry

research in Estonia and multiphoton microscopy in France. In 2006, Tkaczyk won the International Biomedical Optics Society Best Student Paper Award at the Photonics West conference.  Estonia and multiphoton microscopy in France.  In 2007, he won first prize in the Student Paper Competition at the IEEE International Summer School and Symposium on Medical Devices and Biosensors in Cambridge, England.

Eric completed his PhD degree requirements and is expected to complete his MD degree in May 2010.

Eric Tkaczyk's Thesis:

FEMTOSECOND LASER PULSE OPTIMIZATION FOR MULTIPHOTON CYTOMETRY AND CONTROL OF FLUORESCENCE

May 1, 2008, 2pm, Duderstadt 1180

Chair: Theodore B. Norris

Other committee members are: Gary Luker, Jennifer Ogilvie, and Duncan Steel.

This body of work encompasses optimization of near infrared femtosecond laser pulses both for enhancement of flow cytometry as well as adaptive pulse shaping to control fluorescence. A two-photon system for in vivo flow cytometry is demonstrated, which allows noninvasive quantification of circulating cell populations in a single live mouse. We monitor fluorescently-labeled red blood cells for more than two weeks, and are also
able to noninvasively measure circulation times of two distinct populations of breast cancer cells simultaneously in a single mouse. We build a custom laser excitation source in the form of an extended cavity mode-locked oscillator, which enhances the two-photon signal strength several fold relative to a commercial laser. This enables superior detection in whole blood or saline of cell lines expressing fluorescent proteins including the green fluorescent protein (GFP), tdTomato and mPlum. A mathematical model explains unique features of the signals including: sub-square law scaling of unsaturated two-photon signal; a sigmoidal sensitivity curve for detection under varying excitation powers; and uncorrelated signal strengths in two detection channels.

The ability to distinguish different fluorescent species is central to simultaneous measurement of multiple molecular targets in high throughput applications including the multiphoton flow cytometer. We demonstrate that two dyes which are not distinguishable to one-photon measurements can be differentiated and in fact quantified in mixture via phase-shaped two-photon excitation pulses found by a genetic algorithm. We also selectively enhance or suppress two-photon fluorescence of numerous common dyes with tailored pulse shapes. Using a multiplicative (rather than ratiometric) fitness parameter, we are able to control the fluorescence while maintaining a strong signal. The importance of linear chirp and power scaling checks on the adaptive learning process is investigated in detail. With this method, we control the two-photon fluorescence of the blue fluorescent protein (BFP), which is of particular interest in investigations of protein-protein interactions, and has frustrated previous attempts of control. Implementing an acousto-optic interferometer, we use the same experimental setup to measure two-photon excitation cross-sections of dyes and prove that photon-photon interferences are the predominant mechanism of control.

This research establishes the basis for molecularly tailored pulse shaping in multiphoton flow cytometry, which will advance our ability to probe the biology of circulating cells during disease progression and response to therapy.

Awards and Honors
First Prize, Student Paper Competition, 4th IEEE International Summer School 8/07
and Symposium on Medical Devices and Biosensors, Cambridge, England
International Biomedical Optics Society Best Student Paper Award 1/06
Biomedical Optics Symposium, Photonics West 2006
NIH Medical Scientist Training Program Fellowship 8/08 – Present
National Science Foundation Graduate Research Fellowship 8/05 – 8/08
AMD Design Award, Purdue Engineering Projects in Community Service 4/04
Gates Scholar Elect 2/03
Hertz Fellowship Finalist 1/03
IEEE Technical Paper Writing Contest – 1st place 8/01
Tau Beta Pi National Nagel Scholarship 3/01
Two Intel Staff Recognition Awards for contributions to chipset design quality control 7/00, 8/00
Steven C. Beering Presidential Scholarship (full tuition, room/board, stipend) 8/98 – 5/03
Indiana Academy of Science Talent Search Winner 3/97
National Estonian Math Olympiad - 1st place 3/96

Publications

1. Tkaczyk ER, Tkaczyk AH, Mauring K, Ye JY, Baker JR, Norris TB, “Quantitative
differentiation of dyes with overlapping one-photon spectra by femtosecond pulse shaping,” Journal of Luminescence, 130: 29-34, January 2010. http://dx.doi.org/10.1016/j.jlumin.2009.07.014

2. Tkaczyk ER, Tkaczyk AH, Mauring K, Baker JR, Norris TB, “Control of two-photon
fluorescence of common dyes and conjugated dyes,” Journal of Fluorescence, 19(3): 517-533, May 2009 PMID: 19082694

3. Krasnenko V, Tkaczyk AH, Tkaczyk ER, Mauring K, “Physicochemical properties of Blue Fluorescent Protein determined via molecular dynamics simulation,” Biopolymers, 89(12): 1136-1143, December 2008. PMID: 18690664

4. Tkaczyk ER, Mauring K, Tkaczyk AH, Krasnenko V, Baker JR, Norris TB, “Control of the blue fluorescent protein with advanced evolutionary pulse shaping,” Biochemical and Biophysical Research Communications, 376(4): 733-737, November 2008. PMID: 18817751

5. Tkaczyk ER, Tkaczyk AH, Katnik S, Ye JY, Luker KE, Luker GD, Myc A, Baker JR,
Norris TB, “Extended cavity laser enhanced two-photon flow cytometry,” Journal of
Biomedical Optics, 13(4): 041319-1-041319-12, August 2008.
http://dx.doi.org/10.1117/1.2967983 PMID: 19021327

6. Zhong CF, Tkaczyk ER, Thomas TP, Ye JY, Myc A, Bielinska AU, Cao Z, Majoros I,
Keszler B, Baker JR, Norris TB, “Quantitative two-photon flow cytometry - in vitro and in vivo,” Journal of Biomedical Optics, 13(3): 034008-1-034008-19, June 2008.
http://dx.doi.org/10.1117/1.2931077 PMID: 18601553

7. Tkaczyk ER, Zhong CF, Ye JY, Myc A, Thomas TP, Cao Z, Duran-Struuck R, Luker KE, Luker GD, Norris TB, Baker JR, “In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry,” Optics Communications, 281(4), 888-894, February 2008. PMID: 19221581

8. Krasnenko V, Tkaczyk AH, Tkaczyk ER, Farkas Ö, Mauring K,“Vibrations-determined
properties of Green Fluorescent Protein, ” Biopolymers, 78(3), 140-146, June 2005.
PMID: 15759290

9. Roeder RA, Geddes LA, Tkaczyk ER, “Commotio cordis in children, including a translation of the first paper that describes the condition,” Cardiovascular Engineering, 3(4), 147-148, December 2003. http://dx.doi.org/10.1023/B:CARE.0000018828.12754.c9

10. Tkaczyk ER, Ermentrout B, “Pressure hallucinations and patterns in the brain,” Morehead. Electronic Journal of Applied Mathematics, 1(1), 1-26, 2001.

11. Tkaczyk AH, Tkaczyk ER, Norris TB, Takayama S, “Microfluidic droplet consistency
monitoring and cell detection via laser excitation,” Journal of Mechanics and Medicine in Biology (JMMB), submitted 2009

12. Cheng J, Waite A, Tkaczyk ER, Ke K, Richards N, Hunt AJ, Gumucio D, “Kinetic
properties of ASC protein aggregation in epithelial cells,” Journal of Cellular Physiology,
submitted 2009

13. Chang YC, Ye JY, Thomas TP, Cao Z, Kotlyar A, Tkaczyk ER, Baker JR, Norris TB,
“Fiber-optic multiphoton in vivo flow cytometry,” Journal of Biomedical Optics, submitted 2009.

Seungpyo Hong, Ph.D. Assistant Professor Departments of Biopharmaceutical Sciences and Bioengineering University of Illinois at Chicago 833 S. Wood St. Rm 355 Chicago, IL 60612 Office: 312)413-8294 Cell: 734)239-3173 E-mail: sphong@uic.edu

Dr. Seungpyo Hong is currently an assistant professor in the Department of Biopharmaceutical Sciences, College of Pharmacy at the University of Illinois at

Chicago.  His group's research focus lies at the interface of materials science, biology and nanotechnology to develop novel polymer-based nanomaterials for biological analysis, diagnostics and therapeutics. 

He graduated from Hanyang University, Korea with M.S. (2001) and B.S. (1999) degrees in polymer engineering. After working as a researcher at Korea Institute of Science and Technology, he started his Ph.D. study as a Dwight F. Benton Fellow working with his advisors Prof. Mark Banaszak Holl and Prof. James Baker, Jr. at MNiMBS of the University of Michigan.  His research in nanomedicine involved preparation and functionalization of organic nanoparticles such as dendrimers and their interactions with cells, with a particular interest in nanotoxicology and targeted drug delivery.  His thesis tetle was: Interaction of  Synthetic Polymers with Cell Membranes: Cell Penetration of Polycationic Polymers and Multivalent Effects of Targeted Nanodevices.

Seungpyo graduated with his Ph.D. in Macromolecular Science and Engineering in 2006 and joined Prof. Robert Langer's lab at MIT as a postdoc.  At MIT, he continued in nanomedieince and worked on research projects to develop polymer-based devices which can specifically recognize/sort/kill cancer cells or stem cells using cell rolling behavior.  He recently started his assistant professorship at UIC.  The nanomedicine / nanobiology training he received at MNiMBS allowed him to integrate his expertise in multiple disciplines to ultimately become faculty at the College of Pharmacy.  Dr. Hong has published 19 peer-reviewed papers, over 35 abstracts, and 3 issued or pending US patents.  He has received honors from research communities, which includes a 2006 Most-Cited ACS Journal Article, Best Poster Award at the fall 2005 MRS meeting, and the University of Michigan’s Charles G. Overberger Award. 

Publications while at Michigan of a total of 21 overall per November 2009.

The Binding Avidity of a Nanoparticle-Base Multivalent Targeted Drug Delivery Platform.  S. Hong, I. Majoros, B. G. Orr, J. R. Baker, Jr., M. M. Banaszak Holl.  Chemistry and Biology 2007, 14, 107-115.

HPLC analysis of functionalized poly(amidoamine) dendrimers and the interaction between a folate-dendrimer conjugate and folate binding protein.  X. Shi, X. Bi, T.   R. Ganser, S. Hong, L. A. Myc, A. Desai, M. M. Banaszak Holl, J. R. Baker.  The Analyst, 2006, 131, 842-848.

Interaction of Polycationic Polymers with Supported Lipid Bilayers and Cells: Nanoscale Hole Formation and Enhanced Membrane Permeability.  S. Hong, P. R. Leroueil, E. K. Janus, J. L. Peters, M.-M. Kober, M .T. Islam, B. G. Orr, J. R. Baker, Jr., and M. M. Banaszak Holl.  Bioconjugate Chemistry 2006, 17, 728-734. (An ACS Most-Cited Article 2006).

Physical Interactions of Nanoparticles with Biological Membranes:The Observation of Nanoscale Hole Formation.  S. Hong, J. A. Hessler, M. M. Banaszak Holl, P. Leroueil, A. Mecke, B .G. Orr.  Chemical Health and Safety  2006, 13, 16-20.

The Interaction of Polyamidoamine (PAMAM) Dendrimers with Supported Lipid Bilayers and Cells:  Hole Formation and the Relation to Transport.  S. Hong, A. U. Bielinska, A. Mecke, B. Keszler, J. L. Beals, X. Shi, L. Balogh, B. G. Orr, J. R. Baker Jr., and M. M. Banaszak Holl.  Bioconjugate Chemistry 2004, 15, 774-782.

Jessica Blunt, Ph.D. Bachelor of Science Grand Valley State University, 2001 Doctor of Philosphy University of Michigan, 2006 Jessica Hessler Blunt was an undergraduate at Grand Valley State University and graduated with her PhD in Chemistry from Michigan in 2006. She published the first AFM images ever acquired of a cell undergoing pro-grammed cell death (Langmuir 2005, 21, 9280-9286).

After graduation, she worked at the Van Andel Institute in the lab of Craig Webb.  She is now working in the field of intellectual property.

Publications

Cationic Nanoparticles Induce Nanoscale Disruption in Living Cell Plasma MembranesJ. Chen, J. A. Hessler, K. Putchakayala, B. K. Panama, D. P. Khan, S. Hong, D. G. Mullen, S. C. DiMaggio, A. Som, G. N. Tew, A. N. Lopatin, J. R. Baker, M. M. Banaszak Holl, B. G. Orr. Journal of Physical Chemistry B. 2009, 113, 11179-11185.

Design and Implementation of a Studio-based General Chemistry Course at the University of Michigan. A. C. Gottfried, R. D. Sweeder, J. M. Bartolin, J. A. Hessler, B. P. Reynolds, I. C. Stewart, B. P. Coppola, and M. M. Banaszak Holl. J. Chem. Ed. 2007, 84, 265-270.

Physical Interactions of Nanoparticles with Biological Membranes:The Observation of Nanoscale Hole Formation. S. Hong, J. A. Hessler, M. M. Banaszak Holl, P. Leroueil, A. Mecke, B .G. Orr. Chemical Health and Safety 2006, 13, 16-20.

Atomic Force Microscopy Study of Early Morphological Changes During Apoptosis. J. Hessler, A. Budor, K. Putchakayala, A. Mecke, D. Rieger, M. M. Banaszak Holl, B. G. Orr, A. Bielinska, J. Beals, J. Baker. Langmuir 2005, 21, 9280-9286.

A Novel MEA/AFM Platform for the Measurement of Real-time, Nanometric Morphological Alterations of Electrically Stimulated Neuralblastoma Cells. M. B. Shenai, K. G. Putchakayala, J. A. Hessler, B. G. Orr, M. M. Banaszak Holl, and J. R. Baker. IEEE Transactions on Nanobioscience 2004, 3, 111-117.

Tapping Mode Atomic Force Microscopy Investigation of Poly(amidoamine) Core-Shell Tecto(Dendrimers) using Carbon-Nanoprobes. T. A. Betley, J. A. Hessler, A. Mecke, M. M. Banaszak Holl, B. G. Orr, S. Uppulari, D. A. Tomalia, and J. R. Baker Jr. Langmuir 2002, 18, 3127-3133.

Youngseon Choi, Ph.D. Scientist Nano-Bio Chemistry Group Institut Pasteur Korea Seoul, South Korea Dr. Choi earned his B. S. in Industrial Chemistry at Hanyang University, Seoul, South Korea in 1994. He earned his M.S. in Industrial Chemistry at Hanyang University, Seoul, South Korea in 1998. He earned his Ph.D. in Biomedical Engineering at the University of Michigan in Ann Arbor, MI in 2005.

From 2005 – 2006, he worked as a Postdoctoral Associate on the Application of Biomedical Nanotube Technology at the Center for Research at Bio/Nano Interface, Department of Chemistry, University of Florida, Gainesville, FL, where his Advisor was Professor Charles R. Martin.

Since 2006, Dr. Choi is working at the Institut Pasteur Korea at Seoul, Korea where his research focus is on integration of nanotechnology into target identification in drug discovery and drug screening. Specifically:

• in vitro assay development and in vivo cellular imaging based on nanoparticles including quantum dots, dendrimers, iron oxide, and gold nanoparticles;

(2) visualization of subcellular target genes or proteins using QD-based FRET approach;

(3) establishing cellular library expressing specific target proteins.

 
The focus of the work at the Institut Paster Korea in Seoul, Korea is on HIV, HCV, and TB for new drug discovery, and Dr. Choi’s group is supporting the work through a nanobiotechnology approach. 

Youngseon Choi’s thesis:

Nanostructured supramolecular Arrays based on Dendrimers Using DNA: design, synthesis and biological evaluation

Doctoral Committee:

James R. Baker Jr., MD - Co-Chair

Shuichi Takayama, Ph.D. - Co-Chair

Mark M. Banaszak Holl, Ph.D.

Bradford G. Orr, PhD

Jolanta F. Kukowska-Latallo, PhD

This dissertation describes a series of experiments that synthesize and characterize a new type of supramolecular array. These molecules are based on dendrimers linked together with complementary DNA oligonucleotides. The arrays have a good range of applications including cancer cell specific drug targeting. Three specific studies were designed to test the hypothesis that complementary DNA oligonucleotides would self-assemble different single-functional dendrimers to form functional supramolecular clusters.

First, in order to produce a library of functionalized dendrimers, generation 5 polyamidoamine (PAMAM) dendrimers were first partially acetylated, then conjugated with either imaging agents (fluorochromes; FITC, 6-TAMRA, and AF488) or targeting molecules (folic acid). The molecular characteristics of each functionalized dendrimers were characterized by UV-Vis, NMR, GPC, HPLC, and MALDI-TOF MS to ascertain their molar extinction, degree of functionalization, molecular weight and molecular weight distribution.

Secondly, a synthetic method to covalently attach DNA oligonucleotides to the partially acetylated dendrimers was developed based on phosphoramidate chemistry. This chemistry was then applied to attach three sets of complementary oligonucleotides of different lengths (34, 50, 66 bases-long) to partially acetylated generation 7 and 5 PAMAM dendrimers. The size and the shape of the DNA-linked nanostructure were characterized by dynamic light scattering (DLS) and atomic force microscopy (AFM).

Finally, we extended our strategy of DNA-based assembly to the construction of dimers of different single-functionalized dendrimers designed for cancer cell specific targeting. Three different batches of DNA-linked dendrimers consisting of imaging dendrimers (G5-FITC, G5-6-TAMRA, or G5-AF488) coupled to targeting dendrimer (G5-FA) consistently targeted KB cells overexpressing the high affinity folate receptor with maximum binding at 100 nM and an apparent binding affinity of ~ 50 nM and internalized into the cells after 1 hour incubation at 37 oC in vitro. Preliminary in vivo experiments using SCID mice showed that the clusters accumulated in tumor cells expressing folic acid receptors 16 and 48 hours after injection.

These results confirmed our hypothesis and demonstrated our ability to design and produce supramolecular arrays of dendrimers using DNA duplex formation. This DNA-linked dendrimer supramolecular array provides a potential platform for developing combinatorial targeted therapeutics in the future.

Dr. Choi’s Fellowships and Awards

1. World Technology Networks (WTN) Fellow (Materials/Nanotech 2005)
2. ICI Student Award, 2005 Polymeric Materials Science and Engineering (PMSE) Division in American Chemical Society (to be conferred in March 2006)
3. Outstanding Poster Award, 16th Macromolecular Science and Engineering Symposium, University of Michigan, Ann Arbor, 2004 Oct. 17.
4. Rackham Graduate School Travel Grant, University of Michigan, Ann Arbor, 2004 and 2005
5. Fellowship for Graduate School of Advanced Materials and Chemical Engineering, Hanyang University 1997

Dr. Choi’s publications based upon work done while at MNIMBS

Youngseon Choi and James R. Baker Jr. “Nanoparticles in Medical Diagnostics and Therapy”, in Nanotechnology in Biology and Medicine, CRC Press 2007 (Book chapter).

Youngseon Choi and James R. Baker Jr. “Targeting Cancer Cells with DNA-Assembled Dendrimers - a Mix-and-Match Strategy for Cancer”, Cell Cycle, 4, 669 – 671, 2005.

Youngseon Choi, Thommey Thomas, Alina Kotlyar, Mohammad T. Islam, James R. Baker Jr. , Polymeric Materials: Science and Engineering, 92, 556 - 557, 2005.

Youngseon Choi, Thommey Thomas, Alina Kotlyar, Mohammad T. Islam, James R. Baker Jr. “Synthesis and Functional Evaluation of DNA-Assembled Polyamidoamine (PAMAM) Dendrimers for Cancer Cell-Specific Targeting”, Chemistry & Biology, 12, 35- 43, 2005.

Youngseon Choi, Almut Mecke, Bradford G. Orr, Mark Banaszak Holl, James R. Baker Jr. “DNA-Directed Synthesis of Generation 7 and 5 Polyamidoamine (PAMAM) Dendrimer Nanoclusters”, Nano Letters, 2004, 4, 3, 342.

Almut Mecke, Ph.D. Clinical Documentation Specialist F. Hoffmann-La Roche Ltd.   Basel, Switzerland   My career path has taken me from graduate student research in nanotechnology to a job in the pharmaceuticals and biotechnology industry. After completing my Ph.D. degree at the University of Michigan in Physics in 2004 and two years of postdoctoral research, I joined F. Hoffmann-La Roche, a large

healthcare company based in Switzerland. I now work as a medical writer, which means one of my main tasks is to prepare detailed scientific reports about clinical trials designed to establish the safety and efficacy of new drugs. These documents are submitted to national health authorities (e.g. the FDA), who decide, on the basis of the data, whether the drug provides a benefit to patients and should be marketed. A very condensed version of the results of clinical trials is what you find in the leaflet sold with medication.

Although it might seem unusual for a physicist to do this job, in fact, my education at the Nanotechnology Institute at Michigan prepared me well for this position. While there, I was officially enrolled as a graduate student in physics, but I was fortunate to have the opportunity to work with an interdisciplinary group of scientists developing targeted cancer therapeutics. The Nanotechnology Institute brought together this diverse group of researchers from the departments of chemistry, physics, applied physics, biology, engineering and the medical school. It also provided funding, laboratory equipment and expertise. I was able to use my training in physics to contribute to the understanding of the mechanism of how the nanometer-sized polymers we were investigating can enter cancer cells and kill them.

At Roche, much of my work is related to the development of new oncology drugs, although I learn about other disease areas as well. Medical writers do not perform scientific experiments themselves, but we need to be able to interpret data and present large amounts of information in a clear, scientific way. A graduate degree related to life sciences is a requirement, as is a good writing style in English. Most importantly, our work is performed by teams of people with very diverse scientific and personal backgrounds. Drug development draws on many disciplines, such as chemistry, biology, toxicology, statistics, pharmacology, medical science and marketing. Thus, physics at U of M with a nanomedicine application to Roche doesn’t seem like such a big leap any more, does it? 

Publications from work at Michigan

Nanoparticle Interaction with Biological Membranes:  Does Nanotechnology Present a Janus Face? P. R. Leroueil, S. Hong, A. Mecke, J. R. Baker, Jr., B. G. Orr, M. M. Banaszak Holl Accounts of Chemical Research 2007 40 335-342

Nanoparticle-Membrane Interaction: Mechanism for Enhanced Permeability

2007

Physical Interactions of Nanoparticles with Biological Membranes:The Observation of Nanoscale Hole Formation. S. Hong, J. A. Hessler, M. M. Banaszak Holl, P. Leroueil, A. Mecke, B .G. Orr. Chemical Health and Safety 2006, 13, 16-20.

Atomic Force Microscopy Study of Early Morphological Changes During Apoptosis.

J. Hessler, A. Budor, K. Putchakayala, A. Mecke, D. Rieger, M. M. Banaszak Holl, B. G. Orr, A. Bielinska, J. Beals, J. Baker. Langmuir 2005, 21, 9280-9286.

Lipid Bilayer Disruption by Polyamidoamine Dendrimers: The Role of Generation and Capping Group. A. Mecke, B. G. Orr, M. M. Banaszak Holl, J. R. Baker. Langmuir 2005, 21, 10348-10354.

Membrane thinning due to antimicrobial peptide binding -- An AFM study of MSI-78 in DMPC bilayers. A. Mecke, D.-K. Kee, A. Ramamoorthy, B. G.Orr, M. M. Banaszak Holl. Biophysical Journal 2005, 89, 4043-4045.

Synthetic and natural polycationic polymers interact selectively with fluid phase domains of DMPC lipid bilayers. A. Mecke, D.-K. Lee, R. Ramamoorthy, B. G. Orr, and M. M. Banaszak Holl. Langmuir 2005, 21, 8588-8590.

Deformability of Poly(amidoamine) Dendrimers. A. Mecke, I. Lee, J. R. Baker Jr., M. M. Banaszak Holl, B. G. Orr. European Physical Journal E -- Soft Matter 2004, 14, 7-16.

Direct Observation of Lipid Bilayer Disruption by Poly(amidoamine) Dendrimers.

A. Mecke, S. Uppuluri, T. J. Sassanella, B. G. Orr, M. M. Banaszak Holl, J. R. Baker. Chemistry and Physics of Lipids 2004, 132, 3-14.

DNA-directed Synthesis of Generation 7 and 5 PAMAM Dendrimer Nanoclusters. Y. Choi, A. Mecke, B. G. Orr, M. M. Banaszak Holl, and J. R. Baker Jr. Nanoletters 2004, 4, 391-397.

The Interaction of Polyamidoamine (PAMAM) Dendrimers with Supported Lipid Bilayers and Cells: Hole Formation and the Relation to Transport. S. Hong, A. U. Bielinska, A. Mecke, B. Keszler, J. L. Beals, X. Shi, L. Balogh, B. G. Orr, J. R. Baker Jr., and M. M. Banaszak Holl. Bioconjugate Chemistry 2004, 15, 774-782.

Tapping Mode Atomic Force Microscopy Investigation of Poly(amidoamine) Core-Shell Tecto(Dendrimers) using Carbon-Nanoprobes. T. A. Betley, J. A. Hessler, A. Mecke, M. M. Banaszak Holl, B. G. Orr, S. Uppulari, D. A. Tomalia, and J. R. Baker Jr.. Langmuir 2002, 18, 3127-3133.

Mahesh B. Shenai, M.D., M.S.E. Internship in General Surgery (2005-2006) Resident in Neurosurgery (2006-2011) University of Alabama-Birmingham Interests: neural prosthetics, brain-computer interface, functional neurosurgery Mahesh B. Shenai was a Howard Hughes Medical Student Fellow at the MNIMBS between the years of 2002-2003. During his fellowship, he utilized the multidisciplinary resources of the

Institute, to develop a novel platform for the investigation of nanoscale neurophysiology. The MNIMBS was instrumental in allowing him to propose and develop a unique concept, from paper to practice. Dr. Shenai’s work was published in 2003, and is the first documented observation of nanometer level structural changes in electrically stimulated neuronal cells.  The work was also presented at the Howard Hughes Institute, and the Society for Neuroscience  in 2003.  Additionally, Dr. Shenai developed an interest in the nanodesign of microelectrodes, ultimately needed for a successful brain-computer interface.

Currently, Dr. Shenai is a senior resident in neurological surgery, at the University of Alabama-Birmingham.  He is currently involved in the development of brain-computer interfaces, and the analysis of deep brain stimulation.

Publications

1. Shenai MB, Ross DA, Sagher  O,  “The Use of Multiplanar Trajectory Planning in the Stereotactic Placement of Depth Electrodes”, Neurosurgery. 2007 Apr;60(4 Suppl 2):272-6; discussion 276.

2. Xiao Y, Martin DC, Cui X, Shenai M.  “Surface Modification of Neural Probes With Conducting Polymer Poly(hydroxymethylated-3,4- ethylenedioxythiophene) and Its Biocompatibility”  Applied Biochemistry and Biotechnology, February 2006,  128(2):117-130  (ISSN: 0273-2289).

3. Shenai M., Putchakayala K., Hessler J, Orr B, Banaszak Holl M, Baker J Jr., " A Novel MEA/AFM Platform for the Measurement of Real-time Nanometric Morphological Alterations of Electrically Stimulated Neuroblastoma Cells ."  IEEE Transactions in Nanobioscience, June 2004, 3(2):111-117. 

4. Shenai M., "Myocardial ischemia detection: a time-frequency investigation of intra-QRS changes in the endocardial electrogram." MSE thesis, Department of Biomedical Engineering,  The Johns Hopkins University. 2000. Advisor: Dr.Nitish Thakor.

5. Thakral A. and Stein L, Shenai M., Gramatikov B., Thakor N.V., "Effects of anodal versus cathodal pacing on the mechanical performance of the isolated rabbit heart." Journal of Applied Physiology, 89:1159-1164, 2000.

6. Shenai M., Gramatikov B, Thakor N.V., "Computer Models of  Depolarization Alterations Induced by Myocardial Ischemia. The Effect of Superimposed Ischemic Inhomogeneities on Propagation in Space-, Time-, and Time-Frequency Domains." The Journal of Biological Systems, December 1999, pp 1-22.

7. Thakor N.V., Iyer V. and Shenai M, "From Cellular Electrophysiology to Electrocardiography." Modeling and Imaging of Bioelectric Activity: Principles and Applications, Kluwer Academic/Plenum Publishers (February, 2004;  Hardbound, ISBN 0-306-48112-X)  pp 1-42.

Frank Zhong, Ph.D. Optical Engineer Pacific Biosciences Menlo Park, CA Frank Zhong came to University of Michigan in the summer of 2001 and began his graduate study initially at the Physics Department.  While obtaining solid training of fundamental modern physics for the first year, Frank found his interests to explore applications in various branches of engineering.  After

sitting in a presentation given by Prof. Ted Norris, Frank was drawn to a multi-discipline research project ‘in vivo flow cytometry’ to develop an instrument for non-invasive, real-time monitoring of radiation-induced illness in space.  After joining CUOS and MNIMBS, Frank worked closely with a group of scientists and researchers led by Prof. Jim Baker on a nanomedieince application.  During his graduate study, Frank developed a prototype instrument, which enabled real-time monitoring of biosensors tagged blood cells zipping through blood capillaries near the skin for the first time.  Currently, the prototype instruments are used by doctors in the medical school to investigate the dynamics of circulating cells in cancer and other important diseases.  The broad research experience at MNIMBS prepared Frank well not only in nanotechnology and engineering competence, but in truly in-depth collaboration with multi-discipline groups as well.

After receiving his Ph.D. degree in 2005, Frank Zhong joined an early stage startup biotechnology company, Pacific Biosciences, based in Menlo Park, CA as an optical engineer.  He has been working with a team of scientists and engineers in the development of a transformative single-molecule, real-time (SMRT) DNA sequencing technology, which enables, for the first time, the observation of natural DNA synthesis by a DNA polymerase as it occurs.  Most recently, Frank is working on the commercialization of the SMRT technology, with a goal to eventually enable sequencing of individual genomes as part of routine medical care. 

Publications resulting from work done at Michigan

J. Eid, C.F. Zhong, et al. Single-Molecule, Real-Time DNA Sequencing.  Science 2 January 2009: Vol. 323. no. 5910, pp. 133 - 138

P. Lundquist, C.F. Zhong, et al. Parallel confocal detection of single molecules in real time.  Opt. Lett. 33, 1026-1028 (2008)

C.F. Zhong, et al. Quantitative two-photon flow cytometry – in vitro and in vivo. Journal of Biomedical Optics, 13(3), 2008, p.034008

E.R. Tkaczyk, C.F. Zhong, et al. In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry. Optics Communications. Vol.281, 2008, p.888-894

E.R. Tkaczyk, C.F. Zhong, et al. Two-photon, two-color in vivo flow cytometry to noninvasively monitor multiple circulating cell lines.  Proc. SPIE, Vol.6631, 66310T (2007)

C.F. Zhong, et al. Two-photon Flow Cytometry. Proceedings of SPIE Volume: 5700, Mar 2005 p. 78-89