This project targets the development of a powerful new approach to examine the pharmacokinetics (PK) and pharmacodynamics (PD) of novel drug candidates. Using a 3D printed fluidic device, cell cultures will be dynamically dosed with therapeutics. The 3D printed device offers substantial improvements over current technology, as it contains porous membranes to allow both dosing and clearance of the drugs. In the initial development phase, 3D colon cancer cell cultures, known as spheroids, will be treated with well-characterized chemotherapies. Molecular changes to the spheroids will be monitored via Matrix Assisted Laser/Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS). As both the drugs and their metabolites have defined masses, the penetration and distribution of these species can be mapped throughout the spheroids with MALDI-IMS. The public health benefits of the project lie in the promise of a powerful new tool to characterize the PK/PD of new drugs in a non-invasive, dynamic in vitro context. This approach will make it possible for researchers to build a coherent picture of the molecular changes that underlie the metabolism of new drugs, thus helping to devise more effective treatments, and improve patient outcomes. The project is constructed around three sets of activities. First, the 3D printed fluidic devices will be designed and optimized to dose the 3D cell cultures. The completed end- user friendly device will enable loading of a test-drug molecule and manipulation of its clearance half-life using a simple gradient-pumping scheme. Second, growth and dosing of the spheroids will be optimized in the 3D printed device. Finally, spheroids will be dosed and imaging via MALDI-IMS in a time course experiment. Data will be analyzed via principal component analysis. As an initial proof-of-concept study, spheroids will first be treated with the well-characterized drug irinotecan. Further studies will expand to more complicated therapeutic cocktails, such as an abbreviated simulation of the clinical regime FOLFIRI.

Public Health Relevance

With this research, we have developed a powerful in vitro method to identify both the effect of a new drug on human cells and the response of the cells to the drug without using live organisms. The public health benefits include improved testing of new therapeutics in a non-invasive fashion on cultured human cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM110406-01
Application #
8674206
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Sheeley, Douglas
Project Start
2014-07-01
Project End
2019-03-31
Budget Start
2014-07-01
Budget End
2015-03-31
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Notre Dame
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Boyce, Matthew W; LaBonia, Gabriel J; Hummon, Amanda B et al. (2017) Assessing chemotherapeutic effectiveness using a paper-based tumor model. Analyst 142:2819-2827
Andrews, William T; Skube, Susan B; Hummon, Amanda B (2017) Magnetic bead-based peptide extraction methodology for tissue imaging. Analyst 143:133-140
Bailey, Karen A; Klymenko, Yuliya; Feist, Peter E et al. (2017) Chemical Analysis of Morphological Changes in Lysophosphatidic Acid-Treated Ovarian Cancer Cells. Sci Rep 7:15295
Schunter, Alissa J; Yue, Xiaoshan; Hummon, Amanda B (2017) Phosphoproteomics of colon cancer metastasis: comparative mass spectrometric analysis of the isogenic primary and metastatic cell lines SW480 and SW620. Anal Bioanal Chem 409:1749-1763
Pinger, C W; Entwistle, K E; Bell, T M et al. (2017) C-Peptide replacement therapy in type 1 diabetes: are we in the trough of disillusionment? Mol Biosyst 13:1432-1437
Schroll, Monica M; LaBonia, Gabriel J; Ludwig, Katelyn R et al. (2017) Glucose Restriction Combined with Autophagy Inhibition and Chemotherapy in HCT 116 Spheroids Decreases Cell Clonogenicity and Viability Regulated by Tumor Suppressor Genes. J Proteome Res 16:3009-3018
Feist, Peter E; Sidoli, Simone; Liu, Xin et al. (2017) Multicellular Tumor Spheroids Combined with Mass Spectrometric Histone Analysis To Evaluate Epigenetic Drugs. Anal Chem 89:2773-2781
Liu, Xin; Flinders, Colin; Mumenthaler, Shannon M et al. (2017) MALDI Mass Spectrometry Imaging for Evaluation of Therapeutics in Colorectal Tumor Organoids. J Am Soc Mass Spectrom :
Lukowski, Jessica K; Weaver, Eric M; Hummon, Amanda B (2017) Analyzing Liposomal Drug Delivery Systems in Three-Dimensional Cell Culture Models Using MALDI Imaging Mass Spectrometry. Anal Chem 89:8453-8458
Ludwig, Katelyn R; Hummon, Amanda B (2017) Mass spectrometry for the discovery of biomarkers of sepsis. Mol Biosyst 13:648-664

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