Molecular Therapy-Methods & Clinical Development最新文献_第4页

Assessment of adeno-associated virus purity by capillary electrophoresis-based western 用毛细管电泳法评估腺相关病毒的纯度

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-07 DOI: 10.1016/j.omtm.2024.101332

Julyana Acevedo, Yiling Bi, Jessica Gee, Santoshkumar L. Khatwani

引用次数: 0

Gene Therapy and kidney diseases 基因治疗与肾脏疾病

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-06 DOI: 10.1016/j.omtm.2024.101333

Nadia Tavakolidakhrabadi, Wen Y. Ding, Moin A. Saleem, Gavin I. Welsh, Carl May

引用次数: 0

Longitudinal characterization of sub-retinal pigment epithelium deposit formation in a primary porcine tissue culture model of dry age-related macular degeneration 干性老年性黄斑变性原代猪组织培养模型中视网膜下色素上皮沉积形成的纵向特征描述

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-31 DOI: 10.1016/j.omtm.2024.101331

Erika M. Shaw, Alexander J. Tate, Ramesh Periasamy, Daniel M. Lipinski

{"title":"Longitudinal characterization of sub-retinal pigment epithelium deposit formation in a primary porcine tissue culture model of dry age-related macular degeneration","authors":"Erika M. Shaw, Alexander J. Tate, Ramesh Periasamy, Daniel M. Lipinski","doi":"10.1016/j.omtm.2024.101331","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101331","url":null,"abstract":"Age-related macular degeneration (AMD) affects millions of individuals worldwide and is a leading cause of blindness in the elderly. In dry AMD, lipoproteinaceous deposits called drusen accumulate between the retinal pigment epithelium (RPE) and Bruch’s membrane, leading to impairment of oxygen and nutrient trafficking to the neural retina, and degeneration of the overlying photoreceptor cells. Owing to key differences in human and animal ocular anatomy and the slowly progressing nature of the disease, AMD is not easily modeled In this study, we further characterize a “drusen-in-a-dish” primary porcine RPE model system by employing vital lipid staining to monitor sub-RPE deposition over time in monolayers of cells cultured on porous transwell membranes. We demonstrate for the first time using a semi-automated image analysis pipeline that the number and size of sub-RPE deposits increases gradually but significantly over time and confirm that sub-RPE deposits grown in culture immunostain positive for multiple known components found in human drusen. As a result, we propose that drusen-in-a-dish cell culture models represent a high-throughput and cost-scalable alternative to animal models in which to study the pathobiology of drusen accumulation and may serve as useful tools for screening novel therapeutics aimed at treating dry AMD.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"19 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synergy between Lactobacillus murinus and anti-PcrV antibody delivered in the airways to boost protection against Pseudomonas aeruginosa 鼠乳杆菌与在气道中递送的抗 PcrV 抗体之间的协同作用可增强对铜绿假单胞菌的抵抗力

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-30 DOI: 10.1016/j.omtm.2024.101330

Thomas Sécher, Mélanie Cortes, Chloé Boisseau, Marie-Thérèse Barba Goudiaby, Aubin Pitiot, Christelle Parent, Muriel Thomas, Nathalie Heuzé-Vourc’h

{"title":"Synergy between Lactobacillus murinus and anti-PcrV antibody delivered in the airways to boost protection against Pseudomonas aeruginosa","authors":"Thomas Sécher, Mélanie Cortes, Chloé Boisseau, Marie-Thérèse Barba Goudiaby, Aubin Pitiot, Christelle Parent, Muriel Thomas, Nathalie Heuzé-Vourc’h","doi":"10.1016/j.omtm.2024.101330","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101330","url":null,"abstract":"<p>Therapeutic antibodies (Ab) have revolutionized the management of multiple illnesses including respiratory tract infections (RTIs). However, anti-infectious Ab displayed several limitations including antigen restrictiveness, narrowed therapeutic windows, and limited dose in the vicinity of the target when delivered by parenteral routes. Strategies enhancing further Ab-dependent containment of infection are currently needed. Here we showed that a combination of inhaled anti-infectious Ab and probiotics is an efficient formulation to protect against lung infection. Using a mouse model of <em>Pseudomonas aeruginosa</em>-induced pneumonia, we demonstrated a synergistic effect reducing both bacterial burden and pro-inflammatory response affording protection against primary and secondary infections. This is the first study showing that the local combination in the airways of anti-infective Ab and probiotics subverts suboptimal potency of Ab monotherapy and provides protection against respiratory pathogen.</p>","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"879 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Resolving hidden subpopulations of filled AAVs by probing capsid integrity 通过探测囊壳完整性解析填充型 AAV 隐藏亚群

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-29 DOI: 10.1016/j.omtm.2024.101322

Weida Wu, Georgios Katsikis

引用次数: 0

Unlocking DOE potential by selecting the most appropriate design for rAAV optimization 为 rAAV 优化选择最合适的设计，释放 DOE 潜能

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-26 DOI: 10.1016/j.omtm.2024.101329

Konstantina Tzimou, David Catalán-Tatjer, Lars K. Nielsen, Jesús Lavado-García

{"title":"Unlocking DOE potential by selecting the most appropriate design for rAAV optimization","authors":"Konstantina Tzimou, David Catalán-Tatjer, Lars K. Nielsen, Jesús Lavado-García","doi":"10.1016/j.omtm.2024.101329","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101329","url":null,"abstract":"Producing recombinant adeno-associated virus (rAAV) for gene therapy via triple transfection is an intricate process involving many cellular interactions. Each of the different elements encoded in the three required plasmids—pHelper, pRepCap, and pGOI—plays a distinct role, affecting different cellular pathways when producing rAAVs. The required expression balance emphasizes the critical need to fine-tune the concentration of all these different elements. The use of design of experiments (DOE) to find optimal ratios is a powerful method to streamline the process. However, the choice of the DOE method and design construction is crucial to avoid misleading results. In this work, we examined and compared four distinct DOE approaches: rotatable central composite design (RCCD), Box-Behnken design (BBD), face-centered central composite design (FCCD), and mixture design (MD). We compared the abilities of the different models to predict optimal ratios and interactions among the plasmids and the transfection reagent. Our findings revealed that blocking is essential to reduce the variability caused by uncontrolled random effects and that MD coupled with FCCD outperformed all other approaches, improving volumetric productivity 109-fold. These outcomes underscore the importance of selecting a model that can effectively account for the biological context, ultimately yielding superior results in optimizing rAAV production.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"27 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197291","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Linker-specific monoclonal antibodies present a simple and reliable detection method for scFv-based CARNK cells 连接子特异性单克隆抗体为基于 scFv 的 CARNK 细胞提供了一种简单可靠的检测方法

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-22 DOI: 10.1016/j.omtm.2024.101328

Katharina Schindler, Katharina Eva Ruppel, Claudia Müller, Ulrike Koehl, Stephan Fricke, Dominik Schmiedel

{"title":"Linker-specific monoclonal antibodies present a simple and reliable detection method for scFv-based CARNK cells","authors":"Katharina Schindler, Katharina Eva Ruppel, Claudia Müller, Ulrike Koehl, Stephan Fricke, Dominik Schmiedel","doi":"10.1016/j.omtm.2024.101328","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101328","url":null,"abstract":"Chimeric antigen receptor (CAR) T cell therapies have demonstrated significant successes in treating cancer. Currently, there are six approved CAR T cell products available on the market that target different malignancies of the B cell lineage. However, to overcome the limitations of CAR T cell therapies, other immune cells are being investigated for CAR-based cell therapies. CAR natural killer (NK) cells can be applied as allogeneic cell therapy, providing an economical, safe, and efficient alternative to autologous CAR T cells. To improve CAR research and future in-patient monitoring of cell therapeutics, a simple, reliable, and versatile CAR detection reagent is crucial. As most existing CARs contain a single-chain variable fragment (scFv) with either a Whitlow or a G4S linker site, linker-specific monoclonal antibodies (mAbs) can detect a broad range of CARs. This study demonstrates that these linker-specific mAbs can detect different CAR NK cells , spiked in whole blood, and within patient-derived tumor spheroids with high specificity and sensitivity, providing an effective and almost universal alternative for scFv-based CAR detection. Additionally, we confirm that linker-specific antibodies can be used for functional testing and enrichment of CAR NK cells, thereby providing a useful research tool to fast-track the development of novel CAR-based therapies.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"11 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantification of circulating TCR-engineered T cells targeting a human endogenous retrovirus post-adoptive transfer using nanoplate digital PCR 利用纳米平板数字 PCR 对采用人类内源性逆转录病毒的循环 TCR 工程 T 细胞进行定量分析

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-20 DOI: 10.1016/j.omtm.2024.101324

Stefan Barisic, Elena Cherkasova, Rosa Nadal, Xin Tian, Long Chen, Angelina Parrizzi, Robert N. Reger, Gina M. Scurti, Michael I. Nishimura, Richard W. Childs

{"title":"Quantification of circulating TCR-engineered T cells targeting a human endogenous retrovirus post-adoptive transfer using nanoplate digital PCR","authors":"Stefan Barisic, Elena Cherkasova, Rosa Nadal, Xin Tian, Long Chen, Angelina Parrizzi, Robert N. Reger, Gina M. Scurti, Michael I. Nishimura, Richard W. Childs","doi":"10.1016/j.omtm.2024.101324","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101324","url":null,"abstract":"expansion of genetically modified T cells in cancer patients following adoptive transfer has been linked to both anti-tumor activity and T cell-mediated toxicities. The development of digital PCR has improved the accuracy in quantifying the status of adoptively infused T cells compared to qPCR or flow cytometry. Here, we developed and evaluated the feasibility and performance of nanoplate-based digital PCR (ndPCR) to quantify adoptively infused T cells engineered with a T cell receptor (TCR) that recognizes a human endogenous retrovirus type E (HERV-E) antigen. Analysis of blood samples collected from patients with metastatic kidney cancer following the infusion of HERV-E TCR-transduced T cells established the limit of detection of ndPCR to be 0.3 transgene copies/μL of reaction. The lower limit of quantification for ndPCR was one engineered T cell per 10,000 PBMCs, which outperformed both qPCR and flow cytometry by 1 log. High inter-test and test-retest reliability was confirmed by analyzing blood samples collected from multiple patients. In conclusion, we demonstrated the feasibility of ndPCR for detecting and monitoring the fate of TCR-engineered T cells in adoptive cell therapy.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"182 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142225186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Lentiviral vectors for precise expression to treat X-linked lymphoproliferative disease 用于精确表达的慢病毒载体治疗 X 连锁淋巴细胞增生症

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-20 DOI: 10.1016/j.omtm.2024.101323

Paul G. Ayoub, Julia Gensheimer, Lindsay Lathrop, Colin Juett, Jason Quintos, Kevin Tam, Jack Reid, Feiyang Ma, Curtis Tam, Grace E. McAuley, Devin Brown, Xiaomeng Wu, Ruixue Zhang, Kathryn Bradford, Roger P. Hollis, Gay M. Crooks, Donald B. Kohn

{"title":"Lentiviral vectors for precise expression to treat X-linked lymphoproliferative disease","authors":"Paul G. Ayoub, Julia Gensheimer, Lindsay Lathrop, Colin Juett, Jason Quintos, Kevin Tam, Jack Reid, Feiyang Ma, Curtis Tam, Grace E. McAuley, Devin Brown, Xiaomeng Wu, Ruixue Zhang, Kathryn Bradford, Roger P. Hollis, Gay M. Crooks, Donald B. Kohn","doi":"10.1016/j.omtm.2024.101323","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101323","url":null,"abstract":"X-linked lymphoproliferative disease (XLP1) results from gene mutations affecting the SLAM-associated protein (SAP). A regulated lentiviral vector (LV), XLP-SMART LV, designed to express SAP at therapeutic levels in T, NK, and NKT cells, is crucial for effective gene therapy. We experimentally identified 34 genomic regulatory elements of the gene and designed XLP-SMART LVs to emulate the lineage and stage-specific control of SAP. We screened them for their on-target enhancer activity in T, NK, and NKT cells and their off-target enhancer activity in B cell and myeloid populations. In combination, three enhancer elements increased SAP promoter expression up to 4-fold in on-target populations . NSG-Tg(Hu-IL15) xenograft studies with XLP-SMART LVs demonstrated up to 7-fold greater expression in on-target cells over a control EFS-LV, with no off-target expression. The XLP-SMART LVs exhibited stage-specific T and NK cell expression in peripheral blood, bone marrow, spleen, and thymic tissues (mimicking expression patterns of SAP). Transduction of XLP1 patient CD8+ T cells or BM CD34+ cells with XLP-SMART LVs restored restimulation-induced cell death and NK cytotoxicity to wild-type levels, respectively. These data demonstrate that it is feasible to create a lineage and stage-specific LV to restore the XLP1 phenotype by gene therapy.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"2 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197292","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Predictive power of deleterious single amino acid changes to infer on AAV2 and AAV2-13 capsids fitness 单个氨基酸的有害变化对推断 AAV2 和 AAV2-13 外壳适应性的预测能力

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-20 DOI: 10.1016/j.omtm.2024.101327

Tiziana La Bella, Bérangère Bertin, Ante Mihaljevic, Justine Nozi, Patrice Vidal, Sandrine Imbeaud, Jean-Charles Nault, Jessica Zucman-Rossi, Giuseppe Ronzitti

{"title":"Predictive power of deleterious single amino acid changes to infer on AAV2 and AAV2-13 capsids fitness","authors":"Tiziana La Bella, Bérangère Bertin, Ante Mihaljevic, Justine Nozi, Patrice Vidal, Sandrine Imbeaud, Jean-Charles Nault, Jessica Zucman-Rossi, Giuseppe Ronzitti","doi":"10.1016/j.omtm.2024.101327","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101327","url":null,"abstract":"Adeno-associated virus (AAV) is the most widely used vector for gene transfer. A major limitation of capsid engineering is the incomplete understanding of the consequences of multiple amino acid variations on AAV capsid stability resulting in high frequency of non-viable capsids. In this context, the study of natural AAV variants can provide valuable insights into capsid regions that exhibit greater tolerance to mutations. Here, the characterization of AAV2 variants and the analysis of two public capsid libraries highlighted common features associated with deleterious mutations, suggesting that the impact of mutations on capsid viability is strictly dependent on their 3D location within the capsid structure. We developed a novel prediction method to infer the fitness of AAV2 variants containing multiple amino acid variations with 98% sensitivity, 98% accuracy, and 95% specificity. This novel approach might streamline the development of AAV vector libraries enriched in viable capsids, thus accelerating the identification of therapeutic candidates among engineered capsids.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"28 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0