BioE PhD Proposal- Adriana Mulero- Russe

Advisor:
Andrés García, Ph.D., School of Mechanical Engineering, Georgia Institute of Technology

Committee Members:

Michael A. Helmrath, M.D., Division of Pediatric General and Thoracic Surgery, Cincinnati Children’s Hospital Medical Center

Hang Lu, Ph.D. School of Chemical & Biomolecular Engineering, Georgia Institute of Technology

Asma Nusrat, M.D., Department of Pathology University of Michigan

Johnna S. Temenoff, Ph.D., School of Biomedical Engineering Georgia Institute of Technology

Engineered Synthetic Platform for Human Intestinal Organoid Generation and Delivery 

Human intestinal organoids (HIOs) are three-dimensional (3D) multicellular structures, derived from either adult intestinal stem cells or human pluripotent stem cells (hPSCs), that recapitulate human intestinal tissue architecture. HIOs are a promising cell source for intestinal tissue repair, disease modeling, and drug screening. Previous work has demonstrated that HIOs engraft to the injured intestinal wall in vivo, however, these approaches are significantly limited by the lack of an appropriate delivery vehicle to drive HIO engraftment. HIO generation from hPSCs is a multi-stage directed differentiation process comprising three stages: (I) a definitive endoderm 2D monolayer, (II) self-organized 3D aggregates (human intestinal spheroids, HIS), and (III) intestinal specification into HIOs within a 3D extracellular matrix. This in vitro culture process spans a 2D growth substrate (stage I and II) to a 3D matrix (stage III). The growth stages (2D and 3D) are supported by Matrigel, a murine tumor-derived basement membrane extract with ill-defined composition, lot-to-lot variability, and limited clinical translation potential presenting a major roadblock to HIOs clinical translation. Another roadblock to HIO technologies is the low yield and consistency of HIS differentiation in HIOs. The objectives of this project are to (1) engineer a synthetic hydrogel platform with independent control of the biochemical and biophysical cues guiding the entire in vitro differentiation of hPSCs into HIOs, and (2) deliver HIOs in a synthetic coating to intestinal injuries in vivo. The central hypothesis of this work is that engineering a PEG-based synthetic matrix to support HIO in vitro generation and in vivo delivery will increase the reproducibility, yield, and clinical translatability of this transformative organoid technology.