Synthetic Monopartite Peptide That Enables the Nuclear Import of Genes Delivered by the Neurotensin-Polyplex Vector

Francisco E. Lopez-Salas, Rasajna Nadella, Minerva Maldonado-Berny, Maria L. Escobedo-Sanchez, Rosana Fiorentino-Pérez, Bismark Gatica-García, Manuel A. Fernandez-Parrilla, Moreno Mario Gil, David Reyes-Corona, Ubaldo García, Carlos E. Orozco-Barrios, Maria E. Gutierrez-Castillo, Daniel Martinez-Fong

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/β pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/β pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy.

Original languageEnglish
Pages (from-to)4572-4588
Number of pages17
JournalMolecular Pharmaceutics
Volume17
Issue number12
DOIs
StatePublished - 7 Dec 2020

Keywords

  • cancer
  • fusion proteins
  • gene therapy
  • importins
  • nonviral vectors
  • Parkinson's disease

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