Development of Organelle Replacement Therapy Using a Stearyl-Polyhistidine Peptide against Lysosomal Storage Disease Cells

Abstract

We previously reported on a polyhistidine peptide, His16 peptide, as a new cell-penetrating peptide. This peptide is anticipated to be a new carrier for drug delivery systems (DDSs) for targeting intracellular lysosomes because it can transport macromolecules (e.g., liposomes) into these organelles. In the present study, we examined the application of His16 peptide as a DDS carrier against lysosomal storage disease (LSD) cells. LSDs are metabolic disorders caused by loss of specific lysosomal enzymes. For the treatment of LSD cells, we devised a system designated organelle replacement therapy (ORT). ORT is a strategy for transporting exogenous lysosomes containing all kinds of lysosomal enzymes from normal cells into endogenous lysosomes in LSD cells using His16 peptide. To develop the ORT system, we prepared His16 peptide-modified healthy lysosomes (His16-Lyso) by insertion of a stearyl-His16 peptide into a hydrophobic region in the lysosomal membrane. His16-Lyso showed cellular uptake and localization to endogenous lysosomes in LSD cells. His16-Lyso also restored the proliferation of LSD cells, which otherwise showed slower proliferation than normal cells. These results suggested that His16-Lyso replenished deficient lysosomal enzymes in LSD cells. The results further suggest that His16-Lyso are promising candidates as a treatment tool for LSD cells and to establish a foundation for ORT.

Reduced α-galactosidase A activity in zebrafish (Danio rerio) mirrors distinct features of Fabry nephropathy phenotype

“…4 A). Similar findings have been reported in human Fabry nephropathy [68] , [69] , [70] , [71] but not in other FD model [29] , [72] .…”

Zebra-Sphinx: Modeling Sphingolipidoses in Zebrafish

“…Accordingly, microscopic analysis of the kidney revealed an increased glomerular size, dilated capillary loop, and thinner Bowman's space. In contrast with the results obtained in other animal models of Fabry disease that do not show renal abnormalities [155], these data are in keeping with the nephropathy that occurs in Fabry patients [156]. Notably, the absence of a Gb3 synthase encoding gene ortholog in zebrafish provides the unique opportunity to identify pathogenic processes that may work in concert with Gb3 accumulation in Fabry disease [131].…”

Emerging Landscape of Cell-Penetrating Peptide-Mediated Organelle Restoration and Replacement

“…To date, the only reported method of ORT is peptide-based lysosome replacement, which is still in the early stages of research. Cell-permeable H16-modified liposomes were applied as carriers of intercellular lysosome transfer in a model experiment by using human fibrosarcoma HT1080 cells as non-LSD cells (Figure ). Healthy lysosomes were first extracted from human fibrosarcoma HT1080 cells, which have been used to produce recombinant lysosomes for ERT. − Stearyl-His16 was inserted to the hydrophobic region of the lysosomal membrane, designated as His16-Lyso.…”

“…Research in CPP-mediated organelle targeting and organelle replacement is still relatively new, and two potential scenarios of this field can be considered depending on the degree of organelle damage. , One scenario is to achieve precise-targeting CPP-conjugated exogenous cargos (e.g., enzymes, DNA/RNA, antisense oligonucleotides (ASO), or peptide nucleic acids (PNA)) at the organelle level, including lysosomes, mitochondria, − endoplasmic reticulum, extracellular vesicles, exosomes, and Golgi apparatus. Another scenario such as injury, damage, disease, or dysfunction leads to the majority of organelles within eukaryotic cells becoming irreparable; therefore, direct replacement of organelles (mitochondria, − lysosomes, and exosomes , ) may be the last way to restore cellular function. All of this progress has started to offer valuable insights and provides a therapeutic avenue of CPP-based organelle targeting and replacement precision medicine.…”