Clinical & Translational Immunology最新文献

{"title":"Optimising the IgG-degrading enzyme treatment regimen for enhanced adeno-associated virus transduction in the presence of neutralising antibodies","authors":"Irene Ros-Ga?án,&nbsp;Mirja Hommel,&nbsp;Laia Trigueros-Motos,&nbsp;Blanche Tamarit,&nbsp;Estefanía Rodríguez-García,&nbsp;David Salas,&nbsp;Guiomar Pérez,&nbsp;Anne Douar,&nbsp;Jean Philippe Combal,&nbsp;Bernard Benichou,&nbsp;Veronica Ferrer,&nbsp;Gloria González-Aseguinolaza","doi":"10.1002/cti2.1375","DOIUrl":"https://doi.org/10.1002/cti2.1375","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objective</h3>\u0000 \u0000 <p>Pre-existing neutralising antibodies (NAbs) to adeno-associated viruses (AAVs) remain an impediment for systemically administered AAV-mediated gene therapy treatment in many patients, and various strategies are under investigation to overcome this limitation. Here, IgG-degrading enzymes (Ides) derived from bacteria of the genus <i>Streptococcus</i> were tested for their ability to cleave human IgG and allow AAV-mediated transduction in individuals with pre-existing NAbs.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Cleavage activity of three different Ides was evaluated <i>in vitro</i> in serum from different species. Passively immunised mice or non-human primates (NHP) with naturally occurring anti-AAV NAbs were used to define the optimal IdeS dose and administration window for AAVAnc80 and AAV8 vectors in mice and AAV3B in NHPs.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The selected candidate, IdeS, was found to be highly efficient at cleaving human IgG, less efficient against NHP IgG and inefficient against mouse IgG. <i>In vivo</i>, we observed differences in how IdeS affected liver transduction in the presence of NAbs depending on the AAV serotype. For AAVAnc80 and AAV3B, the best transduction levels were achieved when the vector was administered after IgG digestion products were cleared from circulation. However, for AAV8 we only observed a modest and transient inhibition of transduction by IdeS cleavage products.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Preconditioning with IdeS represents a unique treatment opportunity for patients primarily excluded from participation in gene therapy clinical trials because of elevated circulating anti-AAV NAb levels. However, careful determination of the optimal IdeS dose and timing for the administration of each AAV serotype is essential for optimal transduction.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1375","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5787812","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}

{"title":"Retraction statement: Dual targeting of RANKL and PD-1 with a bispecific antibody improves anti-tumor immunity","authors":"","doi":"10.1002/cti2.1378","DOIUrl":"https://doi.org/10.1002/cti2.1378","url":null,"abstract":"<p>Dougall WC, Roman Aguilera A, Smyth MJ (2019), Dual targeting of RANKL and PD-1 with a bispecific antibody improves anti-tumor immunity. <i>Clin Transl Immunol</i>, 8: e01081. https://doi.org/10.1002/cti2.1081. The above article published online on 27 September 2019 in Wiley Online Library (wileyonlinelibrary.com) has been retracted by agreement between the journal's Editor-in-Chief, Rajiv Khanna, the Australian and New Zealand Society for Immunology Inc., and John Wiley and Sons Australia, Ltd. The retraction has been agreed following an investigation by the QIMR Berghofer Medical Research Institute into research published by the senior author, Professor Mark J Smyth. The investigation concluded that at least two figures (3b and 4b) generated by the senior author Professor Mark Smyth are likely fabricated. As a result, the conclusions are considered unreliable, and the article has been retracted.</p>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1378","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5997867","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}

{"title":"Epigenetic regulation of inflammation by microRNAs in post-infectious bronchiolitis obliterans","authors":"Ruth P Duecker,&nbsp;Ines De Mir Messa,&nbsp;Silvija-Pera Jerkic,&nbsp;Annalena Kochems,&nbsp;Gabriele Gottwald,&nbsp;Antonio Moreno-Galdó,&nbsp;Martin Rosewich,&nbsp;Lucia Gronau,&nbsp;Stefan Zielen,&nbsp;Andreas Geburtig-Chiocchetti,&nbsp;Hermann Kreyenberg,&nbsp;Ralf Schubert","doi":"10.1002/cti2.1376","DOIUrl":"https://doi.org/10.1002/cti2.1376","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Post-infectious bronchiolitis obliterans (PiBO) is a rare, chronic disease initiated by severe infection and followed by perpetuating inflammation and obliteration of the small airways. MicroRNAs (miRNAs) have been proposed to play a central role as epigenetic regulators, which control resolution and prevent the uncontrolled progress of inflammation. The aim of this study was to define biomarkers on the level of post-transcriptional gene regulation in order to characterise PiBO.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A total of 39 patients with well-defined PiBO and 31 controls from two centres, Barcelona, Spain, and Frankfurt, Germany, were analysed by next-generation sequencing (NGS). The evaluation of the biological targets of the miRNAs was performed by pathway enrichment analysis and protein–protein interaction network analysis respectively.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Patients with PiBO had significantly lower lung function values and increased airway inflammation in induced sputum as indicated by total cell counts, neutrophils, IL-1β, IL-6, IL-8 and TGF-β compared to controls.</p>\u0000 \u0000 <p>Next-generation sequencing analysis revealed a total of 22 dysregulated miRNAs, which passed significance threshold for <i>P</i>adj ≤ 0.001 with 17 being upregulated and 5 being downregulated. Of these dysregulated miRNAs, miR-335-5p, miR-186-5p, miR-30b-5p and miR-30c-5p were further validated using qRT-PCR. Interestingly, these miRNAs are functionally implicated in cytokine–cytokine receptor interaction, TGF-β signalling and FoxO signalling pathway and significantly correlated with lung function values (FEV1).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Our results demonstrate an aberrant miRNA expression profile in PiBO, which impacts pathways responsible for the regulation of inflammation and fibrosis. The defined miRNAs are useful biomarkers and should be assessed as potential target in the field of miRNA therapeutics.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1376","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5732373","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}

{"title":"Inflammatory marker trajectories associated with frailty and ageing in a 20-year longitudinal study","authors":"Leonard Dani?l Samson,&nbsp;Anne-Marie Buisman,&nbsp;José A Ferreira,&nbsp;H Susan J Picavet,&nbsp;W M Monique Verschuren,&nbsp;Annemieke MH Boots,&nbsp;Peter Engelfriet","doi":"10.1002/cti2.1374","DOIUrl":"https://doi.org/10.1002/cti2.1374","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objective</h3>\u0000 \u0000 <p>The aim of this exploratory study was to investigate the development of low-grade inflammation during ageing and its relationship with frailty.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The trajectories of 18 inflammatory markers measured in blood samples, collected at 5-year intervals over a period of 20 years from 144 individuals aged 65–75 years at the study endpoint, were related to the degree of frailty later in life.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>IFN-γ-related markers and platelet activation markers were found to change in synchrony. Chronically elevated levels of IL-6 pathway markers, such as CRP and sIL-6R, were associated with more frailty, poorer lung function and reduced physical strength. Being overweight was a possible driver of these associations. More and stronger associations were detected in women, such as a relation between increasing sCD14 levels and frailty, indicating a possible role for monocyte overactivation. Multivariate prediction of frailty confirmed the main results, but predictive accuracy was low.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>In summary, we documented temporal changes in and between inflammatory markers in an ageing population over a period of 20 years, and related these to clinically relevant health outcomes.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6162622","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}

{"title":"Allogeneic CD20-targeted γδ T cells exhibit innate and adaptive antitumor activities in preclinical B-cell lymphoma models","authors":"Kevin P Nishimoto,&nbsp;Taylor Barca,&nbsp;Aruna Azameera,&nbsp;Amani Makkouk,&nbsp;Jason M Romero,&nbsp;Lu Bai,&nbsp;Mary M Brodey,&nbsp;Jackie Kennedy-Wilde,&nbsp;Hui Shao,&nbsp;Stephanie Papaioannou,&nbsp;Amy Doan,&nbsp;Cynthia Masri,&nbsp;Ngoc T Hoang,&nbsp;Hayden Tessman,&nbsp;Vidhya Dhevi Ramanathan,&nbsp;Ana Giner-Rubio,&nbsp;Frank Delfino,&nbsp;Kriti Sharma,&nbsp;Kevin Bray,&nbsp;Matthew Hoopes,&nbsp;Daulet Satpayev,&nbsp;Ranjita Sengupta,&nbsp;Marissa Herrman,&nbsp;Stewart E Abbot,&nbsp;Blake T Aftab,&nbsp;Zili An,&nbsp;Swapna Panuganti,&nbsp;Sandra M Hayes","doi":"10.1002/cti2.1373","DOIUrl":"https://doi.org/10.1002/cti2.1373","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Autologous chimeric antigen receptor (CAR) αβ T-cell therapies have demonstrated remarkable antitumor efficacy in patients with haematological malignancies; however, not all eligible cancer patients receive clinical benefit. Emerging strategies to improve patient access and clinical responses include using premanufactured products from healthy donors and alternative cytotoxic effectors possessing intrinsic tumoricidal activity as sources of CAR cell therapies. γδ T cells, which combine innate and adaptive mechanisms to recognise and kill malignant cells, are an attractive candidate platform for allogeneic CAR T-cell therapy. Here, we evaluated the manufacturability and functionality of allogeneic peripheral blood-derived CAR⁺ Vδ1 γδ T cells expressing a second-generation CAR targeting the B-cell-restricted CD20 antigen.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Donor-derived Vδ1 γδ T cells from peripheral blood were <i>ex vivo</i>-activated, expanded and engineered to express a novel anti-CD20 CAR. <i>In vitro</i> and <i>in vivo</i> assays were used to evaluate CAR-dependent and CAR-independent antitumor activities of CD20 CAR⁺ Vδ1 γδ T cells against B-cell tumors.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Anti-CD20 CAR⁺ Vδ1 γδ T cells exhibited innate and adaptive antitumor activities, such as <i>in vitro</i> tumor cell killing and proinflammatory cytokine production, in addition to <i>in vivo</i> tumor growth inhibition of B-cell lymphoma xenografts in immunodeficient mice. Furthermore, CD20 CAR⁺ Vδ1 γδ T cells did not induce xenogeneic graft-versus-host disease in immunodeficient mice.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>These preclinical data support the clinical evaluation of ADI-001, an allogeneic CD20 CAR⁺ Vδ1 γδ T cell, and a phase 1 study has been initiated in patients with B-cell malignancies (NCT04735471).</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1373","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6051935","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}

{"title":"Inhibition of renal fibrosis with a human CXCL9-derived glycosaminoglycan-binding peptide","authors":"Fariba Poosti,&nbsp;Mohammad Ayodhia Soebadi,&nbsp;Helena Crijns,&nbsp;Alexandra De Zutter,&nbsp;Mieke Metzemaekers,&nbsp;Nele Berghmans,&nbsp;Vincent Vanheule,&nbsp;Maarten Albersen,&nbsp;Ghislain Opdenakker,&nbsp;Jo Van Damme,&nbsp;Ben Sprangers,&nbsp;Paul Proost,&nbsp;Sofie Struyf","doi":"10.1002/cti2.1370","DOIUrl":"https://doi.org/10.1002/cti2.1370","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Renal fibrosis accompanies all chronic kidney disorders, ultimately leading to end-stage kidney disease and the need for dialysis or even renal replacement. As such, renal fibrosis poses a major threat to global health and the search for effective therapeutic strategies to prevent or treat fibrosis is highly needed. We evaluated the applicability of a highly positively charged human peptide derived from the COOH-terminal domain of the chemokine CXCL9, namely CXCL9(74–103), for therapeutic intervention. Because of its high density of net positive charges at physiological pH, CXCL9(74–103) competes with full-length chemokines for glycosaminoglycan (GAG) binding. Consequently, CXCL9(74–103) prevents recruitment of inflammatory leucocytes to sites of inflammation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>CXCL9(74–103) was chemically synthesised and tested <i>in vitro</i> for anti-fibrotic properties on human fibroblasts and <i>in vivo</i> in the unilateral ureteral obstruction (UUO) mouse model.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>CXCL9(74–103) significantly reduced the mRNA and/or protein expression of connective tissue growth factor (CTGF), alpha-smooth muscle actin (α-SMA) and collagen III by transforming growth factor (TGF)-β1-stimulated human fibroblasts. In addition, administration of CXCL9(74–103) inhibited fibroblast migration towards platelet-derived growth factor (PDGF), without affecting cell viability. In the UUO model, CXCL9(74–103) treatment significantly decreased renal α-SMA, vimentin, and fibronectin mRNA and protein expression. Compared with vehicle, CXCL9(74–103) attenuated mRNA expression of TGF-β1 and the inflammatory markers/mediators MMP-9, F4/80, CCL2, IL-6 and TNF-α. Finally, CXCL9(74–103) treatment resulted in reduced influx of leucocytes in the UUO model and preserved tubular morphology. The anti-fibrotic and anti-inflammatory effects of CXCL9(74–103) were mediated by competition with chemokines and growth factors for GAG binding.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Our findings provide a scientific rationale for targeting GAG–protein interactions in renal fibrotic disease.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1370","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6051930","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}

{"title":"Quantitative measurement of IgG to SARS-CoV-2 antigens using monoclonal antibody-based enzyme-linked immunosorbent assays","authors":"Ingrid Sander,&nbsp;Sabine Kespohl,&nbsp;Eva Zahradnik,&nbsp;Philipp G?cke,&nbsp;Ingolf Hosbach,&nbsp;Burkhard L Herrmann,&nbsp;Thomas Brüning,&nbsp;Monika Raulf","doi":"10.1002/cti2.1369","DOIUrl":"https://doi.org/10.1002/cti2.1369","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objective</h3>\u0000 \u0000 <p>Standardised quantitative analysis of the humoral immune response to SARS-CoV-2 antigens may be useful for estimating the extent and duration of immunity. The aim was to develop enzyme-linked immunosorbent assays (ELISAs) for the quantification of human IgG antibodies against SARS-CoV-2 antigens.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Enzyme-linked immunosorbent assays were developed based on monoclonal antibodies against human IgG and recombinant SARS-CoV-2 antigens (Spike-S1 and Nucleocapsid). The WHO 67/086 immunoglobulin and WHO 20/136 SARS-CoV-2 references were used for standardisation. Sera of a study group of COVID-19-positive subjects (<i>n</i> = 144), pre-pandemic controls (<i>n</i> = 135) and individuals vaccinated with BioNTech–Pfizer BNT162b2 vaccine (<i>n</i> = 48) were analysed. The study group sera were also tested using EuroImmun SARS-CoV-2-ELISAs and a quantitative S1-specific fluorescence enzyme immunoassay (FEIA) from Thermo Fisher.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The ELISA results were repeatable and traceable to international units because of their parallelism to both WHO references. In the study group, median anti-S1-IgG concentrations were 102 BAU mL⁻¹, compared to 100 and 1457 BAU mL⁻¹ in the vaccination group after first and second vaccination, respectively. The ELISAs achieved an area under the curve (AUC) of 0.965 (S1) and 0.955 (Nucleocapsid) in receiver operating characteristic (ROC) analysis, and a specificity of 1 (S1) and 0.963 (Nucleocapsid) and sensitivity of 0.903 (S1) and 0.833 (Nucleocapsid) at the maximum Youden index. In comparison, the commercial assays (S1-FEIA, S1 and Nucleocapsid ELISA EuroImmun) achieved sensitivities of 0.764, 0.875 and 0.882 in the study group, respectively.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The quantitative ELISAs to measure IgG binding to SARS-CoV-2 antigens have good analytical and clinical performance characteristics and units traceable to international standards.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 2","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1369","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5782976","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}

{"title":"DROSHA but not DICER is required for human haematopoietic stem cell function","authors":"Karen Gu,&nbsp;Carina Walpole,&nbsp;Shayarana Gooneratne,&nbsp;Xin Liu,&nbsp;Oscar L Haigh,&nbsp;Kristen J Radford,&nbsp;Mark MW Chong","doi":"10.1002/cti2.1361","DOIUrl":"https://doi.org/10.1002/cti2.1361","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>DROSHA and DICER have central roles in the biogenesis of microRNAs (miRNAs). However, we previously showed that in the murine system, DROSHA has an alternate function where it directly recognises and cleaves protein-coding messenger (m)RNAs and this is critical for safeguarding the pluripotency of haematopoietic stem cells (HSCs). Maintenance of murine HSC function is dependent on DROSHA-mediated cleavage of two mRNAs, <i>Myl9</i> and <i>Todr1</i>. The goal of this study is to determine whether this pathway is conserved in human HSCs.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>DROSHA and DICER were knocked down in human cord blood CD34⁺ HSCs with short hairpin RNAs. The function of HSCs was analysed <i>in vitro</i> and in humanised mice. Analysis of mRNA cleavage was performed by capture of 5′ phosphorylated RNAs.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Consistent with murine HSCs, DROSHA knockdown impaired the differentiation of human HSCs <i>in vitro</i> and engraftment into humanised mice, whereas DICER knockdown had no impact. DROSHA cleaves the MYL9 mRNA in human HSCs and DROSHA deficiency resulted in the accumulation of the mRNA. However, ectopic expression of MYL9 did not impair human HSC function. We were unable to identify a human homolog of Todr1.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>A miRNA-independent function of DROSHA is critical for the function of human HSCs. DROSHA directly recognises and degrades mRNAs in humans HSCs. However, unlike in murine HSCs, the degradation of the MYL9 mRNA alone is not critical for human HSC function. Therefore, DROSHA must be inhibiting other targets and/or has another miRNA-independent function that is essential for safeguarding the pluripotency of human HSCs.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 1","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1361","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5763800","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}

{"title":"Genome-wide off-target analyses of CRISPR/Cas9-mediated T-cell receptor engineering in primary human T cells","authors":"Theresa Kaeuferle,&nbsp;Tanja A Stief,&nbsp;Stefan Canzar,&nbsp;Nayad N Kutlu,&nbsp;Semjon Willier,&nbsp;Dana Stenger,&nbsp;Paulina Ferrada-Ernst,&nbsp;Nicola Habjan,&nbsp;Annika E Peters,&nbsp;Dirk H Busch,&nbsp;Tobias Feuchtinger","doi":"10.1002/cti2.1372","DOIUrl":"https://doi.org/10.1002/cti2.1372","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Exploiting the forces of human T cells for treatment has led to the current paradigm of emerging immunotherapy strategies. Genetic engineering of the T-cell receptor (TCR) redirects specificity, ablates alloreactivity and brings significant progress and off-the-shelf options to emerging adoptive T-cell transfer (ACT) approaches. Targeted CRISPR/Cas9-mediated double-strand breaks in the DNA enable knockout or knock-in engineering.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Here, we perform CRISPR/Cas9-mediated TCR knockout using a therapeutically relevant ribonucleoprotein (RNP) delivery method to assess the safety of genetically engineered T-cell products. Whole-genome sequencing was performed to analyse whether CRISPR/Cas9-mediated DNA double-strand break at the TCR locus is associated with off-target events in human primary T cells.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>TCRα chain and TCRβ chain knockout leads to high on-target InDel frequency and functional knockout. None of the predicted off-target sites could be confirmed experimentally, whereas whole-genome sequencing and manual Integrative Genomics Viewer (IGV) review revealed 9 potential low-frequency off-target events genome-wide. Subsequent amplification and targeted deep sequencing in 7 of 7 evaluable loci did not confirm these low-frequency InDels. Therefore, off-target events are unlikely to be caused by the CRISPR/Cas9 engineering.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The combinatorial approach of whole-genome sequencing and targeted deep sequencing confirmed highly specific genetic engineering using CRISPR/Cas9-mediated TCR knockout without potentially harmful exonic off-target effects.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 1","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1372","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"6000178","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}

{"title":"TGF-β-activated kinase-1 inhibitor LL-Z1640-2 reduces joint inflammation and bone destruction in mouse models of rheumatoid arthritis by inhibiting NLRP3 inflammasome, TACE, TNF-α and RANKL expression","authors":"Hirofumi Tenshin,&nbsp;Jumpei Teramachi,&nbsp;Mohannad Ashtar,&nbsp;Masahiro Hiasa,&nbsp;Yusuke Inoue,&nbsp;Asuka Oda,&nbsp;Kotaro Tanimoto,&nbsp;So Shimizu,&nbsp;Yoshiki Higa,&nbsp;Takeshi Harada,&nbsp;Masahiro Oura,&nbsp;Kimiko Sogabe,&nbsp;Tomoyo Hara,&nbsp;Ryohei Sumitani,&nbsp;Tomoko Maruhashi,&nbsp;Mayu Sebe,&nbsp;Rie Tsutsumi,&nbsp;Hiroshi Sakaue,&nbsp;Itsuro Endo,&nbsp;Toshio Matsumoto,&nbsp;Eiji Tanaka,&nbsp;Masahiro Abe","doi":"10.1002/cti2.1371","DOIUrl":"https://doi.org/10.1002/cti2.1371","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Aberrant NLRP3 inflammasome activation has been demonstrated in rheumatoid arthritis (RA), which may contribute to debilitating inflammation and bone destruction. Here, we explored the efficacy of the potent TGF-β-activated kinase-1 (TAK1) inhibitor LL-Z1640-2 (LLZ) on joint inflammation and bone destruction in collagen-induced arthritis (CIA).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>LL-Z1640-2 was administered every other day in CIA mice. Clinical and histological evaluation was performed. Priming and activation of NLRP3 inflammasome and osteoclastogenic activity were assessed.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>NLRP3 inflammasome formation was observed in synovial macrophages and osteoclasts (OCs) in CIA mice. TACE and RANKL were also overexpressed in synovial macrophages and fibroblasts, respectively, in the CIA joints. Treatment with LLZ mitigated all the above changes. As a result, LLZ markedly suppressed synovial hypertrophy and pannus formation to alleviate pain and inflammation in CIA mice. LLZ could block the priming and activation of NLRP3 inflammasome in RAW264.7 macrophage cell line, primary bone marrow macrophages and OCs upon treatment with LPS followed by ATP, thereby suppressing their IL-1β production. LLZ also suppressed LPS-induced production of TACE and TNF-α in bone marrow macrophages and abolished IL-1β-induced production of MMP-3, IL-6 and RANKL in synovial fibroblasts. In addition, LLZ directly inhibits RANKL-mediated OC formation and activation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>TAK1 inhibition with LLZ may become a novel treatment strategy to effectively alleviate inflammasome-mediated inflammation and RANKL-induced osteoclastic bone destruction in joints alongside its potent suppression of TNF-α and IL-6 production and proteinase-mediated pathological processes in RA.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"11 1","pages":""},"PeriodicalIF":5.8,"publicationDate":"2022-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.1371","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5729190","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}