Mechanobiology of Breast Cancer Cells

Shea Thompson Author

04/04/2021 Added

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Investigation on how shear flow affects breast cancer cell migration in a three-dimensional space with obstructions

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[00:00:00.900]Hi everyone. My name is Shea Thompson.
[00:00:03.630]I am a junior in biological systems engineering and my research
[00:00:08.430]is on mechanical biology of breast cancer cells.
[00:00:12.330]I have done my research under the guidance of Dr. Jung Yul Lim.
[00:00:17.760]I want to start this presentation off by providing some background on my
[00:00:21.030]project.
[00:00:22.140]So mechanobiology is the study of how cells respond to mechanical loading.
[00:00:27.060]And this mechanical loading can be a sheer flow against the cells or stretching
[00:00:30.660]and compression of the cells.
[00:00:33.900]Breast cancer cells are exposed to flow upon leaving the tumor site and as cells
[00:00:38.610]migrate,
[00:00:39.330]they have to move around obstructions like extra-cellular matrices in the body.
[00:00:44.910]So this raises the question,
[00:00:46.650]how does shear flow affect breast cancer cell migration in a three-dimensional
[00:00:50.760]space with obstructions?
[00:00:54.030]In order to test this hypothesis,
[00:00:55.770]I first needed to convert the parallel flow chamber through a three-dimensional
[00:00:59.820]flow chamber using the glass light extension piece.
[00:01:03.840]Then I designed a maze to make my experiments similar to a cell might experience
[00:01:08.070]in the body.
[00:01:09.690]I'd be using a very aggressive breast cancer cell line for this experiment to
[00:01:14.040]get some results. With an empty maze,
[00:01:17.490]I would hold one trial with and one without flow as my control
[00:01:22.350]group, then I would insert my maze for my experimental group.
[00:01:28.380]After applying a natural flow rate to this cells,
[00:01:32.400]I can observe their movements by an imaging microscope using developed
[00:01:36.780]software. I can determine the speed, the arrest coefficient,
[00:01:40.740]the cell group displacement, and the individual cell displacement.
[00:01:44.550]And by comparing these to the control,
[00:01:46.380]I can determine a flow promotes or inhibits migration in a three-dimensional
[00:01:50.520]space with obstructions.
[00:01:53.640]So my project comes with two parts, the engineering part and the biology part.
[00:01:58.500]So first I'm going to cover the engineering part
[00:02:03.120]in order to do my experiment with cells.
[00:02:05.310]I first needed to convert the 2d parallel flow chamber
[00:02:10.320]to 3d flow chamber and to do that,
[00:02:13.950]I needed to extend the vacuum seal on the device.
[00:02:18.750]So last year I designed a glass light extension piece to create 3d
[00:02:23.580]space.
[00:02:24.780]I use SolidWorks to design it as well as MATLAB to determine the
[00:02:29.250]height. As you can see,
[00:02:31.200]my design changed over time as I optimize the device.
[00:02:35.940]So this year I designed an interchangeable piece
[00:02:39.630]I call the maze. At first,
[00:02:42.330]when I thought of making a maze I thought of walls and rectangles. However,
[00:02:46.710]I decided that columns are more realistic.
[00:02:50.010]My next issue was determining the size.
[00:02:52.860]The average breast cancer cell is about 20 micrometers.
[00:02:57.390]So that being said,
[00:02:58.320]I only had to a 3d printer that could only print about
[00:03:03.520]300 micrometers accurately.
[00:03:06.130]So I decided to use the smallest I could print and went with 300 micro meter
[00:03:10.990]columns with 300 micrometers in between each column.
[00:03:15.760]My final obstacle was determining which truss system to use to
[00:03:20.500]connect the columns as well as withstand the flow subjected to it.
[00:03:25.030]After applying different weights to each truss system,
[00:03:28.480]I concluded that just a straight rectangle was more efficient and that's how I
[00:03:33.190]got my final design for the maze.
[00:03:37.270]So the two pieces together would look like this.
[00:03:40.480]I wanted the middle piece to be interchangeable so that I,
[00:03:43.960]and anyone else could test different scenarios. However,
[00:03:47.770]I'm still working out the kinks in my design.
[00:03:50.950]I was unable to test the extension piece before the pandemic closed my lab.
[00:03:55.390]So I continued to the maze step under the assumption that the extension piece
[00:04:00.010]worked.
[00:04:01.330]I was finally able to return to the lab the spring semester of 2021 and low and
[00:04:06.220]behold, my extension piece does not work correctly.
[00:04:11.860]My problem with my extension piece is that the vacuum seal leaks
[00:04:16.780]in order to ever come, that I've tried sanding the extension piece.
[00:04:20.290]I've tried multiple silicon glue gaskets,
[00:04:23.950]even multiple pumps to increase the suction force.
[00:04:27.640]So I got the extension piece to completely seal from the outside,
[00:04:32.860]but the water with in the flow chamber is still getting sucked into the
[00:04:37.570]vacuum seal. And this would completely ruin my experiment.
[00:04:41.980]If I put cells in there,
[00:04:43.840]the cells would get sucked up and then I wouldn't see anything in the
[00:04:47.740]microscope. So from here on out,
[00:04:50.980]I have a couple more options to consider.
[00:04:53.650]I can alter the thickness of the extension piece so that the
[00:04:58.390]vacuum seal doesn't have to travel as far or I can try to
[00:05:03.370]use a clamp to try to force the vacuum seal. And if those don't work,
[00:05:08.290]I would probably just have to redesign my extension piece.
[00:05:12.490]So since my extension piece is not quite working correctly,
[00:05:16.300]I can not proceed to the biology part of my experiment that I would report
[00:05:20.500]results on. That being said,
[00:05:23.620]I have made significant progress on my extension piece.
[00:05:26.620]And once I get it working,
[00:05:28.360]I'll be ready for multiple types of experiments with cells.
[00:05:34.030]One of those experiments would be looking into the role of nesprin in flow
[00:05:37.480]induced breast cancer
[00:05:38.500]Cell migration in 3d spaces. Nesprin helps with cellular
[00:05:43.390]movement and acts as a transducer for a mechanical signals between the
[00:05:48.040]cytoskeleton and the nuclear membrane.
[00:05:51.680]Therefore mechanical signals may influence cells via nesprin.
[00:05:56.410]I am planning see how flow affects cells with nesprin versus without nesprin
[00:06:01.180]next year. Now I want to finish by saying,
[00:06:05.510]understanding how breast cancer cells migrate is essential to preventing the
[00:06:09.860]spread of cancer. By studying the cells response to certain mechanical
[00:06:14.420]loading,
[00:06:14.960]We could potentially inhibit those responses and slope cancer progression.
[00:06:20.150]Thank you.

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Mechanobiology of Breast Cancer Cells

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