Untangling Intercellular Communication Using Optical Manipulation in 3D Models of Tumor Microenvironment

Persistent Link:
http://hdl.handle.net/10150/325421
Title:
Untangling Intercellular Communication Using Optical Manipulation in 3D Models of Tumor Microenvironment
Author:
Orsinger, Gabriel V.
Issue Date:
2014
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Embargo:
Release 18-Jul-2015
Abstract:
The tumor microenvironment is a tangled web of multiple cell types, extracellular matrix components, and a multitude of cell signaling pathways frequently contribute to poor outcomes, which make cancer the second leading killer in the United States. A better understanding of how these constituents interact will inevitably facilitate development of novel cancer therapeutics and diagnostics. To advance scientific discovery towards this goal, innovative experimental techniques are required. In this dissertation, new research methods for probing cell communication at a single to multi cell level within 3D models of the tumor microenvironment are presented. Optical trapping, composite nanocapsules (i.e., gold-coated liposomes), and 3D cell culture models were the foundation for the development of these research tools. The first aim of this dissertation was to optimize our ability to optically manipulate gold-coated liposomes for the purpose of delivering molecular content to cells. The second aim was to apply optical manipulation of gold-coated liposomes to quantitatively deliver signaling molecules into a single cell to activate communication. The third aim was to develop a 3D model of the tumor microenvironment and demonstrate cell communication within this physiologically accurate architecture. The basis for this work was gold-coated liposomes' strong plasmon resonance with visible to near infrared (NIR) wavelengths of light, which enabled photo-thermal conversion and optical trapping. To identify preferred conditions for optical manipulation of gold-coated liposomes for delivering content into cells, gold-coated liposomes made with different dielectric properties were optically trapped under various laser modulation schemes and thoroughly characterized, enabled by high speed (kHz) imaging. Application of this technique was realized by precise delivery of molecular agents into a single cell (i.e., optical injection). As a demonstration of optical injection, the NIR trapping beam was utilized to propel gold-coated liposomes encapsulating inositol trisphosphate (IP3) into a single cell to initiate calcium (Ca²⁺) signaling. In another method for intracellular delivery, cells were preloaded with similar gold-coated liposomes, internalized by macropinocytosis, and then exposed to on-resonant laser light to trigger on-demand release of IP3 to activate Ca²⁺ signaling. Lastly, a 3D cell culture model of ovarian cancer microenvironment was developed as a platform for interrogating cell signaling. The in vitro model comprised human ovarian cancerous epithelial cells grown upon a collagen and human fibroblast stroma recapitulating architecture of human tissue. Gold-coated liposomes encapsulating signaling molecules, optical manipulation, and a 3D model of ovarian cancer, a trio of versatile experimental tools opens new opportunities for studying the tumor microenvironment.
Type:
text; Electronic Dissertation
Keywords:
cell signaling; liposomes; optical trap; plasmon resonance; tumor microenvironment; Biomedical Engineering; calcium
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Graduate College; Biomedical Engineering
Degree Grantor:
University of Arizona
Advisor:
Romanowski, Marek

Full metadata record

DC FieldValue Language
dc.language.isoen_USen
dc.titleUntangling Intercellular Communication Using Optical Manipulation in 3D Models of Tumor Microenvironmenten_US
dc.creatorOrsinger, Gabriel V.en_US
dc.contributor.authorOrsinger, Gabriel V.en_US
dc.date.issued2014-
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.releaseRelease 18-Jul-2015en_US
dc.description.abstractThe tumor microenvironment is a tangled web of multiple cell types, extracellular matrix components, and a multitude of cell signaling pathways frequently contribute to poor outcomes, which make cancer the second leading killer in the United States. A better understanding of how these constituents interact will inevitably facilitate development of novel cancer therapeutics and diagnostics. To advance scientific discovery towards this goal, innovative experimental techniques are required. In this dissertation, new research methods for probing cell communication at a single to multi cell level within 3D models of the tumor microenvironment are presented. Optical trapping, composite nanocapsules (i.e., gold-coated liposomes), and 3D cell culture models were the foundation for the development of these research tools. The first aim of this dissertation was to optimize our ability to optically manipulate gold-coated liposomes for the purpose of delivering molecular content to cells. The second aim was to apply optical manipulation of gold-coated liposomes to quantitatively deliver signaling molecules into a single cell to activate communication. The third aim was to develop a 3D model of the tumor microenvironment and demonstrate cell communication within this physiologically accurate architecture. The basis for this work was gold-coated liposomes' strong plasmon resonance with visible to near infrared (NIR) wavelengths of light, which enabled photo-thermal conversion and optical trapping. To identify preferred conditions for optical manipulation of gold-coated liposomes for delivering content into cells, gold-coated liposomes made with different dielectric properties were optically trapped under various laser modulation schemes and thoroughly characterized, enabled by high speed (kHz) imaging. Application of this technique was realized by precise delivery of molecular agents into a single cell (i.e., optical injection). As a demonstration of optical injection, the NIR trapping beam was utilized to propel gold-coated liposomes encapsulating inositol trisphosphate (IP3) into a single cell to initiate calcium (Ca²⁺) signaling. In another method for intracellular delivery, cells were preloaded with similar gold-coated liposomes, internalized by macropinocytosis, and then exposed to on-resonant laser light to trigger on-demand release of IP3 to activate Ca²⁺ signaling. Lastly, a 3D cell culture model of ovarian cancer microenvironment was developed as a platform for interrogating cell signaling. The in vitro model comprised human ovarian cancerous epithelial cells grown upon a collagen and human fibroblast stroma recapitulating architecture of human tissue. Gold-coated liposomes encapsulating signaling molecules, optical manipulation, and a 3D model of ovarian cancer, a trio of versatile experimental tools opens new opportunities for studying the tumor microenvironment.en_US
dc.typetexten
dc.typeElectronic Dissertationen
dc.subjectcell signalingen_US
dc.subjectliposomesen_US
dc.subjectoptical trapen_US
dc.subjectplasmon resonanceen_US
dc.subjecttumor microenvironmenten_US
dc.subjectBiomedical Engineeringen_US
dc.subjectcalciumen_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineBiomedical Engineeringen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorRomanowski, Mareken_US
dc.contributor.committeememberRomanowski, Mareken_US
dc.contributor.committeememberLynch, Ronalden_US
dc.contributor.committeememberUtzinger, Ursen_US
dc.contributor.committeememberYoon, Jeong-Yeolen_US
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