Détails sur le projet
Description
PROJECT SUMMARY/ABSTRACT
Ligament injuries are compounded by the drastically increased risks of reinjury and eventual osteoarthritis. These
risks result from a failure of differentiation; torn mammalian ligaments reform with fibrous scar tissue rather than
with true ligamentocytes, altering the biomechanical properties of the repaired ligament and destabilizing the
nearby joint. While the current models (mice and rabbits) used to study ligament repair recapitulate ligament
scarring, they are unlikely to yield novel insights into targets for regenerative therapeutics. To address this
shortcoming, our lab has developed the highly regenerative zebrafish as a model for craniofacial ligament
regeneration. Zebrafish have the remarkable capacity to regenerate their ligaments without scarring in under a
month. Our preliminary data demonstrate that the jaw joint-supporting interopercle (IOP) ligament in zebrafish
shows identical morphology to the uninjured ligament as soon as 28 days after transection. During this time, the
injured ligament is replaced with a dense mesenchyme which deposits a complex extracellular matrix to remake
a functional ligament. Additionally, preliminary lineage tracing experiments show that both the uninjured ligament
and the regenerative mesenchyme are cranial neural crest-derived. This project aims to identify the source of
the mesenchymal cells which reform the ligament, as well as the genetic and epigenetic changes which promote
ligamentocyte fate. In Aim 1, I will use enhancer peaks with increased accessibility after ligament injury to firstly
trace the lineage of the regenerative mesenchyme to the regenerated ligament, and secondly trace the lineage
of the periosteum through regeneration. In Aim 2, I will analyze my single-nuclei multiomic (gene expression
and chromatin accessibility for each cell) data from jaw joints through IOP ligament regeneration to generate a
short list of candidate transcription factors using a selection funnel of motif accessibility, gene expression, and
transcription factor binding. I will then assess in vivo if these transcription factors are expressed before
ligamentocyte fate is adopted, and generate knockout zebrafish lines to assess if each transcription factor is
necessary for ligamentocyte fate. Through these aims, we will unveil a novel population of cells capable of
regenerating ligaments, as well as the transcription factors necessary for ligamentocyte differentiation.
The training plan outlined through this proposal will develop the techniques, mentorship, and communication
skills necessary to establish my future career as an independent scientist studying craniofacial regeneration.
Through the mentorship of Dr. Joanna Smeeton (Sponsor), Dr. Stavros Thomopoulos (Co-sponsor), and my
thesis advisory committee, I will be well prepared for my transition to a postdoctoral fellowship at a leading
biomedical institution. Dr. Smeeton has extensive knowledge of the zebrafish as a model for craniofacial
regeneration, and Dr. Thomopoulos is an expert in translational mouse tendon development with an outstanding
mentorship record. Under their combined sponsorship, I will learn cutting-edge computational and in vivo
genetics to develop a productive career in regenerative craniofacial biology.
Statut | Terminé |
---|---|
Date de début/de fin réelle | 9/1/22 → 8/31/23 |
Keywords
- Biotecnología
- Genética
Empreinte numérique
Explorer les sujets de recherche abordés dans ce projet. Ces étiquettes sont créées en fonction des prix/bourses sous-jacents. Ensemble, ils forment une empreinte numérique unique.