Article
Chemistry, Multidisciplinary
Adriana Coll De Pena, Lucy Masto, James Atwood, Anubhav Tripathi
Summary: Adeno-associated virus (AAV) is highly promising for gene therapy due to its low immunogenicity and ability to provide long-term gene expression. This study proposes a method that can estimate the composition of a given AAV sample independent of its input concentration. The method combines protein and ssDNA information relative to a standard to determine the percentage of full capsids in the sample.
Article
Biochemical Research Methods
Jack P. Ryan, Marius M. Kostelic, Chih-Chieh Hsieh, Joshua Powers, Craig Aspinwall, James N. Dodds, John E. Schiel, Michael T. Marty, Erin S. Baker
Summary: This article introduces a method that combines different analytical workflows to evaluate the composition and ratio of AAV capsids. The method includes two steps, each for evaluating the intact capsids and viral protein subunits. By applying this method, a better understanding of the characteristics and functions of AAV capsids can be achieved.
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
(2023)
Article
Chemistry, Analytical
Marius M. Kostelic, Jack P. Ryan, Levi S. Brown, Tyler W. Jackson, Chih-Chieh Hsieh, Ciara K. Zak, Henry M. Sanders, Yang Liu, Victor Shugui Chen, Michael Byrne, Craig A. Aspinwall, Erin S. Baker, Michael T. Marty
Summary: In this study, we combined charge detection-mass spectrometry with a variable temperature electrospray source to develop a method for better measuring the thermal stability of AAVs. With this approach, we successfully measured the thermal stability and dissociation pathways of different AAV subpopulations, and found that filled AAVs ejected their cargo first and formed intermediate empty capsids.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Felipe Guapo, Florian Fussl, Lisa Strasser, Jonathan Bones
Summary: This study developed a pH-gradient anion exchange chromatography method for separating full and empty capsid species in AAV-based therapeutics, allowing for subsequent mass spectrometric detection. The demonstrated generic applicability and mass spectrometric compatibility enable a more sophisticated analysis of AAV-based gene therapy, paving the way for future developments in the field.
ANALYTICAL METHODS
(2023)
Article
Biotechnology & Applied Microbiology
Shantoshini Dash, David M. Sharon, Alaka Mullick, Amine A. Kamen
Summary: This study reveals that only a small proportion of cells are capable of producing measurable levels of assembled AAV capsids despite high transfection efficiencies and nominal vector yields. Increasing the proportion of productive cells could significantly increase vector titer. The flow cytometry assay used in this study may serve as a useful metric for optimizing transfection-based AAV vector manufacturing platforms.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Chemistry, Multidisciplinary
Yu Ri Nam, Helen H. Ju, Jeehee Lee, Daiheon Lee, Yoojin Kim, Sung Jin Lee, Hong Kee Kim, Jae-Hyung Jang, Haeshin Lee
Summary: Recently, AAV therapy for incurable diseases has been introduced due to its safety properties and neutralizing antibodies. However, the mixture of empty and full capsids in AAV production increases viral doses to humans. TEM, the primary tool for distinguishing between empty and full capsids, creates research limitations. This study demonstrates that AFM can be an alternative tool to TEM, allowing the distinct height and morphological differences between empty and full AAV particles.
Article
Chemistry, Multidisciplinary
Amelie Bauer, Matteo Puglisi, Dennis Nagl, Joel A. Schick, Thomas Werner, Andreas Klingl, Jihad El Andari, Veit Hornung, Horst Kessler, Magdalena Goetz, Dirk Grimm, Ruth Brack-Werner
Summary: Astrocytes play crucial roles in the central nervous system and are involved in various CNS diseases. This study focuses on astrocyte-centered diseases and the need for effective and well-characterized gene transfer vectors. The findings highlight the use of a synthetic peptide-displaying AAV9 variant, rAAV9P1, which was shown to efficiently target and transduce human astrocytes. The study also identifies the specific receptors and intracellular pathways utilized by rAAV9P1 for astrocyte targeting. These results enhance our understanding of synthetic AAV vectors for astrocytes and their potential applications in research and medicine.
Article
Medicine, Research & Experimental
Christopher R. Barnes, Hyuncheol Lee, David S. Ojala, Kazuomori K. Lewis, Prajit Limsirichai, David Schaffer
Summary: The study presents a genome-wide screening strategy to identify target genes for enhancing AAV vector production, leading to the discovery of gain-of-function targets such as SKA2 and ITPRIP. Modulation of these targets can significantly increase AAV manufacturing capacity.
MOLECULAR THERAPY-NUCLEIC ACIDS
(2021)
Article
Medicine, General & Internal
Dezhuang Ye, Jinyun Yuan, Yaoheng Yang, Yimei Yue, Zhongtao Hu, Siaka Fadera, Hong Chen
Summary: This study investigates the potential of focused ultrasound-mediated intranasal delivery (FUSIN) in delivering adeno-associated viral (AAV) vectors to the brain. The results show that FUSIN achieves safe and efficient delivery to spatially targeted brain locations and is comparable to established delivery methods.
Article
Medicine, Research & Experimental
Santoshkumar L. Khatwani, Anna Pavlova, Zhu Pirot
Summary: Gene therapy has made significant progress in treating severe and rare diseases, thanks to better scientific understanding and improved manufacturing processes.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2021)
Review
Biochemistry & Molecular Biology
Lyubava Belova, Alexander Lavrov, Svetlana Smirnikhina
Summary: Cellular 3D structures, such as organoids, are important for biomedical research. However, there are challenges in delivering transgenes to these structures. This review explores ways to improve the efficiency of transgene delivery.
Article
Medicine, Research & Experimental
Andrew D. Marques, Michael Kummer, Oleksandr Kondratov, Arunava Banerjee, Oleksandr Moskalenko, Sergei Zolotukhin
Summary: Machine learning (ML) is used to train algorithms on data from adeno-associated virus (AAV) capsid libraries to predict the assembly of unknown capsid variants into viable virus-like structures. By utilizing artificial neural networks (ANNs) and support vector machines (SVMs), significant mutation patterns in AAV2-derived capsids, such as N495, G546, and I554, are identified. The study demonstrates the predictive power of ML in vector design through the creation and validation of comparative libraries.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2021)
Article
Biochemistry & Molecular Biology
Ah Young Kim, Felix Michael Duerr, Jennifer Phillips, Richard Jude Samulski, Josh C. Grieger, Laurie R. Goodrich
Summary: A formal screening of self-complementary adeno-associated virus (scAAV) vector serotypes in canine joint tissues has not been performed to date. This study aims to identify the most optimal scAAV vector serotype that maximizes transduction efficiencies in canine cell cultures and tissue explant cultures. The results show that scAAV2 and 2.5 have the highest transduction efficiencies among the serotypes, and a dose-response relationship is observed. Further evaluation of the safety and efficacy of scAAV2.5 vector with an appropriate therapeutic gene in vivo is recommended.
Article
Biotechnology & Applied Microbiology
Michelle Mendiola Pla, Yuting Chiang, Antonio Roki, Chunbo Wang, Franklin H. Lee, Matthew F. Smith, Ryan T. Gross, Jun-Neng Roan, Muath Bishawi, Amy Evans, Lynden E. Gault, Sam Ho, Carolyn Glass, Jacob N. Schroder, Paul Lezberg, Carmelo A. Milano, Dawn E. Bowles
Summary: This study demonstrates the feasibility and efficiency of delivering genes to porcine cardiac allografts utilizing adeno-associated viral vectors during ex vivo perfusion. Durable and dose-dependent transgene expression was achieved in the allografts, supporting the idea of gene therapy interventions to enhance transplantation outcomes.
HUMAN GENE THERAPY
(2023)
Article
Biochemical Research Methods
Aurelien Jacob, Laurie Brun, Paloma Jimenez Gil, Lucie Menard, Mohammed Bouzelha, Frederic Broucque, Aline Roblin, Luk H. Vandenberghe, Oumeya Adjali, Cecile Robin, Achille Francois, Veronique Blouin, Magalie Penaud-Budloo, Eduard Ayuso
Summary: Manufacturing viral vectors for gene delivery is challenging, with a scalable baculovirus-insect cell platform being used for producing rAAV vectors. A faster and safer method based on homologous recombination (HR) has been investigated to avoid technical constraints and safety concerns associated with the Tn7 transposition method. The HR system offers increased stability and avoids undesirable bacterial genes, ensuring the generation of safer rAAV biotherapeutic products.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Biochemistry & Molecular Biology
Alexander E. Yarawsky, Lance R. English, Steven T. Whitten, Andrew B. Herr
JOURNAL OF MOLECULAR BIOLOGY
(2017)
Article
Biochemistry & Molecular Biology
Alexander E. Yarawsky, Stefanie L. Johns, Peter Schuck, Andrew B. Herr
JOURNAL OF BIOLOGICAL CHEMISTRY
(2020)
Article
Biochemistry & Molecular Biology
Alexander E. Yarawsky, Andrew B. Herr
JOURNAL OF BIOLOGICAL CHEMISTRY
(2020)
Article
Allergy
Marat Khodoun, Suzanne C. Morris, Wen-Hai Shao, Crystal Potter, Elizabeth Angerman, Artem Kiselev, Alexander E. Yarawsky, Andrew B. Herr, Katja Klausz, Anna Otte, Matthias Peipp, Fred D. Finkelman
Summary: Monovalent anti-huFc epsilon RI alpha mAbs have advantages in safety over divalent mAbs, with weaker abilities to induce allergic reactions and deplete IgE, but still effectively suppress IgE-mediated diseases.
JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Svetlana Zamakhaeva, Catherine T. Chaton, Jeffrey S. Rush, Sowmya Ajay Castro, Cameron W. Kenner, Alexander E. Yarawsky, Andrew B. Herr, Nina M. van Sorge, Helge C. Dorfmueller, Gregory Frolenkov, Konstantin Korotkov, Natalia Korotkova
Summary: In ovoid-shaped, Gram-positive bacteria, the major cell separation autolysin AtlA was identified as an SCC-binding protein. GroP-deficient immature SCCs were found to localize at cell poles and equators, potentially leading to AtlA mislocalization, dysregulated cellular autolysis, and morphological abnormalities. This indicates a model where maturation of a cell wall polysaccharide plays a role in recruiting cell division machinery for proper daughter cell separation and FtsZ-ring positioning.
NATURE CHEMICAL BIOLOGY
(2021)
Article
Multidisciplinary Sciences
Greg Davis, Allen J. York, Willis Clark Bacon, Suh-Chin Lin, Monica Malone McNeal, Alexander E. Yarawsky, Joseph J. Maciag, Jeanette L. C. Miller, Kathryn C. S. Locker, Michelle Bailey, Rebecca Stone, Michael Hall, Judith Gonzalez, Alyssa Sproles, E. Steve Woodle, Kristen Safier, Kristine A. Justus, Paul Spearman, Russell E. Ware, Jose A. Cancelas, Michael B. Jordan, Andrew B. Herr, David A. Hildeman, Jeffery D. Molkentin
Summary: This study investigated the SARS-CoV-2 infection rates in Greater Cincinnati area from August to December 2020. Different rates of infection were observed among different age groups, genders, and ethnicities. Geographic analysis revealed higher infection rates on the West side of Cincinnati and the lowest rates in the adjoining region of Kentucky.
Article
Multidisciplinary Sciences
Marta T. Borowska, Christoph Drees, Alexander E. Yarawsky, Mayuri Viswanathan, Sean M. Ryan, Jeffrey J. Bunker, Andrew B. Herr, Albert Bendelac, Erin J. Adams
Summary: Research has identified immunoglobulin-binding proteins (Ibp) A and B in commensal microorganisms that can activate B cells and potentially cluster full-length IgA antibodies in vivo. In vitro experiments suggest that binding of Ibp HCBD alone is sufficient to activate responsive murine B cell receptors.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Article
Biochemistry & Molecular Biology
Alexander E. Yarawsky, Jesse B. Hopkins, Leonie Chatzimagas, Jochen S. Hub, Andrew B. Herr
Summary: This study investigates the structural characteristics of B-repeat mutant constructs of Aap from S. epidermidis using multiple techniques. The resulting models provide insights into the mechanisms of functional amyloid fibril formation by Aap, highlighting an important contributing factor to recurrent staphylococcal infections.
JOURNAL OF MOLECULAR BIOLOGY
(2022)
Article
Multidisciplinary Sciences
John J. Erickson, Stephanie Archer-Hartmann, Alexander E. Yarawsky, Jeanette L. C. Miller, Stephanie Seveau, Tzu-Yu Shao, Ashley L. Severance, Hilary Miller-Handley, Yuehong Wu, Giang Pham, Brian R. Wasik, Colin R. Parrish, Yueh-Chiang Hu, Joseph T. Y. Lau, Parastoo Azadi, Andrew B. Herr, Sing Sing Way
Summary: In this study, researchers discovered that pregnancy-induced antibody modification can protect neonates from intracellular pathogen Listeria monocytogenes infection. Maternal antibodies play a crucial role in vertically transferred immunity, and the protective function of antibodies depends on sialic acid esterase for deacetylation. These findings reveal the protective mechanism of the maternal-fetal immune unit and fine-tuned adaptations in host defense during pregnancy and early life.
Article
Biophysics
Alexander E. Yarawsky, Vlad Dinu, Stephen E. Harding, Andrew B. Herr
Summary: A recent investigation used SEC-MALS-SAXS to study the structural behavior of a highly extended protein. The study discovered viscous fingering, typically observed at high concentrations, occurring at lower concentrations for this protein. The non-ideal behavior and its correlation with intrinsic viscosity were explored for different proteins using SEC, AUC, and viscosity measurements. The importance of considering such non-ideal behavior in the investigation of extended macromolecules is discussed.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
(2023)
Article
Biochemistry & Molecular Biology
Alexander E. Yarawsky, Andrea L. Ori, Lance R. English, Steven T. Whitten, Andrew B. Herr
Summary: Staphylococcus epidermidis and Staphylococcus aureus are problematic bacteria in hospitals due to their ability to form biofilms, which resist antibiotic treatment. Cell wall-anchored (CWA) proteins are important for biofilm formation and infection. Recent study found that the stalk region of S. epidermidis Aap remains highly extended, consistent with its expected function of projecting adhesive domains away from the cell surface. This study evaluates the compaction resistance of stalk regions from various staphylococcal CWA proteins.
Article
Endocrinology & Metabolism
Emily C. Kappes, Chandramohan Kattamuri, Magdalena Czepnik, Alexander E. Yarawsky, Emilie Brule, Ying Wang, Luisina Ongaro, Andrew B. Herr, Kelly L. Walton, Daniel J. Bernard, Thomas B. Thompson
Summary: This study characterized the inhibin A:follistatin complex and found that it can effectively antagonize activin signaling, even though it forms a substoichiometric complex and readily dissociates.