A multipurpose microfluidic device designed to mimic microenvironment gradients and develop targeted cancer therapeutics

Walsh, Colin; Babin, Brett M.; Kasinskas, Rachel W.; Foster, Jean A.; McGarry, Marissa J.; Forbes, Neil S.

doi:10.1039/b810571e

Cited by 104 publications

(116 citation statements)

References 48 publications

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“…Microfluidic devices have become a versatile platform to provide precise concentration gradient control for understanding various biological systems and controlling the population size (8)(9)(10). Gradient-generating devices can be classified as (i) static generators, which are based solely on diffusion (11,12), and (ii) constant-flow generators (13)(14)(15)(16).…”

Section: Resultsmentioning

confidence: 99%

Cell motility and drug gradients in the emergence of resistance to chemotherapy

Loutherback

Lambert

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The emergence of resistance to chemotherapy by cancer cells, when combined with metastasis, is the primary driver of mortality in cancer and has proven to be refractory to many efforts. Theory and computer modeling suggest that the rate of emergence of resistance is driven by the strong selective pressure of mutagenic chemotherapy and enhanced by the motility of mutant cells in a chemotherapy gradient to areas of higher drug concentration and lower population competition. To test these models, we constructed a synthetic microecology which superposed a mutagenic doxorubicin gradient across a population of motile, metastatic breast cancer cells (MDA-MB-231). We observed the emergence of MDA-MB-231 cancer cells capable of proliferation at 200 nM doxorubicin in this complex microecology. Individual cell tracking showed both movement of the MDA-MB-231 cancer cells toward higher drug concentrations and proliferation of the cells at the highest doxorubicin concentrations within 72 h, showing the importance of both motility and drug gradients in the emergence of resistance. Cancer cells evolve drug resistance to chemotherapy within the tumor microenvironment. Although it is widely accepted that the tumor microenvironment provides a sequential selective pressure for preexisting mutants within the population (1-3), an additional contribution to rapid cancer evolution is mutagenic stress response followed by the emergence of adaptive phenotypes (4, 5). Further, mutagenic drug gradients in the tumor microenvironment lead to a spatially dependent fitness landscape of the cancer cells and can further accelerate the evolution of drug resistance if the cells are motile across the gradient (5, 6). We recently demonstrated using a bacteria model how a spatial gradient of antibiotic concentration in a metapopulation accelerated the evolution of antibiotic resistance (7). We would expect similar processes to occur in cancer cell metapopulations as well. Because cancer cells have a much longer doubling time (∼1 d) compared with that of bacteria (∼30 min), similar experiments with cancer cells take an order of magnitude more time (days vs. hours) than those for bacteria. This presents two experimental challenges: (i) creation of a drug stable gradient and (ii) creation of an environment hospitable for healthy cell growth. Once these conditions are established, it is possible to probe in an in vitro system the complex driving forces of resistance in systems that are in vivo. ResultsMicrofluidic devices have become a versatile platform to provide precise concentration gradient control for understanding various biological systems and controlling the population size (8-10). Gradient-generating devices can be classified as (i) static generators, which are based solely on diffusion (11, 12), and (ii) constant-flow generators (13-16). In this paper we adopt the constant-flow approach because it is capable of creating timeindependent stable gradients. However, to date, it has been challenging to grow mammalian cells in such platforms (17,18)...

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Section: Resultsmentioning

confidence: 99%

Cell motility and drug gradients in the emergence of resistance to chemotherapy

Loutherback

Lambert

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…1e). In another setup, two channels sprout in to multiple channels; in this way, a gradient is build up between the components in channel 1 and 2, mimicking the tumourenvironment gradient [42,87,112]. Microfluidics reduce the sample volume, enabling upscaling for screening applications and control of the spatio-temporal dynamics of the environment.…”

Section: Three-dimensional Biomimetic Approaches To Better Understandmentioning

confidence: 98%

Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

et al. 2015

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Cancer-associated fibroblasts (CAFs) are drivers of tumour progression and are considered as a target and a tool in cancer diagnostic and therapeutic applications. An increased abundance of CAFs or CAF signatures are recognized as a bad prognostic marker in several cancer types. Tumour-environment biomimetics strongly improve our understanding of the communication between CAFs, cancer cells and other host cells. Several experimental drugs targeting CAFs are in clinical trials for multiple tumour entities; alternatively, CAFs can be exploited as a tool to characterize the functionality of circulating tumour cells or to capture them as a tool to prevent metastasis. The continuous interaction between tissue engineers, biomaterial experts and cancer researchers creates the possibility to biomimic the tumour-environment and provides new opportunities in cancer diagnostics and management.

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“…In addition, significant advances have been made in the development of microfluidic systems for cell biology (30)(31)(32)(33). Of particular importance to this work, precise fluid handling on microfluidic devices provides more control over the microenvironment of the cells (34,35). The integration of automated valves and pumps on-chip has enabled the addition of sample preparation and analysis on one device (36)(37)(38).…”

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confidence: 99%

Short-Stalked Prosthecomicrobium hirschii Cells Have a Caulobacter-Like Cell Cycle

et al. 2016

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The dimorphic alphaproteobacterium Prosthecomicrobium hirschii has both short-stalked and long-stalked morphotypes. Notably, these morphologies do not arise from transitions in a cell cycle. Instead, the maternal cell morphology is typically reproduced in daughter cells, which results in microcolonies of a single cell type. In this work, we further characterized the shortstalked cells and found that these cells have a Caulobacter-like life cycle in which cell division leads to the generation of two morphologically distinct daughter cells. Using a microfluidic device and total internal reflection fluorescence (TIRF) microscopy, we observed that motile short-stalked cells attach to a surface by means of a polar adhesin. Cells attached at their poles elongate and ultimately release motile daughter cells. Robust biofilm growth occurs in the microfluidic device, enabling the collection of synchronous motile cells and downstream analysis of cell growth and attachment. Analysis of a draft P. hirschii genome sequence indicates the presence of CtrA-dependent cell cycle regulation. This characterization of P. hirschii will enable future studies on the mechanisms underlying complex morphologies and polymorphic cell cycles. IMPORTANCEBacterial cell shape plays a critical role in regulating important behaviors, such as attachment to surfaces, motility, predation, and cellular differentiation; however, most studies on these behaviors focus on bacteria with relatively simple morphologies, such as rods and spheres. Notably, complex morphologies abound throughout the bacteria, with striking examples, such as P. hirschii, found within the stalked Alphaproteobacteria. P. hirschii is an outstanding candidate for studies of complex morphology generation and polymorphic cell cycles. Here, the cell cycle and genome of P. hirschii are characterized. This work sets the stage for future studies of the impact of complex cell shapes on bacterial behaviors. The Alphaproteobacteria comprise a diverse group of bacteria, including important pathogens of animals (Brucella spp. and Rickettsia spp.) and plants (Agrobacterium spp.), plant symbionts (Rhizobium spp., Sinorhizobium spp., and Mesorhizobium spp.), photosynthetic bacteria (Rhodobacter spp.), freshwater bacteria (Caulobacter spp.), and marine bacteria (Hyphomonas spp.). Despite the diverse habitats and lifestyles of these bacteria, many alphaproteobacterial species have a characteristic life cycle that culminates in asymmetric cell division (1-4). Caulobacter crescentus has served as a model bacterial system for the study of cell cycle regulation, and decades of research have provided insights into the mechanism underlying the precise cell cycle control that allows

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A multipurpose microfluidic device designed to mimic microenvironment gradients and develop targeted cancer therapeutics

Cited by 104 publications

References 48 publications

Cell motility and drug gradients in the emergence of resistance to chemotherapy

Cell motility and drug gradients in the emergence of resistance to chemotherapy

Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

Short-Stalked Prosthecomicrobium hirschii Cells Have a Caulobacter-Like Cell Cycle

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