Organizational metrics of interchromatin speckle factor domains: Integrative classifier for stem cell adhesion & lineage signaling

Sebastián L. Vega, Anandika Dhaliwal, Varun Arvind, Parth J. Patel, Nick R.M. Beijer, Jan De Boer, N. Sanjeeva Murthy, Joachim Kohn, Prabhas V. Moghe

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Stem cell fates on biomaterials are influenced by the complex confluence of microenvironmental cues emanating from soluble growth factors, cell-to-cell contacts, and biomaterial properties. Cell-microenvironment interactions influence the cell fate by initiating a series of outside-in signaling events that traverse from the focal adhesions to the nucleus via the cytoskeleton and modulate the sub-nuclear protein organization and gene expression. Here, we report a novel imaging-based framework that highlights the spatial organization of sub-nuclear proteins, specifically the splicing factor SC-35 in the nucleoplasm, as an integrative marker to distinguish between minute differences of stem cell lineage pathways in response to stimulatory soluble factors, surface topologies, and microscale topographies. This framework involves the high resolution image acquisition of SC-35 domains and imaging-based feature extraction to obtain quantitative nuclear metrics in tandem with machine learning approaches to generate a predictive cell state classification model. The acquired SC-35 metrics led to >90% correct classification of emergent human mesenchymal stem cell (hMSC) phenotypes in populations of hMSCs exposed for merely 3 days to basal, adipogenic, or osteogenic soluble cues, as well as varying levels of dexamethasone-induced alkaline phosphatase (ALP) expression. Early osteogenic cellular responses across a series of surface patterns, fibrous scaffolds, and micropillars were also detected and classified using this imaging-based methodology. Complex cell states resulting from inhibition of RhoGTPase, β-catenin, and FAK could be classified with >90% sensitivity on the basis of differences in the SC-35 organizational metrics. This indicates that SC-35 organization is sensitively impacted by adhesion-related signaling molecules that regulate osteogenic differentiation. Our results show that diverse microenvironment cues affect different attributes of the SC-35 organizational metrics and lead to distinct emergent organizational patterns. Taken together, these studies demonstrate that the early organization of SC-35 domains could serve as a "fingerprint" of the intracellular mechanotransductive signaling that governs growth factor- and topography-responsive stem cell states. This journal is

Original languageEnglish (US)
Pages (from-to)435-446
Number of pages12
JournalIntegrative Biology (United Kingdom)
Volume7
Issue number4
DOIs
StatePublished - Apr 1 2015

Fingerprint

Cell adhesion
Speckle
Stem cells
Cell Adhesion
Classifiers
Stem Cells
Biocompatible Materials
Cues
Nuclear Proteins
Imaging techniques
Topography
Intercellular Signaling Peptides and Proteins
Adhesion
Catenins
Protein Splicing
Organizations
Image acquisition
Cellular Microenvironment
Gene expression
Scaffolds

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry

Cite this

Vega, Sebastián L. ; Dhaliwal, Anandika ; Arvind, Varun ; Patel, Parth J. ; Beijer, Nick R.M. ; De Boer, Jan ; Murthy, N. Sanjeeva ; Kohn, Joachim ; Moghe, Prabhas V. / Organizational metrics of interchromatin speckle factor domains : Integrative classifier for stem cell adhesion & lineage signaling. In: Integrative Biology (United Kingdom). 2015 ; Vol. 7, No. 4. pp. 435-446.
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abstract = "Stem cell fates on biomaterials are influenced by the complex confluence of microenvironmental cues emanating from soluble growth factors, cell-to-cell contacts, and biomaterial properties. Cell-microenvironment interactions influence the cell fate by initiating a series of outside-in signaling events that traverse from the focal adhesions to the nucleus via the cytoskeleton and modulate the sub-nuclear protein organization and gene expression. Here, we report a novel imaging-based framework that highlights the spatial organization of sub-nuclear proteins, specifically the splicing factor SC-35 in the nucleoplasm, as an integrative marker to distinguish between minute differences of stem cell lineage pathways in response to stimulatory soluble factors, surface topologies, and microscale topographies. This framework involves the high resolution image acquisition of SC-35 domains and imaging-based feature extraction to obtain quantitative nuclear metrics in tandem with machine learning approaches to generate a predictive cell state classification model. The acquired SC-35 metrics led to >90{\%} correct classification of emergent human mesenchymal stem cell (hMSC) phenotypes in populations of hMSCs exposed for merely 3 days to basal, adipogenic, or osteogenic soluble cues, as well as varying levels of dexamethasone-induced alkaline phosphatase (ALP) expression. Early osteogenic cellular responses across a series of surface patterns, fibrous scaffolds, and micropillars were also detected and classified using this imaging-based methodology. Complex cell states resulting from inhibition of RhoGTPase, β-catenin, and FAK could be classified with >90{\%} sensitivity on the basis of differences in the SC-35 organizational metrics. This indicates that SC-35 organization is sensitively impacted by adhesion-related signaling molecules that regulate osteogenic differentiation. Our results show that diverse microenvironment cues affect different attributes of the SC-35 organizational metrics and lead to distinct emergent organizational patterns. Taken together, these studies demonstrate that the early organization of SC-35 domains could serve as a {"}fingerprint{"} of the intracellular mechanotransductive signaling that governs growth factor- and topography-responsive stem cell states. This journal is",
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Organizational metrics of interchromatin speckle factor domains : Integrative classifier for stem cell adhesion & lineage signaling. / Vega, Sebastián L.; Dhaliwal, Anandika; Arvind, Varun; Patel, Parth J.; Beijer, Nick R.M.; De Boer, Jan; Murthy, N. Sanjeeva; Kohn, Joachim; Moghe, Prabhas V.

In: Integrative Biology (United Kingdom), Vol. 7, No. 4, 01.04.2015, p. 435-446.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Organizational metrics of interchromatin speckle factor domains

T2 - Integrative classifier for stem cell adhesion & lineage signaling

AU - Vega, Sebastián L.

AU - Dhaliwal, Anandika

AU - Arvind, Varun

AU - Patel, Parth J.

AU - Beijer, Nick R.M.

AU - De Boer, Jan

AU - Murthy, N. Sanjeeva

AU - Kohn, Joachim

AU - Moghe, Prabhas V.

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