Multivalent polyrotaxane vectors as adaptive cargo complexes for gene therapy

Significance 

The need to boost the number of delivered nucleotides in gene therapy has encouraged the utilization of non-viral vectors as reliable carriers. A brief flashback shows us that until 2012, the Food & Drug Administration had approved only 1843 clinical trials implying DNA and RNA vectorization for gene therapy purposes. Close to two-thirds of those clinical trials were noted to have utilized viral vectors, despite their deficiencies such as low carrying capacity and immunological demerits. Recently, dynamic supermolecular bio-carrier conjugates, alternatively termed as cargo-complexes, emerged as excellent tool to deliver highly large amounts of sophisticated sensitive pharmaco- and bio-active species. The key components of the cargo-complexes are the carrier molecules, which must display selective affinity towards the molecules in transit, and be endowed with molecular segments active towards both the biological targets and the biochemical “assailants and hindrances”. To function in a reproducible manner, carriers must be themselves reproducibly produced. An important number of recently published studies investigate the transfection ability of non-viral vectors produced by the techniques of constitutional dynamic chemistry. Unfortunately, incentive results were obtained in both cases, but the versatility remained conditional.

To this note, a team of researchers led by Professor Mariana Pinteala from the Centre of Advanced Research in Bionano-conjugates and Biopolymers, “Petru Poni” Institute of Macromolecular Chemistry in Romania established a new procedure to construct carriers of cyclodextrin-based polyrotaxane-type. The research team demonstrated their ability to conjunctively cooperate to generate cargo-complexes with plasmid DNA, able to efficiently transfect cells in culture. Their work is currently published in the research journal, Polymer Chemistry.

The research technique employed commenced by obtaining polyrotaxane carriers having an axle of precise length, which allowed the threading of nine functionalized cyclodextrin units, and end capped with silatrane molecules. Next, in order to reduce the carrier cytotoxicity and to increase their physical–chemical versatility in loading nucleic acids, two supplemental ways to functionalize the CD units were investigated: first, with a mixture of polyethylenimines and short linear poly(ethyleneglycol) molecules, and second, by post-decorating polyethylenimines branches with ionizable small molecules: guanidine and arginine. Lastly, the experimental DNA binding capacity of these carriers in relationship with size, morphology and electrical charge was evaluated.

The authors observed that both alternatives numerically diminish the positive charges supported beared by the carrier, the first of them to a greater extent, and thus both reduce the packaging capacity of the carriers, in favor of cytocompatibiliy. The researchers also noted that due to the precise synthesis pathway, highly reproducible carriers were obtained.

Rodinel Ardeleanu and colleagues study has presented an efficient design and synthesis, of a new class of polyrotaxane carriers that were able to collaboratively generate cargo-complexes with nucleic acids. The scholars here demonstrated the cargo-complexes’ ability to transfect HeLa cells with a high efficiency. Altogether, the in vitro tests, showing no cytotoxicity and good transfection efficiency of the investigated carriers, provided new information on gene vector design and prove their excellent attributes and wide applicability. In vivo testing will further corroborate the utility of these new gene vectors for gene therapy.

About the author

Rodinel ARDELEANU obtained his B.Sc.in 1984 at the “Gheorghe Asachi” Technical University from Iasi and the Ph.D. in 1998 from The Romanian Academy. He has conducted research in the field of inorganic polymers based on silicon. Currently he is a senior scientist and his work is focused on organic/inorganic materials for gas storage/separation with fluorescent properties and supramolecular chemistry for the design of gene therapy non-viral vectors.

Email: [email protected];

About the author

Andrei DASCALU received his Ph.D. degree in chemistry in 2016 from The Romanian Academy, Chemical Sciences Section, “Petru Poni” Institute of Macromolecular Chemistry, Iași, currently a research assistant in the same institute.

His research interests include chemical functionalization of fullerene, cyclodextrins and metal nanoparticles for biomedical application (drug/gene delivery) and more recently the design of organic ligands and metal-organic frameworks for gas storage/separation.
Email: [email protected];

About the author

Andrei NEAMTU studied physics at „Al. I. Cuza” University and medicine at „Gr.T. Popa” University of Medicine and Pharmacy (UMF) of Iași. He has a Master’s degree in Molecular Genetics at the Biology Faculty of „Al. I. Cuza” University. He received his PhD in 2007 at the University of Medicine and Pharmacy of Iași, România in the field of controlled release of active principles from nanoparticulate transporters. He followed a post-doctoral program focused on biomaterials interaction with host tissues, at “P. Poni” Institute for Macromolecular Chemistry of Iasi. At present, he occupies an associate professor position at the Physiology Department of UMF Iași, where he worked since 1998. Scientific fields of interests include molecular modelling and simulations of: membrane proteins, nucleic acids and glycosaminoglycans, inclusion complexes of cyclodextrins/modified cyclodextrins with different drugs, artificial membrane structures, biomaterials.

He has published 19 papers in ISI journals and more than 30 papers in peer reviewed journals indexed in international databases. He is the author of 5 books, 2 book chapters and many electronic materials for teaching. He has been the coordinator of 1 national research project and member in other 5 projects. He received five awards at national/international scientific meetings.

About the author

Dragos PEPTANARIU is a junior research scientist in the Centre of Advanced Research in Bionanoconjugates and Biopolymers (IntelCentre) at “Petru Poni” Institute of Macromolecular Chemistry in Iasi. After obtaining his medical degree from “Gr. T. Popa” University of Medicine and Pharmacy in 2003 he completed an internship in neurosurgery at Emergency Hospital “Prof. Dr. Nicolae Oblu”. Following an one-year stage in the department of Molecular Biology and Biochemistry at Wesleyan University in Middletown, CT, he obtained an M.Sc. from “Al.I.Cuza” University, and a Ph.D. in Medical Sciences from “Gr. T. Popa” University of Medicine and Pharmacy in 2012.

His research activity has an interdisciplinary nature and includes designing drug carriers, vectors for gene transfection, and novel approaches developing biomaterials used in tissue engineering and regenerative medicine. He is a member of the Romanian Regenerative Medicine Association and of the Romanian Chemical Society.

Email: [email protected];

About the author

Alina NICOLESCU is currently scientific researcher and manager of the NMR facilities in the “Petru Poni” Institute of Macromolecular Chemistry, Iasi, Romania. She graduated the Polytechnic University of Bucharest and she holds an MSc degree from the “Gheorghe Asachi” Technical University Iasi and a PhD in chemistry from the Romanian Academy. She has 12 years’ experience in structure elucidation of organic and organometallic compounds by NMR, IR and NIR spectroscopies and extensive expertise in NMR based metabolomics of human, animal and plant biofluids.

Her research activities include: 2D NMR experiments and pulse sequences for complete assignment of signals and stereochemical analysis of organic (terpenoids, octahydroacridine, azulene, quinoxalines, benzimidazoles, etc.) organometallic and macromolecular compounds, specific techniques for 15N NMR spectroscopy at natural abundance, and applications of NMR spectroscopy in the study of biological samples and foodstuff.

She was visiting-researcher in the Institute of Biodiagnostics, Winnipeg, Canada (biomedical NMR group of Prof. Ian C. P. Smith) and in the “Max Planck” Institute for Polymer Research, Mainz, Germany (solid state NMR group of Prof. Hans W. Spiess). She published over 90 papers in peer-reviewed journals. In 2009 she was awarded the “Costin D. Nenitescu” Medal of the Romanian Chemical Society for “exceptional merits in the advancement of chemistry in Romania” and in 2012 she was awarded the “Researcher of the Year Trophy for Women in Science” by UNESCO and L’Oreal Romania.

Email: [email protected];

About the author

Cristina URITU is a research scientist in the Advanced Research and Development Center in Experimental Medicine (CEMEX), “Gr. T. Popa” University of Medicine and Pharmacy Iasi. After obtaining her bachelor and M.Sc. degrees in Biomedical Engineering at “Gr. T. Popa” University of Medicine and Pharmacy Iasi in 2009 she has completed the Ph.D. studies in 2015 in the Centre of Advanced Research in Bionanoconjugates and Biopolymers (IntelCentre) at “Petru Poni” Institute of Macromolecular Chemistry in Iasi. Holding a previous degree in Chemical Engineering it was a good opportunity to connect the medical and chemistry knowledge in the interdisciplinary research area endorseded by IntelCentre (www.intelcentru.ro).

Her research activity has emerged in the field at the border between chemistry and biology contributing to the development of new systems capable to bind, carry and release biological active substances such as drugs or nucleic acids. She is currently also involved in the development radiopharmaceuticals, aimed to accumulate in target tissues that can be used in molecular and functional imaging. She is a member of the Romanian Chemical Society.

About the author

Stelian S. MAIER is a chemical engineer with a Ph.D. degree in chemical processes automation (1997), and a M.Sc. degree in medical biotechnology (2015). Currently he is working in the field of large scale protein extraction and purification, and biomacromolecular-based structures for tissue engineering. He also works as a consultant for technological troubleshooting in the bio-pharma industry. He coordinated six national grants as project director, and eleven as scientific officer, mainly in the field of applied biomacromolecular chemistry and medical devices;

Email: [email protected].

About the author

Bogdan C. SIMIONESCU graduated the “Gheorghe Asachi” Technical University of Iasi (TUI), Romania in 1971 (Polymer science and technology) and received his PhD in Polymer sci-ence at the Catholic University of Leuven, Belgium in 1975. Currently, he is Professor of Poly-mer science at the “Gheorghe Asachi” Technical University of Iasi and senior researcher and Head of Department at the “Petru Poni” Institute of Macromolecular Chemistry of Iasi. He is Vice-President of Romanian Academy (since 2014), President of the National Council of Scien-tific Research and Director of the “High performance polymer materials for medicine, pharmacy, microelectronics, energy/information storage, environmental protection” Training and Interdisci-plinary Research Platform and of “Polymer” Center of Excellence at TUI.

Professor Simionescu was Director of the “Petru Poni” Institute of Macromolecular Chemistry of Iasi (an Institute of excellence of Romanian Academy), 2000 – 2014 and President of the Iasi Branch of Romanian Academy (2012 – 2014).

He co-authored more than 350 papers in international journals and more than 20 book chapters in international editing houses. He was invited lecturer at more than 100 international conferences and organized/co-organized over 40 international congresses/symposia;

Email: [email protected];

About the author

Mariana PINTEALA studied chemical engineering and received her PhD at “Gheorghe Asachi” Technical University, Iasi, Romania, in 1995. She activates in “Petru Poni” Institute of Macromolecular Chemistry since 1987 and from 2009 is the Head of the “IntelCenter” integrated into institute. She has published more than 100 papers, 1 book and 9 book chapters, is evaluator for national projects in the chemistry field. The scientific and managerial activities include the successful coordination of two EU projects, 8 former national projects and 4 bilateral projects, as well as the participation to a former NSF grant and others. In addition she has implemented the IntelCentre infrastructure project (5.0 M€) based on EU Structural Funds (2009-2012), and is currently coordinator of a project funded from the European Union’s Horizon 2020 research and innovation programme under grant agreement No667387 H2020, financed with 2.4 M€.

The research interests are focused on the development of polymer-based biomaterials, supramolecular assemblies of hydrophobic and hydrophylic (co)polymers; pseudo- and polyrotaxanes, (co)polymers containing cyclodextrins, and more recently on biomedical-oriented nanotechnologies (non-viral vectors for gene and drug delivery systems; nanoparticles based on metal and metal oxides for tumors imaging and therapy; development of fullerene, cyclodextrin, and polymer derivatives for medical applications; design of cyclodextrin inclusion complexes with different drugs as potential drug delivery systems);

Email: [email protected];

Acknowledgement

European Union’s Horizon 2020 Advances in EngineeringThis project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 667387 WIDESPREAD 2-2014 SupraChem Lab

 

Reference

Rodinel Ardeleanu, Andrei I. Dascalu, Andrei Neamtu, Dragos Peptanariu, Cristina M. Uritu, Stelian S. Maier, Alina Nicolescu, Bogdan C. Simionescu, Mihail Barboiu and Mariana Pinteala. Multivalent polyrotaxane vectors as adaptive cargo complexes for gene therapy. Polymer Chemistry, 2018, volume 9, page 845

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