William Zamboni Lab

Dr. Zamboni’s research program is part of the Division of Pharmacotherapy and Experimental Therapeutics in the UNC Eshelman School of Pharmacy. The research program is also part of UNC Lineberger Comprehensive Cancer Center (LCCC) and Carolina Institute of Nanomedicine (CIN). He considers himself as a translational pharmacologist where he applies standard and novel analytical chemistry, pharmacology, pharmacokinetic, pharmacodynamic, and biomarker methods to preclinical, translational, and clinical development of drugs, especially anticancer agents.

Dr. Zamboni has been involved in translational development and pharmacology studies of small molecule drugs, nanoparticles, conjugates, biologics, and implantable agents for over 25 years. A majority of his work and interests have been on anticancer agents. His research interests focus on the application of pharmacokinetic, pharmacodynamic, phenotypic 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, allow for the rational design of therapeutic regimens, and permit individualization of treatment via precision medicine approaches.

One focus of Dr. Zamboni’s research is evaluating the processes and mechanisms associated with the delivery and distribution of anticancer agents into solid tumors. He is especially interested in identifying barriers to the delivery of agents into solid tumors and developing novel methods to measure these barriers. In addition, he is extremely interested in the development of novel technologies and modulators to overcome these barriers and increase the delivery and efficacy of anticancer agents in the treatment of solid tumors.

A second focus of his research is on the development of complex agents, such as nanoparticles, liposomes, conjugates, biologics, antibodies, and antibody drug conjugates (ADCs). As part of these studies, the Zamboni group has developed methods and technologies to differentiate between the inactive-conjugated and active-released forms of these agents in blood, tumor, and tissues. The Zamboni lab also focuses on evaluating the bi-directional interaction between these agents and the mononuclear phagocyte system (MPS), which part of the innate immune system (IIS), and is the primary clearance pathway for these agents. They have developed biomarkers of the IIS/MPS, which can be used to predict the pharmacokinetics, pharmacodynamics, and the potential for drug-drug and drug-disease interactions of these complex agents, especially immune-oncology agents, antibodies, and nanoparticles. The IIS/MPS biomarkers are also being evaluated as a method to optimize the dose and regimen of these complex agents, in special populations, such as in obesity, inflammatory diseases, and COVID-19 infection.

The clinical relevance of Dr. Zamboni’s research is underscored by the need to optimize the selection of the best agent, dose, regimen, and combination therapies for the treatment of cancer and other diseases as a path to increase efficacy and reduce toxicities.

To learn more about Dr. Zamboni’s major research interests and accomplishments, click here.

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:

GLP validated:

Docetaxel – total and protein unbound
Irinotecan (CPT-11)
SN-38
DTPA and C2E5

Non-GLP validated:

Docetaxel
Paclitaxel
nab-paclitaxel (Abraxane)
Irinotecan (CPT-11)
SN-38 and glucuronidated SN-38 (SN-38G)
Camptothecin (CPT)
Topotecan
Gemcitabine
Doxorubicin – total and DNA-bound
PEGylated liposomal doxorubicin (e.g. Doxil and Lipodox) – total, encapsulated, released
5-fluorouracil
Carboplatin – total and protein unbound
Cisplatin – total and protein unbound
Oxaliplatin – total and protein unbound
Curcumin
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)

The Zamboni lab regularly works with the following UNC Core Facilities:

Nanomedicines Characterization

UNC Gillings School of Global Public Health Biomarker Mass Spectrometry

Flow Cytometry

Animal Studies

Animal Histopathology

Translational Pathology Laboratory

Tissue Culture

William Zamboni, PhD

(919) 843-6665

zamboni@email.unc.edu

William Zamboni, Pharm.D., Ph.D., is a professor in the Division of Pharmacotherapy and Experimental Therapeutics and UNC Lineberger Comprehensive Cancer Center. He is the director of the UNC Advanced Translational Pharmacology and Analytical Chemistry (ATPAC) Lab and Recharge Center in the Eshelman School of Pharmacy, the UNC Lineberger Comprehensive Cancer Center (LCCC), and the Carolina Institute of Nanomedicine. His research interests focus on the application of pharmacokinetic, pharmacodynamic, and pharmacogenetic principles in the optimization of the chemotherapeutic treatment of cancer.

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.