PMID- 34440250 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20210830 IS - 2227-9059 (Print) IS - 2227-9059 (Electronic) IS - 2227-9059 (Linking) VI - 9 IP - 8 DP - 2021 Aug 19 TI - Novel Orthogonally Hydrocarbon-Modified Cell-Penetrating Peptide Nanoparticles Mediate Efficient Delivery of Splice-Switching Antisense Oligonucleotides In Vitro and In Vivo. LID - 10.3390/biomedicines9081046 [doi] LID - 1046 AB - Splice-switching therapy with splice-switching oligonucleotides (SSOs) has recently proven to be a clinically applicable strategy for the treatment of several mis-splice disorders. Despite this, wider application of SSOs is severely limited by the inherently poor bioavailability of SSO-based therapeutic compounds. Cell-penetrating peptides (CPPs) are a class of drug delivery systems (DDSs) that have recently gained considerable attention for improving the uptake of various oligonucleotide (ON)-based compounds, including SSOs. One strategy that has been successfully applied to develop effective CPP vectors is the introduction of various lipid modifications into the peptide. Here, we repurpose hydrocarbon-modified amino acids used in peptide stapling for the orthogonal introduction of hydrophobic modifications into the CPP structure during peptide synthesis. Our data show that alpha,alpha-disubstituted alkenyl-alanines can be successfully utilized to introduce hydrophobic modifications into CPPs to improve their ability to formulate SSOs into nanoparticles (NPs), and to mediate high delivery efficacy and tolerability both in vitro and in vivo. Conclusively, our results offer a new flexible approach for the sequence-specific introduction of hydrophobicity into the structure of CPPs and for improving their delivery properties. FAU - Bazaz, Safa AU - Bazaz S AUID- ORCID: 0000-0002-6440-7577 AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. AD - Department of General Biology, Cihan University-Erbil, Kurdistan Region, Erbil 44001, Iraq. FAU - Lehto, Tonis AU - Lehto T AUID- ORCID: 0000-0003-1683-9604 AD - Institute of Technology, University of Tartu, Nooruse 1, 50411 Tartu, Estonia. FAU - Tops, Rahel AU - Tops R AUID- ORCID: 0000-0001-9023-879X AD - Institute of Technology, University of Tartu, Nooruse 1, 50411 Tartu, Estonia. FAU - Gissberg, Olof AU - Gissberg O AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Gupta, Dhanu AU - Gupta D AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Bestas, Burcu AU - Bestas B AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Bost, Jeremy AU - Bost J AUID- ORCID: 0000-0002-3804-0521 AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Wiklander, Oscar P B AU - Wiklander OPB AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Sork, Helena AU - Sork H AD - Institute of Technology, University of Tartu, Nooruse 1, 50411 Tartu, Estonia. FAU - Zaghloul, Eman M AU - Zaghloul EM AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. AD - Department of Pharmaceutics, Faculty of Pharmacy, Alexandria University, El-Khartoum Square, Azareeta, Alexandria 21521, Egypt. FAU - Mamand, Doste R AU - Mamand DR AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. AD - Department of Biomedical Sciences, Cihan University-Erbil, Erbil 44001, Kurdistan Region, Iraq. FAU - Hallbrink, Mattias AU - Hallbrink M AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Sillard, Rannar AU - Sillard R AD - PeptiSystems AB, Virdings Alle 22, 75450 Uppsala, Sweden. FAU - Saher, Osama AU - Saher O AUID- ORCID: 0000-0003-3286-5706 AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. AD - Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt. FAU - Ezzat, Kariem AU - Ezzat K AUID- ORCID: 0000-0003-4186-0675 AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Smith, C I Edvard AU - Smith CIE AUID- ORCID: 0000-0003-1907-3392 AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Andaloussi, Samir El AU - Andaloussi SE AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. FAU - Lehto, Taavi AU - Lehto T AD - Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186 Huddinge, Sweden. AD - Institute of Technology, University of Tartu, Nooruse 1, 50411 Tartu, Estonia. LA - eng PT - Journal Article DEP - 20210819 PL - Switzerland TA - Biomedicines JT - Biomedicines JID - 101691304 PMC - PMC8392223 OTO - NOTNLM OT - cell-penetrating peptides OT - cellular uptake OT - endocytosis OT - oligonucleotide delivery OT - splice-switching oligonucleotides COIS- D.G., O.P.B.W., and T.L (Taavi Lehto) are consultants for Evox Therapeutics. S.E.A is a co-founder and shareholder in Evox Therapeutics. Other authors declare no conflict of interest. EDAT- 2021/08/28 06:00 MHDA- 2021/08/28 06:01 PMCR- 2021/08/19 CRDT- 2021/08/27 01:06 PHST- 2021/07/20 00:00 [received] PHST- 2021/08/09 00:00 [revised] PHST- 2021/08/11 00:00 [accepted] PHST- 2021/08/27 01:06 [entrez] PHST- 2021/08/28 06:00 [pubmed] PHST- 2021/08/28 06:01 [medline] PHST- 2021/08/19 00:00 [pmc-release] AID - biomedicines9081046 [pii] AID - biomedicines-09-01046 [pii] AID - 10.3390/biomedicines9081046 [doi] PST - epublish SO - Biomedicines. 2021 Aug 19;9(8):1046. doi: 10.3390/biomedicines9081046.