@article{EngineeringExVivoModelsforProbingTumor:5231,
      recid = {5231},
      author = {Weathered, Rachel},
      title = {Engineering Ex Vivo Models for Probing Tumor, Lymphatic,  and Immune Interactions in Metastasis and Immunotherapy},
      publisher = {University of Chicago},
      school = {Ph.D.},
      address = {2022-12},
      pages = {124},
      abstract = {My thesis work has been based on developing a model of the  tumor, lymphatic, and immune landscape. I've utilized  unconventional materials, such as paper, recycled pipette  tip holders, and double-sided tape, to fabricate a system  that is both easy to build and easy to adapt. In this  effort, I've modeled several biological processes that  would otherwise be difficult to observe using traditional  in vitro methods.

In chapter 1, I provide an overview of  the complexities of the tumor-immune microenvironment. I  describe a complicated relationship between vascular  endothelial growth factor C (VEGFC), metastasis, and immune  education. I also describe current methods for modeling  biological interactions in vitro and the limitations of  these methods.

In chapter 2, I describe the design and  optimization of two devices for the culture of ex vivo  tissue. In the vertical flow design, we see long-term  viability of several cell lines and demonstrate how this  device may be used for studying drug resistance. In the  horizontal flow design, we have the ability to include  upstream and downstream cell populations. In both devices,  we utilize materials that can be easily modified and  translated to other labs.

In chapter 3, we use the  horizontal flow design to study the formation of a  pre-metastatic niche. Here, we culture lymph nodes that  have been pre-educated in vivo by tumors either with or  without VEGFC expression. We find that wild-type melanoma  cells survive better in lymph nodes pre-educated by the  VEGFC expressing tumor. With the addition of  collagen-binding IL-12, we see a dampened pro-survival  effect in the VEGFC educated tumor-draining lymph node.  Finally, we visualize migration towards a pre-metastatic  niche. Tumor draining lymph nodes pre-educated by VEGFC  expressing tumors promote the migration of cancer cells ex  vivo.

In chapter 4, we probe the effects of combinatorial  immunotherapy, checkpoint inhibitors anti-PD-1 and  anti-CTLA-4 with collagen-binding IL-12, on tumor draining  lymph nodes and spleens pre-educated by VEGFC expressing  tumors. In both the lymph node and the spleen we observe a  stronger immunotherapy effect when a tumor expresses VEGFC.  This suggests VEGFC may act to potentiate immunotherapy  through the pre-education of secondary lymphoid organs with  tumor specific antigens. 

In chapter 5, we explore the  impact that lymphatic transport has in another disease  indication, lipedema. Here, I present a continuation of  work performed in the Swartz Lab, where we purified  extracellular vesicles from lipedema patient lipoaspirate.  We find 31 differentially expressed miRNAs in stage 3  lipedema patients. I also describe early efforts to  characterize changes in lipedema skin lymphatic vessel  structure, fibrin clotting, and NETosis.

Finally, in  chapter 6, I offer a brief overview of my findings. I also  suggest future directions that should be further explored  in modeling tumor, lymphatic, and immune interactions.},
      url = {http://knowledge.uchicago.edu/record/5231},
      doi = {https://doi.org/10.6082/uchicago.5231},
}