Date of Award

January 2012

Document Type

Thesis

Degree Name

Medical Doctor (MD)

Department

Medicine

First Advisor

W M. Saltzman

Subject Area(s)

Surgery, Biomedical engineering

Abstract

SYNTHESIS AND DELIVERY OF ULTRA-SMALL POLYMER NANOPARTICLES FOR TREATMENT OF GLIOBLASTOMA MULTIFORME IN THE RAT CAUDATE.

Whitney Sheen, Toral Patel, Jiangbing Zhou, and W. Mark Saltzman. Department of Biomedical Engineering and Department of Cellular and Molecular Physiology, Yale University, School of Medicine, New Haven, Connecticut.

Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor in humans with a dismal prognosis, largely due to its highly infiltrative nature. To apply and study a novel glioblastoma multiforme treatment approach via delivery of chemotherapeutic nanoparticles, our research team aimed to 1) confirm in vitro inhibition of glioblastoma multiforme tumor cells with selected chemotherapeutic agent, anisomycin, via MTT assay, 2) create an in vivo model of glioblastoma multiforme in the rat, 3) deliver chemotherapeutic nanoparticles for treatment of glioblastoma multiforme tumor cells in our in vivo model, and 4) measure volume of distribution of coumarin-6 (C6) loaded nanoparticles in naïve and tumor-laden rat brains. We hypothesized that 1) anisomycin would show inhibitory activity against PS11 and GS5 human glioblastoma multiforme tumor cells in vitro, 2) we would be able to successfully transplant PS11 human glioblastoma multiforme tumor cells in the caudate of rats for creation of an in vivo model of glioblastoma multiforme, 3) we would be able to deliver anisomycin nanoparticles for the treatment of PS11 tumor cells in our in vivo model, and 4) we would be able to successfully measure volume of distribution of C6 loaded nanoparticles following convection-enhanced delivery in naïve and tumor-laden rat brain. In vitro PS11 and GS5 tumor cells were plated and subjected to MTT assays to assess tumor cell viability 24 hrs post treatment with anisomycin. PS11 tumor cells were inoculated into right rat caudate for creation of an in vivo animal model of glioblastoma multiforme. Animal surgeries for the treatment of PS11 tumor cells via convection-enhanced delivery of anisomycin-loaded nanoparticles occurred 10 days post-tumor inoculation. To study distribution of nanoparticles in rat brain, C6 nanoparticles were infused into the caudate of tumor-laden and tumor free rat brains. Thirty minutes following C6 nanoparticle infusion, animals were sacrificed and brains were removed for histological analysis. Anisomycin dose response curves were generated for the treatment of GS5 and PS11 tumor cell lines with IC50s of 2.40 uM and 2.70 uM respectively. Implantation and growth of PS11 tumor cells in our in vivo animal model was confirmed via histological sectioning 10 days post inoculation. PS11 tumor cells were observed to grow in the brains of animals used for the C6 nanoparticle distribution study. In the two rats infused with PS11 tumor cells, C6 volume of distribution was 80.86 mm3 and 62.01 mm3. In the naïve rat without tumor, volume of distribution of C6 nanoparticles was 24.41 mm3. In conclusion, this work suggests that convection-enhanced delivery of chemotherapeutic nanoparticles should be investigated as an approach for the treatment of tumor cells, particularly infiltrating tumor cells, in patients with glioblastoma multiforme brain tumors.

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