Page 15 - Q. Neuroscience

Intraneuronal Delivery Of Parkin Protects Neurons
By Replacing Damaged Mitochondria

Jiae Park, Eunna Chung, Wonheum Nah, Joonno Lee, Soyoung Park, Shinyoung Kang, Jaehyung Park and Daewoong Jo
Drug Development Team, Cellivery R&D Institute, Cellivery Therapeutics, Inc., Seoul 03929, Korea.

BACKGROUND AIM
Parkinson’s disease (PD) is a neurodegenerative disorder that causes The goal of this study is to develop aMTD-fused Parkin
abnormal movement due to the selective loss of dopaminergic neurons in recombinant protein as a Parkinson’s disease (PD) therapeutic
brain. Parkin is an E3 ubiquitin ligase that plays a critical role in the agent. A combination of aMTD524 with a solubilization domain
ubiquitination process to fix damaged mitochondria. Improved Cell-Permeable (SD) shows the highest solubility, yield and cell-permeability of
(iCP) Parkin fused to an advanced macromolecule transduction domain Parkin protein, which is called improved cell-permeable (iCP)
(aMTD), a recombinant protein, has been developed in E. coli system. Parkin.
METHODS
Improved cell-permeable (iCP) Parkin has been developed and optimized by fusing human Parkin sequence to advanced macromolecule
transduction domain (aMTD) and a solubilization domain. In addition, to investigate biological activity of iCP-Parkin, 1) auto-ubiquitination
assay, 2) immunoprecipitation of PINK1 and iCP-Parkin, 3) mitophagy assay and 4) expression levels of mitochondrial membrane protein were
performed in vitro.
RESULTS

Figure 1. iCP-Parkin Is Phosphorylated by Figure 2. iCP-Parkin Does Not Require Figure 3. iCP-Parkin Is Constitutively Active
PINK1 in Tube & Cell Phosphorylation of Ser-78 To Be Activated Regardless of Phosphorylation of Ser-78

A B A

B

Figure 4. iCP-Parkin Accumulation In
Damaged Mitochondria

A
B C
Figure 7. iCP-Parkin Induces Mitochondria
Biogenesis By Expression Of Mitochondria Proteins
A

Figure 5. iCP-Parkin Ubiquitinates Various Substrates Figure 6. iCP-Parkin Induces
for Mitophagy & Mitochondral Biogenesis Mitophagy
B
A B A
B

C
C D

CONCLUSION REFERENCES Contact information

These results suggest that iCP-Parkin Chung et al. (2020) Science Advances, 6: eaba 1193 Minyong Jung
recovers mitochondria functionality by Lim et al. (2013) Clinical Cancer Research, 19: 680-690 New Drug & Business Development
utilizing its ubiquitin E3 ligase activity, and Lim et al. (2013) Biomaterials, 34: 6261-6271 Cellivery Therapeutics, Inc.
rescues neurons under PD-like pathological Lim et al. (2012) Molecular Therapy, 20: 1540-1549 jungmy@cellivery.com
condition. Jo et al. (2005) Nature Medicine, 11: 892-898 +82-2-3151-8900
Jo et al. (2001) Nature Biotechnology, 19: 929-933