Review
Pharmacology & Pharmacy
Robert G. Wallace, Keith D. Rochfort, Peter Barabas, Timothy M. Curtis, Hironori Uehara, Balamurali K. Ambati, Philip M. Cummins
Summary: The Angiopoietin-1/2 system shows potential for therapeutic intervention in a wide range of vascular pathologies, particularly through its association with endothelium. Engineered COMP-Ang1 has demonstrated greater potency in activating the Tie2 pathway compared to native Ang1, with multiple benefits reported including promotion of skeletal muscle regeneration, wound healing and angiogenesis.
VASCULAR PHARMACOLOGY
(2021)
Article
Cell & Tissue Engineering
Hyun-Jaung Sim, Govinda Bhattarai, Min-Hye Kim, Han-Sol So, Sher Bahadur Poudel, Eui-Sic Cho, Sung-Ho Kook, Jeong-Chae Lee
Summary: Excessive Ang1 leads to bone marrow impairment, HSC senescence, and erythropoietic defect. Through experiments in transgenic mice models, we found that the effects of genetic overexpression on angiogenesis varied in terms of growth and lifespan. Inhibition of Tie2 and GATA-1 expression can restore platelet production.
Article
Biotechnology & Applied Microbiology
Pan Liu, Michael Ryczko, Xinfang Xie, Jason Baardsnes, Simon Lord-Dufour, Yves Duroche, Emily Anne Hicks, Aftab Taiyab, Heather Sheardown, Susan E. Quaggin, Jing Jin
Summary: The study introduces a new soluble ANG1 mimetic, Hepta-ANG1, which effectively reduces vascular leakage by activating the Tie2 signaling pathway. This fusion protein displays remarkable stability and high multimericity, making it a potential candidate for treating inflammatory vascular leak disorders.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biochemical Research Methods
Aleksandra Stec, Monika Targonska, Edyta Karkosinska, Monika Slowik, Agata Ploska, Leszek Kalinowski, Bartosz Wielgomas, Krzysztof Waleron, Jacek Jasiecki, Szymon Dziomba
Summary: The overproduction of proteins in Chinese hamster ovary (CHO) cells can induce the secretion of extracellular vesicles (EVs), potentially leading to the packaging of overexpressed proteins into exosomes. These findings are crucial for the manufacturing of therapeutic drugs in CHO expression systems, as there is a risk of product loss during downstream processing of culture medium and the use of exosomes as nanocarriers for therapeutic proteins.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2023)
Review
Biotechnology & Applied Microbiology
Melanie Nguyen, Aline Zimmer
Summary: Chinese hamster ovary (CHO) cells are widely used in the biopharmaceutical industry for producing recombinant proteins. The proteomic studies on CHO cells have provided valuable insights into the protein clusters, regulatory networks, and pathways governing phenotypic changes, which can be utilized to optimize CHO-based bioprocesses.
BIOTECHNOLOGY ADVANCES
(2023)
Review
Biotechnology & Applied Microbiology
Jun-He Zhang, Lin-Lin Shan, Fan Liang, Chen-Yang Du, Jing-Jing Li
Summary: Recombinant antibodies are a rapidly developing class of therapeutic agents, but high-level expression and production of these antibodies remain a major challenge. This review provides a comprehensive overview of various strategies for optimizing gene sequences, expression vectors, host cell lines, and glycosylation modifications to improve the efficiency and quality of recombinant antibody production.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Azam Rahimpour, Es'hagh Pourmaleki, Forough Shams, Zahra Payandeh, Navid Pourzardosht, Mojtaba Didehdar, Milad Gholami
Summary: This study constructed a novel vector system using the Ccnb1ip1 gene insulator element for the expression of an anti-CD52 mAb in CHO cells. The expression of mAb in cells created with the insulator-containing vector was found to be increased by 10.9% compared to the control vector.
MOLECULAR BIOLOGY REPORTS
(2022)
Article
Biotechnology & Applied Microbiology
Thu M. Cao, Dayue Chen, Gavin C. Barnard, Amy Shen
Summary: This study explores the potential of using CHO cells for rAAV production and identifies cap protein expression as a limiting factor. By increasing cap protein expression, rAAV titers in CHO cells were comparable to those in HEK-derived rAAV, providing a foundation for potential rAAV production in CHO cells.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Eleftheria Skrika-Alexopoulos, C. Mark Smales
Summary: Exosomes were isolated from CHO cells using a novel method and characterized to determine their protein and RNA content. These findings contribute to the future research and improvement of exosome manufacturing from CHO cells.
BIOTECHNOLOGY LETTERS
(2023)
Article
Biotechnology & Applied Microbiology
Ioscani Jimenez del Val, Sarantos Kyriakopoulos, Simone Albrecht, Henning Stockmann, Pauline M. M. Rudd, Karen M. M. Polizzi, Cleo Kontoravdi
Summary: Metabolic modeling is important for characterizing biopharmaceutical cell culture processes. However, current metabolic models of CHO cells face challenges in terms of size and interpretation. To address these challenges, the CHOmpact model was developed as a reduced metabolic network with improved interpretability and physiological consistency. This model provides a platform for developing dynamic metabolic models to optimize biopharmaceutical cell culture processes.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Review
Biotechnology & Applied Microbiology
Weifeng Li, Zhenlin Fan, Yan Lin, Tian-Yun Wang
Summary: This review highlights the importance of serum-free medium (SFM) in the production of recombinant therapeutic proteins (RTPs) using Chinese hamster ovary (CHO) cells, with nearly 70% of RTPs currently produced in this manner. It discusses the history, key components, preparation, and experimental design of SFM, as well as the potential benefits of small molecule compound additives in improving RTP yield and quality. Lastly, it explores the future prospects of using SFM with CHO cells for RTP production.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2021)
Article
Multidisciplinary Sciences
Richard Karlsson, Pradeep Chopra, Apoorva Joshi, Zhang Yang, Sergey Y. Vakhrushev, Thomas Mandel Clausen, Chelsea D. Painter, Gergo P. Szekeres, Yen-Hsi Chen, Daniel R. Sandoval, Lars Hansen, Jeffrey D. Esko, Kevin Pagel, Douglas P. Dyer, Jeremy E. Turnbull, Henrik Clausen, Geert-Jan Boons, Rebecca L. Miller
Summary: The study integrated various approaches to reveal differences in 3-O-sulfation profiles and their impact on cellular functions, providing new insights for tailored synthesis of next-generation heparins.
Review
Biochemistry & Molecular Biology
Katja Glinsek, Kristof Bozovicar, Tomaz Bratkovic
Summary: CRISPR technology has made significant progress in CHO cell line engineering, improving protein production and product quality. It allows for glycosylation modulation, productivity enhancement, tackling adventitious agents, elimination of problematic host cell proteins, development of antibiotic-free selection systems, site-specific transgene integration, and CRISPR-mediated gene activation and repression. The review highlights the potential of CRISPR technology in CHO cell line genome editing and epigenetic engineering for more efficient and cost-effective development of biopharmaceuticals, ensuring the safety and quality of the final product.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Cell Biology
Aziza R. Alrafiah
Summary: This study found dysregulation of angiogenesis in monocyte subsets in ischemic stroke patients, which could be used as an early diagnostic marker of neurovascular damage and may require corresponding treatment to prevent further damage to blood vessels.
HISTOLOGY AND HISTOPATHOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Kexue Liang, Hongzhen Luo, Qi Li
Summary: In this study, the effects of different perfusion culture strategies on process stability, product titer, and product quality were investigated. Continuous perfusion was found to be more effective in maintaining high levels of cell density, product titer, and quality compared to intermittent perfusion culture. Furthermore, the study optimized the perfusion culture conditions by adjusting perfusion rates and culture temperature. The optimized strategy resulted in significantly higher product titer and maintained excellent product quality.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Editorial Material
Biochemistry & Molecular Biology
Kyung S. Lee, Michel O. Steinmetz
CURRENT OPINION IN STRUCTURAL BIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Ashwani Sharma, Natacha Olieric, Michel O. Steinmetz
Summary: Centrioles are microtubule-based structures crucial for cell division and ciliogenesis. Proteins controlling centriole length can be categorized into CEAs and CEIs, with antagonistic activities on centriolar microtubules. The review discusses the structure-function relationship and open questions in the field, suggesting potential directions for future studies.
CURRENT OPINION IN STRUCTURAL BIOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Tobias Muehlethaler, Dario Gioia, Andrea E. Prota, May E. Sharpe, Andrea Cavalli, Michel O. Steinmetz
Summary: Using a combined computational and crystallographic approach, researchers identified 27 distinct binding sites in tubulin, including 11 previously unknown sites. They also observed an intricate pocket communication network and identified 56 chemically diverse fragments that bound to 10 distinct tubulin sites. These results provide a structural basis for developing novel small molecules as tubulin modulators and lay down a framework that may aid in discovering new pockets in other pharmaceutically important targets.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Chemistry, Medicinal
Cassandra Yong, Shane M. Devine, Anne-Catherine Abel, Stefan D. Tomlins, Divya Muthiah, Xuexin Gao, Richard Callaghan, Michel O. Steinmetz, Andrea E. Prota, Ben Capuano, Peter J. Scammells
Summary: Extensive research has been conducted over the past two decades to discover noscapine derivatives with improved potency. This study focused on modifications to the 1,3-benzodioxole moiety of noscapine analogues, resulting in the discovery of potent cytotoxic agents against various types of cancer. Additionally, X-ray crystallography studies provided insight into the interactions between these noscapinoids and tubulin, which will aid in the development of future noscapine derivatives with enhanced properties.
Article
Oncology
Finith Jernigan, Arthur Branstrom, John D. Baird, Liangxian Cao, Mandar Dali, Bansri Furia, Min Jung Kim, Kylie O'Keefe, Ronald Kong, Oscar L. Laskin, Joseph M. Colacino, Mark Pykett, Anna Mollin, Josephine Sheedy, Melissa Dumble, Young-Choon Moon, Richard Sheridan, Tobias Muehlethaler, Robert J. Spiegel, Andrea E. Prota, Michel O. Steinmetz, Marla Weetall
Summary: PTC596 is an orally bioavailable tubulin-binding agent with broad-spectrum anticancer activity. In preclinical mouse models of leiomyosarcomas and glioblastoma, PTC596 demonstrated efficacy as a monotherapy and showed additive or synergistic efficacy in combination. Additionally, PTC596 exhibited efficacy in an orthotopic model of glioblastoma under conditions where standard treatment was ineffective.
MOLECULAR CANCER THERAPEUTICS
(2021)
Article
Biochemistry & Molecular Biology
Anil Kumar, Sandro M. Meier, Ana-Maria Farcas, Cristina Manatschal, Yves Barral, Michel O. Steinmetz
Summary: The study reveals the crystal structure of Kar9's folded domain, showing spectrin repeats similar to other +TIPs. Mutations disrupting spectrin-repeat-mediated dimerization of Kar9 affects its distribution to microtubule tips and spindle positioning. Phosphomimetic substitution at Cdk1 sites inhibits Kar9 dimerization, displaces Kar9 from microtubules, and affects its interaction with the myosin motor Myo2.
Article
Chemistry, Multidisciplinary
Tobias Muhlethaler, Lampros Milanos, Jose Antonio Ortega, Thorsten B. Blum, Dario Gioia, Bibhas Roy, Andrea E. Prota, Andrea Cavalli, Michel O. Steinmetz
Summary: This study developed an antitubulin small molecule, Todalam, through two rounds of chemical synthesis based on a crystallographic fragment screen. Todalam binds to a novel tubulin site, disrupts microtubule networks, induces cell death, and synergizes with vinblastine. It destabilizes microtubules by inhibiting the conformational switch in alpha-tubulin and sequestering tubulin dimers. This work demonstrates the importance of using tubulin-binding fragments for innovative antitubulin drug and chemical probe discovery campaigns.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Multidisciplinary
Li Gao, Joyce C. M. Meiring, Adam Varady, Iris E. Ruider, Constanze Heise, Maximilian Wranik, Cecilia D. Velasco, Jennifer A. Taylor, Beatrice Terni, Tobias Weinert, Jorg Standfuss, Clemens C. Cabernard, Artur Llobet, Michel O. Steinmetz, Andreas R. Bausch, Martin Distel, Julia Thorn-Seshold, Anna Akhmanova, Oliver Thorn-Seshold
Summary: Photoswitchable reagents have great potential in high-precision studies in cell biology. However, their in vivo application remains largely unrealized. In this study, we optimized the potency and solubility of photoswitchable reagents and successfully achieved high-precision research in microtubule dynamics, network architecture, and microtubule-dependent processes. The design of these reagents can also inspire the development of optical reagents for other cytosolic protein targets, advancing the field of in vivo photopharmacology.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2022)
Article
Chemistry, Medicinal
Noelia Montel de la Roche, Tobias Muehlethaler, Rita Maria Concetta Di Martino, Jose Antonio Ortega, Dario Gioia, Bibhas Roy, Andrea E. Prota, Michel O. Steinmetz, Andrea Cavalli
Summary: This study identified tubulin-binding fragments through computational and crystallographic screening, and used a rational design strategy to develop a new series of 2-aminobenzimidazole derivatives that destabilize microtubules. The synthesized compound showed submicromolar activity in inhibiting cell viability and demonstrated a similar mechanism of action as classical colchicine-binding site ligands.
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Chemistry, Medicinal
Marilia Barreca, Virginia Span, Roberta Rocca, Roberta Bivacqua, Anne -Catherine Abel, Annalisa Maruca, Alessandra Montalbano, Maria Valeria Raimondi, Chiara Tarantelli, Eugenio Gaudio, Luciano Cascione, Andrea Rinaldi, Ruoli Bai, Michel O. Steinmetz, Andrea E. Prota, Stefano Alcaro, Ernest Hamel, Francesco Bertoni, Paola Barraja
Summary: Lymphomas are common cancers and require more effective therapeutic approaches. This study identified a series of oxazolo isoindoles as potential candidates for the treatment of refractory lymphomas due to their ability to bind to tubulin.
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Cell Biology
Sandro M. Meier, Ana-Maria Farcas, Anil Kumar, Mahdiye Ijavi, Robert T. Bill, Joerg Stelling, Eric R. Dufresne, Michel O. Steinmetz, Yves Barral
Summary: This study investigates the interactions and functions of the Kar9 network in yeast cells, finding that it forms a liquid condensate at selected microtubule ends, serving as a mechanical coupling between microtubules and actin cables.
NATURE CELL BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Juan Estevez-Gallego, Beatriz Alvarez-Bernard, Benet Pera, Christoph Wullschleger, Olivier Raes, Dirk Menche, Juan Carlos Martinez, Daniel Lucena-Agell, Andrea E. Prota, Francesca Bonato, Katja Bargsten, Jelle Cornelus, Juan Francisco Gimenez-Abian, Peter Northcote, Michel O. Steinmetz, Shinji Kamimura, Karl-Heinz Altmann, Ian Paterson, Federico Gago, Johan Van der Eycken, J. Fernando Diaz, Maria Angela Oliva
Summary: Taxanes are microtubule-stabilizing agents used in cancer treatment, but they have side effects on the peripheral nervous system. This study investigates the stabilization mechanism of laulimalide and peloruside, which bind to a different site and can modulate the diameter of microtubules. Some compounds with low cytotoxicity are found to induce stabilization without compromising the native structure of microtubules, suggesting potential applications beyond cancer treatment.
Article
Biology
Charlotte Guyomar, Clement Bousquet, Siou Ku, John M. Heumann, Gabriel Guilloux, Natacha Gaillard, Claire Heichette, Laurence Duchesne, Michel O. Steinmetz, Romain Gibeaux, Denis Chretien, Julie P. Welburn
Summary: Microtubules, structures with a diameter of about 25 nm, play critical roles in cellular functions such as motility, compartmentalization, and division. This study reveals that microtubules consist of variable numbers of protofilaments and tubulin subunit helical-starts, as well as variable numbers of seams. Interestingly, the number and location of seams vary along individual microtubules, leading to the formation of holes within their lattices. These findings demonstrate that the organization of microtubules is an intrinsic property of tubulin and requires unique lateral interactions without longitudinal ones.
Article
Biochemistry & Molecular Biology
Li Gao, Joyce C. M. Meiring, Yvonne Kraus, Maximilian Wranik, Tobias Weinert, Stefanie D. Pritzl, Rebekkah Bingham, Evangelia Ntouliou, Klara Jansen, Natacha Olieric, Jorg Standfuss, Lukas C. Kapitein, Theobald Lohmuller, Julia Ahlfeld, Anna Akhmanova, Michel O. Steinmetz, Oliver Thorn-Seshold
Summary: Researchers have designed a photoswitchable SBT scaffold to create metabolically stable and fully GFP/YFP-orthogonal SBTub photopharmaceuticals for precise control of microtubule dynamics and organization. These photopharmaceuticals have great potential applications in cell biology and are favorable for intracellular biological control in in vivo settings. The robustness and imaging orthogonality of the SBT scaffold may inspire further derivatizations for photocontrol of a range of other biological targets.
CELL CHEMICAL BIOLOGY
(2021)
Meeting Abstract
Biophysics
Joseph Atherton, Jessica J. Hummel, Natacha Olieric, Michel O. Steinmetz, Anna Akhmanova, Casper Hoogenraad, Carolyn A. Moores
BIOPHYSICAL JOURNAL
(2021)
