Electrostatic Coating of Viral Particles for Gene Delivery Applications in Muscular Dystrophies: Influence of Size on Stability and Antibody Protection

2021 ◽  
pp. 1-11
Author(s):  
Marta Guerra-Rebollo ◽  
María Stampa ◽  
Miguel Ángel Lázaro ◽  
Anna Cascante ◽  
Cristina Fornaguera ◽  
...  
Keyword(s):  
Electrostatic Interaction ◽  
Viral Vectors ◽  
Dystrophin Gene ◽  
Muscular Dystrophies ◽  
Antibody Neutralization ◽  
Adeno Associated Virus ◽  
In Vivo Biodistribution ◽  
Biodistribution Studies ◽  
Viral Particles

Background: Duchenne Muscular Dystrophy (DMD) is one of the most common muscular dystrophies, caused by mutated forms of the dystrophin gene. Currently, the only treatment available is symptoms management. Novel approximations are trying to treat these patients with gene therapy, namely, using viral vectors. However, these vectors can be recognized by the immune system decreasing their therapeutic activity and making impossible a multidose treatment due to the induction of the humoral immunity following the first dose. Objective: Our objective is to demonstrate the feasibility of using a hybrid vector to avoid immune clearance, based on the electrostatic coating of adeno-associated virus (AAVs) vectors with our proprietary polymers. Methods: We coated model adeno-associated virus vectors by electrostatic interaction of our cationic poly (beta aminoester) polymers with the viral anionic capsid and characterized biophysical properties. Once the nanoformulations were designed, we studied their in vivo biodistribution by bioluminescence analysis and we finally studied the capacity of the polymers as potential coatings to avoid antibody neutralization. Results: We tested two polymer combinations and we demonstrated the need for poly(ethylene glycol) addition to avoid vector aggregation after coating. In vivo biodistribution studies demonstrated that viral particles are located in the liver (short times) and also in muscles (long times), the target organ. However, we did not achieve complete antibody neutralization shielding using this electrostatic coating. Conclusions: The null hypothesis stands: although it is feasible to coat viral particles by electrostatic interaction with a proprietary polymer, this strategy is not appropriate for AAVs due to their small size, so other alternatives are required as a novel treatment for DMD patients.

In vitro binding and in vivo biodistribution studies of the neuroprotective peptide humanin using [125I]humanin derivatives

Peptides ◽  
2009 ◽  
Vol 30 (12) ◽  
pp. 2409-2417 ◽  
Author(s):  
Alexandra Evangelou ◽  
Christos Zikos ◽  
Dimitra Benaki ◽  
Maria Pelecanou ◽  
Penelope Bouziotis ◽  
...  
Keyword(s):  
In Vitro Binding ◽  
In Vivo Biodistribution ◽  
Biodistribution Studies ◽  
Vitro Binding

scholarly journals In vivo biodistribution studies and ex vivo lymph node imaging using heavy metal-free quantum dots

Biomaterials ◽  
2016 ◽  
Vol 104 ◽  
pp. 182-191 ◽  
Author(s):  
Elnaz Yaghini ◽  
Helen D. Turner ◽  
Alix M. Le Marois ◽  
Klaus Suhling ◽  
Imad Naasani ◽  
...  
Keyword(s):  
Heavy Metal ◽  
Quantum Dots ◽  
Lymph Node ◽  
Ex Vivo ◽  
Metal Free ◽  
Lymph Node Imaging ◽  
In Vivo Biodistribution ◽  
Biodistribution Studies

scholarly journals Weighing the DNA content of Adeno-Associated Virus vectors with zeptogram precision using nanomechanical resonators

2021 ◽  
Author(s):  
Georgios Katsikis ◽  
Iris E Hwang ◽  
Wade Wang ◽  
Vikas S Bhat ◽  
Nicole L McIntosh ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Quality Control ◽  
Viral Vectors ◽  
Analytical Ultracentrifugation ◽  
Vaccine Development ◽  
Turnaround Time ◽  
Charge Detection ◽  
Adeno Associated Virus ◽  
Nanomechanical Resonators

Quantifying the composition of viral vectors used in vaccine development and gene therapy is critical for assessing their functionality. Adeno-Associated Virus (AAV) vectors, which are the most widely used viral vectors for in-vivo gene therapy, are typically characterized using PCR, ELISA, and Analytical Ultracentrifugation which require laborious protocols or hours of turnaround time. Emerging methods such as Charge-Detection Mass Spectroscopy, Static Light Scattering, and Mass Photometry offer turnaround times of minutes for measuring AAV mass, but mostly require purified AAV-based reference materials for calibration. Here, we demonstrate a method for using Suspended Nanomechanical Resonators (SNR) to directly measure both AAV mass and aggregation from a few microliters of sample within minutes. We achieve a resolution near 10 zeptograms which corresponds to 1% of the genome holding capacity of the AAV capsid. Our results show the potential of our method for providing real-time quality control of viral vectors during biomanufacturing.

scholarly journals 124I-Radiolabeling of a Au(III)-NHC Complex for in Vivo Biodistribution Studies

2020 ◽  
Author(s):  
Federica Guarra ◽  
Alessio Terenzi ◽  
Christine Pirker ◽  
Rossana Passannante ◽  
Dina Baier ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Direct Evidence ◽  
Heterocyclic Carbenes ◽  
Emission Tomography ◽  
Direct Oxidation ◽  
In Vivo Biodistribution ◽  
Biodistribution Studies ◽  
Positron Emission

Au(III) complexes with N-Heterocyclic Carbenes (NHCs) ligands have shown remarkable potential as anticancer agents, yet their fate in vivo has not been thoroughly examined and understood. Herein we report on the synthesis of new Au(III)-NHC complexes via direct oxidation with radioactive [124I]I2 as a valuable strategy to monitor the in vivo biodistribution of this class of compounds using positron emission tomography (PET) and, in combination with in vitro analyses, to provide direct evidence of the importance of Au(III)-to-Au(I) reduction for achieving full anticancer activity.

scholarly journals 124 I Radiolabeling of a Au III ‐NHC Complex for In Vivo Biodistribution Studies

2020 ◽  
Vol 59 (39) ◽  
pp. 17130-17136 ◽  
Author(s):  
Federica Guarra ◽  
Alessio Terenzi ◽  
Christine Pirker ◽  
Rossana Passannante ◽  
Dina Baier ◽  
...  
Keyword(s):  
In Vivo Biodistribution ◽  
Biodistribution Studies

Silica nanoparticles: Preparation, characterization and in vitro/in vivo biodistribution studies

2015 ◽  
Vol 71 ◽  
pp. 46-55 ◽  
Author(s):  
B.I. Tamba ◽  
A. Dondas ◽  
M. Leon ◽  
A.N. Neagu ◽  
G. Dodi ◽  
...  
Keyword(s):  
Silica Nanoparticles ◽  
In Vivo Biodistribution ◽  
Biodistribution Studies

scholarly journals Enhanced CRISPR-Cas9 correction of Duchenne muscular dystrophy in mice by a self-complementary AAV delivery system

2020 ◽  
Vol 6 (8) ◽  
pp. eaay6812 ◽  
Author(s):  
Yu Zhang ◽  
Hui Li ◽  
Yi-Li Min ◽  
Efrain Sanchez-Ortiz ◽  
Jian Huang ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Genome Editing ◽  
Dystrophin Gene ◽  
Adeno Associated Virus ◽  
Guide Rnas ◽  
Dystrophin Expression ◽  
Cas9 Nuclease ◽  
High Doses

Duchenne muscular dystrophy (DMD) is a lethal neuromuscular disease caused by mutations in the dystrophin gene (DMD). Previously, we applied CRISPR-Cas9–mediated “single-cut” genome editing to correct diverse genetic mutations in animal models of DMD. However, high doses of adeno-associated virus (AAV) are required for efficient in vivo genome editing, posing challenges for clinical application. In this study, we packaged Cas9 nuclease in single-stranded AAV (ssAAV) and CRISPR single guide RNAs in self-complementary AAV (scAAV) and delivered this dual AAV system into a mouse model of DMD. The dose of scAAV required for efficient genome editing were at least 20-fold lower than with ssAAV. Mice receiving systemic treatment showed restoration of dystrophin expression and improved muscle contractility. These findings show that the efficiency of CRISPR-Cas9–mediated genome editing can be substantially improved by using the scAAV system. This represents an important advancement toward therapeutic translation of genome editing for DMD.

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