Application of human pluripotent stem cells and genetic engineering to treating and modeling pancreatic and cardiovascular degenerative diseases
Open Access
Author:
Jiang, Yuqian
Graduate Program:
Bioengineering
Degree:
Doctor of Philosophy
Document Type:
Dissertation
Date of Defense:
April 22, 2022
Committee Members:
Xiaojun Lian, Chair & Dissertation Advisor Daniel Hayes, Major Field Member Daniel Hayes, Program Head/Chair Jian Yang, Major Field Member Weihua Guan, Outside Unit & Field Member Jian Yang, Major Field Member
Human pluripotent stem cells (hPSCs) hold unprecedented promise in providing unlimited cell sources for regenerative therapies, serving as versatile platforms for drug screening, disease modeling and understanding embryonic development. The combination with various genetic engineering tools unfolds acceleratively increasing applications.
In this work, firstly I developed two genetically-modified hPSC models, which enables visualized characterization of hPSC-derived cardiomyocytes (hPSC-CMs). In addition, I established a robust growth-factor-free protocol named GiBi for generating definitive endoderm (DE) and pancreatic progenitor (PP) cells, facilitating scalable production of pancreatic cells for regenerative applications. Further optimization identified the culture medium and coating matrix compatible for our GiBi DE differentiation, enabling adaptation of this small-molecule-mediated DE protocol in different stem cell labs. Moreover, I investigated the possibility of forward programming of hPSCs or pancreatic progenitor cells into pancreatic β cells via manipulation of transcription factors. Evidence was also provided for induced β cell proliferation by activating Wnt signaling pathway. Last but not least, I developed several CRISPR-mediated systems, including two all-in-one PiggyBac vectors that can be utilized for genome or RNA editing in cell platforms and the Solid-State CRISPR-Cas12-Assisted Nanopores (SCAN) system for molecule detection.
