Understanding lineage capacity of blastema-driven craniofacial regeneration

The National Institute of Dental and Craniofacial Research awarded The Trustees of Columbia University in the City of New York $152,166 on September 4, 2026, under the Oral Diseases and Disorders Research program (CFDA 93.121) to conduct research on blastema-driven craniofacial regeneration in zebrafish.

The research investigates how multiple blastemas—regenerative cell clusters—coordinate to restore craniofacial joint tissues following injury. The project uses an adult zebrafish whole joint resection model that creates three distinct blastemas in the craniofacial region, a system unique in regeneration research. Preliminary data demonstrate that zebrafish can regenerate functional lubricated articular joints with complete restoration of bone, ligament, tendon, muscle, and articular cartilage within two weeks of injury. The funded work aims to determine transcriptional regulation during blastema progression and characterize spatial heterogeneity in blastema formation. Aim 1 includes isolating blastemas, culturing them in vitro, and exposing them to chondrogenic, osteogenic, adipogenic, and muscle inductive media to assess each blastema's cell-type bias and lineage capacity.

Work is performed in New York, New York. The period of performance runs from September 4, 2026, through September 3, 2029. The award is classified as a Project Grant under NIDCR's extramural research program supporting basic, translational, and clinical research in dental, oral, and craniofacial health and disease.

Abstract

And abstract a key feature of animals with extraordinary regenerative capacity is the generation of a blastema, or a regenerative cluster of cells that forms at the site(s) of damage shortly after injury. This regenerative cell population is responsible for restoring and repatterning mature tissue types at the damaged location. The majority of blastema regeneration models currently in use involve a singular blastema origin for their research, such as the axolotl limb model or zebrafish FIN model. In the smeeton lab, we have developed a whole joint resection model in the adult zebrafish craniofacial skeleton to assess functional regeneration of synovial joint tissues. Our model is unique within the field as the whole joint resection surgery results in a three-wound surgical site that creates three distinct blastema in the craniofacial region. IT is currently unknown how these three blastema operate in concert to form a single regenerated joint and how unique environmental cues from the cells around each blastema may influence cellular lineage contribution or fate decisions in regeneration. Our preliminary data show that a new functional lubricated articular joint forms in zebrafish following a complete joint resection injury. We find that regenerated joints reestablish all the mature joint cell types within the first 2 weeks of healing, such as a bone, ligament, tendon, muscle, and articular cartilage, followed by maturation and tissue remodeling to integrate these new tissues into the craniofacial skeletal complex. This project aims to determine the transcriptional regulation of crucial time points in blastema progression, and the spatial heterogeneity involved with blastema formation. In aim 1, blastema will be isolated, cultured in vitro, and treated with chondrogenic, osteogenic, adipogenic, and muscle inductive media to test each blastema's bias to forming cell specific fates. Additionally, blastema from transgenic-fluorescent animals will be dissociated into a single cell solution and injected into the corresponding blastema of non-transgenic sibling recipients to trace the contributions of each blastema to the final regenerated joint. In aim 2, i will determine where our blastema meet in a spatiotemporal manner by assessing the relative proliferation of each blastema and discerning when they fuse and integrate with one another. I will perform repeated blastema injuries after whole joint resection to compare the contributions of the three distinct blastema after injury. I will use scrna sequencing to assess the transcriptome of each blastema and their surrounding injury-responsive tissue to identify potential factors responsible for triggering blastema initiation and progression during joint regeneration. I will then contrast these transcriptomic blastema profiles against known non-craniofacial datasets such as the caudal FIN blastema to discern the programs specific to craniofacial regeneration, and generate conditional ablation lines to test the impact of the most highly differentially upregulated genes during craniofacial regeneration. The results of these experiments will elucidate the mechanisms behind multi-blastema integration and define the formation and dynamics of a multi-blastema-driven craniofacial regenerative program.