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Nuclear Deformability Constitutes a Rate-Limiting Step During Cell Migration in 3-D Environments

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Abstract

Cell motility plays a critical role in many physiological and pathological settings, ranging from wound healing to cancer metastasis. While cell migration on 2-dimensional (2-D) substrates has been studied for decades, the physical challenges cells face when moving in 3-D environments are only now emerging. In particular, the cell nucleus, which occupies a large fraction of the cell volume and is normally substantially stiffer than the surrounding cytoplasm, may impose a major obstacle when cells encounter narrow constrictions in the interstitial space, the extracellular matrix, or small capillaries. Using novel microfluidic devices that allow observation of cells moving through precisely defined geometries at high spatial and temporal resolution, we determined nuclear deformability as a critical factor in the cells’ ability to pass through constrictions smaller than the size of the nucleus. Furthermore, we found that cells with reduced levels of the nuclear envelope proteins lamins A/C, which are the main determinants of nuclear stiffness, passed significantly faster through narrow constrictions during active migration and passive perfusion. Given recent reports that many human cancers have altered lamin expression, our findings suggest a novel biophysical mechanism by which changes in nuclear structure and composition may promote cancer cell invasion and metastasis.

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Abbreviations

2-D:

Two-dimensional

3-D:

Three-dimensional

ANOVA:

Analysis of variance

BSA:

Bovine serum albumin

DMEM:

Dulbecco Modified Eagle Medium

FBS:

Fetal bovine serum

GFP:

Green fluorescent protein

LINC:

Linker of nucleoskeleton and cytoskeleton

MEF:

Mouse embryonic fibroblast

MMP:

Matrix metalloproteinase

PBS:

Phosphate buffered saline

PDGF:

Platelet derived growth factor

PDMS:

Polydimethylsiloxane

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Acknowledgments

The authors thank Philipp Isermann for providing plasmids (GFP-LifeAct, mCherry–Histone-4), advice, and some MATLAB image analysis scripts; Drs. Colin Stewart and Tom Glover for providing cells and reagents; Dr. Amy Rowat for design and fabrication of some of the perfusion devices; Dr. Sarah Jandricic for help with statistical analysis; Kathy Zhang and Ileana D’Aloisio for quantification of cell migration assays; and Rachel Gilbert for helpful discussions. This work was performed in part at the Cornell NanoScale Facility, a member of the National Nanotechnology Infrastructure Network, which is supported by the National Science Foundation (Grant ECCS-0335765). This work was supported by National Institutes of Health awards [R01 NS059348 and R01 HL082792]; the Department of Defense Breast Cancer Idea Award [BC102152]; a National Science Foundation CAREER award to Lammerding J [CBET-1254846]; and a Pilot Project Award by the Cornell Center on the Microenvironment & Metastasis through Award Number U54CA143876 from the National Cancer Institute. The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health.

Conflicts of Interest

Patricia M. Davidson, Celine Denais, Maya C. Bakshi, and Jan Lammerding declare that they have no conflicts of interest.

Ethical Standards

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY, 14853, USA

    Patricia M. Davidson, Celine Denais, Maya C. Bakshi & Jan Lammerding

Authors
  1. Patricia M. Davidson
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  2. Celine Denais
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  3. Maya C. Bakshi
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  4. Jan Lammerding
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Corresponding author

Correspondence to Jan Lammerding.

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Associate Editor David Schaffer oversaw the review of this article.

This paper is part of the 2014 Young Innovators Issue.

Dr. Jan Lammerding is an Assistant Professor in the Department of Biomedical Engineering and the Weill Institute for Cell and Molecular Biology at Cornell University. He received a Bachelor of Engineering degree from the Thayer School of Engineering at Dartmouth College, a Diplom Ingenieur degree in Mechanical Engineering from the University of Technology Aachen, Germany, and a Ph.D. in Biological Engineering from the Massachusetts Institute of Technology (MIT). Before joining Cornell University, Dr. Lammerding served as a faculty member at Harvard Medical School/Brigham and Women’s Hospital (BWH) while also teaching in the Department of Biological Engineering at MIT. At Cornell, the Lammerding laboratory is developing novel experimental techniques to investigate the interplay between cellular mechanics and function, with a particular emphasis on the cell nucleus and its response to mechanical forces. Dr. Lammerding has won several prestigious awards, including a National Science Foundation CAREER Award, an American Heart Association Scientist Development Grant, and the BWH Department of Medicine Young Investigator Award. Dr. Lammerding has published over 40 peer-reviewed articles, including in Nature and PNAS. His research is supported by grants from the National Institutes of Health, the National Science Foundation, the Department of Defense Breast Cancer Research Program, the American Heart Association, and the Progeria Research Foundation.

figure a

Electronic supplementary material

Below is the link to the electronic supplementary material.

12195_2014_342_MOESM1_ESM.avi

Supplemental Video 1: Lmna +/+ cell migrating through a narrow constriction. Representative time-lapse video of a wild-type (Lmna +/+) mouse embryonic fibroblast migrating through a 3 × 5 µm2 constriction. To aid the viewer, the nucleus is outlined with a dashed white line at the beginning of the video. The vertical white lines indicate the beginning (left) and end (right) of the constriction, as defined for the migration transit time measurements (compare with Figure 4). Time measurements (indicated in the top left corner of the image) are based relative to the nucleus entering the constriction. In this video, the migration transit time was 3:20 h. The time interval between images was 10 minutes; scale bar: 25 µm. (AVI 629 kb)

12195_2014_342_MOESM2_ESM.avi

Supplemental Video 2: Lmna +/– cell migrating through a narrow constriction. Representative time-lapse video of a Lmna +/– mouse embryonic fibroblast migrating through a 3 × 5 µm2 constriction. To aid the viewer, the nucleus is outlined with a dashed white line at the beginning of the video. The vertical white lines indicate the beginning (left) and end (right) of the constriction, as defined for the migration transit time measurements (compare with Figure 4). Time measurements (indicated in the top left corner of the image) are based relative to the nucleus entering the constriction. In this video, the migration transit time was 1:40 h. The time interval between images was 10 minutes; scale bar: 25 µm. (AVI 594 kb)

12195_2014_342_MOESM3_ESM.avi

Supplemental Video 3: Lmna –/– cell migrating through a narrow constriction. Representative time-lapse video of a lamin A/C-deficient (Lmna –/–) mouse embryonic fibroblast migrating through a 3 × 5 µm2 constriction. To aid the viewer, the nucleus is outlined with a dashed white line at the beginning of the video. The vertical white lines indicate the beginning (left) and end (right) of the constriction, as defined for the migration transit time measurements (compare with Figure 4). Time measurements (indicated in the top left corner of the image) are based relative to the nucleus entering the constriction. In this video, the migration transit time was 1:40 h. The time interval between images was 10 minutes; scale bar: 25 µm. (AVI 675 kb)

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Davidson, P.M., Denais, C., Bakshi, M.C. et al. Nuclear Deformability Constitutes a Rate-Limiting Step During Cell Migration in 3-D Environments. Cel. Mol. Bioeng. 7, 293–306 (2014). https://doi.org/10.1007/s12195-014-0342-y

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