Molecular Therapy. Methods & Clinical Development最新文献

Optimizing lentiviral genomic integrations to cure beta-thalassemia: The least required for success? 优化慢病毒基因组整合以治疗地中海贫血症：成功所需的最少条件？

Molecular Therapy. Methods & Clinical Development Pub Date : 2024-03-05 DOI: 10.1016/j.omtm.2024.101222

Stefano Rivella

引用次数: 0

A gene silencing-based approach to tackle fatty liver disease 基于基因沉默的脂肪肝治疗方法

Molecular Therapy. Methods & Clinical Development Pub Date : 2024-02-09 DOI: 10.1016/j.omtm.2024.101198

Pavel Strnad, Christina Schrader, Nicola Brunetti-Pierri

引用次数: 0

A third dose of the unmodified COVID-19 mRNA vaccine CVnCoV enhances quality and quantity of immune responses. 第三剂未经修饰的COVID-19 mRNA疫苗CVnCoV可提高免疫反应的质量和数量。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-12-08 Epub Date: 2022-10-06 DOI: 10.1016/j.omtm.2022.10.001

Klara Lenart, Fredrika Hellgren, Sebastian Ols, Xianglei Yan, Alberto Cagigi, Rodrigo Arcoverde Cerveira, Inga Winge, Jakub Hanczak, Stefan O Mueller, Edith Jasny, Kim Schwendt, Susanne Rauch, Benjamin Petsch, Karin Loré

{"title":"A third dose of the unmodified COVID-19 mRNA vaccine CVnCoV enhances quality and quantity of immune responses.","authors":"Klara Lenart, Fredrika Hellgren, Sebastian Ols, Xianglei Yan, Alberto Cagigi, Rodrigo Arcoverde Cerveira, Inga Winge, Jakub Hanczak, Stefan O Mueller, Edith Jasny, Kim Schwendt, Susanne Rauch, Benjamin Petsch, Karin Loré","doi":"10.1016/j.omtm.2022.10.001","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.10.001","url":null,"abstract":"A third vaccine dose is often required to achieve potent, long-lasting immune responses. We investigated the effect of three 8-μg doses of CVnCoV, CureVac's severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine candidate containing sequence-optimized unmodified mRNA encoding the spike (S) glycoprotein, administered at 0, 4, and 28 weeks, on immune responses in rhesus macaques. After the third dose, S-specific binding and neutralizing antibodies increased 50-fold compared with post-dose 2 levels, with increased responses also evident in the lower airways and against the SARS-CoV-2 B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), and B.1.617.2 (Delta) variants. Enhanced binding affinity of serum antibodies after the third dose correlated with higher somatic hypermutation in S-specific B cells, corresponding with improved binding properties of monoclonal antibodies expressed from isolated B cells. Administration of low-dose mRNA led to fewer cells expressing antigen in vivo at the injection site and in the draining lymph nodes compared with a 10-fold higher dose, possibly reducing engagement of precursor cells with the antigen and resulting in the suboptimal response observed after two-dose vaccination schedules in phase IIb/III clinical trials of CVnCoV. However, when immune memory is established, a third dose efficiently boosts the immunological responses and improves antibody affinity and breadth.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"309-323"},"PeriodicalIF":4.7,"publicationDate":"2022-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9535876/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33520486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Assessing production variability in empty and filled adeno-associated viruses by single molecule mass analyses. 通过单分子质量分析评估空和填充腺相关病毒的生产变异性。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-11-15 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.11.003

Eduard H T M Ebberink, Alisa Ruisinger, Markus Nuebel, Marco Thomann, Albert J R Heck

{"title":"Assessing production variability in empty and filled adeno-associated viruses by single molecule mass analyses.","authors":"Eduard H T M Ebberink, Alisa Ruisinger, Markus Nuebel, Marco Thomann, Albert J R Heck","doi":"10.1016/j.omtm.2022.11.003","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.11.003","url":null,"abstract":"Adeno-associated viruses (AAVs) are useful vehicles for gene therapy because of their stability, low immunogenicity. and non-pathogenicity. However, disparity in AAV sample preparations (e.g., in capsid composition, DNA packaging, and impurities) gives rise to product heterogeneity, with possibly undesired effects on gene delivery. Ideally, AAV production should be with full control of AAV structure and genetic payload. Therefore, robust, efficient, and low material consuming methods are essential to characterize AAVs. Here, we use two emerging single-molecule techniques, mass photometry and Orbitrap-based charge-detection mass spectrometry, and show how they may efficiently and accurately characterize AAVs. We were able to resolve heterogeneous pools of particles, evaluating AAVs from two different serotypes (AAV8 and AAV2), produced by three independent production platforms, either lacking a genome or packed with a transgene. Together our data confirm that the different AAV production methods result in rather different and diverse AAV particle distributions. Especially for the packed AAVs, frequently additional subspecies were observed, next to the expected packed genome, mostly resulting from under- or overpackaging of genome material and/or residual empty particles. This work further establishes that both these single-particle techniques may become valuable tools in characterizing AAVs before they are used in gene therapy.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"491-501"},"PeriodicalIF":4.7,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9706604/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40557486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Erratum: Intravenous immunoglobulin prevents peripheral liver transduction of intrathecally delivered AAV vectors. 勘误:静脉注射免疫球蛋白阻止鞘内递送的AAV载体的外周肝转导。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-11-12 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.11.001

Makoto Horiuchi, Christian J Hinderer, Jenny A Greig, Cecilia Dyer, Elizabeth L Buza, Peter Bell, Jessica A Chichester, Peter M Hayashi, Hanying Yan, Tamara Goode, James M Wilson

引用次数: 0

Population-wide gene disruption in the murine lung epithelium via AAV-mediated delivery of CRISPR-Cas9 components. 通过aav介导的CRISPR-Cas9组分的递送，在小鼠肺上皮中进行群体范围的基因破坏。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-11-01 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.10.016

Honglin Chen, Steffen Durinck, Hetal Patel, Oded Foreman, Kathryn Mesh, Jeffrey Eastham, Roger Caothien, Robert J Newman, Merone Roose-Girma, Spyros Darmanis, Soren Warming, Annalisa Lattanzi, Yuxin Liang, Benjamin Haley

{"title":"Population-wide gene disruption in the murine lung epithelium via AAV-mediated delivery of CRISPR-Cas9 components.","authors":"Honglin Chen, Steffen Durinck, Hetal Patel, Oded Foreman, Kathryn Mesh, Jeffrey Eastham, Roger Caothien, Robert J Newman, Merone Roose-Girma, Spyros Darmanis, Soren Warming, Annalisa Lattanzi, Yuxin Liang, Benjamin Haley","doi":"10.1016/j.omtm.2022.10.016","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.10.016","url":null,"abstract":"With the aim of expediting drug target discovery and validation for respiratory diseases, we developed an optimized method for in situ somatic gene disruption in murine lung epithelial cells via AAV6-mediated CRISPR-Cas9 delivery. Efficient gene editing was observed in lung type II alveolar epithelial cells and distal airway cells following assessment of single- or dual-guide AAV vector formats, Cas9 variants, and a sequential dosing strategy with combinatorial guide RNA expression cassettes. In particular, we were able to demonstrate population-wide gene disruption within distinct epithelial cell types for separate targets in Cas9 transgenic animals, with minimal to no associated inflammation. We also observed and characterized AAV vector integration events that occurred within directed double-stranded DNA break sites in lung cells, highlighting a complicating factor with AAV-mediated delivery of DNA nucleases. Taken together, we demonstrate a uniquely effective approach for somatic engineering of the murine lung, which will greatly facilitate the modeling of disease and therapeutic intervention.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"431-449"},"PeriodicalIF":4.7,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/8f/29/main.PMC9668740.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40483242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Antibodies against recombinant enzyme in the treatment of Fabry disease: Now you see them, now you don't. 法布里病治疗中抗重组酶的抗体:现在你看到了，现在你看不见了。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-10-29 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.10.007

Sanne Jolien van der Veen, Mirjam Langeveld

引用次数: 1

Development of an insect cell-based adeno-associated virus packaging cell line employing advanced Rep gene expression control system. 采用Rep基因表达控制系统的昆虫细胞型腺相关病毒包装细胞系的建立。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-10-28 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.10.015

Fiona Moreno, Florence Lip, Hugo Rojas, Anggakusuma

{"title":"Development of an insect cell-based adeno-associated virus packaging cell line employing advanced Rep gene expression control system.","authors":"Fiona Moreno, Florence Lip, Hugo Rojas, Anggakusuma","doi":"10.1016/j.omtm.2022.10.015","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.10.015","url":null,"abstract":"The insect cell-based baculovirus expression vector (BEV) system is a leading platform for scalable production of adeno-associated viruses (AAVs). The previously described One-Bac system consists of an insect packaging cell line harboring the AAV Rep and Cap genes and a BEV carrying the transgene and AAV inverted terminal repeats. Here we describe a new system where we successfully translated the molecular design of a double AAV Rep expression cassette to inducible plasmid vectors. These optimized plasmid vectors employ non-canonical late promoters and alternative start codons that alleviate promoter-promoter competition. Because too much Rep expression can be toxic to the host cells, tighter regulation of AAV Rep expression is warranted. This has been achieved by adopting alternate baculovirus homologous region enhancers. Inoculation of the resultant stable insect Rep packaging cell line by a recombinant BEV produced high-titer recombinant AAV (rAAV) preparations (1 × 1011 genome copies/mL). Sequential batch reactor experiments indicate that this system is amenable to large-scale AAV production. We generated an insect packaging cell line that employs an optimized Rep gene control system, ensuring stable and appropriate Rep expression. This platform produces potent and high-yield AAV particles and demonstrates potential for scale up.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"391-403"},"PeriodicalIF":4.7,"publicationDate":"2022-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/db/a1/main.PMC9661643.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40465632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

The seroprevalence of neutralizing antibodies against the adeno-associated virus capsids in Japanese hemophiliacs. 日本血友病患者腺相关病毒衣壳中和抗体的血清流行率。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-10-27 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.10.014

Yuji Kashiwakura, Nemekhbayar Baatartsogt, Shoji Yamazaki, Azusa Nagao, Kagehiro Amano, Nobuaki Suzuki, Tadashi Matsushita, Akihiro Sawada, Satoshi Higasa, Naoya Yamasaki, Teruhisa Fujii, Taemi Ogura, Hideyuki Takedani, Masashi Taki, Takeshi Matsumoto, Jun Yamanouchi, Michio Sakai, Masako Nishikawa, Yutaka Yatomi, Koji Yada, Keiji Nogami, Ryota Watano, Takafumi Hiramoto, Morisada Hayakawa, Nobuhiko Kamoshita, Akihiro Kume, Hiroaki Mizukami, Shizukiyo Ishikawa, Yoichi Sakata, Tsukasa Ohmori

{"title":"The seroprevalence of neutralizing antibodies against the adeno-associated virus capsids in Japanese hemophiliacs.","authors":"Yuji Kashiwakura, Nemekhbayar Baatartsogt, Shoji Yamazaki, Azusa Nagao, Kagehiro Amano, Nobuaki Suzuki, Tadashi Matsushita, Akihiro Sawada, Satoshi Higasa, Naoya Yamasaki, Teruhisa Fujii, Taemi Ogura, Hideyuki Takedani, Masashi Taki, Takeshi Matsumoto, Jun Yamanouchi, Michio Sakai, Masako Nishikawa, Yutaka Yatomi, Koji Yada, Keiji Nogami, Ryota Watano, Takafumi Hiramoto, Morisada Hayakawa, Nobuhiko Kamoshita, Akihiro Kume, Hiroaki Mizukami, Shizukiyo Ishikawa, Yoichi Sakata, Tsukasa Ohmori","doi":"10.1016/j.omtm.2022.10.014","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.10.014","url":null,"abstract":"Adeno-associated virus (AAV) vectors are promising modalities of gene therapy to address unmet medical needs. However, anti-AAV neutralizing antibodies (NAbs) hamper the vector-mediated therapeutic effect. Therefore, NAb prevalence in the target population is vital in designing clinical trials with AAV vectors. Hence, updating the seroprevalence of anti-AAV NAbs, herein we analyzed sera from 100 healthy individuals and 216 hemophiliacs in Japan. In both groups, the overall seroprevalence against various AAV serotypes was 20%-30%, and the ratio of the NAb-positive population increased with age. The seroprevalence did not differ between healthy participants and hemophiliacs and was not biased by the concomitant blood-borne viral infections. The high neutralizing activity, which strongly inhibits the transduction with all serotypes in vitro, was mostly found in people in their 60s or of older age. The multivariate analysis suggested that \"60s or older age\" was the only independent factor related to the high titer of NAbs. Conversely, a large proportion of younger hemophiliacs was seronegative, rendering them eligible for AAV-mediated gene therapy in Japan. Compared with our previous study, the peak of seroprevalences has shifted to older populations, indicating that natural AAV exposure in the elderly occurred in their youth but not during the last decade.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"404-414"},"PeriodicalIF":4.7,"publicationDate":"2022-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c7/70/main.PMC9661668.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40465630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Edging closer to successful gene therapy for Wilson disease. 离威尔森氏症的基因治疗成功又近了一步。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-10-21 eCollection Date: 2022-12-08 DOI: 10.1016/j.omtm.2022.10.005

Thomas Damgaard Sandahl, Valentina Medici

引用次数: 0