As emerging and re-emerging pathogens, filoviruses, especially Ebola virus (EBOV), pose a great threat to public health and require sustained attention and ongoing surveillance. More vaccines and antiviral drugs are imperative to be developed and stockpiled to respond to unpredictable outbreaks. Virus-like vesicles, generated by alphavirus replicons expressing homogeneous or heterogeneous glycoproteins, have demonstrated the capacity of self-propagation and shown great potential in vaccine development. Here, we describe a novel class of Ebola virus-like vesicles (eVLVs) incorporating both EBOV GP and VP40. The eVLVs exhibited similar antigenicity as EBOV. In murine models, eVLVs were highly attenuated and elicited robust GP-specific antibodies with neutralizing activities. Importantly, a single dose of eVLVs conferred complete protection in a surrogate EBOV lethal mouse model. Furthermore, our VLVs strategy was also successfully applied to Marburg virus (MARV), the representative member of the genus Marburgvirus. Taken together, our findings indicate the feasibility of alphavirus-derived VLVs strategy in combating infection of filoviruses represented by EBOV and MARV, which provides further evidence of the potential of this platform for universal live-attenuated vaccine development.
丝状病毒,尤其是埃博拉病毒(EBOV),作为新出现和再次出现的病原体,对公共卫生构成了巨大威胁,需要持续关注和不断监测。必须开发和储备更多疫苗和抗病毒药物,以应对不可预测的疫情爆发。由表达同种或异种糖蛋白的α-病毒复制子生成的病毒样囊泡已证明具有自我繁殖能力,在疫苗开发方面具有巨大潜力。在这里,我们描述了一类新型的埃博拉病毒样囊泡(eVLVs),其中同时含有埃博拉病毒 GP 和 VP40。eVLVs 表现出与 EBOV 相似的抗原性。在小鼠模型中,eVLVs 被高度减毒,并激发出具有中和活性的强效 GP 特异性抗体。重要的是,在EBOV致死小鼠替代模型中,单剂量eVLVs可提供完全保护。此外,我们的 VLVs 策略还成功地应用于马尔堡病毒属的代表成员马尔堡病毒(MARV)。综上所述,我们的研究结果表明,α病毒衍生的VLVs策略在对抗以EBOV和MARV为代表的丝状病毒感染方面是可行的,这进一步证明了该平台在通用减毒活疫苗开发方面的潜力。
{"title":"Rational design of self-amplifying virus-like vesicles with Ebola virus glycoprotein as vaccines.","authors":"Hong-Qing Zhang, Ya-Nan Zhang, Cheng-Lin Deng, Qin-Xuan Zhu, Zhe-Rui Zhang, Xiao-Dan Li, Zhi-Ming Yuan, Bo Zhang","doi":"10.1016/j.ymthe.2024.08.026","DOIUrl":"https://doi.org/10.1016/j.ymthe.2024.08.026","url":null,"abstract":"<p><p>As emerging and re-emerging pathogens, filoviruses, especially Ebola virus (EBOV), pose a great threat to public health and require sustained attention and ongoing surveillance. More vaccines and antiviral drugs are imperative to be developed and stockpiled to respond to unpredictable outbreaks. Virus-like vesicles, generated by alphavirus replicons expressing homogeneous or heterogeneous glycoproteins, have demonstrated the capacity of self-propagation and shown great potential in vaccine development. Here, we describe a novel class of Ebola virus-like vesicles (eVLVs) incorporating both EBOV GP and VP40. The eVLVs exhibited similar antigenicity as EBOV. In murine models, eVLVs were highly attenuated and elicited robust GP-specific antibodies with neutralizing activities. Importantly, a single dose of eVLVs conferred complete protection in a surrogate EBOV lethal mouse model. Furthermore, our VLVs strategy was also successfully applied to Marburg virus (MARV), the representative member of the genus Marburgvirus. Taken together, our findings indicate the feasibility of alphavirus-derived VLVs strategy in combating infection of filoviruses represented by EBOV and MARV, which provides further evidence of the potential of this platform for universal live-attenuated vaccine development.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142109685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-29DOI: 10.1016/j.ymthe.2024.08.020
Hossein A Hamed, Adly Yacoub, Margaret A Park, Patrick J Eulitt, Rupesh Dash, Devanand Sarkar, Igor P Dmitriev, Maciej S Lesniak, Khalid Shah, Steven Grant, David T Curiel, Paul B Fisher, Paul Dent
{"title":"Retraction Notice to: Inhibition of Multiple Protective Signaling Pathways and Ad.5/3 Delivery Enhances mda-7/IL-24 Therapy of Malignant Glioma.","authors":"Hossein A Hamed, Adly Yacoub, Margaret A Park, Patrick J Eulitt, Rupesh Dash, Devanand Sarkar, Igor P Dmitriev, Maciej S Lesniak, Khalid Shah, Steven Grant, David T Curiel, Paul B Fisher, Paul Dent","doi":"10.1016/j.ymthe.2024.08.020","DOIUrl":"10.1016/j.ymthe.2024.08.020","url":null,"abstract":"","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142109686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-27DOI: 10.1016/j.ymthe.2024.08.022
João Fonseca-Gomes, Tiago Costa-Coelho, Mafalda Ferreira-Manso, Sara Inteiro-Oliveira, Sandra H Vaz, Nuno Alemãn-Serrano, Henrique Atalaia-Barbacena, Leonor Ribeiro-Rodrigues, Rita M Ramalho, Rui Pinto, Hugo Vicente Miranda, Sara R Tanqueiro, Carolina de Almeida-Borlido, Maria João Ramalho, Catarina Miranda-Lourenço, Rita F Belo, Catarina B Ferreira, Vera Neves, Diogo M Rombo, Ricardo Viais, Ivo C Martins, André Jerónimo-Santos, António Caetano, Nuno Manso, Petra Mäkinen, Mikael Marttinen, Mari Takalo, Michael Bremang, Ian Pike, Annakaisa Haapasalo, Joana A Loureiro, Maria Carmo Pereira, Nuno C Santos, Tiago F Outeiro, Miguel A R B Castanho, Adelaide Fernandes, Mikko Hiltunen, Carlos B Duarte, Eero Castrén, Alexandre de Mendonça, Ana M Sebastião, Tiago M Rodrigues, Maria José Diógenes
In Alzheimer's disease (AD), amyloid β (Aβ)-triggered cleavage of TrkB-FL impairs Brain-derived neurotrophic factor (BDNF) signaling, thereby compromising neuronal survival, differentiation, as well as synaptic transmission and plasticity. Using cerebrospinal fluid and post-mortem human brain samples, we show that TrkB-FL cleavage occurs from the early stages of the disease and increases as function of pathology severity. To explore the therapeutic potential of this disease mechanism, we designed small TAT-fused peptides and screened their ability to prevent TrkB-FL receptor cleavage. Among these, a TAT-TrkB peptide with a lysine-lysine linker prevented TrkB-FL cleavage both in vitro and in vivo and rescued synaptic deficits induced by oligomeric Aβ in hippocampal slices. Furthermore, this TAT-TrkB peptide improved the cognitive performance, ameliorated synaptic plasticity deficits and prevented Tau pathology progression in vivo in the 5XFAD mouse model of AD. No evidence of liver or kidney toxicity was found. We provide proof-of-concept evidence for the efficacy and safety of this therapeutic strategy and anticipate that this TAT-TrkB peptide has the potential of a disease-modifying drug that can prevent and/or reverse cognitive deficits in patients with AD.
在阿尔茨海默病(AD)中,淀粉样蛋白β(Aβ)触发的TrkB-FL裂解会损害脑源性神经营养因子(BDNF)信号传导,从而影响神经元的存活、分化以及突触传递和可塑性。利用脑脊液和死后人脑样本,我们发现TrkB-FL裂解在疾病早期就会发生,并随着病理严重程度的增加而增加。为了探索这一疾病机制的治疗潜力,我们设计了与 TAT 融合的小肽,并筛选了它们阻止 TrkB-FL 受体裂解的能力。其中,一种带有赖氨酸-赖氨酸连接的TAT-TrkB肽在体外和体内都能阻止TrkB-FL的裂解,并能挽救海马片中寡聚Aβ诱导的突触缺陷。此外,这种TAT-TrkB肽还能改善5XFAD小鼠AD模型的认知能力,改善突触可塑性缺陷,防止体内Tau病理学进展。没有发现肝脏或肾脏毒性的证据。我们为这一治疗策略的有效性和安全性提供了概念性证据,并预计这种TAT-TrkB肽有可能成为一种改变疾病的药物,可以预防和/或逆转AD患者的认知障碍。
{"title":"A Small TAT-TrkB Peptide Prevents BDNF Receptor Cleavage and Restores Synaptic Physiology in Alzheimer's Disease.","authors":"João Fonseca-Gomes, Tiago Costa-Coelho, Mafalda Ferreira-Manso, Sara Inteiro-Oliveira, Sandra H Vaz, Nuno Alemãn-Serrano, Henrique Atalaia-Barbacena, Leonor Ribeiro-Rodrigues, Rita M Ramalho, Rui Pinto, Hugo Vicente Miranda, Sara R Tanqueiro, Carolina de Almeida-Borlido, Maria João Ramalho, Catarina Miranda-Lourenço, Rita F Belo, Catarina B Ferreira, Vera Neves, Diogo M Rombo, Ricardo Viais, Ivo C Martins, André Jerónimo-Santos, António Caetano, Nuno Manso, Petra Mäkinen, Mikael Marttinen, Mari Takalo, Michael Bremang, Ian Pike, Annakaisa Haapasalo, Joana A Loureiro, Maria Carmo Pereira, Nuno C Santos, Tiago F Outeiro, Miguel A R B Castanho, Adelaide Fernandes, Mikko Hiltunen, Carlos B Duarte, Eero Castrén, Alexandre de Mendonça, Ana M Sebastião, Tiago M Rodrigues, Maria José Diógenes","doi":"10.1016/j.ymthe.2024.08.022","DOIUrl":"https://doi.org/10.1016/j.ymthe.2024.08.022","url":null,"abstract":"<p><p>In Alzheimer's disease (AD), amyloid β (Aβ)-triggered cleavage of TrkB-FL impairs Brain-derived neurotrophic factor (BDNF) signaling, thereby compromising neuronal survival, differentiation, as well as synaptic transmission and plasticity. Using cerebrospinal fluid and post-mortem human brain samples, we show that TrkB-FL cleavage occurs from the early stages of the disease and increases as function of pathology severity. To explore the therapeutic potential of this disease mechanism, we designed small TAT-fused peptides and screened their ability to prevent TrkB-FL receptor cleavage. Among these, a TAT-TrkB peptide with a lysine-lysine linker prevented TrkB-FL cleavage both in vitro and in vivo and rescued synaptic deficits induced by oligomeric Aβ in hippocampal slices. Furthermore, this TAT-TrkB peptide improved the cognitive performance, ameliorated synaptic plasticity deficits and prevented Tau pathology progression in vivo in the 5XFAD mouse model of AD. No evidence of liver or kidney toxicity was found. We provide proof-of-concept evidence for the efficacy and safety of this therapeutic strategy and anticipate that this TAT-TrkB peptide has the potential of a disease-modifying drug that can prevent and/or reverse cognitive deficits in patients with AD.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142109683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-26DOI: 10.1016/j.ymthe.2024.08.024
Hector Ribeiro Benatti, Vania Anagnostakou, Toloo Taghian, Erin F Hall, Sarah Nath, Carl B Heilman, Brandon M Beneduce, Anita Leporati, Christopher Raskett, Mark Epshtein, Robert King, Matthew J Gounis, Adel M Malek, Heather L Gray-Edwards
Neurological disorders pose a challenge for targeted therapy due to restricted access of therapeutic agents to the central nervous system (CNS). Current methods are limited by procedure-related risks, invasiveness, and insufficient CNS biodistribution. A novel percutaneous transvenous technology, currently in clinical trials for communicating hydrocephalus, offers a minimally invasive approach by providing endovascular access to the cerebrospinal fluid-filled cerebellopontine angle (CPA) cistern. We hypothesized that drug delivery to the CPA cistern could yield widespread CNS distribution. Using an ovine model, we compared the biodistribution of scAAV9-CB-GFP following CPA cistern infusion with previously reported cisterna magna (CM) administration. Targeting both the CPA cistern and CM in sheep, we employed a lumbar spine-inserted microcatheter under fluoroscopy. CPA delivery of AAV9 demonstrated biodistribution and transduction in the cerebral cortices, striatum, thalamus, midbrain, cerebellum, and spinal cord, with minor liver distribution comparable to CM. The favorable safety profile in humans with hydrocephalus suggests that percutaneous endovascular injection into the CPA could offer a clinically safer and minimally invasive delivery system for CNS gene and cell-based therapies.
{"title":"A minimally invasive endovascular approach to the cerebellopontine angle cistern enables broad CNS biodistribution of scAAV9-CB-GFP.","authors":"Hector Ribeiro Benatti, Vania Anagnostakou, Toloo Taghian, Erin F Hall, Sarah Nath, Carl B Heilman, Brandon M Beneduce, Anita Leporati, Christopher Raskett, Mark Epshtein, Robert King, Matthew J Gounis, Adel M Malek, Heather L Gray-Edwards","doi":"10.1016/j.ymthe.2024.08.024","DOIUrl":"10.1016/j.ymthe.2024.08.024","url":null,"abstract":"<p><p>Neurological disorders pose a challenge for targeted therapy due to restricted access of therapeutic agents to the central nervous system (CNS). Current methods are limited by procedure-related risks, invasiveness, and insufficient CNS biodistribution. A novel percutaneous transvenous technology, currently in clinical trials for communicating hydrocephalus, offers a minimally invasive approach by providing endovascular access to the cerebrospinal fluid-filled cerebellopontine angle (CPA) cistern. We hypothesized that drug delivery to the CPA cistern could yield widespread CNS distribution. Using an ovine model, we compared the biodistribution of scAAV9-CB-GFP following CPA cistern infusion with previously reported cisterna magna (CM) administration. Targeting both the CPA cistern and CM in sheep, we employed a lumbar spine-inserted microcatheter under fluoroscopy. CPA delivery of AAV9 demonstrated biodistribution and transduction in the cerebral cortices, striatum, thalamus, midbrain, cerebellum, and spinal cord, with minor liver distribution comparable to CM. The favorable safety profile in humans with hydrocephalus suggests that percutaneous endovascular injection into the CPA could offer a clinically safer and minimally invasive delivery system for CNS gene and cell-based therapies.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142081013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-26DOI: 10.1016/j.ymthe.2024.08.023
Ragan A Pitner, Jaime L Chao, Noelle P Dahl, Meng-Ni Fan, Xiaohe Cai, Nathan G Avery, Kelsey Roe, P Clint Spiegel, Carol H Miao, Michael Y Gerner, Richard G James, David J Rawlings
Antibody inhibitors pose an ongoing challenge to the treatment of subjects with inherited protein deficiency disorders, limiting the efficacy of both protein replacement therapy and corrective gene therapy. Beyond their central role as producers of serum antibody, B cells also exhibit many unique properties that could be exploited in cell therapy applications, notably including antigen-specific recognition and the linked capacity for antigen presentation. Here we employed CRISPR/Cas9 to demonstrate that ex vivo antigen-primed Blimp1-knockout "decoy" B cells, incapable of differentiation into plasma cells, participated in and downregulated host antigen-specific humoral responses after adoptive transfer. Following ex vivo antigen pulse, adoptively transferred high affinity antigen-specific decoy B cells were diverted into germinal centers en masse, thereby reducing participation by endogenous antigen-specific B cells in T-dependent humoral responses and suppressing both cognate and linked antigen-specific IgG following immunization with conjugated antigen. This effect was dose-dependent and, importantly, did not impact concurrent unrelated antibody responses. We demonstrated the therapeutic potential of this approach by treating factor VIII (FVIII)-knockout mice with antigen-pulsed decoy B cells prior to immunization with a FVIII conjugate protein, thereby blunting the production of serum FVIII-specific IgG by an order of magnitude as well as reducing the proportion of animals exhibiting functional FVIII inhibition by 6-fold.
{"title":"Blunting specific T-dependent antibody responses with engineered 'decoy' B cells.","authors":"Ragan A Pitner, Jaime L Chao, Noelle P Dahl, Meng-Ni Fan, Xiaohe Cai, Nathan G Avery, Kelsey Roe, P Clint Spiegel, Carol H Miao, Michael Y Gerner, Richard G James, David J Rawlings","doi":"10.1016/j.ymthe.2024.08.023","DOIUrl":"https://doi.org/10.1016/j.ymthe.2024.08.023","url":null,"abstract":"<p><p>Antibody inhibitors pose an ongoing challenge to the treatment of subjects with inherited protein deficiency disorders, limiting the efficacy of both protein replacement therapy and corrective gene therapy. Beyond their central role as producers of serum antibody, B cells also exhibit many unique properties that could be exploited in cell therapy applications, notably including antigen-specific recognition and the linked capacity for antigen presentation. Here we employed CRISPR/Cas9 to demonstrate that ex vivo antigen-primed Blimp1-knockout \"decoy\" B cells, incapable of differentiation into plasma cells, participated in and downregulated host antigen-specific humoral responses after adoptive transfer. Following ex vivo antigen pulse, adoptively transferred high affinity antigen-specific decoy B cells were diverted into germinal centers en masse, thereby reducing participation by endogenous antigen-specific B cells in T-dependent humoral responses and suppressing both cognate and linked antigen-specific IgG following immunization with conjugated antigen. This effect was dose-dependent and, importantly, did not impact concurrent unrelated antibody responses. We demonstrated the therapeutic potential of this approach by treating factor VIII (FVIII)-knockout mice with antigen-pulsed decoy B cells prior to immunization with a FVIII conjugate protein, thereby blunting the production of serum FVIII-specific IgG by an order of magnitude as well as reducing the proportion of animals exhibiting functional FVIII inhibition by 6-fold.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142081014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-22DOI: 10.1016/j.ymthe.2024.08.013
Jia Yao, Svetlana Atasheva, Nicole Wagner, Nelson C Di Paolo, Phoebe L Stewart, Dmitry M Shayakhmetov
{"title":"Targeted, safe, and efficient gene delivery to human hematopoietic stem and progenitor cells in vivo using the engineered AVID adenovirus vector platform.","authors":"Jia Yao, Svetlana Atasheva, Nicole Wagner, Nelson C Di Paolo, Phoebe L Stewart, Dmitry M Shayakhmetov","doi":"10.1016/j.ymthe.2024.08.013","DOIUrl":"https://doi.org/10.1016/j.ymthe.2024.08.013","url":null,"abstract":"","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142046926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-22DOI: 10.1016/j.ymthe.2024.08.019
Yuna Jo, Ju A Shim, Jin Woo Jeong, Hyori Kim, So Min Lee, Juhee Jeong, Segi Kim, Sun-Kyoung Im, Donghoon Choi, Byung Ha Lee, Yun Hak Kim, Chi Dae Kim, Chan Hyuk Kim, Changwan Hong
Cytotoxic T lymphocytes (CTLs) play a crucial role in cancer rejection. However, CTLs encounter dysfunction and exhaustion in the immunosuppressive tumor microenvironment (TME). Although the reactive oxygen species (ROS)-rich TME attenuates CTL function, the underlying molecular mechanism remains poorly understood. The nuclear factor erythroid 2-related 2 (Nrf2) is the ROS-responsible factor implicated in increasing susceptibility to cancer progression. Therefore, we examined how Nrf2 is involved in anti-tumor responses of CD8+ T and chimeric antigen receptor (CAR) T cells in the ROS-rich TME. Here, we demonstrated that tumor growth in Nrf2-/- mice was significantly controlled and was reversed by T cell depletion and further confirmed that Nrf2 deficiency in T cells promotes anti-tumor responses using an adoptive transfer model of antigen-specific CD8+ T cells. Nrf2-deficient CTLs are resistant to ROS, and their effector functions are sustained in the TME. Furthermore, Nrf2 knockdown in human CAR-T cells enhanced the survival and function of intratumoral CAR-T cells in a solid tumor xenograft model and effectively controlled tumor growth. ROS-sensing Nrf2 inhibits the anti-tumor T cell responses, indicating that Nrf2 may be a potential target for T cell immunotherapy strategies against solid tumors.
细胞毒性 T 淋巴细胞(CTL)在癌症排斥反应中发挥着至关重要的作用。然而,CTL 在具有免疫抑制作用的肿瘤微环境(TME)中会出现功能障碍和衰竭。虽然富含活性氧(ROS)的肿瘤微环境会削弱 CTL 的功能,但其潜在的分子机制仍鲜为人知。核因子-红细胞 2 相关-2(Nrf2)是 ROS 反应因子,与增加癌症进展的易感性有关。因此,我们研究了在富含 ROS 的 TME 下,Nrf2 如何参与 CD8+ T 细胞和嵌合抗原受体(CAR)-T 细胞的抗肿瘤反应。在这里,我们证明了Nrf2-/-小鼠的肿瘤生长受到显著控制,并通过T细胞耗竭逆转了肿瘤生长,并利用抗原特异性CD8+ T细胞的采纳转移模型进一步证实了T细胞中Nrf2的缺乏促进了抗肿瘤反应。Nrf2缺陷的CTL对ROS有抵抗力,其效应功能在TME中得以维持。此外,在实体瘤异种移植模型中,敲除人CAR-T细胞中的Nrf2可提高瘤内CAR-T细胞的存活率和功能,并有效控制肿瘤生长。ROS传感Nrf2抑制了T细胞的抗肿瘤反应,这表明Nrf2可能是T细胞免疫治疗实体瘤策略的潜在靶点。
{"title":"Targeting ROS-sensing Nrf2 potentiates anti-tumor immunity of intratumoral CD8<sup>+</sup> T and CAR-T cells.","authors":"Yuna Jo, Ju A Shim, Jin Woo Jeong, Hyori Kim, So Min Lee, Juhee Jeong, Segi Kim, Sun-Kyoung Im, Donghoon Choi, Byung Ha Lee, Yun Hak Kim, Chi Dae Kim, Chan Hyuk Kim, Changwan Hong","doi":"10.1016/j.ymthe.2024.08.019","DOIUrl":"10.1016/j.ymthe.2024.08.019","url":null,"abstract":"<p><p>Cytotoxic T lymphocytes (CTLs) play a crucial role in cancer rejection. However, CTLs encounter dysfunction and exhaustion in the immunosuppressive tumor microenvironment (TME). Although the reactive oxygen species (ROS)-rich TME attenuates CTL function, the underlying molecular mechanism remains poorly understood. The nuclear factor erythroid 2-related 2 (Nrf2) is the ROS-responsible factor implicated in increasing susceptibility to cancer progression. Therefore, we examined how Nrf2 is involved in anti-tumor responses of CD8<sup>+</sup> T and chimeric antigen receptor (CAR) T cells in the ROS-rich TME. Here, we demonstrated that tumor growth in Nrf2<sup>-/-</sup> mice was significantly controlled and was reversed by T cell depletion and further confirmed that Nrf2 deficiency in T cells promotes anti-tumor responses using an adoptive transfer model of antigen-specific CD8<sup>+</sup> T cells. Nrf2-deficient CTLs are resistant to ROS, and their effector functions are sustained in the TME. Furthermore, Nrf2 knockdown in human CAR-T cells enhanced the survival and function of intratumoral CAR-T cells in a solid tumor xenograft model and effectively controlled tumor growth. ROS-sensing Nrf2 inhibits the anti-tumor T cell responses, indicating that Nrf2 may be a potential target for T cell immunotherapy strategies against solid tumors.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142018093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-22DOI: 10.1016/j.ymthe.2024.08.015
Vi Pham, Lucas Tricoli, Xinying Hong, Parith Wongkittichote, Carlo Castruccio Castracani, Amaliris Guerra, Lars Schlotawa, Laura A Adang, Amanda Kuhs, Margaret M Cassidy, Owen Kane, Emily Tsai, Maximiliano Presa, Cathleen Lutz, Stefano B Rivella, Rebecca C Ahrens-Nicklas
Multiple sulfatase deficiency (MSD) is a severe, lysosomal storage disorder caused by pathogenic variants in the gene SUMF1, encoding the sulfatase modifying factor formylglycine-generating enzyme. Patients with MSD exhibit functional deficiencies in all cellular sulfatases. The inability of sulfatases to break down their substrates leads to progressive and multi-systemic complications in patients, similar to those seen in single-sulfatase disorders such as metachromatic leukodystrophy and mucopolysaccharidoses IIIA. Here, we aimed to determine if hematopoietic stem cell transplantation with ex vivo SUMF1 lentiviral gene therapy could improve outcomes in a clinically relevant mouse model of MSD. We first tested our approach in MSD patient-derived cells and found that our SUMF1 lentiviral vector improved protein expression, sulfatase activities, and glycosaminoglycan accumulation. In vivo, we found that our gene therapy approach rescued biochemical deficits, including sulfatase activity and glycosaminoglycan accumulation, in affected organs of MSD mice treated post-symptom onset. In addition, treated mice demonstrated improved neuroinflammation and neurocognitive function. Together, these findings suggest that SUMF1 HSCT-GT can improve both biochemical and functional disease markers in the MSD mouse.
{"title":"Hematopoietic stem cell gene therapy improves outcomes in a clinically relevant mouse model of multiple sulfatase deficiency.","authors":"Vi Pham, Lucas Tricoli, Xinying Hong, Parith Wongkittichote, Carlo Castruccio Castracani, Amaliris Guerra, Lars Schlotawa, Laura A Adang, Amanda Kuhs, Margaret M Cassidy, Owen Kane, Emily Tsai, Maximiliano Presa, Cathleen Lutz, Stefano B Rivella, Rebecca C Ahrens-Nicklas","doi":"10.1016/j.ymthe.2024.08.015","DOIUrl":"10.1016/j.ymthe.2024.08.015","url":null,"abstract":"<p><p>Multiple sulfatase deficiency (MSD) is a severe, lysosomal storage disorder caused by pathogenic variants in the gene SUMF1, encoding the sulfatase modifying factor formylglycine-generating enzyme. Patients with MSD exhibit functional deficiencies in all cellular sulfatases. The inability of sulfatases to break down their substrates leads to progressive and multi-systemic complications in patients, similar to those seen in single-sulfatase disorders such as metachromatic leukodystrophy and mucopolysaccharidoses IIIA. Here, we aimed to determine if hematopoietic stem cell transplantation with ex vivo SUMF1 lentiviral gene therapy could improve outcomes in a clinically relevant mouse model of MSD. We first tested our approach in MSD patient-derived cells and found that our SUMF1 lentiviral vector improved protein expression, sulfatase activities, and glycosaminoglycan accumulation. In vivo, we found that our gene therapy approach rescued biochemical deficits, including sulfatase activity and glycosaminoglycan accumulation, in affected organs of MSD mice treated post-symptom onset. In addition, treated mice demonstrated improved neuroinflammation and neurocognitive function. Together, these findings suggest that SUMF1 HSCT-GT can improve both biochemical and functional disease markers in the MSD mouse.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142018090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-22DOI: 10.1016/j.ymthe.2024.08.016
Sabrina Capelletti, Sofía C García Soto, Manuel A F V Gonçalves
The repurposing of RNA-programmable CRISPR systems from genome editing into epigenome editing tools is gaining pace, including in research and development efforts directed at tackling human disorders. This momentum stems from the increasing knowledge regarding the epigenetic factors and networks underlying cell physiology and disease etiology and from the growing realization that genome editing principles involving chromosomal breaks generated by programmable nucleases are prone to unpredictable genetic changes and outcomes. Hence, engineered CRISPR systems are serving as versatile DNA-targeting scaffolds for heterologous and synthetic effector domains that, via locally recruiting transcription factors and chromatin remodeling complexes, seek interfering with loss-of-function and gain-of-function processes underlying recessive and dominant disorders, respectively. Here, after providing an overview about epigenetic drugs and CRISPR-Cas-based activation and interference platforms, we cover the testing of these platforms in the context of molecular therapies for muscular dystrophies. Finally, we examine attributes, obstacles, and deployment opportunities for CRISPR-based epigenetic modulating technologies.
{"title":"On RNA-programmable gene modulation as a versatile set of principles targeting muscular dystrophies.","authors":"Sabrina Capelletti, Sofía C García Soto, Manuel A F V Gonçalves","doi":"10.1016/j.ymthe.2024.08.016","DOIUrl":"10.1016/j.ymthe.2024.08.016","url":null,"abstract":"<p><p>The repurposing of RNA-programmable CRISPR systems from genome editing into epigenome editing tools is gaining pace, including in research and development efforts directed at tackling human disorders. This momentum stems from the increasing knowledge regarding the epigenetic factors and networks underlying cell physiology and disease etiology and from the growing realization that genome editing principles involving chromosomal breaks generated by programmable nucleases are prone to unpredictable genetic changes and outcomes. Hence, engineered CRISPR systems are serving as versatile DNA-targeting scaffolds for heterologous and synthetic effector domains that, via locally recruiting transcription factors and chromatin remodeling complexes, seek interfering with loss-of-function and gain-of-function processes underlying recessive and dominant disorders, respectively. Here, after providing an overview about epigenetic drugs and CRISPR-Cas-based activation and interference platforms, we cover the testing of these platforms in the context of molecular therapies for muscular dystrophies. Finally, we examine attributes, obstacles, and deployment opportunities for CRISPR-based epigenetic modulating technologies.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142018091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-22DOI: 10.1016/j.ymthe.2024.08.014
Nina Volf, Roman Vuerich, Andrea Colliva, Maria Concetta Volpe, Margherita Marengon, Lorena Zentilin, Mauro Giacca, Nadja Anneliese Ruth Ring, Simone Vodret, Luca Braga, Serena Zacchigna
A major obstacle in inducing therapeutic angiogenesis in the heart is inefficient gene transfer to endothelial cells (ECs). Here, we identify compounds able to enhance the permissiveness of cardiac ECs to adeno-associated virus (AAV) vectors, which stand as ideal tools for in vivo gene delivery. We screened a library of >1,500 US Food and Drug Administration (FDA)-approved drugs, in combination with AAV vectors, in cardiac ECs. Among the top drugs increasing AAV-mediated transduction, we found vatalanib, an inhibitor of multiple tyrosine kinase receptors. The increased AAV transduction efficiency by vatalanib was paralleled by induction of the endothelial-to-mesenchymal transition, as documented by decreased endothelial and increased mesenchymal marker expression. Induction of the endothelial-to-mesenchymal transition by other strategies similarly increased EC permissiveness to AAV vectors. In vivo injection of AAV vectors in the heart after myocardial infarction resulted in the selective transduction of cells undergoing the endothelial-to-mesenchymal transition, which is known to happen transiently after cardiac ischemia. Collectively, these results point to the endothelial-to-mesenchymal transition as a mechanism for improving AAV transduction in cardiac ECs, with implications for both basic research and the induction of therapeutic angiogenesis in the heart.
{"title":"Endothelial-to-mesenchymal transition enhances permissiveness to AAV vectors in cardiac endothelial cells.","authors":"Nina Volf, Roman Vuerich, Andrea Colliva, Maria Concetta Volpe, Margherita Marengon, Lorena Zentilin, Mauro Giacca, Nadja Anneliese Ruth Ring, Simone Vodret, Luca Braga, Serena Zacchigna","doi":"10.1016/j.ymthe.2024.08.014","DOIUrl":"10.1016/j.ymthe.2024.08.014","url":null,"abstract":"<p><p>A major obstacle in inducing therapeutic angiogenesis in the heart is inefficient gene transfer to endothelial cells (ECs). Here, we identify compounds able to enhance the permissiveness of cardiac ECs to adeno-associated virus (AAV) vectors, which stand as ideal tools for in vivo gene delivery. We screened a library of >1,500 US Food and Drug Administration (FDA)-approved drugs, in combination with AAV vectors, in cardiac ECs. Among the top drugs increasing AAV-mediated transduction, we found vatalanib, an inhibitor of multiple tyrosine kinase receptors. The increased AAV transduction efficiency by vatalanib was paralleled by induction of the endothelial-to-mesenchymal transition, as documented by decreased endothelial and increased mesenchymal marker expression. Induction of the endothelial-to-mesenchymal transition by other strategies similarly increased EC permissiveness to AAV vectors. In vivo injection of AAV vectors in the heart after myocardial infarction resulted in the selective transduction of cells undergoing the endothelial-to-mesenchymal transition, which is known to happen transiently after cardiac ischemia. Collectively, these results point to the endothelial-to-mesenchymal transition as a mechanism for improving AAV transduction in cardiac ECs, with implications for both basic research and the induction of therapeutic angiogenesis in the heart.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142036358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}