Implementation of Genetic Circuits for Engineered Mesenchymal Stem Cell Chondrogenic Differentiation

Simple item page
dc.contributor.advisorBashor, Caleb J
dc.creatorPiepergerdes, Trenton C
dc.date.accessioned2023-08-09T18:20:16Z
dc.date.created2023-05
dc.date.issued2023-04-20
dc.date.submittedMay 2023
dc.date.updated2023-08-09T18:20:16Z
dc.descriptionEMBARGO NOTE: This item is embargoed until 2024-05-01
dc.description.abstractThe ability to culture stem cells and guide their differentiation has allowed myriad advancements in developmental biology and cell-based therapies. In vitro differentiation protocols have historically taken an “outside-in” approach where bioactive molecules (purified proteins, small molecules, etc.) are added exogenously to culture medium or functionalized onto culture surfaces (plates, scaffolds, etc.) to guide cell differentiation. Despite decades of protocol optimization, “outside-in” approaches result in heterogeneous populations where only a subset of cells matches the desired phenotype, and the presence of aberrant cell states makes the cells ineffective for therapeutic use. These shortcomings are particularly evident in cartilage tissue engineering, where the multipotency of mesenchymal stem cells (MSCs) is harnessed to regenerate cartilage tissue. In these therapies, MSCs produce areas of proper cartilage, but concurrently produce hypertrophic and fibrotic chondrocytes. These cells deposit bone and fibrous tissue, respectively, thus leading to suboptimal tissue properties and limiting clinical translation. The ability to encourage proper chondrogenic phenotypes while preventing undesired hypertrophic and fibrotic ones is thus of great interest to tissue engineers and developmental biologists alike. One promising strategy involves using synthetic gene circuits to precisely control the dose and timing of expression of genes critical to functional chondrogenic differentiation. This “inside-out” approach is inspired by natural cellular differentiation, where it has been demonstrated that precise timing and magnitude of expression of genes in key regulatory networks are responsible for driving differentiation to mature cell states. In this work, I developed a novel engineering platform that enabled this functionality with synthetic genetic circuits. The platform I created includes a novel framework for the quantitative design and implementation of genetic circuits in a variety of cell types paired with an in vitro model and assessment method using single cell RNA sequencing (scRNA-seq) that allows iterative circuit implementation and assessment of the effect of circuit function on cell phenotypes in a model of MSC chondrogenesis. This work represents a significant step forward for the fields of mammalian synthetic biology and tissue engineering by (1) allowing high throughput circuit design, creation, and implementation in mammalian cells and (2) providing an unprecedented description of chondrogenic differentiation trajectories and how to manipulate them.
dc.embargo.lift2024-05-01
dc.embargo.terms2024-05-01
dc.format.mimetypeapplication/pdf
dc.identifier.citationPiepergerdes, Trenton C. "Implementation of Genetic Circuits for Engineered Mesenchymal Stem Cell Chondrogenic Differentiation." (2023) Diss., Rice University. <a href="https://hdl.handle.net/1911/115134">https://hdl.handle.net/1911/115134</a>.
dc.identifier.urihttps://hdl.handle.net/1911/115134
dc.language.isoeng
dc.rightsCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.
dc.subjectsynthetic biology
dc.subjectbioengineering
dc.subjecttissue engineering
dc.subject
dc.titleImplementation of Genetic Circuits for Engineered Mesenchymal Stem Cell Chondrogenic Differentiation
dc.typeThesis
dc.type.materialText
thesis.degree.departmentBioengineering
thesis.degree.disciplineEngineering
thesis.degree.grantorRice University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
PIEPERGERDES-DOCUMENT-2023.pdf
Size:
9.41 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 2 of 2
No Thumbnail Available
Name:
PROQUEST_LICENSE.txt
Size:
5.85 KB
Format:
Plain Text
Description:
Download
No Thumbnail Available
Name:
LICENSE.txt
Size:
2.61 KB
Format:
Plain Text
Description:
Download
Collections
Rice University Electronic Theses and Dissertations