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Development of novel synthetic gene transfer vectors for metabolic liver therapy

Applicant Huwyler Jörg
Number 180257
Funding scheme Sinergia
Research institution Abteilung für pharmazeutische Technologie Departement Pharmazie Universität Basel
Institution of higher education University of Basel - BS
Main discipline Interdisciplinary
Start/End 01.07.2018 - 30.06.2022
Approved amount 1'715'372.00
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All Disciplines (3)

Discipline
Interdisciplinary
Cellular Biology, Cytology
Pharmacology, Pharmacy

Keywords (7)

metabolic liver disesase; ornithine transcarbamylase (OTC) deficiency; phenylketonuria (PKU); DNA based therapeutics; non-viral gene delivery; polymer based nanoparticles; liver gene therapy

Lay Summary (German)

Lead
Viele Stoffwechsel-Erkrankungen der Leber werden durch einen genetischen Defekt verursacht. Dies führt bereits bei Neugeborenen zu beträchtlichen Problemen. Beispielsweise ist der angeborene Defekt der mitochondrialen Ornithintranscarbamylase (OTC) die häufigste Störung des Harnstoffzyklus. Bei der Phenylketonurie (PKU) kann die Aminosäure Phenylalanin nicht abgebaut werden. In beiden Fällen lagern sich deshalb giftige Abbauprodukte im Körper ab. Es besteht die Hoffnung, dass mit Hilfe einer Gentherapie diese Defekte korrigiert werden können. Es wird dabei genetisches Material in Leberzellen eingeschleust um diesen die Möglichkeit zu geben die fehlenden Eiweisse wieder selbst herzustellen.
Lay summary
Inhalt und Ziel des Forschungsprojekts:
Als Alternative zu viralen Gen-Vektoren soll in diesem Projekt ein neuartiges Trägersystem für genetisches Material entwickelt werden. Das genetische Material besteht aus einer sogenannten 'Minicircle DNA'. Dieser DNA Vektor führt zu einer langanhaltenden Expression von Eiweissen in der Zelle (im Gegensatz zu RNA-basierten Verfahren) und ist frei von jeglichen viralen oder bakteriellen Komponenten. Als Träger für das genetische Material werden Polymere verwendet. In ihrem Innern wird die 'Minicircle DNA' eingeschlossen und dem Körper zugeführt. Die Effizienz und Sicherheit dieses Systems soll in experimentellen Tiermodellen gezeigt werden um somit die Grundlagen für eine spätere klinische Erprobung zu schaffen.   

Wissenschaftlicher und gesellschaftlicher Kontext des Forschungsprojekts:
Metabolische Erkrankungen der Leber sind für die Betroffenen mit grossem Leid verbunden. Es besteht bei Neugeborenen die Gefahr von bleibenden Gesundheitsschäden. In späteren Lebensjahren sind betroffene Patienten auf eine strikte und einschränkende Diät angewiesen. Eine frühzeitige und nachhaltige Behandlung dieser Erkrankungen wäre eine grosse Hilfe für die Betroffenen und würde unser Gesundheitssystem entlasten.


4.6.2018
Direct link to Lay Summary Last update: 04.06.2018

Responsible applicant and co-applicants

Employees

Project partner

Publications

Publication
Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis, Characterization, and Evaluation
Nicolle Laura, Casper Jens, Willimann Melanie, Journot Céline M. A., Detampel Pascal, Einfalt Tomaž, Grisch-Chan Hiu Man, Thöny Beat, Gerber-Lemaire Sandrine, Huwyler Jörg (2021), Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis, Characterization, and Evaluation, in International Journal of Molecular Sciences, 22(8), 3828-3828.
Improvement of DNA Vector Delivery of DOTAP Lipoplexes by Short-Chain Aminolipids
Buck Jonas, Mueller Dennis, Mettal Ute, Ackermann Miriam, Grisch-Chan Hiu Man, Thöny Beat, Zumbuehl Andreas, Huwyler Jörg, Witzigmann Dominik (2020), Improvement of DNA Vector Delivery of DOTAP Lipoplexes by Short-Chain Aminolipids, in ACS Omega, 5, 24724-24732.
Selection of Water-Soluble Chitosan by Microwave-Assisted Degradation and pH-Controlled Precipitation
Journot Céline M. A., Nicolle Laura, Lavanchy Yann, Gerber-Lemaire Sandrine (2020), Selection of Water-Soluble Chitosan by Microwave-Assisted Degradation and pH-Controlled Precipitation, in Polymers, 12(6), 1274-1274.
State-of-the-Art 2019 on Gene Therapy for Phenylketonuria
Grisch-Chan Hiu Man, Schwank Gerald, Harding Cary O., Thöny Beat (2019), State-of-the-Art 2019 on Gene Therapy for Phenylketonuria, in Human Gene Therapy, 30(10), 1274-1283.
Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads
Kulkarni Jayesh A., Witzigmann Dominik, Leung Jerry, van der Meel Roy, Zaifman Josh, Darjuan Maria M., Grisch-Chan Hiu Man, Thöny Beat, Tam Yuen Yi C., Cullis Pieter R. (2019), Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads, in Nanoscale, 11(18), 9023-9031.

Datasets

Selection of Water-Soluble Chitosan by Microwave-Assisted Degradation and pH Controlled Precipitation

Author Journot, Céline M. A.; Nicolle, Laura; Lavanchy, Yann; Gerber-Lemaire, Sandrine
Publication date 07.08.2020
Persistent Identifier (PID) 10.5281/zenodo.3975898
Repository Zenodo
Abstract
Dataset for "Selection of Water-Soluble Chitosan by Microwave-Assisted Degradation and pH Controlled Precipitation" Polymers (2020) 12, 1274. DOI: 10.3390/polym12061274.Raw data for NMR spectra; raw GPC data; experimental descriptions; metadata file

Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads

Author Kulkarni, Jayesh A.; Witzigmann, Dominik; Leung, Jerry; van der Meel, Roy; Zaifman, Josh; Darjuan, Maria M.; Grisch-Chan, Hiu Man; Thöny, Beat; Tam, Yuen Yi C.; Cullis, Pieter R.
Publication date 09.05.2019
Persistent Identifier (PID) 31021343
Repository Zenodo
Abstract
Neutralization of the pH (and ionizable lipid) drives the fusion of precursor vesicles into the electron-dense core structures attributed to lipid nanoparticles.

Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis, Characterization, and Evaluation

Author Nicole, Laura; Casper, Jens; Willimann, Melanie; Journot, Céline M.A.; Detampel, Pascal; Einfalt, Tomaž; Grisch-Chan, Hiu Man; Thöny, Beat; Gerber-Lemaire, Sandrine; Huwyler, Jörg
Publication date 16.04.2021
Persistent Identifier (PID) 10.5281/zenodo.4697428
Repository zenodo


Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Fall Meeting of the Swiss Chemical Society Poster Synthesis of chitosan-based nanomaterials for condensation of DNA in gene therapy of liver diseases 06.09.2019 Zurich, Switzerland Nicolle Laura; Journot Céline;
SSIEM Annual Symposium 2019 Poster Various posters 03.09.2019 Rotterdam, Netherlands Thöny Beat;
14th International Symposium on Macrocyclic and Supramolecular Chemistry (ISMSC2019) Poster Synthesis of cross-linked polymeric nanomaterials for condensation of DNA in gene therapy of liver diseases 02.06.2019 Lecce, Italy Nicolle Laura;
ASGCT Annual Cogress 2019 Poster various posters 29.04.2019 Washington, DC, United States of America Thöny Beat; Willimann Melanie; Grisch Hiu Man;
XXVI ESGCT Annual Congress 2018 Poster 3 Posters 16.10.2018 Swisstech Lausanne, Switzerland Grisch Hiu Man; Deplazes Sereina; Thöny Beat;
SSIEM Annual Symposium 2018 Poster various posters 04.09.2018 Athens, Greece Grisch Hiu Man; Thöny Beat; Deplazes Sereina;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Crossing Norway for a cure 2020 International 2020

Associated projects

Number Title Start Funding scheme
198265 Development of a microfluidic platform for real-time detection of SARS-CoV-2 virus based on multifunctional silica membrane biosensors 01.10.2020 NRP 78 Covid-19
162547 Evaluation of novel treatment approches for acute and chronic hyperammonemia in an OTC deficient porcine model 01.10.2015 Project funding (Div. I-III)
186741 NanoOrph2.0 - Restoring Enzyme Function in Lysosomal Storage Diseases Using Lipid Nanoparticle Mediated Gene Therapy 01.01.2020 Postdoc.Mobility
164250 New standardized and functionalized microcapsules: Applications to Type 1 Diabetes cell therapy 01.02.2016 Project funding (Div. I-III)
173057 Drug Targeting to Hepatocytes: Gene Delivery using Myrcludex B Coupled Lipid Nanoparticles 01.05.2017 Project funding (Div. I-III)
182366 A feasibility study on salvage radiotherapy with adjuvant hyperthermia delivered by MR-guided Transperineal Ultrasound (MRgTUS) for recurrent prostate cancer after radical prostatectomy 01.01.2019 Project funding (Div. I-III)

Abstract

Therapeutic vectors for gene delivery remain the currently most challenging factor for human gene therapy. The translation from in vitro to in vivo applications remains a major hurdle for most nucleic acid delivery systems since there is an inherent lack of both efficient and safe carrier systems. For liver targeting of postmitotic hepatocytes, adeno-associated virus (AAV) derived vectors are thought to have the greatest potential despite concerns about a future routine clinical use. The major hurdles of AAV vectors for long-term treatment of pediatric patients are the risk of chromosomal integration and development of hepatocellular carcinoma, immune responses to viral vectors, limited loading capacity, and the difficulty to treat neonates which likely would require subsequent further injections. Consequently, the development of non-viral gene delivery systems has gained much attention due to their versatility, safety, and ease of manufacturing. During the last decades, a wide range of nanoparticle based gene delivery systems were developed and remarkable results in the field of RNA therapeutics were achieved. However, the induced pharmacological effects obtained by these siRNA or mRNA delivery strategies are short-lived and thus weekly administrations of therapeutic formulations are necessary. The use of DNA-based therapeutics would offer a favorable option for the induction of long-term therapeutic effects without need for insertion into the genome. Here, we propose an alternative approach to overcome the challenges of viral vectors or RNA-based therapeutics by developing novel nanoparticles for delivery of non-integrating, so-called minicircle (MC) vectors lacking any viral or bacterial components for liver-directed gene therapy. The successful use of MC vectors to treat genetic (metabolic) liver defects is based on the experience of one application partner with naked DNA-vectors delivered in an experimental setting by hydrodynamic pressure to either target pericentral or periportal hepatocytes to treat two classical defects in mouse models for human diseases, phenylketonuria (PKU) and ornithine transcarbamylase (OTC) deficiency, respectively. Such MC vectors exhibited persistent expression combined with basically no DNA size limitation, which made it possible to use natural promoters/enhancers in combination with introns to mimic “physiological” expression. While MC vectors bear almost ideal properties with great potential for liver gene therapy, delivery of naked DNA solely by hydrodynamic pressure is not applicable in a clinical setting. In an interdisciplinary approach, we want to develop multifunctional polymeric nanoparticles encapsulating MC vectors for non-viral gene delivery specifically to the key pathogenic cell type, i.e. hepatocytes. In order to optimize gene delivery efficiency, a novel library of polymer-peptide hybrids will be created, formulations strategies will be optimized and resulting nanoparticles will be validated in vivo using various animal models, i.e. transgenic mice, xenotransplanted mice with human liver or pig models. The combination of this novel class of polymer-peptide hybrids with a reproducible and scalable nanoparticle formulation technique (i.e. microfluidics) is expected to greatly impact further optimization of the synthetic gene delivery system for clinical applications. The overall aim of this translational project is the development of an alternative approach to AAV vectors with the potential of a breakthrough for liver gene therapy and thus a paradigm shift from potentially harmful viral vectors to safe, efficient and completely synthetic non-viral vectors.
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