PROJECT SUMMARY/ABSTRACT
This application proposes a tailored research training plan that is designed to promote my development into an
independent clinician-scientist. The plan includes a rigorous laboratory training experience in multiple
techniques as well as a customized professional and career development plan. Furthermore, the training plan
is supported by outstanding institutional resources, including a strong and interwoven research community and
a supportive mentorship team. My research area of interest is translationally-relevant biomimetic approaches
to tissue repair. In the field of tissue engineering, there is a need for therapeutic tools that target specific
inflammatory and regenerative pathways. The NLRP3 inflammasome pathway is activated in several systemic
diseases, such as type I and type II diabetes. This pathway also contributes to chronic inflammation and
directly impairs repair and regeneration of various tissues within the body including bone and craniofacial
tissues. Diabetes-related chronic inflammation and poor craniofacial bone repair are significant oral healthcare
problems. Mesenchymal stem cell-derived extracellular vesicles (MSC EVs) and their miRNA cargo have high
therapeutic potential for immunomodulation in dental diseases and craniofacial tissue regeneration. Several
miRNAs within MSC EV cargo have been identified to regulate and suppress the NLRP3 pathway. However,
there is a gap in knowledge regarding the lack of mechanistic approaches to direct pathway-specific
manipulation of inflammation using EVs and to achieve controlled release of EVs to employ their therapeutic
effects. The proposed research will address this gap in knowledge through developing a controlled release
system for engineered anti-inflammatory EVs at wound sites. This release system will use an alginate-based
hydrogel platform to deliver engineered EVs. The alginate-based hydrogel can be functionally modified by
incorporating cell/EV binding motifs to the alginate backbone. The results of this proposal will work towards the
long-term goal of maximizing the therapeutic potential of EVs and enhancing tissue repair. Aim 1 will utilize
miRNA-based EV engineering to generate engineered MSC EVs to target the NLRP3 inflammasome pathway
using miRNA-22-3p as the candidate. Aim 2 entails developing a photocrosslinkable hydrogel for the controlled
release of engineered EVs using tissue resident metalloproteases abundant at wound sites as a trigger for EV
release. Aim 3 is focused on assessing the efficacy of the engineered EVs and the delivery system in vivo
using a diabetic mouse calvarial defect model. Overall, successful completion of these aims will provide me
with a comprehensive training in EV biology and related techniques, stem cell biology, molecular biology,
immunology, biomaterial design, imaging, animal handling and related histology and immunohistochemical
techniques. Results from this work will directly address oral healthcare issues such as diabetes-related chronic
inflammation and poor craniofacial bone repair.