Functionalization of nanotextured substrates for enhanced identification of metastatic breast cancer cells

Metastasis is the major cause of low survival rates among cancer patients. Once cancer cells metastasize, it is extremely difficult to contain the disease. We report on a nanotextured platform for enhanced detection of metastatic cells. We captured metastatic (MDA-MDB-231) and non-metastatic (MCF-7) breast cancer cells on anti-EGFR aptamer modified plane and nanotextured substrates. Metastatic cells were seen to change their morphology at higher rates when captured on nanotextured substrates than on plane substrates. Analysis showed statistically different morphological behaviors of metastatic cells that were very pronounced on the nanotextured substrates. Several distance matrices were calculated to quantify the dissimilarity of cell shape change. Nanotexturing increased the dissimilarity of the metastatic cells and as a result the contrast between metastatic and non-metastatic cells increased. Jaccard distance measurements found that the shape change ratio of the non-metastatic and metastatic cells was enhanced from 1:1.01 to 1:1.81, going from plane to nanotextured substrates. The shape change ratio of the non-metastatic to metastatic cells improved from 1:1.48 to 1:2.19 for the Hausdorff distance and from 1:1.87 to 1:4.69 for the Mahalanobis distance after introducing nanotexture. Distance matrix analysis showed that nanotexture increased the shape change ratios of non-metastatic and metastatic cells. Hence, the detectability of metastatic cells increased. These calculated matrices provided clear and explicit measures to discriminate single cells for their metastatic state on functional nanotextured substrates.

[1]  K E Healy,et al.  Kinetics of bone cell organization and mineralization on materials with patterned surface chemistry. , 1996, Biomaterials.

[2]  C. Parent,et al.  A cell's sense of direction. , 1999, Science.

[3]  Tejal A Desai,et al.  Biomimetic nanowire coatings for next generation adhesive drug delivery systems. , 2009, Nano letters.

[4]  A. Wells,et al.  Epidermal growth factor receptor-mediated cell motility: phospholipase C activity is required, but mitogen-activated protein kinase activity is not sufficient for induced cell movement , 1994, The Journal of cell biology.

[5]  L. Terstappen,et al.  Characterization of circulating tumor cells by fluorescence in situ hybridization , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[6]  Y. Wan,et al.  Synthesis of nano-textured biocompatible scaffolds from chicken eggshells , 2012, Nanotechnology.

[7]  Paul Nealey,et al.  Characterization of endothelial basement membrane nanotopography in rhesus macaque as a guide for vessel tissue engineering. , 2009, Tissue engineering. Part A.

[8]  G. Carpenter,et al.  EPIDERMAL GROWTH FACTOR * , 1982, The Journal of biological chemistry.

[9]  Andrew D Ellington,et al.  One-step tumor detection from dynamic morphology tracking on aptamer-grafted surfaces. , 2015, Technology.

[10]  Bharat Bhushan,et al.  Biologically Inspired Surfaces: Broadening the Scope of Roughness** , 2008 .

[11]  K. Anselme,et al.  Osteoblast adhesion on biomaterials. , 2000, Biomaterials.

[12]  Swee Jin Tan,et al.  Microdevice for the isolation and enumeration of cancer cells from blood , 2009, Biomedical microdevices.

[13]  G. Oster,et al.  Cell motility driven by actin polymerization. , 1996, Biophysical journal.

[14]  D. F. Far,et al.  Neutrophil F-actin and myosin but not microtubules functionally regulate transepithelial migration induced by interleukin 8 across a cultured intestinal epithelial monolayer. , 1999, European cytokine network.

[15]  L. Liotta,et al.  Metastatic potential correlates with enzymatic degradation of basement membrane collagen , 1980, Nature.

[16]  Thomas J Webster,et al.  Endothelial and vascular smooth muscle cell function on poly(lactic-co-glycolic acid) with nano-structured surface features. , 2004, Biomaterials.

[17]  William Shain,et al.  Preferential glial cell attachment to microcontact printed surfaces , 1997, Journal of Neuroscience Methods.

[18]  A. Wells,et al.  Cell movement elicited by epidermal growth factor receptor requires kinase and autophosphorylation but is separable from mitogenesis , 1994, The Journal of cell biology.

[19]  J. Peterse,et al.  Breast cancer metastasis: markers and models , 2005, Nature Reviews Cancer.

[20]  Y. Wan,et al.  Micro+nanotexturing of substrates to enhance ligand-assisted cancer cell isolation , 2014, Nanotechnology.

[21]  Thomas J Webster,et al.  Nano-structured polymers enhance bladder smooth muscle cell function. , 2003, Biomaterials.

[22]  Katherine A Vallis,et al.  Relationship Between Induction of Phosphorylated H2AX and Survival in Breast Cancer Cells Exposed to 111In-DTPA-hEGF , 2008, Journal of Nuclear Medicine.

[23]  P. Mahalanobis.  On the generalized distance in statistics , 1936 .

[24]  Andrew D Ellington,et al.  Nanotextured substrates with immobilized aptamers for cancer cell isolation and cytology , 2012, Cancer.

[25]  A. Wells.  EGF receptor. , 1999, The international journal of biochemistry & cell biology.

[26]  S. McGuire.  World Cancer Report 2014. Geneva, Switzerland: World Health Organization, International Agency for Research on Cancer, WHO Press, 2015. , 2016, Advances in nutrition.

[27]  Subra Suresh,et al.  Biomechanics and biophysics of cancer cells. , 2007, Acta biomaterialia.

[28]  J. Mendelsohn.  EGF receptors as a target for cancer therapy. , 2004, Transactions of the American Clinical and Climatological Association.

[29]  Kwanghee Kim,et al.  Novel streptavidin-functionalized silicon nanowire arrays for CD4+ T lymphocyte separation. , 2010, Nano letters.

[30]  C. R. Howlett,et al.  Titanium substrata composition influences osteoblastic phenotype: In vitro study. , 1999, Journal of biomedical materials research.

[31]  Andrew D. Ellington,et al.  Surface-immobilized aptamers for cancer cell isolation and microscopic cytology. , 2010, Cancer research.

[32]  I. Macdonald,et al.  Metastasis: Dissemination and growth of cancer cells in metastatic sites , 2002, Nature Reviews Cancer.

[33]  A. Wells,et al.  A role for gelsolin in actuating epidermal growth factor receptor- mediated cell motility , 1996, The Journal of cell biology.

[34]  Isabelle Robert,et al.  Translational up-regulation of the EGFR by tumor hypoxia provides a nonmutational explanation for its overexpression in human cancer , 2007, Proceedings of the National Academy of Sciences.

[35]  M. Plummer,et al.  International agency for research on cancer. , 2020, Archives of pathology.

[36]  B. Alberts,et al.  Molecular Biology of the Cell (4th Ed) , 2002 .

[37]  Daniel P. Huttenlocher,et al.  Comparing Images Using the Hausdorff Distance , 1993, IEEE Trans. Pattern Anal. Mach. Intell..

[38]  A D Ellington,et al.  Aptamers as therapeutic and diagnostic reagents: problems and prospects. , 1997, Current opinion in chemical biology.

[39]  David A. Russell,et al.  Self-assembled monolayers: a versatile tool for the formulation of bio-surfaces , 2000 .

[40]  L. Tanoue,et al.  Detection of Mutations in EGFR in Circulating Lung-Cancer Cells , 2010 .

[41]  Paul I. Okagbare,et al.  Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. , 2008, Journal of the American Chemical Society.

[42]  Andrew D. Ellington,et al.  Electrical Profiling and Aptamer Functionalized Nanotextured Surface in a Single Biochip for the Detection of Tumor Cells , 2016 .

[43]  P. Jaccard.  Distribution de la flore alpine dans le bassin des Dranses et dans quelques régions voisines , 1901 .

[44]  S. Digumarthy,et al.  Isolation of rare circulating tumour cells in cancer patients by microchip technology , 2007, Nature.

[45]  John G. Collard,et al.  Rac regulates phosphorylation of the myosin-II heavy chain, actinomyosin disassembly and cell spreading , 1999, Nature Cell Biology.

[46]  D. Bunka,et al.  Aptamers come of age – at last , 2006, Nature Reviews Microbiology.

[47]  Stefan Schinkinger,et al.  Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. , 2005, Biophysical journal.

[48]  I. Fidler,et al.  The pathogenesis of cancer metastasis: the 'seed and soil' hypothesis revisited , 2003, Nature Reviews Cancer.

[49]  B. Sullenger,et al.  Emerging clinical applications of RNA , 2002, Nature.

[50]  Samir M. Iqbal,et al.  Effects of Nanotexture on Electrical Profiling of Single Tumor Cell and Detection of Cancer from Blood in Microfluidic Channels , 2015, Scientific Reports.

[51]  J. Käs,et al.  Neuronal growth: a bistable stochastic process. , 2006, Physical review letters.

[52]  C. R. Howlett,et al.  Effect of surface chemical modification of bioceramic on phenotype of human bone-derived cells. , 1999, Journal of biomedical materials research.

[53]  Sandra Downes,et al.  Protein adsorption and human osteoblast-like cell attachment and growth on alkylthiol on gold self-assembled monolayers. , 2002, Journal of biomedical materials research.