CRANIOFACIAL CHONDROGENESIS IN ORGANOIDS FROM HUMAN STEM CELLDERIVED NEURAL CREST CELLS

Author

• Nagashree Krishna Avabhrath

Year of Award

2026

Document Type

Dissertation

Degree Type

Doctor of Philosophy (PhD)

Degree Name

Cellular, Molecular and Microbial Biology

Department or School/College

Division of Biological Sciences

Committee Chair

Mark L. Grimes

Commitee Members

Ekaterina Voronina, Jesse Hay, Brent Ryckman, Monica Serban, Aleksander Skardal

Keywords

Cell Signaling, Craniofacial Cartilage, Neural Crest, Organoids, Stem Cells

Abstract

Knowledge of cell signaling pathways that drive human neural crest differentiation into craniofacial chondrogenesis is incomplete, yet essential for using stem cells to regenerate craniomaxillofacial structures. To accelerate translational progress, we developed a differentiation protocol that generated self-organizing craniofacial cartilage organoids from human embryonic stem cell-derived neural crest cells. Histological staining of cartilage organoids revealed tissue architecture and staining typical of elastic cartilage. Protein and post-translational modifications (PTM) mass spectrometry and snRNA-seq data showed that chondrocyte organoids expressed robust levels of cartilage extracellular matrix (ECM) components; many collagens, aggrecan, perlecan, proteoglycans, and elastic fibers. We identified two populations of chondroprogenitor cells, mesenchyme cells and nascent chondrocytes, and the growth factors involved in paracrine signaling between them. We show that ECM components secreted by chondrocytes not only create a structurally resilient matrix that defines cartilage but also play a pivotal autocrine cell signaling role in determining chondrocyte fate. The organoids also served as a model to understand the disease pathogenesis of congenital arrhinia, a rare syndrome where the babies are born without noses. Arrhinia patient-derived induced pluripotent cells (iPSCs) when differentiated into craniofacial cartilage organoids showed reduced cartilage. Mass spectrometry showed less expression of cartilage markers such as COL2A1, ELN and COMP. Spatial transcriptomics of the organoids and snRNA-seq analysis of cells at early time-point showed that the mutant organoids produced chondroprogenitors but the cells didn’t mature into cartilage.

Recommended Citation

Avabhrath, Nagashree Krishna, "CRANIOFACIAL CHONDROGENESIS IN ORGANOIDS FROM HUMAN STEM CELLDERIVED NEURAL CREST CELLS" (2026). Graduate Student Theses, Dissertations, & Professional Papers. 12671.
https://scholarworks.umt.edu/etd/12671