Molecular Therapy-Methods & Clinical Development最新文献_第3页

Transcriptomics-informed pharmacology identifies epigenetic and cell cycle regulators that enhance AAV production. 转录组学告知药理学鉴定表观遗传和细胞周期调节增强AAV的生产。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-18 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101384

Joshua Tworig, Francis Grafton, Kaylin Fisher, Markus Hörer, Christopher A Reid, Mohammad A Mandegar

{"title":"Transcriptomics-informed pharmacology identifies epigenetic and cell cycle regulators that enhance AAV production.","authors":"Joshua Tworig, Francis Grafton, Kaylin Fisher, Markus Hörer, Christopher A Reid, Mohammad A Mandegar","doi":"10.1016/j.omtm.2024.101384","DOIUrl":"10.1016/j.omtm.2024.101384","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) is a widely used viral vector for gene therapy. However, these vectors have limited availability due to manufacturing challenges with productivity and quality. These challenges can be addressed by better understanding the mechanisms that influence cellular responses during rAAV production. In this study, we aimed to identify targets that may enhance rAAV production using transcriptomic analyses of five cell lines with variable capacities for rAAV production. Using an intersectional approach, we measured the transcriptional responses of these cells during rAAV production and compared transcriptional profiles between high and base producers to identify possible targets for enhancing production. During rAAV production, we found transcriptional differences in cell cycle and nucleosome components contributed to proliferative capacity and DNA replication. We also saw upregulation of several core functions, including transcription, stress response, and Golgi and endoplasmic reticulum organization. Conversely, we saw consistent downregulation of other factors, including inhibitors of DNA-binding proteins and mitochondrial components. With a drug-connectivity analysis, we identified five classes of drugs that were predicted to enhance rAAV production. We also validated the efficacy of histone deacetylase and microtubule inhibitors. Our data uncover novel and previously identified pathways that may enhance rAAV production and quality to expand availability of rAAV for gene therapies.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101384"},"PeriodicalIF":4.6,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11647610/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142839660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantitative proteomic analysis of residual host cell protein retention across adeno-associated virus affinity chromatography. 通过腺相关病毒亲和层析对宿主细胞蛋白残留进行定量蛋白质组学分析。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-18 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101383

Thomas M Leibiger, Lie Min, Kelvin H Lee

{"title":"Quantitative proteomic analysis of residual host cell protein retention across adeno-associated virus affinity chromatography.","authors":"Thomas M Leibiger, Lie Min, Kelvin H Lee","doi":"10.1016/j.omtm.2024.101383","DOIUrl":"10.1016/j.omtm.2024.101383","url":null,"abstract":"To better understand host cell protein (HCP) retention in adeno-associated virus (AAV) downstream processes, sequential window acquisition of all theoretical fragment ion mass spectra (SWATH-MS) was used to quantitatively profile residual HCPs for four AAV serotypes (AAV2, -5, -8, and -9) produced with HEK293 cells and purified using POROS CaptureSelect AAVX affinity chromatography. A broad range of residual HCPs were detected in affinity eluates after purification (N total = 2,746), and HCP profiles showed universally present species (N universal = 1,117) and species unique to one or more AAV serotype. SWATH-MS revealed that HCP persistence was dominated by high-abundance conserved species (HACS), which appeared across all serotype conditions studied. Due to the notable contribution of these species to overall residual HCP levels, physical and functional characteristics of HACS were examined to determine trends that coincide with persistence. Subnetwork interaction mapping and Gene Ontology function enrichment analysis revealed extensive physical interactions between these proteins and significant enrichment for biological processes, molecular functions, and reactome pathways related to protein folding, nucleic acid binding, and cellular stress. The abundant and conserved nature of these HCPs and their functions offers a new perspective for mechanistic evaluations of impurity retention for AAV downstream processes.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101383"},"PeriodicalIF":4.6,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11650319/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142848055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-affinity chimeric antigen receptor signaling induces an inflammatory program in human regulatory T cells. 高亲和力嵌合抗原受体信号诱导人类调节性 T 细胞的炎症程序。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-18 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101385

Russell W Cochrane, Rob A Robino, Bryan Granger, Eva Allen, Silvia Vaena, Martin J Romeo, Aguirre A de Cubas, Stefano Berto, Leonardo M R Ferreira

{"title":"High-affinity chimeric antigen receptor signaling induces an inflammatory program in human regulatory T cells.","authors":"Russell W Cochrane, Rob A Robino, Bryan Granger, Eva Allen, Silvia Vaena, Martin J Romeo, Aguirre A de Cubas, Stefano Berto, Leonardo M R Ferreira","doi":"10.1016/j.omtm.2024.101385","DOIUrl":"10.1016/j.omtm.2024.101385","url":null,"abstract":"Regulatory T cells (Tregs) are promising cellular therapies to induce immune tolerance in organ transplantation and autoimmune disease. The success of chimeric antigen receptor (CAR) T cell therapy for cancer has sparked interest in using CARs to generate antigen-specific Tregs. Here, we compared CAR with endogenous T cell receptor (TCR)/CD28 activation in human Tregs. Strikingly, CAR Tregs displayed increased cytotoxicity and diminished suppression of antigen-presenting cells and effector T (Teff) cells compared with TCR/CD28-activated Tregs. RNA sequencing revealed that CAR Tregs activate Teff cell gene programs. Indeed, CAR Tregs secreted high levels of inflammatory cytokines, with a subset of FOXP3+ CAR Tregs uniquely acquiring CD40L surface expression and producing IFN-γ. Interestingly, decreasing CAR antigen affinity reduced Teff cell gene expression and inflammatory cytokine production by CAR Tregs. Our findings showcase the impact of engineered receptor activation on Treg biology and support tailoring CAR constructs to Tregs for maximal therapeutic efficacy.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101385"},"PeriodicalIF":4.6,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11647616/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142840383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

mRNA vaccines encoding membrane-anchored RBDs of SARS-CoV-2 mutants induce strong humoral responses and can overcome immune imprinting. 编码膜锚定SARS-CoV-2突变体rbd的mRNA疫苗可诱导强烈的体液反应，并可克服免疫印迹。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-15 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101380

Hareth A Al-Wassiti, Stewart A Fabb, Samantha L Grimley, Ruby Kochappan, Joan K Ho, Chinn Yi Wong, Chee Wah Tan, Thomas J Payne, Asuka Takanashi, Chee Leng Lee, Rekha Shandre Mugan, Horatio Sicilia, Serena L Y Teo, Julie McAuley, Paula Ellenberg, James P Cooney, Kathryn C Davidson, Richard Bowen, Marc Pellegrini, Steven Rockman, Dale I Godfrey, Terry M Nolan, Lin-Fa Wang, Georgia Deliyannis, Damian F J Purcell, Colin W Pouton

{"title":"mRNA vaccines encoding membrane-anchored RBDs of SARS-CoV-2 mutants induce strong humoral responses and can overcome immune imprinting.","authors":"Hareth A Al-Wassiti, Stewart A Fabb, Samantha L Grimley, Ruby Kochappan, Joan K Ho, Chinn Yi Wong, Chee Wah Tan, Thomas J Payne, Asuka Takanashi, Chee Leng Lee, Rekha Shandre Mugan, Horatio Sicilia, Serena L Y Teo, Julie McAuley, Paula Ellenberg, James P Cooney, Kathryn C Davidson, Richard Bowen, Marc Pellegrini, Steven Rockman, Dale I Godfrey, Terry M Nolan, Lin-Fa Wang, Georgia Deliyannis, Damian F J Purcell, Colin W Pouton","doi":"10.1016/j.omtm.2024.101380","DOIUrl":"10.1016/j.omtm.2024.101380","url":null,"abstract":"We investigated mRNA vaccines encoding a membrane-anchored receptor-binding domain (RBD), each a fusion of a variant RBD, the transmembrane (TM) and cytoplasmic tail fragments of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. In naive mice, RBD-TM mRNA vaccines against SARS-CoV-2 variants induced strong humoral responses against the target RBD. Multiplex surrogate viral neutralization (sVNT) assays revealed broad neutralizing activity against a range of variant RBDs. In the setting of a heterologous boost, against the background of exposure to ancestral whole-spike vaccines, sVNT studies suggested that BA.1 and BA.5 RBD-TM vaccines had the potential to overcome the detrimental effects of immune imprinting. A subsequent heterologous boost study using XBB.1.5 booster vaccines was evaluated using both sVNT and authentic virus neutralization. Geometric mean XBB.1.5 neutralization values after third-dose RBD-TM or whole-spike XBB.1.5 booster vaccines were compared with those after a third dose of ancestral spike booster vaccine. Fold-improvement over ancestral vaccine was just 1.3 for the whole-spike XBB.1.5 vaccine, similar to data published using human serum samples. In contrast, the fold-improvement achieved by the RBD-TM XBB.1.5 vaccine was 16.3, indicating that the RBD-TM vaccine induced the production of antibodies that neutralize the XBB.1.5 variant despite previous exposure to ancestral spike protein.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101380"},"PeriodicalIF":4.6,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11646785/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142840385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Implementing a robust platform analytical procedure for measuring adeno-associated virus vector genome titer. 实施稳健的平台分析程序，测量腺相关病毒载体基因组滴度。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-15 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101381

Thomas W Powers, Courtney D K Sloan, Don Stano, Brad Evans, Kang Liu, Shawn Mariani, Jessica A Campbell, Thomas F Lerch, Jim J Mo

{"title":"Implementing a robust platform analytical procedure for measuring adeno-associated virus vector genome titer.","authors":"Thomas W Powers, Courtney D K Sloan, Don Stano, Brad Evans, Kang Liu, Shawn Mariani, Jessica A Campbell, Thomas F Lerch, Jim J Mo","doi":"10.1016/j.omtm.2024.101381","DOIUrl":"10.1016/j.omtm.2024.101381","url":null,"abstract":"The vector genome (vg) titer measurement, which is used to control patient dosing and ensure control over drug product manufacturing, is essential for the development of recombinant adeno-associated virus (AAV) gene therapy products. While qPCR and droplet digital PCR technologies are commonly implemented for measuring vg titer, chromatographic techniques with UV detectors represent promising future approaches, in line with traditional biotherapeutics. Here, we introduce a novel vg titer measurement approach using size-exclusion high-performance liquid chromatography with UV detection, which achieves excellent method precision (<2% relative SD), demonstrates linearity across a range of concentrations and varied particle content, is stability indicating, and can be bridged with existing vg titer methods. As there is no bias between this procedure and existing vg titer procedures, such as qPCR, this method can be implemented even at late stages during pharmaceutical development. The procedure was demonstrated to be applicable across serotypes and transgenes, enabling the approach to be used as a platform method for AAV. Given the method performance and criticality of vg titer measurements for AAV, this approach represents a beneficial technology for AAV therapeutics.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101381"},"PeriodicalIF":4.6,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11634990/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142820161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Patient-specific responses to SMN2 splice-modifying treatments in spinal muscular atrophy fibroblasts. 脊髓性肌萎缩成纤维细胞对SMN2剪接修饰治疗的患者特异性反应

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-13 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101379

Ilaria Signoria, Maria M Zwartkruis, Lotte Geerlofs, Elena Perenthaler, Kiterie M E Faller, Rachel James, Harriet McHale-Owen, Jared W Green, Joris Kortooms, Sophie H Snellen, Fay-Lynn Asselman, Thomas H Gillingwater, Gabriella Viero, Renske I Wadman, W Ludo van der Pol, Ewout J N Groen

{"title":"Patient-specific responses to SMN2 splice-modifying treatments in spinal muscular atrophy fibroblasts.","authors":"Ilaria Signoria, Maria M Zwartkruis, Lotte Geerlofs, Elena Perenthaler, Kiterie M E Faller, Rachel James, Harriet McHale-Owen, Jared W Green, Joris Kortooms, Sophie H Snellen, Fay-Lynn Asselman, Thomas H Gillingwater, Gabriella Viero, Renske I Wadman, W Ludo van der Pol, Ewout J N Groen","doi":"10.1016/j.omtm.2024.101379","DOIUrl":"10.1016/j.omtm.2024.101379","url":null,"abstract":"The availability of three therapies for the neuromuscular disease spinal muscular atrophy (SMA) highlights the need to match patients to the optimal treatment. Two of these treatments (nusinersen and risdiplam) target splicing of SMN2, but treatment outcomes vary from patient to patient. An incomplete understanding of the complex interactions among SMA genetics, SMN protein and mRNA levels, and gene-targeting treatments, limits our ability to explain this variability and identify optimal treatment strategies for individual patients. To address this, we analyzed responses to nusinersen and risdiplam in 45 primary fibroblast cell lines. Pre-treatment SMN2-FL, SMN2Δ7 mRNA, and SMN protein levels were influenced by SMN2 copy number, age, and sex. After treatment, SMN and mRNA levels were more heterogeneous. In 43% of patients, response to both therapies was similar, but in 57% one treatment led to a significantly higher SMN increase than the other treatment. Younger age, higher SMN2 copy number, and higher SMN levels before treatment predicted better in vitro efficacy. These findings showcase patient-derived fibroblasts as a tool for identifying molecular predictors for personalized treatment.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101379"},"PeriodicalIF":4.6,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11626024/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142803329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

E2A, VA RNA I, and L4-22k adenoviral helper genes are sufficient for AAV production in HEK293 cells. E2A、VA RNA I和L4-22k腺病毒辅助基因足以在HEK293细胞中产生AAV。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-13 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101376

Jiten Doshi, Emma Couto, Jillian Staiti, Luk H Vandenberghe, Nerea Zabaleta

{"title":"E2A, VA RNA I, and L4-22k adenoviral helper genes are sufficient for AAV production in HEK293 cells.","authors":"Jiten Doshi, Emma Couto, Jillian Staiti, Luk H Vandenberghe, Nerea Zabaleta","doi":"10.1016/j.omtm.2024.101376","DOIUrl":"10.1016/j.omtm.2024.101376","url":null,"abstract":"The replication-defective adeno-associated virus (AAV) is extensively utilized as a research tool or vector for gene therapy. The production process of AAV remains intricate, expensive, and mechanistically underexplored. With the aim of enhancing AAV manufacturing efficiencies in mammalian cells, we revisited the questions and optimization surrounding the requirement of the various adenoviral helper genes in enabling AAV production. First, we refined the minimal set of adenoviral genes in HEK293 AAV production to E2A, L4-22 K /33 K, and VA RNA I. These findings challenge the previously accepted necessity of adenoviral E4orf6 in AAV production. In addition, we identified L4-22 K genes as crucial helpers for AAV production. Next, a revised minimal adenoviral helper plasmid comprising E2A, L4-22 K, and VA RNA I genes was designed and demonstrated to yield high titer and potent AAV preps in HEK293 transient transfection. Lastly, stable packaging cells harboring inducible E2A and L4-22 K genes were shown to maintain AAV production yields comparable to transient transfection over a culture period of ∼10 weeks. Combined, these findings further our understanding of adenoviral helper function in mammalian AAV production and provide novel plasmid and cell-line reagents with an improved safety profile for potential broad applicability in the research and gene therapy community.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101376"},"PeriodicalIF":4.6,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11635002/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142820145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

αFAP-specific nanobodies mediate a highly precise retargeting of modified AAV2 capsids thereby enabling specific transduction of tumor tissues. αFAP特异性纳米抗体可高度精确地再靶向修饰过的AAV2包囊，从而实现对肿瘤组织的特异性转导。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-12 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101378

Olaniyi Olarewaju, Franziska Held, Pamela Curtis, Cynthia Hess Kenny, Udo Maier, Tadas Panavas, Francois du Plessis

{"title":"αFAP-specific nanobodies mediate a highly precise retargeting of modified AAV2 capsids thereby enabling specific transduction of tumor tissues.","authors":"Olaniyi Olarewaju, Franziska Held, Pamela Curtis, Cynthia Hess Kenny, Udo Maier, Tadas Panavas, Francois du Plessis","doi":"10.1016/j.omtm.2024.101378","DOIUrl":"10.1016/j.omtm.2024.101378","url":null,"abstract":"Due to the refractiveness of tumor tissues to adeno-associated virus (AAV) transduction, AAV vectors are poorly explored for cancer therapy delivery. Here, we aimed to engineer AAVs to target tumors by enabling the specific engagement of fibroblast activation protein (FAP). FAP is a cell surface receptor distinctly upregulated in the reactive tumor stroma, but rarely expressed in healthy tissues. Thus, targeting FAP presents an opportunity to selectively transduce tumor tissues. To achieve this, we modified the capsid surface of AAV2 with an αFAP nanobody to retarget the capsid to engage FAP receptor. Following transduction, we observed a 23- to 80-fold increase in the selective transduction of FAP+ tumor cells in vitro, and greater than 5-fold transduction of FAP+ tumor tissues in vivo. Subsequent optimization of the VP1-nanobody expression cassette further enhanced the transduction efficiency of the modified capsids. Due to the limited αFAP nanobodies repertoires, we broadened the versatility of this high-fidelity platform by screening a naive VHH yeast display library, leading to the identification of several novel αFAP nanobody candidates (KD = 0.1 to >100 nM). Hence, our study offers new opportunity for the application of AAV vectors for highly selective delivery of therapeutics to the tumor stroma.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101378"},"PeriodicalIF":4.6,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11655695/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142866294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CAR-NK cells derived from cord blood originate mainly from CD56^-CD7⁺CD34^-HLA-DR^-Lin^- NK progenitor cells. 来源于脐带血的CAR-NK细胞主要来源于CD56-CD7+CD34-HLA-DR-Lin- NK祖细胞。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-12 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101374

Tansri Wibowo, Yosuke Kogue, Shunya Ikeda, Moto Yaga, Mana Tachikawa, Makiko Suga, Shuhei Kida, Kumi Shibata, Kazuhito Tsutsumi, Hiraku Murakami, Yasutaka Ueda, Hisashi Kato, Kentaro Fukushima, Jiro Fujita, Tomoaki Ueda, Shinsuke Kusakabe, Akihisa Hino, Michiko Ichii, Chihaya Imai, Daisuke Okuzaki, Atsushi Kumanogoh, Naoki Hosen

{"title":"CAR-NK cells derived from cord blood originate mainly from CD56-CD7+CD34-HLA-DR-Lin- NK progenitor cells.","authors":"Tansri Wibowo, Yosuke Kogue, Shunya Ikeda, Moto Yaga, Mana Tachikawa, Makiko Suga, Shuhei Kida, Kumi Shibata, Kazuhito Tsutsumi, Hiraku Murakami, Yasutaka Ueda, Hisashi Kato, Kentaro Fukushima, Jiro Fujita, Tomoaki Ueda, Shinsuke Kusakabe, Akihisa Hino, Michiko Ichii, Chihaya Imai, Daisuke Okuzaki, Atsushi Kumanogoh, Naoki Hosen","doi":"10.1016/j.omtm.2024.101374","DOIUrl":"10.1016/j.omtm.2024.101374","url":null,"abstract":"Cord blood (CB)-derived chimeric antigen receptor (CAR)-natural killer (NK) cells targeting CD19 have been shown to be effective against B cell malignancies. While human CD56+ NK cells can be expanded in vitro, NK cells can also be differentiated from hematopoietic progenitor cells. It is still unclear whether CAR-NK cells originate from mature NK cells or NK progenitor cells in CB. Here, we determined that CAR-NK cells were predominantly derived from CD56- NK progenitor cells. We first found that substantial numbers of CD19 CAR-NK cells were produced from CD56- CB mononuclear cells after in vitro culture for 2 weeks. Single-cell RNA sequencing analysis of CD56-CD3-CD14-CD19- CB mononuclear cells revealed that these cells could be subdivided into three subpopulations based on the expression of CD34 and human leukocyte antigen (HLA)-DR. NK cells originated primarily from CD34-HLA-DR- cells. In addition, among the CD34-HLA-DR- cells, only CD7+ cells could differentiate into NK cells. These results indicate that CD56-CD7+CD34-HLA-DR- lineage marker (Lin)- cells are the major origin of human CB-derived CAR-NK cells, indicating the importance of developing methods to enhance the quality and quantity of NK cells produced from these NK progenitor cells.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101374"},"PeriodicalIF":4.6,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11629225/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142807719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Preclinical efficacy and safety of adeno-associated virus 5 alpha-galactosidase: A gene therapy for Fabry disease. 腺相关病毒 5 α-半乳糖苷酶的临床前疗效和安全性：法布里病的基因疗法。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-12 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101375

Jolanda M P Liefhebber, Giso Brasser, Elisabeth A Spronck, Roelof Ottenhoff, Lieke Paerels, Maria J Ferraz, Lukas K Schwarz, Nikoleta Efthymiopoulou, Chi-Lin Kuo, Paula S Montenegro-Miranda, Melvin M Evers, Johannes M F G Aerts, Ying Poi Liu

{"title":"Preclinical efficacy and safety of adeno-associated virus 5 alpha-galactosidase: A gene therapy for Fabry disease.","authors":"Jolanda M P Liefhebber, Giso Brasser, Elisabeth A Spronck, Roelof Ottenhoff, Lieke Paerels, Maria J Ferraz, Lukas K Schwarz, Nikoleta Efthymiopoulou, Chi-Lin Kuo, Paula S Montenegro-Miranda, Melvin M Evers, Johannes M F G Aerts, Ying Poi Liu","doi":"10.1016/j.omtm.2024.101375","DOIUrl":"10.1016/j.omtm.2024.101375","url":null,"abstract":"We developed a novel adeno-associated virus 5 gene therapy (AAV5-GLA) expressing human alpha-galactosidase A (GLA) under the control of a novel, small and strong, liver-restricted promoter. We assessed the preclinical potential of AAV5-GLA for treating Fabry disease, an X-linked hereditary metabolic disorder resulting from mutations in the gene encoding GLA that lead to accumulation of the substrates globotriaosylceramide and globotriaosylsphingosine, causing heart, kidney, and central nervous system dysfunction. Effects of intravenously administered AAV5-GLA were evaluated in (1) GLA-knockout mice aged 7-8 weeks (early in disease) and 20 weeks (nociception phenotype manifestation) and (2) cynomolgus macaques during an 8-week period. In both species, AAV5-GLA was observed as safe, generated detectable vector DNA and mRNA levels in liver, and produced stable enzyme activity in liver and plasma. In mice, dose-dependent transgene enzyme activity, cross-correction (substrate reduction) in kidney and heart, and improved nociception lasted over 6 months. Moreover, after delayed administration when animals displayed the nociception phenotype, target organ enzyme activity was present, and accumulated substrates were reduced. Given the strong, durable expression of active GLA with this promoter and favorable profile of adeno-associated virus 5-based gene therapy in humans, AAV5-GLA warrants further investigation in clinical trials for Fabry disease.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101375"},"PeriodicalIF":4.6,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11646755/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142839256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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