Permeability transition‐independent release of mitochondrial cytochrome c induced by valinomycin

International Journal of Pharmaceutics

Shinohara

Kakudo

et al. 2005

Self Cite

Mastoparan (MP), a potent facilitator of mitochondrial permeability transition (PT), could be used as an antitumor agent, if it were encapsulated in a tumor selective delivery system. We recently developed transferrin-modified liposomes (Tf-L) with a pH-sensitive fusogenic peptide (GALA), which delivers an encapsulated fluorescent marker into cytosol efficiently. In this study, we encapsulated MP into Tf-L with GALA for the selective delivery to mitochondria of tumor cells. The MP showed potent PT activity at concentrations above 25 M in a homogenate of K 562 cells as well as in isolated mitochondria in the presence of phosphate. Tf-L equipped with cholesteryl GALA can release encapsulated sulforhodamine B, while Tf-L failed, as evidenced by confocal laser scanning microscopy. The MP, which was delivered with Tf-L with GALA, released cytochrome c (cyt c) from mitochondria to the cytosol, while free MP released cyt c not only to the cytosol but also extracellulary. These results demonstrate the utility of MP in Tf-L with GALA for cancer therapy.

Section: Methodsmentioning

confidence: 99%

Mitochondrial delivery of mastoparan with transferrin liposomes equipped with a pH-sensitive fusogenic peptide for selective cancer therapy

International Journal of Pharmaceutics

Shinohara

Kakudo

et al. 2005

Self Cite

“…In this experiment, the livers were harvested following systemic injection and homogenized, and the mitochondria-enriched fraction was then isolated from the homogenate by differential centrifugation, as described in a previous report [11,23]. Samples of the homogenate and mitochondria-enriched fraction were subjected to Western blotting using primary antibodies against COX IV, Histone H3 and GAPDH, as mitochondrial, nuclear and cytosolic fraction markers ( Figure 2B).…”

Section: Distribution Of the Mito-porter In Liver Tissue And Mitochonmentioning

confidence: 99%

Mitochondrial delivery of Coenzyme Q10 via systemic administration using a MITO-Porter prevents ischemia/reperfusion injury in the mouse liver

Journal of Controlled Release

Nakamura

Abe

et al. 2015

We herein report on a mitochondrial therapeutic effect based on the delivery of coenzyme Q10 (CoQ10), an anti-oxidant, to in vivo mitochondria using a MITO-Porter, a liposome-based mitochondrial delivery system that functions via membrane fusion. To evaluate the effects, we used a mouse liver ischemia/reperfusion injury (I/R injury) model, in which mitochondrial reactive oxygen species are overexpressed. We packaged CoQ10 in the lipid phase of a MITO-Porter and optimized the mitochondrial fusogenic activities to produce the CoQ10-MITO-Porter. A histological observation of the carriers in the liver by confocal laser scanning microscopy was done and the accumulation of the carrier labeled with a radio isotope in the liver confirmed that the CoQ10-MITO-Porter was delivered to liver mitochondria via systemic injection. These analytical results permitted us to optimize the compositions of the CoQ10-MITO-Porter so as to permit it to efficiently accumulate in mouse liver mitochondria.Finally, we applied the optimized CoQ10-MITO-Porter to mice via tail vein injection, and hepatic I/R injury was then induced, followed by measuring serum alanine aminotransferase (ALT) levels, a marker of liver injury. We confirmed that the use of the CoQ10-MITO-Porter resulted in a significant decrease in serum ALT levels, indicating that in vivo mitochondrial delivery of the CoQ10 via MITO-Porter prevents I/R injury in mice livers. This provides a demonstration of the potential use of such a delivery system in mitochondrial therapies.

“…To verify the mitochondrial delivery of pDNA, the purity of the isolated mitochondria needs to be verified. In this experiment, the crural muscles were harvested following HLV injection and homogenized, and a mitochondria-enriched fraction was then isolated from the homogenate by differential centrifugation, as described in a previous report [27,28]. Samples of the homogenate and mitochondria-enriched fraction were subjected to western blotting using primary antibodies against the COX IV, Lamin A+C and GAPDH, as mitochondrial, nuclear and cytosolic fraction markers (Fig.…”

Section: Validation Of Mitochondrial Delivery Of Pdna Targeted To Skementioning

confidence: 99%

Localization of exogenous DNA to mitochondria in skeletal muscle following hydrodynamic limb vein injection

Yasuzaki

Journal of Controlled Release

Kanefuji

et al. 2013

ABSTRACT.Mitochondrial genetic disorders are a major cause of mitochondrial diseases. It is therefore likely that mitochondrial gene therapy will be useful for the treatment of such diseases. Here, we report on the possibility of mitochondrial gene delivery in skeletal muscle using hydrodynamic limb vein (HLV) injection. The HLV injection procedure, a useful method for transgene expression in skeletal muscle, involves the rapid injection of a large volume of naked plasmid DNA (pDNA) into the distal vein of a limb. We hypothesized that the technique could be used to deliver pDNA not only to nuclei but also mitochondria, since cytosolic pDNA that is internalized by the method may be able to overcome mitochondrial membrane. We determined if pDNA could be delivered to myofibrillar mitochondria by HLV injection by PCR analysis. Mitochondrial toxicity assays showed that the HLV injection had no influence on mitochondrial function. These findings indicate that HLV injection promises to be a useful technique for in vivo mitochondrial gene delivery.