Pub Date : 2024-08-01DOI: 10.1016/j.jcyt.2024.03.008
Background aims
Rheumatoid arthritis (RA) is characterized by an overactive immune system, with limited treatment options beyond immunosuppressive drugs or biological response modifiers. Human embryonic stem cell–derived mesenchymal stromal cells (hESC-MSCs) represent a novel alternative, possessing diverse immunomodulatory effects. In this study, we aimed to elucidate the therapeutic effects and underlying mechanisms of hESC-MSCs in treating RA.
Methods
MSC-like cells were differentiated from hESC (hESC-MSCs) and cultured in vitro. Cell proliferation was assessed using Cell Counting Kit-8 assay and Ki-67 staining. Flow cytometry was used to analyze cell surface markers, T-cell proliferation and immune cell infiltration. The collagen-induced arthritis (CIA) mouse model and bleomycin-induced model of lung fibrosis (BLE) were established and treated with hESC-MSCs intravenously for in vivo assessment. Pathological analyses, reverse transcription-quantitative polymerase chain reaction and Western blotting were conducted to evaluate the efficacy of hESC-MSCs treatment.
Results
Intravenous transplantation of hESC-MSCs effectively reduced inflammation in CIA mice in this study. Furthermore, hESC-MSC administration enhanced regulatory T cell infiltration and activation. Additional findings suggest that hESC-MSCs may reduce lung fibrosis in BLE mouse models, indicating their potential to mitigate complications associated with RA progression. In vitro experiments revealed a significant inhibition of T-cell activation and proliferation during co-culture with hESC-MSCs. In addition, hESC-MSCs demonstrated enhanced proliferative capacity compared with traditional primary MSCs.
Conclusions
Transplantation of hESC-MSCs represents a promising therapeutic strategy for RA, potentially regulating T-cell proliferation and differentiation.
背景目的：类风湿性关节炎（RA）的特点是免疫系统过度活跃，除免疫抑制剂或生物反应调节剂外，治疗方案有限。人胚胎干细胞衍生的间充质基质细胞（hESC-MSCs）是一种新的选择，具有多种免疫调节作用。在这项研究中，我们旨在阐明hESC-间充质干细胞治疗RA的疗效和内在机制：方法：从hESC分化出间充质干细胞（hESC-MSCs）并在体外培养。采用细胞计数试剂盒-8测定法和Ki-67染色法评估细胞增殖。流式细胞术用于分析细胞表面标志物、T细胞增殖和免疫细胞浸润。建立了胶原诱导的关节炎（CIA）小鼠模型和博莱霉素诱导的肺纤维化（BLE）模型，并静脉注射 hESC-间充质干细胞进行体内评估。通过病理分析、逆转录-定量聚合酶链反应和 Western 印迹分析来评估 hESC-间充质干细胞的疗效：结果：在本研究中，静脉移植 hESC-间充质干细胞可有效减轻 CIA 小鼠的炎症反应。此外，hESC-间充质干细胞还能增强调节性 T 细胞的浸润和活化。其他研究结果表明，hESC-间充质干细胞可减少BLE小鼠模型的肺纤维化，这表明它们具有减轻与RA进展相关的并发症的潜力。体外实验显示，在与 hESC-MSCs 共同培养过程中，T 细胞的活化和增殖受到明显抑制。此外，与传统的原代间充质干细胞相比，hESC-间充质干细胞的增殖能力更强：结论：hESC-间充质干细胞移植是一种很有前景的治疗 RA 的策略，有可能调节 T 细胞的增殖和分化。
{"title":"Human embryonic stem cell–derived mesenchymal stromal cells suppress inflammation in mouse models of rheumatoid arthritis and lung fibrosis by regulating T-cell function","authors":"","doi":"10.1016/j.jcyt.2024.03.008","DOIUrl":"10.1016/j.jcyt.2024.03.008","url":null,"abstract":"<div><h3>Background aims</h3><p>Rheumatoid arthritis (RA) is characterized by an overactive immune system, with limited treatment options beyond immunosuppressive drugs or biological response modifiers. Human embryonic stem cell–derived mesenchymal stromal cells (hESC-MSCs) represent a novel alternative, possessing diverse immunomodulatory effects. In this study, we aimed to elucidate the therapeutic effects and underlying mechanisms of hESC-MSCs in treating RA.</p></div><div><h3>Methods</h3><p>MSC-like cells were differentiated from hESC (hESC-MSCs) and cultured <em>in vitro</em>. Cell proliferation was assessed using Cell Counting Kit-8 assay and Ki-67 staining. Flow cytometry was used to analyze cell surface markers, T-cell proliferation and immune cell infiltration. The collagen-induced arthritis (CIA) mouse model and bleomycin-induced model of lung fibrosis (BLE) were established and treated with hESC-MSCs intravenously for <em>in vivo</em> assessment. Pathological analyses, reverse transcription-quantitative polymerase chain reaction and Western blotting were conducted to evaluate the efficacy of hESC-MSCs treatment.</p></div><div><h3>Results</h3><p>Intravenous transplantation of hESC-MSCs effectively reduced inflammation in CIA mice in this study. Furthermore, hESC-MSC administration enhanced regulatory T cell infiltration and activation. Additional findings suggest that hESC-MSCs may reduce lung fibrosis in BLE mouse models, indicating their potential to mitigate complications associated with RA progression. <em>In vitro</em> experiments revealed a significant inhibition of T-cell activation and proliferation during co-culture with hESC-MSCs. In addition, hESC-MSCs demonstrated enhanced proliferative capacity compared with traditional primary MSCs.</p></div><div><h3>Conclusions</h3><p>Transplantation of hESC-MSCs represents a promising therapeutic strategy for RA, potentially regulating T-cell proliferation and differentiation.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 8","pages":"Pages 930-938"},"PeriodicalIF":3.7,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140195007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-01DOI: 10.1016/j.jcyt.2024.03.012
Background
Deficits in T cell immunity translate into increased risk of severe viral infection in recipients of solid organ and hematopoietic cell transplants. Thus, therapeutic strategies that employ the adoptive transfer of virus-specific T cells are being clinically investigated to treat and prevent viral diseases in these highly immunocompromised patients. Posoleucel is an off-the-shelf multivirus-specific T cell investigational product for the treatment and prevention of infections due to adenovirus, BK virus, cytomegalovirus, Epstein–Barr virus, human herpesvirus 6 or JC virus.
Methods
Herein we perform extensive characterization of the phenotype and functional profile of posoleucel to illustrate the cellular properties that may contribute to its in vivo activity.
Results and Conclusions
Our results demonstrate that posoleucel is enriched for central and effector memory CD4+ and CD8+ T cells with specificity for posoleucel target viruses and expressing a broad repertoire of T cell receptors. Antigen-driven upregulation of cell-surface molecules and production of cytokine and effector molecules indicative of proliferation, co-stimulation, and cytolytic potential demonstrate the specificity of posoleucel and its potential to mount a broad, polyfunctional, and effective Th1-polarized antiviral response upon viral exposure. We also show the low risk for off-target and nonspecific effects as evidenced by the enrichment of posoleucel in memory T cells, low frequency of naive T cells, and lack of demonstrated alloreactivity in vitro. The efficacy of posoleucel is being explored in four placebo-controlled clinical trials in transplant recipients to treat and prevent viral infections (NCT05179057, NCT05305040, NCT04390113, NCT04605484).
T 细胞免疫缺陷会增加接受实体器官移植和造血细胞移植者感染严重病毒的风险。因此，临床上正在研究采用病毒特异性 T 细胞收养性转移的治疗策略，以治疗和预防这些免疫功能高度低下患者的病毒性疾病。Posoleucel 是一种现成的多病毒特异性 T 细胞研究产品，用于治疗和预防由腺病毒、BK 病毒、巨细胞病毒、爱泼斯坦-巴氏病毒、人类疱疹病毒 6 或 JC 病毒引起的感染。在此，我们对 posoleucel 的表型和功能特征进行了广泛的描述，以说明可能有助于其体内活性的细胞特性。我们的研究结果表明，posoleucel富含对posoleucel靶病毒具有特异性的中枢和效应记忆CD4和CD8 T细胞，并表达广泛的T细胞受体。抗原驱动的细胞表面分子上调以及细胞因子和效应分子的产生表明了posoleucel的增殖、协同刺激和细胞溶解潜力，这证明了posoleucel的特异性及其在病毒暴露时启动广泛、多功能和有效的Th1极化抗病毒反应的潜力。我们还发现，posoleucel 在记忆 T 细胞中的富集、幼稚 T 细胞的低频率以及体外异体活性的缺乏都证明了它产生脱靶和非特异性效应的风险很低。目前正在四项安慰剂对照临床试验中探讨posoleucel在移植受者中治疗和预防病毒感染的疗效（NCT05179057、NCT05305040、NCT04390113、NCT04605484）。
{"title":"Phenotypic and functional characterization of posoleucel, a multivirus-specific T cell therapy for the treatment and prevention of viral infections in immunocompromised patients","authors":"","doi":"10.1016/j.jcyt.2024.03.012","DOIUrl":"10.1016/j.jcyt.2024.03.012","url":null,"abstract":"<div><h3>Background</h3><p>Deficits in T cell immunity translate into increased risk of severe viral infection in recipients of solid organ and hematopoietic cell transplants. Thus, therapeutic strategies that employ the adoptive transfer of virus-specific T cells are being clinically investigated to treat and prevent viral diseases in these highly immunocompromised patients. Posoleucel is an off-the-shelf multivirus-specific T cell investigational product for the treatment and prevention of infections due to adenovirus, BK virus, cytomegalovirus, Epstein–Barr virus, human herpesvirus 6 or JC virus.</p></div><div><h3>Methods</h3><p>Herein we perform extensive characterization of the phenotype and functional profile of posoleucel to illustrate the cellular properties that may contribute to its in vivo activity.</p></div><div><h3>Results and Conclusions</h3><p>Our results demonstrate that posoleucel is enriched for central and effector memory CD4<sup>+</sup> and CD8<sup>+</sup> T cells with specificity for posoleucel target viruses and expressing a broad repertoire of T cell receptors. Antigen-driven upregulation of cell-surface molecules and production of cytokine and effector molecules indicative of proliferation, co-stimulation, and cytolytic potential demonstrate the specificity of posoleucel and its potential to mount a broad, polyfunctional, and effective Th1-polarized antiviral response upon viral exposure. We also show the low risk for off-target and nonspecific effects as evidenced by the enrichment of posoleucel in memory T cells, low frequency of naive T cells, and lack of demonstrated alloreactivity <em>in vitro</em>. The efficacy of posoleucel is being explored in four placebo-controlled clinical trials in transplant recipients to treat and prevent viral infections (NCT05179057, NCT05305040, NCT04390113, NCT04605484).</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 8","pages":"Pages 869-877"},"PeriodicalIF":3.7,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140199790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-01DOI: 10.1016/j.jcyt.2024.03.484
CAR-T cell therapies have been successful in treating numerous hematologic malignancies as the T cell can be engineered to target a specific antigen associated with the disease. However, translating CAR-T cell therapies for solid cancers is proving more challenging due to the lack of truly tumor-associated antigens and the high risk of off-target toxicities. To combat this, numerous synthetic biology mechanisms are being incorporated to create safer and more specific CAR-T cells that can be spatiotemporally controlled with increased precision. Here, we seek to summarize and analyze the advancements for CAR-T cell therapies with respect to clinical implementation, from the perspective of synthetic biology and immunology. This review should serve as a resource for further investigation and growth within the field of personalized cellular therapies.
CAR-T 细胞疗法已成功治疗了多种血液系统恶性肿瘤，因为 T 细胞可被设计为靶向与疾病相关的特定抗原。然而，由于缺乏真正的肿瘤相关抗原以及脱靶毒性的高风险，将CAR-T细胞疗法应用于实体瘤证明更具挑战性。为了解决这一问题，人们正在采用多种合成生物学机制来制造更安全、更具特异性的CAR-T细胞，这些细胞可以在时空上得到更精确的控制。在此，我们试图从合成生物学和免疫学的角度总结和分析 CAR-T 细胞疗法在临床应用方面的进展。这篇综述将为个性化细胞疗法领域的进一步研究和发展提供资源。
{"title":"Synthetic biology approaches for enhancing safety and specificity of CAR-T cell therapies for solid cancers","authors":"","doi":"10.1016/j.jcyt.2024.03.484","DOIUrl":"10.1016/j.jcyt.2024.03.484","url":null,"abstract":"<div><p>CAR-T cell therapies have been successful in treating numerous hematologic malignancies as the T cell can be engineered to target a specific antigen associated with the disease. However, translating CAR-T cell therapies for solid cancers is proving more challenging due to the lack of truly tumor-associated antigens and the high risk of off-target toxicities. To combat this, numerous synthetic biology mechanisms are being incorporated to create safer and more specific CAR-T cells that can be spatiotemporally controlled with increased precision. Here, we seek to summarize and analyze the advancements for CAR-T cell therapies with respect to clinical implementation, from the perspective of synthetic biology and immunology. This review should serve as a resource for further investigation and growth within the field of personalized cellular therapies.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 8","pages":"Pages 842-857"},"PeriodicalIF":3.7,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1465324924005760/pdfft?md5=8dc944a8230a4ed5c9789e7300ad25fc&pid=1-s2.0-S1465324924005760-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140399740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-30DOI: 10.1016/j.jcyt.2024.07.014
Thong Teck Tan, Ruenn Chai Lai, Wei Kian Sim, Bin Zhang, Sai Kiang Lim
Mesenchymal stem/stromal cells (MSC) have displayed promising therapeutic potential. Nonetheless, no United States Food and Drug Administration (FDA)-approved MSC product exists due largely to the absence of a reliable potency assay based on the mechanisms of action to ensure consistent efficacy. MSCs are now thought to exert their effects primarily by releasing small extracellular vesicles (sEVs) of 50-200 nm. While non-living MSC-sEV drugs offer distinct advantages over larger, living MSC drugs, elucidating their mechanism of action to develop robust potency assays remains a challenge. A pivotal prelude to elucidating the mechanism of action for MSC-sEVs is how extracellular vesicles (EVs) engage their primary target cells. Given the inherent inefficiencies of processes such as endocytosis, endosomal escape and EV uncoating during cellular internalization, we propose an alternative EV-cell engagement: EMCEV (Extracellular Modulation of Cells by EV). This approach involves extracellular modulation by EV attributes to generate signaling/inhibitory molecules that have the potential to affect many cells within the vicinity, thereby eliciting a more widespread tissue response.
间充质干细胞/基质细胞（MSC）已显示出良好的治疗潜力。然而，美国食品和药物管理局（FDA）尚未批准间充质干细胞产品，主要原因是缺乏基于作用机制的可靠药效检测方法来确保疗效的一致性。目前认为间充质干细胞主要通过释放 50-200 纳米的细胞外小泡（sEVs）来发挥药效。虽然非活体间充质干细胞-sEV 药物与较大的活体间充质干细胞药物相比具有明显优势，但阐明其作用机制以开发可靠的药效测定方法仍是一项挑战。阐明间充质干细胞-sEV 作用机制的关键前奏是细胞外囊泡 (EV) 如何与主要靶细胞结合。鉴于细胞内化过程中的内吞、内逸和EV解包裹等过程固有的低效率，我们提出了另一种EV-细胞啮合方式：EMCEV（EV 对细胞的胞外调制）。这种方法涉及通过 EV 属性进行细胞外调节，以产生信号/抑制分子，这些分子有可能影响附近的许多细胞，从而引起更广泛的组织反应。
{"title":"Enhancing EV-cell communication through \"External Modulation of Cell by EV\" (EMCEV).","authors":"Thong Teck Tan, Ruenn Chai Lai, Wei Kian Sim, Bin Zhang, Sai Kiang Lim","doi":"10.1016/j.jcyt.2024.07.014","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.07.014","url":null,"abstract":"<p><p>Mesenchymal stem/stromal cells (MSC) have displayed promising therapeutic potential. Nonetheless, no United States Food and Drug Administration (FDA)-approved MSC product exists due largely to the absence of a reliable potency assay based on the mechanisms of action to ensure consistent efficacy. MSCs are now thought to exert their effects primarily by releasing small extracellular vesicles (sEVs) of 50-200 nm. While non-living MSC-sEV drugs offer distinct advantages over larger, living MSC drugs, elucidating their mechanism of action to develop robust potency assays remains a challenge. A pivotal prelude to elucidating the mechanism of action for MSC-sEVs is how extracellular vesicles (EVs) engage their primary target cells. Given the inherent inefficiencies of processes such as endocytosis, endosomal escape and EV uncoating during cellular internalization, we propose an alternative EV-cell engagement: EMCEV (Extracellular Modulation of Cells by EV). This approach involves extracellular modulation by EV attributes to generate signaling/inhibitory molecules that have the potential to affect many cells within the vicinity, thereby eliciting a more widespread tissue response.</p>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142037608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The preclinical efficacy of mesenchymal stem cell (MSC) therapy after intravenous infusion has been promising, but clinical studies have yielded only modest results. Although most preclinical studies have focused solely on the ischemic lung, it is crucial to evaluate both lungs after ischemia-reperfusion injury, considering the various mechanisms involved. This study aimed to bridge this gap by assessing the acute effects of bone marrow MSC(BM) infusion before ischemic insult and evaluating both ischemic and non-ischemic lungs after reperfusion.
Methods
Eighteen male Wistar rats (403 ± 23 g) were anesthetized and mechanically ventilated using a protective strategy. After baseline data collection, the animals were randomized to 3 groups (n = 6/group): (1) SHAM; (2) ischemia-reperfusion (IR), and (3) intravenous MSC(BM) infusion followed by IR. Ischemia was induced by complete clamping of the left hilum, followed by 1 h of reperfusion after clamp removal. At the end of the experiment, the right and left lungs (non-ischemic and ischemic, respectively) were collected for immunohistochemistry and molecular biology analysis.
Results
MSC(BM)s reduced endothelial cell damage and apoptosis markers and improved markers associated with endothelial cell integrity in both lungs. In addition, gene expression of catalase and nuclear factor erythroid 2-related factor 2 increased after MSC(BM) therapy. In the ischemic lung, MSC(BM) therapy mitigated endothelial cell damage and apoptosis and increased gene expression associated with endothelial cell integrity. Conversely, in the non-ischemic lung, apoptosis gene expression increased in the IR group but not after MSC(BM) therapy.
Conclusion
This study demonstrates distinct effects of MSC(BM) therapy on ischemic and non-ischemic lungs after ischemia-reperfusion injury. The findings underscore the importance of evaluating both lung types in ischemia-reperfusion studies, offering insights into the therapeutic potential of MSC(BM) therapy in the context of lung injury.
{"title":"Distinct effects of intravenous bone marrow-derived mesenchymal stem cell therapy on ischemic and non-ischemic lungs after ischemia-reperfusion injury","authors":"Julia Radicetti-Silva , Milena Oliveira , Camila Machado Baldavira , Cassia Lisboa Braga , Renata Trabach Santos , Nathane Santanna Felix , Adriana Lopes Silva , Vera Luiza Capelozzi , Fernanda Ferreira Cruz , Patricia Rieken Macedo Rocco , Pedro Leme Silva","doi":"10.1016/j.jcyt.2024.07.009","DOIUrl":"10.1016/j.jcyt.2024.07.009","url":null,"abstract":"<div><h3>Background</h3><div>The preclinical efficacy of mesenchymal stem cell (MSC) therapy after intravenous infusion has been promising, but clinical studies have yielded only modest results. Although most preclinical studies have focused solely on the ischemic lung, it is crucial to evaluate both lungs after ischemia-reperfusion injury, considering the various mechanisms involved. This study aimed to bridge this gap by assessing the acute effects of bone marrow MSC(BM) infusion before ischemic insult and evaluating both ischemic and non-ischemic lungs after reperfusion.</div></div><div><h3>Methods</h3><div>Eighteen male Wistar rats (403 ± 23 g) were anesthetized and mechanically ventilated using a protective strategy. After baseline data collection, the animals were randomized to 3 groups (<em>n</em> = 6/group): (1) SHAM; (2) ischemia-reperfusion (IR), and (3) intravenous MSC(BM) infusion followed by IR. Ischemia was induced by complete clamping of the left hilum, followed by 1 h of reperfusion after clamp removal. At the end of the experiment, the right and left lungs (non-ischemic and ischemic, respectively) were collected for immunohistochemistry and molecular biology analysis.</div></div><div><h3>Results</h3><div>MSC(BM)s reduced endothelial cell damage and apoptosis markers and improved markers associated with endothelial cell integrity in both lungs. In addition, gene expression of catalase and nuclear factor erythroid 2-related factor 2 increased after MSC(BM) therapy. In the ischemic lung, MSC(BM) therapy mitigated endothelial cell damage and apoptosis and increased gene expression associated with endothelial cell integrity. Conversely, in the non-ischemic lung, apoptosis gene expression increased in the IR group but not after MSC(BM) therapy.</div></div><div><h3>Conclusion</h3><div>This study demonstrates distinct effects of MSC(BM) therapy on ischemic and non-ischemic lungs after ischemia-reperfusion injury. The findings underscore the importance of evaluating both lung types in ischemia-reperfusion studies, offering insights into the therapeutic potential of MSC(BM) therapy in the context of lung injury.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1505-1513"},"PeriodicalIF":3.7,"publicationDate":"2024-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141841613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-25DOI: 10.1016/j.jcyt.2024.07.013
Madeline P. Lauener , Erin Tanaka , Ao Mei , Sayeh Abdossamadi , Elena Ostroumov , Ramon I. Klein Geltink , Subra Malarkannan , Kirk R. Schultz
Background: Chronic graft-versus-host disease (cGvHD) is a major cause of morbidity and mortality after Hematopoietic Stem Cell Transplantation (HSCT). Previously, in large patient cohorts, we identified increased numbers of CD56brightPerforin− regulatory-like NK cells (NKreg-like) associated with cGvHD suppression. Thus, we hypothesized that NKreg-like cells may be a potential candidate for cGvHD cell therapy. Aim: To expand NKreg-like cells while maintaining regulatory phenotype and function. Methods: Total NK cells were first expanded with IL-2, which was then combined with rapamycin, Transforming Growth Factor Beta 1 (TGF-β1), NECA (Adenosine A2A receptor (A2AR) agonist), metformin, or dexamethasone, to prevent change in cell phenotype/function. The functional characteristics were evaluated via T cell suppression assays and the phenotype was measured using flow cytometry. The optimal expansion protocol was compared in terms of function and metabolism for three NK expansion media, and cells from cord vs. peripheral blood. Further, expanded NKreg-like cell gene expression was characterized using bulk RNA sequencing. Finally, NKreg-like cells were differentiated from CD34+ hematopoietic stem and progenitor cells (HSPCs) and compared in terms of proliferation and function. Results: The expansion of total NK cells found the addition of TGF-β1 and/or NECA with the pulsing of rapamycin in IL-2 containing media to prevent NKreg-like differentiation (up to 200-fold expansion). Expanded NKreg-like cells maintained a phenotype, transcriptome, and T cell suppression similar to freshly isolated NKreg-like cells. NKreg-like expansion was greatest in the Immunocult media (up to 300-fold), and NKreg-like cells from peripheral blood demonstrated significantly greater proliferation than cells isolated from cord blood (65-fold). The metabolic profile of NKreg-like and cytolytic NK cells appeared similar at baseline, though rapamycin induced a shift to oxidative over glycolytic metabolism. Further, we demonstrated that suppressive NKreg-like cells may alternatively be expanded from CD34+ cells isolated from cord blood, reaching an average 340-fold expansion. Conclusions: In conclusion, our studies have optimized two alternative expansion approaches for deriving functional NKreg-like cells. Additionally, evaluating the transcriptomic and metabolic characteristics provides useful information regarding NKreg-like cell function and differentiation. With further optimization and in vivo validation, we may work towards preparing these cells as a therapy for cGvHD.
背景：慢性移植物抗宿主疾病（cGvHD）是造血干细胞移植（HSCT）后发病和死亡的主要原因。此前，我们在大型患者队列中发现，CD56bright穿刺素调节样NK细胞（NKreg-like）数量的增加与cGvHD抑制有关。因此，我们推测NKreg样细胞可能是cGvHD细胞疗法的潜在候选者。目的：扩增 NKreg 样细胞，同时保持调节表型和功能。方法：首先用一种新的 NK 细胞扩增试剂盒扩增总 NK 细胞：首先用IL-2扩增总NK细胞，然后与雷帕霉素、转化生长因子β1（TGF-β1）、NECA（腺苷A2A受体（A2AR）激动剂）、二甲双胍或地塞米松联合使用，以防止细胞表型/功能改变。功能特征通过 T 细胞抑制实验进行评估，表型则通过流式细胞术进行测量。比较了三种 NK 扩增培养基的功能和代谢，以及脐带血和外周血细胞的最佳扩增方案。此外，还使用大量 RNA 测序鉴定了扩增的 NKreg 样细胞基因表达。最后，从 CD34+ 造血干细胞和祖细胞（HSPCs）分化出 NKreg 样细胞，并对其增殖和功能进行比较。结果在扩增总 NK 细胞时发现，在含有 IL-2 的培养基中加入 TGF-β1 和/或 NECA 以及雷帕霉素脉冲可防止 NKreg 样分化（最高扩增 200 倍）。扩增的 NKreg 样细胞保持了与新鲜分离的 NKreg 样细胞相似的表型、转录组和 T 细胞抑制。NKreg样细胞在免疫培养基中的扩增最大（高达300倍），外周血中的NKreg样细胞的增殖明显高于从脐带血中分离的细胞（65倍）。NKreg 样细胞和细胞溶解 NK 细胞的新陈代谢特征在基线时相似，但雷帕霉素诱导的新陈代谢转向氧化，而不是糖酵解。此外，我们还证明，抑制性 NKreg 样细胞也可以从脐带血中分离的 CD34+ 细胞中扩增，平均扩增倍数为 340 倍。结论总之，我们的研究优化了获得功能性 NKreg 样细胞的两种替代扩增方法。此外，对转录组和代谢特征的评估提供了有关 NKreg 样细胞功能和分化的有用信息。通过进一步优化和体内验证，我们可能会将这些细胞作为治疗 cGvHD 的一种疗法。
{"title":"Expansion and characterization of immune suppressive CD56(bright)Perforin(-) regulatory-like natural killer cells in chronic graft-versus-host disease","authors":"Madeline P. Lauener , Erin Tanaka , Ao Mei , Sayeh Abdossamadi , Elena Ostroumov , Ramon I. Klein Geltink , Subra Malarkannan , Kirk R. Schultz","doi":"10.1016/j.jcyt.2024.07.013","DOIUrl":"10.1016/j.jcyt.2024.07.013","url":null,"abstract":"<div><div>Background: Chronic graft-versus-host disease (cGvHD) is a major cause of morbidity and mortality after Hematopoietic Stem Cell Transplantation (HSCT). Previously, in large patient cohorts, we identified increased numbers of CD56<sup>bright</sup>Perforin<sup>−</sup> regulatory-like NK cells (NK<sub>reg</sub>-like) associated with cGvHD suppression. Thus, we hypothesized that NK<sub>reg</sub>-like cells may be a potential candidate for cGvHD cell therapy. Aim: To expand NK<sub>reg</sub>-like cells while maintaining regulatory phenotype and function. Methods: Total NK cells were first expanded with IL-2, which was then combined with rapamycin, Transforming Growth Factor Beta 1 (TGF-β1), NECA (Adenosine A2A receptor (A2AR) agonist), metformin, or dexamethasone, to prevent change in cell phenotype/function. The functional characteristics were evaluated via T cell suppression assays and the phenotype was measured using flow cytometry. The optimal expansion protocol was compared in terms of function and metabolism for three NK expansion media, and cells from cord vs. peripheral blood. Further, expanded NK<sub>reg</sub>-like cell gene expression was characterized using bulk RNA sequencing. Finally, NK<sub>reg</sub>-like cells were differentiated from CD34<sup>+</sup> hematopoietic stem and progenitor cells (HSPCs) and compared in terms of proliferation and function. Results: The expansion of total NK cells found the addition of TGF-β1 and/or NECA with the pulsing of rapamycin in IL-2 containing media to prevent NK<sub>reg</sub>-like differentiation (up to 200-fold expansion). Expanded NK<sub>reg</sub>-like cells maintained a phenotype, transcriptome, and T cell suppression similar to freshly isolated NK<sub>reg</sub>-like cells. NK<sub>reg</sub>-like expansion was greatest in the Immunocult media (up to 300-fold), and NK<sub>reg</sub>-like cells from peripheral blood demonstrated significantly greater proliferation than cells isolated from cord blood (65-fold). The metabolic profile of NK<sub>reg</sub>-like and cytolytic NK cells appeared similar at baseline, though rapamycin induced a shift to oxidative over glycolytic metabolism. Further, we demonstrated that suppressive NK<sub>reg</sub>-like cells may alternatively be expanded from CD34<sup>+</sup> cells isolated from cord blood, reaching an average 340-fold expansion. Conclusions: In conclusion, our studies have optimized two alternative expansion approaches for deriving functional NK<sub>reg</sub>-like cells. Additionally, evaluating the transcriptomic and metabolic characteristics provides useful information regarding NK<sub>reg</sub>-like cell function and differentiation. With further optimization and <em>in vivo</em> validation, we may work towards preparing these cells as a therapy for cGvHD.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1472-1483"},"PeriodicalIF":3.7,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141841086","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-25DOI: 10.1016/j.jcyt.2024.07.011
Bin Xue , Yifan Liu , Jie Zhou , Lili Zhou , Shiguang Ye , Yan Lu , Wenjun Zhang , Bing Xiu , Aibin Liang , Ping Li , Ying Lu , Wenbin Qian , Xiu Luo
Object
Autologous CD19 chimeric antigen receptor T-cell therapy (CAR-T) significantly modifies the natural course of chemorefractory diffuse large B-cell lymphoma (DLBCL). However, 25% to 50% of patients with relapsed/refractory DLBCL still do not achieve remission. Therefore, investigating new molecular prognostic indicators that affect the effectiveness of CAR-T for DLBCL and developing novel combination therapies are crucial.
Methods
Data from 73 DLBCL patients who received CD19 CAR-T (Axi-cel or Relma-cel) were retrospectively collected from Shanghai Tongji Hospital of Tongji University, The Second Affiliated Hospital Zhejiang University School of Medicine, and The Affiliated People's Hospital of Ningbo University. Prior to CD19 CAR-T-cell transfusions, the patients received fludarabine and cyclophosphamide chemotherapy regimen.
Results
Our study revealed that relapsed/refractory diffuse large B-cell lymphoma (r/r DLBCL) patients with both Double-expression (MYC > 40% and BCL2 > 50%) and TP53 alterations tend to have a poorer clinical prognosis after CAR-T therapy, even when CAR-T therapy is used in combination with other therapies. However, CAR-T therapy was found to be effective in patients with only TP53 alterations or DE status, suggesting that their prognosis is in line with that of patients without TP53 alterations or DE status.
Conclusions
Our study suggests that r/r DLBCL patients with both DE status and TP53 alterations treated with CAR-T therapy are more likely to have a poorer clinical prognosis. However, CAR-T therapy has the potential to improve the prognosis of patients with only TP53 alterations or DE status to be similar to that of patients without these abnormalities.
{"title":"CD19 CAR-T treatment shows limited efficacy in r/r DLBCL with double expression and TP53 alterations","authors":"Bin Xue , Yifan Liu , Jie Zhou , Lili Zhou , Shiguang Ye , Yan Lu , Wenjun Zhang , Bing Xiu , Aibin Liang , Ping Li , Ying Lu , Wenbin Qian , Xiu Luo","doi":"10.1016/j.jcyt.2024.07.011","DOIUrl":"10.1016/j.jcyt.2024.07.011","url":null,"abstract":"<div><h3>Object</h3><div>Autologous CD19 chimeric antigen receptor T-cell therapy (CAR-T) significantly modifies the natural course of chemorefractory diffuse large B-cell lymphoma (DLBCL). However, 25% to 50% of patients with relapsed/refractory DLBCL still do not achieve remission. Therefore, investigating new molecular prognostic indicators that affect the effectiveness of CAR-T for DLBCL and developing novel combination therapies are crucial.</div></div><div><h3>Methods</h3><div>Data from 73 DLBCL patients who received CD19 CAR-T (Axi-cel or Relma-cel) were retrospectively collected from Shanghai Tongji Hospital of Tongji University, The Second Affiliated Hospital Zhejiang University School of Medicine, and The Affiliated People's Hospital of Ningbo University. Prior to CD19 CAR-T-cell transfusions, the patients received fludarabine and cyclophosphamide chemotherapy regimen.</div></div><div><h3>Results</h3><div>Our study revealed that relapsed/refractory diffuse large B-cell lymphoma (r/r DLBCL) patients with both Double-expression (MYC > 40% and BCL2 > 50%) and TP53 alterations tend to have a poorer clinical prognosis after CAR-T therapy, even when CAR-T therapy is used in combination with other therapies. However, CAR-T therapy was found to be effective in patients with only TP53 alterations or DE status, suggesting that their prognosis is in line with that of patients without TP53 alterations or DE status.</div></div><div><h3>Conclusions</h3><div>Our study suggests that r/r DLBCL patients with both DE status and TP53 alterations treated with CAR-T therapy are more likely to have a poorer clinical prognosis. However, CAR-T therapy has the potential to improve the prognosis of patients with only TP53 alterations or DE status to be similar to that of patients without these abnormalities.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1465-1471"},"PeriodicalIF":3.7,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141838430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-24DOI: 10.1016/j.jcyt.2024.07.010
Jing Zhang, Dong Lin, Huihua Hu, Haipeng Xu
Objective
The currently available biomarkers are insufficient to accurately predict the immunotherapy response in patients. This work attempted to investigate effects of PD-1/PD-L1 interaction score combined with NKT-like cell infiltration level in tumor microenvironment on predicting immunotherapy efficacy.
Methods
24 non-small cell lung cancer (NSCLC) patients who underwent immunotherapy were analyzed using multiplex immunofluorescence to quantitatively assess positive cells of target biomarkers and their spatial localization. Correlation between PD-1/PD-L1 interaction score in combination with NKT-like cell infiltration level and immunotherapy response was analyzed. The predictive performance of two individual biomarkers and combined novel biomarkers in immunotherapy efficacy was assessed through receiver operating characteristic curve analysis. Relationships between these factors and patient survival prognosis were analyzed using Kaplan–Meier curves.
Results
Among responders, PD-1/PD-L1 interaction score and NKT-like cell infiltration level were significantly higher than nonresponders (P < 0.05), and PD-1/PD-L1 interaction score and NKT-like cell infiltration level could effectively identify the population with immunotherapy response, with area under the curves (AUCs) of 0.7571 and 0.8643, respectively. Combination of the two had the best performance in predicting the efficacy of immunotherapy (AUC = 0.9070). High PD-1/PD-L1 interaction scores and high levels of NKT-like cell infiltration significantly improved progression-free survival (HR = 0.2544, P = 0.0053) and overall survival (HR = 0.2820, P = 0.0053) in patients.
Conclusions
Combination of PD-1/PD-L1 interaction score and NKT-like cell infiltration level had favorable performance in predicting immunotherapy response in NSCLC patients, contributing to accurately identify patients who may benefit from immunotherapy.
{"title":"PD-1/PD-L1 interaction score and NKT-like cell infiltration predict immunotherapy efficacy in non-small cell lung cancer patients","authors":"Jing Zhang, Dong Lin, Huihua Hu, Haipeng Xu","doi":"10.1016/j.jcyt.2024.07.010","DOIUrl":"10.1016/j.jcyt.2024.07.010","url":null,"abstract":"<div><h3>Objective</h3><div>The currently available biomarkers are insufficient to accurately predict the immunotherapy response in patients. This work attempted to investigate effects of PD-1/PD-L1 interaction score combined with NKT-like cell infiltration level in tumor microenvironment on predicting immunotherapy efficacy.</div></div><div><h3>Methods</h3><div>24 non-small cell lung cancer (NSCLC) patients who underwent immunotherapy were analyzed using multiplex immunofluorescence to quantitatively assess positive cells of target biomarkers and their spatial localization. Correlation between PD-1/PD-L1 interaction score in combination with NKT-like cell infiltration level and immunotherapy response was analyzed. The predictive performance of two individual biomarkers and combined novel biomarkers in immunotherapy efficacy was assessed through receiver operating characteristic curve analysis. Relationships between these factors and patient survival prognosis were analyzed using Kaplan–Meier curves.</div></div><div><h3>Results</h3><div>Among responders, PD-1/PD-L1 interaction score and NKT-like cell infiltration level were significantly higher than nonresponders (<em>P</em> < 0.05), and PD-1/PD-L1 interaction score and NKT-like cell infiltration level could effectively identify the population with immunotherapy response, with area under the curves (AUCs) of 0.7571 and 0.8643, respectively. Combination of the two had the best performance in predicting the efficacy of immunotherapy (AUC = 0.9070). High PD-1/PD-L1 interaction scores and high levels of NKT-like cell infiltration significantly improved progression-free survival (HR = 0.2544, <em>P</em> = 0.0053) and overall survival (HR = 0.2820, <em>P</em> = 0.0053) in patients.</div></div><div><h3>Conclusions</h3><div>Combination of PD-1/PD-L1 interaction score and NKT-like cell infiltration level had favorable performance in predicting immunotherapy response in NSCLC patients, contributing to accurately identify patients who may benefit from immunotherapy.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1484-1490"},"PeriodicalIF":3.7,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141846436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-24DOI: 10.1016/j.jcyt.2024.07.012
Xiaohua Wang , Jing Zang , Yinxiang Yang , Ke Li , Dou Ye , Zhaoyan Wang , Qian Wang , Youjia Wu , Zuo Luan
Aims
Cerebral palsy (CP) is the most common physical disability in children, yet lacks an ideal animal model or effective treatment. This study aimed to develop a reliable CP model in neonatal rats and explore the effectiveness and underlying mechanisms of human neural stem cells (hNSCs) transplantation during the sequelae phase of CP.
Methods
Vasoconstrictor endothelin-1 (ET-1) was administered intracranially to the motor cortex and striatum of rats on postnatal day 5 to establish a CP model. hNSCs (5 × 105/5 μL) pretreated with hypoxia (5% O2 for 24 h) were transplanted near the infarct 3 weeks after ET-1 injury (the sequelae phase). The distribution and differentiation of hNSCs were observed after transplantation. Changes in neurotrophic factors, neurogenesis, angiogenesis, axonal plasticity, and motor function were analyzed.
Results
Neurobehavioral tests showed poor muscle strength and postural control in young ET-1 rats. Motor deficits of the left forelimb and gait abnormalities persisted into adulthood. Histopathological findings and MRI indicated the atrophy of the cortex, striatum, and adjacent corpus callosum in ET-1 rats. At 56 days after transplantation, hNSCs were widely distributed in the ipsilateral hemisphere, and differentiated into neurons, oligodendrocytes and astrocytes. Transplantation of hNSCs increased BDNF and VEGF expression, EdU+ cell number in the SVZ area, RECA-1+ vessel density and GAP-43 intensity around the lesion in ET-1 rats. The cylinder test revealed a significant increase in the left forelimb motor function from 28 days after transplantation, and the staircase and CatWalk tests showed improvements in fine motor function and gait parameters.
Conclusions
Intracerebral injection of ET-1 modelled key functional and histopathological features of CP. hNSCs transplanted during the sequelae phase of CP resulted in long-term improvement in motor performance, possibly attributed to its capacity to stimulate neurotrophic factors, facilitate neurogenesis, angiogenesis, and promote axonal plasticity.
{"title":"Human neural stem cells transplanted during the sequelae phase alleviate motor deficits in a rat model of cerebral palsy","authors":"Xiaohua Wang , Jing Zang , Yinxiang Yang , Ke Li , Dou Ye , Zhaoyan Wang , Qian Wang , Youjia Wu , Zuo Luan","doi":"10.1016/j.jcyt.2024.07.012","DOIUrl":"10.1016/j.jcyt.2024.07.012","url":null,"abstract":"<div><h3>Aims</h3><div>Cerebral palsy (CP) is the most common physical disability in children, yet lacks an ideal animal model or effective treatment. This study aimed to develop a reliable CP model in neonatal rats and explore the effectiveness and underlying mechanisms of human neural stem cells (hNSCs) transplantation during the sequelae phase of CP.</div></div><div><h3>Methods</h3><div>Vasoconstrictor endothelin-1 (ET-1) was administered intracranially to the motor cortex and striatum of rats on postnatal day 5 to establish a CP model. hNSCs (5 × 10<sup>5</sup>/5 μL) pretreated with hypoxia (5% O<sub>2</sub> for 24 h) were transplanted near the infarct 3 weeks after ET-1 injury (the sequelae phase). The distribution and differentiation of hNSCs were observed after transplantation. Changes in neurotrophic factors, neurogenesis, angiogenesis, axonal plasticity, and motor function were analyzed.</div></div><div><h3>Results</h3><div>Neurobehavioral tests showed poor muscle strength and postural control in young ET-1 rats. Motor deficits of the left forelimb and gait abnormalities persisted into adulthood. Histopathological findings and MRI indicated the atrophy of the cortex, striatum, and adjacent corpus callosum in ET-1 rats. At 56 days after transplantation, hNSCs were widely distributed in the ipsilateral hemisphere, and differentiated into neurons, oligodendrocytes and astrocytes. Transplantation of hNSCs increased BDNF and VEGF expression, EdU<sup>+</sup> cell number in the SVZ area, RECA-1<sup>+</sup> vessel density and GAP-43 intensity around the lesion in ET-1 rats. The cylinder test revealed a significant increase in the left forelimb motor function from 28 days after transplantation, and the staircase and CatWalk tests showed improvements in fine motor function and gait parameters.</div></div><div><h3>Conclusions</h3><div>Intracerebral injection of ET-1 modelled key functional and histopathological features of CP. hNSCs transplanted during the sequelae phase of CP resulted in long-term improvement in motor performance, possibly attributed to its capacity to stimulate neurotrophic factors, facilitate neurogenesis, angiogenesis, and promote axonal plasticity.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1491-1504"},"PeriodicalIF":3.7,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141840003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-14DOI: 10.1016/j.jcyt.2024.07.008
Pauline De Berdt , Elodie Deltour , Eric Pauly , Noelia Gordillo , Frédéric Lin , Etienne Sokal , Mustapha Najimi
Human allogeneic liver-derived progenitor cells (HALPCs) display advanced ability to differentiate into hepatocyte-like cells and exhibit potent immunomodulatory, anti-inflammatory, and anti-fibrotic properties. HALPCs have been successfully manufactured under good manufacturing practice (GMP) and are currently in clinical development. A previous phase 2a trial demonstrated the safety of peripheral intravenous infusions of HALPCs and preliminary evidence of the cells’ properties to restore liver function in patients with acute-on-chronic liver failure (ACLF), thus potentially improving their survival. A phase 2b trial is currently ongoing across multiple centers (NCT04229901) to obtain proof-of-concept on efficacy and additional safety. HALPCs are currently manufactured using fetal bovine serum (FBS), which can reveal qualitative and quantitative variations between batches. The use of serum-free medium (SFM) represents an alternative means to overcome this variability while also complying fully with regulations. The aim of this study was to compare current FBS-containing culture conditions with two industry-available GMP-compliant SFMs: StemMACS (Miltenyi Biotec, Bergisch Gladbach, Germany) and PRIME-XV (FUJIFILM Irvine Scientific, Santa Ana, California, USA).
The proliferation of HALPCs was significantly stimulated by both SFMs, which shortened both their emergence period and population doubling time. This effect was correlated with a significant improvement in their genetic stability as analyzed by conventional karyotyping. The expression profile (identity and purity) and functionality of HALPCs cultured in SFM were maintained, as demonstrated by flow cytometry and enzyme-linked immunoassay (ELISA), respectively. Their potency, evaluated via prostaglandin E2 (PGE2) secretion, showed a similar effect on CD4+ T-cell proliferation in FBS and SFM conditions. Furthermore, a greater proportion of HALPCs cultured in SFM showed enhanced expression of tissue factor (CD142) compared with the FBS condition.
Altogether, SFM conditions enabled consistent HALPC quality to be achieved without altering their expression and functional profiles.
{"title":"Expansion of human allogeneic liver-derived progenitor cells for liver regenerative therapy in serum-free culture conditions","authors":"Pauline De Berdt , Elodie Deltour , Eric Pauly , Noelia Gordillo , Frédéric Lin , Etienne Sokal , Mustapha Najimi","doi":"10.1016/j.jcyt.2024.07.008","DOIUrl":"10.1016/j.jcyt.2024.07.008","url":null,"abstract":"<div><div>Human allogeneic liver-derived progenitor cells (HALPCs) display advanced ability to differentiate into hepatocyte-like cells and exhibit potent immunomodulatory, anti-inflammatory, and anti-fibrotic properties. HALPCs have been successfully manufactured under good manufacturing practice (GMP) and are currently in clinical development. A previous phase 2a trial demonstrated the safety of peripheral intravenous infusions of HALPCs and preliminary evidence of the cells’ properties to restore liver function in patients with acute-on-chronic liver failure (ACLF), thus potentially improving their survival. A phase 2b trial is currently ongoing across multiple centers (NCT04229901) to obtain proof-of-concept on efficacy and additional safety. HALPCs are currently manufactured using fetal bovine serum (FBS), which can reveal qualitative and quantitative variations between batches. The use of serum-free medium (SFM) represents an alternative means to overcome this variability while also complying fully with regulations. The aim of this study was to compare current FBS-containing culture conditions with two industry-available GMP-compliant SFMs: StemMACS (Miltenyi Biotec, Bergisch Gladbach, Germany) and PRIME-XV (FUJIFILM Irvine Scientific, Santa Ana, California, USA).</div><div>The proliferation of HALPCs was significantly stimulated by both SFMs, which shortened both their emergence period and population doubling time. This effect was correlated with a significant improvement in their genetic stability as analyzed by conventional karyotyping. The expression profile (identity and purity) and functionality of HALPCs cultured in SFM were maintained, as demonstrated by flow cytometry and enzyme-linked immunoassay (ELISA), respectively. Their potency, evaluated via prostaglandin E2 (PGE2) secretion, showed a similar effect on CD4<sup>+</sup> T-cell proliferation in FBS and SFM conditions. Furthermore, a greater proportion of HALPCs cultured in SFM showed enhanced expression of tissue factor (CD142) compared with the FBS condition.</div><div>Altogether, SFM conditions enabled consistent HALPC quality to be achieved without altering their expression and functional profiles.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"26 12","pages":"Pages 1571-1578"},"PeriodicalIF":3.7,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141694883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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