Download Document 8919103

Undergraduate
Category: Health Sciences
Degree Level: Bachelors
Abstract ID# 1152
Author: Dylan Vance
Bispecific Antibody Targeted Drug Delivery of and Intracellular Trafficking study of Polymer Drug Conjugate Chemotherapeutic Drugs
Abstract
Data
Cancer is a leading cause of death in the United States of America. In 2014 it resulted in
585,720 deaths. Many cancers are treated using chemotherapeutic drugs including
Doxorubicin, Paclitaxel, and 5-Fluorouracil that have saved countless of lives. However,
these drugs are associated with having major toxicity that can even result in death. A
novel method of treating cancer using bispecific antibodies to pre-target cancer cells and
treating the cancer cells with a polymer-drug conjugate has shown to be effective in
killing the cancer cells as well as reducing the toxicity associated with chemotherapeutic
drugs. The delivery system is also able to target and kill cancer cells resistant to
chemotherapeutic drugs. The mechanism behind how the polymer drug conjugate is
internalized in the cancer cell when pre-targeted with bispecific antibodies is currently
unknown. The goal of this project is to understand internalization of polymer drug
conjugates when pre-targeted with bispecific antibodies. In order to achieve this (1) two
novel bispecific antibodies will be created to be used in the internalization. (2) The
cytotoxicity of these novel bispecific antibodies will be determined. (3) Images of the
internalization will be obtained using fluorescence microscopy. Understanding the
internalization of polymer drug conjugates can lead to the development of better
bispecific antibodies and PDCs in order to get the most efficient response form the
cancer cells.
Goal
The goal of the is to obtain images of the mechanism of targeted delivery
using bispecific antibodies with polymer-drug conjugates (PDCs) in cancer
cells.
Method
In order to achieve the imaging of the internalization of the PDCs by this
delivery system, slides will be first sterilized using UV light for thirty
minutes, and then have 40,000 cells added to the slides from our stock of
cells. The cells would then attach to the slide over the next twenty-four hours
and after twenty-four hours, the bispecific antibodies will be added to the
slide for a one hour incubation period. After one hour the PDC will be added
for different time points including 15 minutes, 30 minutes, 45 minutes, 1
hour, 2 hour, and 6 hour. Once the time point is reached the slides will be
fixed using an eight percent paraformaldehyde solution and then had HOST
added to the cells to stain the nucleus. The slides were then sealed with
flouro-mount and then imaged using a fluorescence microscope that can
detect red and blue fluorescence.
Acknowledgements
I would like to thank Dr. Khaw and Prashant for the last three years in the
lab. It has been an eye opening and an amazing experience.
Top left corner,, 1 hour bi-specific antibody, Top right DPGADOX alone, Bottom left 6 hour bispecific
antibody, and Bottom Left DPGADOX alone
Conclusion
So far we have begun to see the story of the internalization of the polymer drug with bispecific antibody. We have begun to see the route of cell membrane to endosome to nucleus
for the paradigm for the mechanism behind the internalization. We are still continuing the
imaging and are looking to finish imaging the entire mechanism.
References
Khaw, B., Gada, K., Patil, V., Panwar, R., Mandapati, S., Hatefi, A., Majewski, S., and Weisenberger, A. 2014. Bispecific antibody complex
pre-targeting and targeted delivery of polymer drug conjugates for imaging and therapy in dual human mammary cancer xenografts:
targeted polymer drug conjugates for cancer diagnosis and therapy. Eur J Nucl Med Mol Imaging. 2014 Aug; 41(8):1603-16.
Wong HL, Bendayan R, Rauth AM, Xue HY, Babakhanian K, Wu XY. A mechanistic study of enhanced doxorubicin uptake and retention
in multidrug resistant breast cancer cells using a polymer-lipid hybrid nanoparticle system. J Pharmacol Exp Ther. 2006
Jun;317(3):1372-81