THE ROLE OF THE MECHANICAL ENVIRONMENT ON CANCER CELL TRANSMIGRATION AND MRNA LOCALIZATION

Most cancer-related deaths are due to metastasis formation, the ability of cancer cells to break away from the primary tumor site, transmigrate through the endothelium, and form secondary tumors in distant areas. Many studies have identified links between the mechanical properties of the cellular microenvironment and the behavior of cancer cells. Cells may experience heterogeneous microenvironments of varying stiffness during tumor progression, transmigration, and invasion into the basement membrane. In addition to mechanical factors, the localization of RNAs to lamellipodial regions has been proposed to play an important part in metastasis. This dissertation provides a quantitative evaluation of the biophysical effects on cancer cell transmigration and RNA localization. In the first part of this dissertation, we sought to compare cancer cell and leukocyte transmigration and investigate the impact of matrix stiffness on transmigration process. We found that cancer cell transmigration includes an additional step, ‘incorporation’, into the endothelial cell (EC) monolayer. During this phase, cancer cells physically displace ECs and spread into the monolayer. Furthermore, the effects of subendothelial matrix stiffness and endothelial activation on cancer cell incorporation are cell-specific, a notable difference from the process by which leukocytes transmigrate. Collectively, our results provide mechanistic insights into tumor cell extravasation and demonstrate that incorporation into the endothelium is one of the earliest steps. In the next part of this work, we investigated how matrix stiffness impacts RNA localization and its relevance to cancer metastasis. In migrating cells, the tumor suppressor protein, adenomatous polyposis coli (APC) targets RNAs to cellular protrusions. We observed that increasing stiffness promotes the peripheral localization of these APC-dependent RNAs and that cellular contractility plays a role in regulating this pathway. We next investigated the mechanism underlying the effect of substrate stiffness and cellular contractility. We found that contractility drives localization of RNAs to protrusions through modulation of detyrosinated microtubules, a network of modified microtubules that associate with, and are required for localization of APC-dependent RNAs. These results raise the possibility that as the matrix environment becomes stiffer during tumor progression, it promotes the localization of RNAs and ultimately induces a metastatic phenotype.

Regenerative Stormwater Conveyance: Design Implications of an Urban Case Demonstration in Baltimore, Maryland

This research-design thesis explores the implementation of Regenerative Stormwater Conveyance (RSC) as a retrofit of an existing impervious drainage system in a small catchment in the degraded Jones Falls watershed in Baltimore City. An introduction to RSC is provided, placing its development within a theoretical context of novel ecosystems, biomimicry and Nassauer and Opdam’s (2008) model of landscape innovation. The case site is in Baltimore’s Hampden neighborhood on City-owned land adjacent to rowhomes, open space and an access point to a popular wooded trail along a local stream. The design proposal employs RSC to retrofit an ill-performing stormwater system, simultaneously providing a range of ecological, social and economic services; water quantity, water quality and economic performance of the proposed RSC are quantified. While the proposed design is site-specific the model is adaptable for retrofitting other small-scale impervious drainage systems, providing a strategic tool in addressing Baltimore City’s stormwater challenges.

Quantifying the Resilience of an Urban Traffic-Electric Power Coupled System

Transportation system resilience has been the subject of several recent studies. To assess the resilience of a transportation network, however, it is essential to model its interactions with and reliance on other lifelines. In this work, a bi-level, mixed-integer, stochastic program is presented for quantifying the resilience of a coupled traffic-power network under a host of potential natural or anthropogenic hazard-impact scenarios. A two-layer network representation is employed that includes details of both systems. Interdependencies between the urban traffic and electric power distribution systems are captured through linking variables and logical constraints. The modeling approach was applied on a case study developed on a portion of the signalized traffic-power distribution system in southern Minneapolis. The results of the case study show the importance of explicitly considering interdependencies between critical infrastructures in transportation resilience estimation. The results also provide insights on lifeline performance from an alternative power perspective.