Drug Resistance Immunotherapy for Cancer Treatment Using a Chimeric Antigen Receptor Approach Open Access

Tylawsky, Daniel Eugene (2015)

Permanent URL: https://etd.library.emory.edu/concern/etds/xs55mc445?locale=en


The toxicity of nucleoside analogue chemotherapeutics on rapidly proliferating cells is a well documented and understood side effect of cancer treatment. One of the most consequential trade-offs of this off-tumor cytotoxicity is that it acts on immune system cells that have a natural capacity to fight tumors. As such, making immunocompetent cells resistant to a chemotherapeutic, so that they can stay alive during it's administration, is a desirable approach to enhance cancer treatment with chemotherapy. This is the basis for drug resistant immunotherapy (DRI), which we implement in our studies. We think that by combining novel immunotherapy strategies with currently used chemotherapy protocols, a potent and enduring cancer treatment can be achieved. In this study, we investigated the ability of overexpressed cytidine deaminase to confer resistance to the chemotherapeutic Ara-C and showed that this could be achieved in MV4-11 cells. For the immunotherapy arm of our studies, we experimented with engineering a chimeric antigen receptor (CAR) to enhance the specificity and activation of immunocompetent cells. We took a novel approach in constructing a CAR by utilizing a variable lymphocyte receptor (VLR) derived from lampreys, to function as the specific antigen-binding portion of the construct. This was done because the independently evolved adaptive immune system of jawless fish, which utilizes VLRs, can potentially recognize epitopes distinct from those recognized by mammalian antibodies. In this study, we experimented with an anti-CD5 VLR-CAR construct. We observed that Jurkat cells transduced with anti-CD5 VLR-CAR encoded lentivirus, were in fact successfully activated in the presence of CD5+ Jurkat cells. This proved that not only can VLRs be effectively used in a CAR construct, but also, for the first time, can activate T cells against a therapeutically relevant antigen.

Table of Contents

Introduction 1

Methods 9

Results 15

Discussion 21

References 27

Tables and Figures

Table 1 31

Table 2 37

Figure 1 29

Figure 2 30

Figure 3 31

Figure 4 32

Figure 5 33

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Figure 7 35

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Figure 9 37

Figure 10 39

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