Enhanced gene expression by a novel designed leucine zipper endosomolytic peptide

Aqeel Ahmad; Kirsi Rilla; Jing Zou; Weikai Zhang; Ilmari Pyykkö; Paavo Kinnunen; Sanjeev Ranjan

doi:10.1016/j.ijpharm.2021.120556

Enhanced gene expression by a novel designed leucine zipper endosomolytic peptide

Aqeel Ahmad
, Kirsi Rilla
, Jing Zou
, Weikai Zhang
, Ilmari Pyykkö
, Paavo Kinnunen
, Sanjeev Ranjan^*

^*Corresponding author for this work

Computing Sciences

Research output: Contribution to journal › Article › Scientific › peer-review

7 Citations (Scopus)

12 Downloads (Pure)

Abstract

An endosomal trap is a major barrier in gene therapy. We have designed an endosomolytic peptide based on the leucine zipper sequence and characterized it both structurally and functionally. The results illustrated that leucine zipper endosomolytic peptide (LZEP) exhibited appreciable hemolysis of human red blood cells (hRBCs) at pH 5.0, but negligible hemolysis at pH 7.4. Calcein release experiment indicated that only at pH 5.0 but not at pH 7.4, LZEP was able to permeabilize hRBCs. LZEP revealed significant self-assembly as well as peptide induced α-helical structure at pH 5.0. Unlike at pH 5.0, LZEP failed to self-assemble and showed a random coil structure at pH 7.4. Transfection data depicted that lipoplexes modified with LZEP resulted in significantly higher gene expression as compared to lipoplexes without LZEP. Interestingly, the transfection efficacy of LZEP modified lipid nanoparticles reached the levels of Lipofectamine 2000 (LF 2000), without any cellular toxicity as observed by MTT assay. The results suggest a novel approach for designing endosomolytic peptides by employing the leucine zipper sequence and simultaneously the designed peptides could be utilized for enhancing gene delivery into mammalian cells.

Original language	English
Article number	120556
Journal	International Journal of Pharmaceutics
Volume	601
DOIs	https://doi.org/10.1016/j.ijpharm.2021.120556
Publication status	Published - 15 May 2021
Publication type	A1 Journal article-refereed

Funding

This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug & gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation. This paper is dedicated to the memory of the late Prof. Paavo Kinnunen. This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug & gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation.

Keywords

Endosomal escape
Endosomolytic peptides
Gene delivery
Gene expression
Leucine zipper
Liposome nanoparticles.

Publication forum classification

Publication forum level 2

ASJC Scopus subject areas

Pharmaceutical Science

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10.1016/j.ijpharm.2021.120556Licence: CC BY-NC-ND

1-s2.0-S0378517321003616-mainFinal published version, 3.24 MBLicence: CC BY-NC-ND

http://urn.fi/URN:NBN:fi:tuni-202105265459Licence: CC BY-NC-ND

Cite this

@article{38915e7dcc2e4f6686fff0e77ea26b0f,

title = "Enhanced gene expression by a novel designed leucine zipper endosomolytic peptide",

abstract = "An endosomal trap is a major barrier in gene therapy. We have designed an endosomolytic peptide based on the leucine zipper sequence and characterized it both structurally and functionally. The results illustrated that leucine zipper endosomolytic peptide (LZEP) exhibited appreciable hemolysis of human red blood cells (hRBCs) at pH 5.0, but negligible hemolysis at pH 7.4. Calcein release experiment indicated that only at pH 5.0 but not at pH 7.4, LZEP was able to permeabilize hRBCs. LZEP revealed significant self-assembly as well as peptide induced α-helical structure at pH 5.0. Unlike at pH 5.0, LZEP failed to self-assemble and showed a random coil structure at pH 7.4. Transfection data depicted that lipoplexes modified with LZEP resulted in significantly higher gene expression as compared to lipoplexes without LZEP. Interestingly, the transfection efficacy of LZEP modified lipid nanoparticles reached the levels of Lipofectamine 2000 (LF 2000), without any cellular toxicity as observed by MTT assay. The results suggest a novel approach for designing endosomolytic peptides by employing the leucine zipper sequence and simultaneously the designed peptides could be utilized for enhancing gene delivery into mammalian cells.",

keywords = "Endosomal escape, Endosomolytic peptides, Gene delivery, Gene expression, Leucine zipper, Liposome nanoparticles.",

author = "Aqeel Ahmad and Kirsi Rilla and Jing Zou and Weikai Zhang and Ilmari Pyykk{\"o} and Paavo Kinnunen and Sanjeev Ranjan",

note = "Funding Information: This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug \& gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation. Funding Information: This paper is dedicated to the memory of the late Prof. Paavo Kinnunen. This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug \& gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

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month = may,

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doi = "10.1016/j.ijpharm.2021.120556",

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volume = "601",

journal = "International Journal of Pharmaceutics",

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T1 - Enhanced gene expression by a novel designed leucine zipper endosomolytic peptide

AU - Ahmad, Aqeel

AU - Rilla, Kirsi

AU - Zou, Jing

AU - Zhang, Weikai

AU - Pyykkö, Ilmari

AU - Kinnunen, Paavo

AU - Ranjan, Sanjeev

N1 - Funding Information: This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug & gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation. Funding Information: This paper is dedicated to the memory of the late Prof. Paavo Kinnunen. This study was conducted in the European Community 6th framework program; contract NMP4-CT-2006-026556, NANOEAR, aimed at targeted drug & gene delivery into the cells of the inner ear. This work was also financially supported by the KR personal grant (project number 64814), which was financed by the Jane and Aatos Erkko Foundation. Publisher Copyright: © 2021 The Author(s)

PY - 2021/5/15

Y1 - 2021/5/15

N2 - An endosomal trap is a major barrier in gene therapy. We have designed an endosomolytic peptide based on the leucine zipper sequence and characterized it both structurally and functionally. The results illustrated that leucine zipper endosomolytic peptide (LZEP) exhibited appreciable hemolysis of human red blood cells (hRBCs) at pH 5.0, but negligible hemolysis at pH 7.4. Calcein release experiment indicated that only at pH 5.0 but not at pH 7.4, LZEP was able to permeabilize hRBCs. LZEP revealed significant self-assembly as well as peptide induced α-helical structure at pH 5.0. Unlike at pH 5.0, LZEP failed to self-assemble and showed a random coil structure at pH 7.4. Transfection data depicted that lipoplexes modified with LZEP resulted in significantly higher gene expression as compared to lipoplexes without LZEP. Interestingly, the transfection efficacy of LZEP modified lipid nanoparticles reached the levels of Lipofectamine 2000 (LF 2000), without any cellular toxicity as observed by MTT assay. The results suggest a novel approach for designing endosomolytic peptides by employing the leucine zipper sequence and simultaneously the designed peptides could be utilized for enhancing gene delivery into mammalian cells.

AB - An endosomal trap is a major barrier in gene therapy. We have designed an endosomolytic peptide based on the leucine zipper sequence and characterized it both structurally and functionally. The results illustrated that leucine zipper endosomolytic peptide (LZEP) exhibited appreciable hemolysis of human red blood cells (hRBCs) at pH 5.0, but negligible hemolysis at pH 7.4. Calcein release experiment indicated that only at pH 5.0 but not at pH 7.4, LZEP was able to permeabilize hRBCs. LZEP revealed significant self-assembly as well as peptide induced α-helical structure at pH 5.0. Unlike at pH 5.0, LZEP failed to self-assemble and showed a random coil structure at pH 7.4. Transfection data depicted that lipoplexes modified with LZEP resulted in significantly higher gene expression as compared to lipoplexes without LZEP. Interestingly, the transfection efficacy of LZEP modified lipid nanoparticles reached the levels of Lipofectamine 2000 (LF 2000), without any cellular toxicity as observed by MTT assay. The results suggest a novel approach for designing endosomolytic peptides by employing the leucine zipper sequence and simultaneously the designed peptides could be utilized for enhancing gene delivery into mammalian cells.

KW - Endosomal escape

KW - Endosomolytic peptides

KW - Gene delivery

KW - Gene expression

KW - Leucine zipper

KW - Liposome nanoparticles.

U2 - 10.1016/j.ijpharm.2021.120556

DO - 10.1016/j.ijpharm.2021.120556

M3 - Article

C2 - 33798688

AN - SCOPUS:85104569180

SN - 0378-5173

VL - 601

JO - International Journal of Pharmaceutics

JF - International Journal of Pharmaceutics

M1 - 120556

ER -

Enhanced gene expression by a novel designed leucine zipper endosomolytic peptide

Abstract

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