William Zamboni Lab
William Zamboni Lab
Dr. Zamboni is an expert in translational studies of anticancer agents. The Zamboni lab, in the Genetic Medicine Research Building, is a drug development and clinical pharmacology lab that focuses on the translational development of drugs, anticancer agents, and nanoparticles. Dr. Zamboni also supervises the Good Laboratory Practice Analytical Facility in Kerr Hall, which supports the development of newly discovered drugs and medical testing procedures.
The goals of the Translational Oncology and Nanoparticle Drug Development Initiative (TOND2I) Laboratory in the Genetic Medicine Building are to use analytical chemistry and pharmacologic infrastructure, methodologies and expertise to support the translational development of small molecule, nanoparticle, and biological anticancer agents. The lab develops and validates analytical assays from biological matrices, analyzes samples, performs pharmacokinetic (PK) and pharmacodynamic (PD) analyses of data and provides specialized methods to evaluate the pharmacology of carrier-mediated agents (e.g. nanoparticle, conjugates, antibody drug conjugates). Increased demand for applied pharmacology services have been created by the expansion of the Molecular Therapeutics Program, development of preclinical models to evaluate anticancer agents, and opening of the new North Carolina Cancer Hospital’s (NCCH) Clinical Trials Unit (CTU). Our lab’s capabilities expand research horizons and funding by providing state of the art support for sample analysis and performing pharmacology studies, and for developing novel methods to meet the needs for sample information.
Examples of this work includes:
1. A series of studies on the development of Particle Replication in Nonwetting Templates (PRINT®) nanoparticles in collaboration with Dr. DeSimone;
- Merkel TJ, Chen K, Jones SW, Pandya AA, Tian S, Napier ME, Zamboni WE, DeSimone JM. The effect of particle size on the biodistribution of low-modulus hydrogel PRINT particles. J Control Release. 2012 Aug 20;162(1):37-44.
- Chu KS, Hasan W, Rawal S, Walsh MD, Enlow EM, Luft JC, Bridges AS, Kuijer JL, Napier ME, Zamboni WC, DeSimone JM. Plasma, tumor and tissue pharmacokinetics of Docetaxel delivered via nanoparticles of different sizes and shapes in mice bearing SKOV-3 human ovarian carcinoma xenograft. Nanomedicine. 2013 Jul;9(5):686-93.
- Sambade M, Deal A, Schorzman A, Luft JC, Bowerman C, Chu K, Karginova O, Swearingen AV, Zamboni W, DeSimone J, Anders CK. Efficacy and pharmacokinetics of a modified acid-labile docetaxel-PRINT(®) nanoparticle formulation against non-small-cell lung cancer brain metastases. Nanomedicine (Lond). 2016 Aug;11(15):1947-55.
- Bowerman CJ, Byrne JD, Chu KS, Schorzman AN, Keeler AW, Sherwood CA, Perry JL, Luft JC, Darr DB, Deal AM, Napier ME, Zamboni WC, Sharpless NE, Perou CM, DeSimone JM. Docetaxel-Loaded PLGA Nanoparticles Improve Efficacy in Taxane-Resistant Triple-Negative Breast Cancer. Nano Lett. 2017 Jan 11;17(1):242-248.
2. Local iontophoretic (IO) administration of cytotoxic therapies to solid tumors in collaboration with Dr. Yeh and DeSimone;
- Byrne JD, Jajja MR, O’Neill AT, Bickford LR, Keeler AW, Hyder N, Wagner K, Deal A, Little RE, Moffitt RA, Stack C, Nelson M, Brooks CR, Lee W, Luft JC, Napier ME, Darr D, Anders CK, Stack R, Tepper JE, Wang AZ, Zamboni WC, Yeh JJ, DeSimone JM. Local iontophoretic administration of cytotoxic therapies to solid tumors. Sci Transl Med. 2015 Feb 4;7(273):273ra14.
- Byrne JD, Jajja MR, Schorzman AN, Keeler AW, Luft JC, Zamboni WC, DeSimone JM, Yeh JJ. Iontophoretic device delivery for the localized treatment of pancreatic ductal adenocarcinoma. Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):2200-5.
3. The evaluation of nanoparticle treatment of intracranial malignancies with Dr. Anders.
- Anders CK, Adamo B, Karginova O, Deal AM, Rawal S, Darr D, Schorzman A, Santos C, Bash R, Kafri T, Carey L, Miller CR, Perou CM, Sharpless N, Zamboni WC. Pharmacokinetics and efficacy of PEGylated liposomal doxorubicin in an intracranial model of breast cancer. PLoS One. 2013 May 1;8(5):e61359.
- Karginova O, Siegel MB, Van Swearingen AE, Deal AM, Adamo B, Sambade MJ, Bazyar S, Nikolaishvili-Feinberg N, Bash R, O’Neal S, Sandison K, Parker JS, Santos C, Darr D, Zamboni W, Lee YZ, Miller CR, Anders CK. Efficacy of Carboplatin Alone and in Combination with ABT888 in Intracranial Murine Models of BRCA-Mutated and BRCA-Wild-Type Triple-Negative Breast Cancer. Mol Cancer Ther. 2015 Apr;14(4):920-30.
Example of previously funded grants and studies includes:
- Pre-clinical Studies of Targeted Nanoparticle Delivery Agents
- Studies Evaluating Measures of the Reticuloendothelial System as Predictors of Doxil Pharmacokinetic and Pharmacodynamic Disposition in Patients with Refractory Ovarian Cancer, sponsored by the University Cancer Research Fund (UCRF) and Carolina Center for Cancer Nanotechnology Excellence (C-CCNE) Pilot Grant
- Study evaluating Relationship between Mononuclear Phagocytic System in Tumors and Tumor Delivery and Efficacy of Nanoparticle Anticancer Agents in Genetically Engineered Mouse Models of Breast Cancer, sponsored by NC Translational and Clinical Sciences Institute
- Precision Engineering of Ultrasonically-Targeted Drug Delivery Vehicles, sponsored by NC State University.
- A New Dimension in Renal Clearance Design Criteria for Dendrimer Nanostructures, sponsored by NIH/NIDDKD.
- Organismal and Genetic Networks in Drug Reward and Reinforcement, sponsored by NIH.
- STTR Phase II Grant: Blockage of NF-Kappa B for Prevention/Treatment of GVHD, sponsored by NIH/NCI P42 Grant.
- Evaluating the Role of Genotype in Tamoxifen Therapy for Breast Cancer, sponsored by UNC Hospitals.
- A Two Arm Phase I Study of Sorafenib with Carboplatin/Pemetrexed and Cisplatin/Etoposide, sponsored by UNC Hospitals.
- Phase I study utilizing an intravenous busulfan test dose to prospectively target and determine the maximum tolerated systemic exposure (MTSE) of a continuous intravenous infusion of busulfan, sponsored by UNC Hospitals.
- Evaluating the Effect of Aprepitant on Cyclophosphamide Pharmacokinetics, sponsored by UNC Hospitals.
- Solid phase separation of encapsulated and released fractions of novel nanoparticle agents in samples: Dr. Zamboni’s lab has developed a solid phase separation (SPS) method that differentiates the inactive nano-carrier encapsulate and the active ‘released’ forms of many common nanomedicines. This process can be applied to any solution or plasma sample and combined with an instrumental assay to determine concentrations of encapsulated, released, and total drug in a given sample. SPS has been used in the lab to separate the encapsulated and released forms of nano-formulations of platinum analogues, camptothecins, taxanes, and anthracyclines, as well as liposomal topotecan and pegylated liposomal doxorubicin (PLD/Doxil). These are the only known sample processing methods that can directly measure the encapsulated and released forms of a nanomedicine agent.
- Biological sample processing: The lab is proficient in sample processing development for novel nano agents in tumor, tissues, and plasma and other fluids. Processing can involve centrifugation, liquid/liquid extraction, evaporation and reconstituting in solvent, solid phase extraction, or a combination of these and other techniques.
- HPLC or LC-MS/MS analysis: Capabilities include high performance liquid chromatography (HPLC) with fluorescence detection (all HPLC equipment by Shimadzu) and HPLC coupled with mass spectrometry (MS). MS capabilities available include triple quad fragmentation analysis (Thermo Quantum Ultra LC-MS/MS), orbitrap full scan analysis for very high resolution and mass accuracy (Thermo Exactive LC-MS), and a hybrid orbitrap-linear ion trap instrument that combines the strengths of both triple quad and orbitrap analysis (Thermo LTQ Orbitrap Discovery).
- Determination and verification of physico-chemical properties
- Validation of chemical composition, molecular structure and mass
- Qualitative and quantitative detection
- Development of in vivo pharmacokinetic and pharmacodynamic studies: The lab has extensive experience evaluating the ADME, pharmacokinetic, toxicology, and efficacy of nano-carrier and non-nano-carrier anticancer agents in preclinical models. This includes the development of sampling strategies and schedules, sampling processing methods, and analytical assays for ADME and pharmacokinetic studies of nano agents in blood, plasma, tissues, tumor, urine, and bile. The lab works closely with university animal facilities for study management and harvesting and has access to an animal facility within the same building. The Animal Studies Core, affiliated with the UNC Lineberger Comprehensive Cancer Center, provides full access to a range of cancer cell lines and animal models, including genetically modified mouse models of human cancers. Consultation is available for assistance with or management of study design and data analysis.
- Pharmacokinetic/pharmacodynamic analysis (PK/PD) :Data collected at multiple timepoints after the administration of a nanomedicine in animal models allows for a complete pharmacokinetic and pharmacodynamic analysis of the nano agent in vivo. PK/PD assessments are performed using WinNonlin data management, a statistics, modeling, and visualization tool for PK data analysis. Parameters typically provided for a given nanomedicine include drug concentration over time, area under the curve (AUC), elimination half life, clearance, and time of maximum concentration.
- Efficacy studies: Studies may be designed that will evaluate a nano agent’s effects in vivo over an extended period of time (usually several weeks). Dose administration at one or several timepoints can be used to evaluate overall toxicity in the animal model and any effects on tumor cells and growth. Efficacy can be measured via survival rate, tumor growth, and quantitative analysis of drug in the tumor.
- Pheno-GLO High-throughput Screening Platform (HTSP): Preliminary studies by the lab suggest that the pharmacokinetic and pharmacodynamic disposition of nanomedicines is related to the function of the immune system, particularly the monocytes and dendritic cells of the mononuclear phagocyte system (MPS). These cells, as well as tissue macrophages, located primarily in the liver and spleen, lymph nodes and general circulation, serve as a potential clearance pathway for nanoparticles. Pheno-GLO HTSP is a process developed internally that uses cellular function assays and flow cytometry to evaluate the relationship between nano agents and the MPS. This relationship can indicate the activity and level of stimulation that a given nanomedicine may have within the immune system. Profiling these interactions builds upon information gained from initial physico-chemical measurements to provide a clearer picture of a nanomedicine’s behavior as defined by its properties.
The following analytical assays are available to be performed by the Zamboni lab:
- Docetaxel – total and protein unbound
- Irinotecan (CPT-11)
- DTPA and C2E5
- nab-paclitaxel (Abraxane)
- Irinotecan (CPT-11)
- SN-38 and glucuronidated SN-38 (SN-38G)
- Camptothecin (CPT)
- Doxorubicin – total and DNA-bound
- PEGylated liposomal doxorubicin (e.g. Doxil and Lipodox) – total, encapsulated, released
- Carboplatin – total and protein unbound
- Cisplatin – total and protein unbound
- Oxaliplatin – total and protein unbound
- Cocaine and metabolites
Instrumentation in the Zamboni Lab
- Shimazdu 20 series High Performance Liquid Chromatograph (HPLC) systems with fluorescence detection
- Thermo TSQ Quantum Ultra triple quad Mass Spectrometer with attached Shimazdu 20 series HPLC
- Thermo TSQ Quantum Access triple quad Mass Spectrometer with attached Shimadzu 10 series HPLC
- Thermo LTQ Orbitrap Discovery Fourier Transform orbitrap/ion trap Mass Spectrometer with attached Shimazdu 20 series HPLC
- Thermo Exactive orbitrap Mass Spectrometer with attached Shimazdu 20 series HPLC
- Agilent 7700 Inductively-Coupled Plasma Mass Spectrometer (ICP-MS)
My research program is part of the Division of Pharmacotherapy and Experimental Therapeutics (DPET) in the Eshelman School of Pharmacy at the University of North Carolina (UNC) and the UNC Lineberger Comprehensive Cancer Center (LCCC). I have been involved in translational studies of anticancer agents for several years. My research interests focus on the application of pharmacokinetic, pharmacodynamic, and pharmacogenetic principles in the optimization of the chemotherapeutic treatment of cancer. Information obtained from preclinical and clinical translational studies can greatly add to the understanding of the pharmacology of anticancer agents, permit individualization of chemotherapeutic treatment based on pharmacokinetic, pharmacodynamic, and pharmacogenetic principles, and allow for the rational design of therapeutic regimens.
A second focus of my research is on the development of liposomal and nanoparticle anticancer agents and evaluating the relationship between the disposition of these agents and the mononuclear phagocyte (or reticuloendothelial) system. As part of these studies, I have used microdialysis to evaluate the tumor extracellular fluid disposition of anticancer agents and factors affecting the delivery and removal of anticancer agents. I have also developed methods and technologies to differentiate between the inactive-encapsulated and active-released forms of nanoparticles drugs. We are evaluating potential phenotypic probes for the pharmacokinetic and pharmacodynamic disposition of liposomal and nanoparticle agents. The clinical relevance of studies is underscored by the need to treat solid tumors using anticancer agents with high tumor penetration, develop methods to increase the tumor delivery of liposomal and nanoparticle agents, and generate administration schedules to enhance selective tumor uptake.