PI: Siyang Zheng
University: Carnegie Mellon University

Intracellular enzymes deficiencies contribute significantly to the child morbidity and mortality in inherited metabolic disorders (IMD). It is projected that at least 28,000 children will be born with IMD every year globally. However, current enzyme replace therapy (ERT) is based on the delivery of native enzymes into the circulation and has many drawbacks, which limits its wide adoption.

To meet this urgent and unmet clinical challenge, in this proposal, we propose to develop an extracellular vesicle (EV)-metal organic framework (MOF)-protein (EMP) nanoparticles platform for intracellular enzyme delivery for IMD treatment. The EMP nanoparticle leverages the fundamental characteristics of biomimetic MOF for caging guest proteins/enzymes with high efficiency and loading capacity. Furthermore, EMP nanoparticle leverages the fundamental structural characteristic of EVs, their lipid membrane, to enable systemic targeted enzyme delivery with significantly longer circulation time.

In our preliminary study, we have demonstrated that guest proteins are caged in the matrix of MOFs with high efficiency (up to ∼94%) and high loading capacity, and the nanoparticles are further enveloped with the EV membrane with high efficiency of ∼97%. Importantly, assisted by EMP nanoparticles, intracellular delivery of the therapeutic protein gelonin significantly inhibits tumor growth in vivo and increased therapeutic efficacy by 14-fold. Herein, we propose to develop the EMP nanoparticles with MOF loaded with alanine glyoxylate aminotransferase (AGT) and coated with cells-derived EV membranes for targeted intracellular enzyme delivery of AGT into the liver cells for a proof-of-concept study of hyperoxaluria type 1 (PH 1) treatment.

Aim one will develop a highly efficient EV generation method. Aim two will synthesize EMP nanoparticles caged with AGT with high efficiency and high loading capacity. Aim three will characterize EMP for intracellular AGT delivery to HepG2 cells with AGXT gene knockout. Future studies will focus on the in vivo characterization of EMP nanoparticles with AGT.