JOURNAL/nrgr/04.03/01300535-202509000-00026/figure1/v/2024-11-05T132919Z/r/image-tiff Neuronal cell death and the loss of connectivity are two of the primary pathological mechanisms underlying Alzheimer’s disease. The accumulation of amyloid-β peptides, a key hallmark of Alzheimer’s disease, is believed to induce neuritic abnormalities, including reduced growth, extension, and abnormal growth cone morphology, all of which contribute to decreased connectivity. However, the precise cellular and molecular mechanisms governing this response remain unknown. In this study, we used an innovative approach to demonstrate the effect of amyloid-β on neurite dynamics in both two-dimensional and three-dimensional culture systems, in order to provide more physiologically relevant culture geometry. We utilized various methodologies, including the addition of exogenous amyloid-β peptides to the culture medium, growth substrate coating, and the utilization of human-induced pluripotent stem cell technology, to investigate the effect of endogenous amyloid-β secretion on neurite outgrowth, thus paving the way for potential future applications in personalized medicine. Additionally, we also explore the involvement of the Nogo signaling cascade in amyloid-β-induced neurite inhibition. We demonstrate that inhibition of downstream ROCK and RhoA components of the Nogo signaling pathway, achieved through modulation with Y-27632 (a ROCK inhibitor) and Ibuprofen (a Rho A inhibitor), respectively, can restore and even enhance neuronal connectivity in the presence of amyloid-β. In summary, this study not only presents a novel culture approach that offers insights into the biological process of neurite growth and inhibition, but also proposes a specific mechanism for reduced neural connectivity in the presence of amyloid-β peptides, along with potential intervention points to restore neurite growth. Thereby, we aim to establish a culture system that has the potential to serve as an assay for measuring preclinical, predictive outcomes of drugs and their ability to promote neurite outgrowth, both generally and in a patient-specific manner.

2025-01-01·ARCHIVES OF ORAL BIOLOGY

The important role of the Wnt/β-catenin signaling pathway in small molecules mediated gingival mesenchymal stem cells transdifferentiate into neuron-like cells

Article

作者: Xu, Wenan ; Zhang, Chuhan ; Lv, Peiyi ; Huang, Yongmao ; Liang, Qiuying ; Zhao, Hang ; Jiang, Shan

OBJECTIVE:

Given their neural crest origin, gingival mesenchymal stem cells (GMSCs) possess high neurogenic potential, which makes them suitable for cell replacement therapy against neurodegenerative diseases. This study investigated whether GMSCs can be transdifferentiated into neurons in vitro using a protocol involving small molecules VCRFY (VPA, CHIR99021, Repsox, Forskolin, and Y-27632). The regulatory mechanisms of key signaling pathways were also investigated.

METHODS:

Neuronal induction of GMSCs was conducted using a small molecules-based protocol over 7 days, which included the evaluation of cell morphology, proliferation, expressions of neurogenic markers, and intracellular calcium oscillation. The activation of canonical the Wnt signaling pathway was assessed by examining the protein content and subcellular localization of β-catenin.

RESULTS:

Small molecules-treated GMSCs displayed neuronal morphology and increased expression of neurogenic markers, including class III beta-tubulin (TUJ1), neuron-specific enolase (NSE), microtube-associated protein 2 (MAP2), and neurofilament medium (NFM), verified through RT-qPCR, western blotting, and immunocytochemistry. Based on the results of Fluo-4 AM calcium flux assay, small molecules-treated GMSCs exhibited enhanced electrophysiological activity. GMSC proliferation halted after 2 days of treatment. Among the small molecules, CHIR99021 exhibited the highest neuronal induction efficiency. Furthermore, activation of the Wnt/β-catenin signaling pathway augmented neuronal differentiation.

CONCLUSIONS:

Small molecule-based cellular reprogramming can efficiently generate neurons from GMSCs, with Wnt/β-catenin signaling to play a critical role in neuronal induction.

2024-12-01·TOXICOLOGY

Bisphenol A disrupts the neuronal F-actin cytoskeleton by activating the RhoA/ROCK/LIMK pathway in Neuro-2a cells

Article

作者: Wang, Chong ; Liu, Qiling ; Zhang, Xuyuan ; Wang, Yuxin ; Ren, Jiajia ; Li, Qian ; Guo, Yi

Bisphenol A (BPA) is an environmental endocrine disruptor that is widely present in the environment and has been reported to affect neuronal cytoskeleton and neural function. However, the exact molecular mechanisms remain unclear. In the present study, the effects of BPA on cytoskeleton rearrangement were examined, and the associated signaling pathways, which were influenced by the RhoA/ROCK/LIMK pathway in Neuro-2a cells in vitro, were identified. Specifically, Neuro-2a cells were exposed to BPA, and the effects of BPA exposure on the cytoskeleton of neuronal cells and on the activation or nonactivation of the RhoA/ROCK signaling pathway were evaluated using Cell Counting Kit-8 (CCK8), phalloidin staining, western blot, and real-time PCR. A RhoA inhibitor (Rhosin hydrochloride) and a ROCK inhibitor (Y-27632) were then used to elucidate the precise function of the pathway. The results demonstrated that 50-100 μM BPA exposure inhibited Neuro-2a cell viability and caused the formation of aberrantly polymerized F-actin and stress fibers. In addition, the RhoA/ROCK pathway was activated, and the expression levels of the pathway-related molecules-RhoA, ROCK2, LIMK1, Cofilin, Profilin, p-MLC2, and F-actin were dramatically elevated. The addition of Rhosin and Y-27632 resulted in a decrease in F-actin polymerization in the Neuro-2a cells, the disassembly of stress fibers, and a noteworthy drop in the levels of molecular proteins related to the RhoA/ROCK pathway affected by BPA. Together, these new findings indicated that BPA exposure thus activated the RhoA/ROCK signaling pathway and caused an abnormal accumulation of F-actin in the Neuro-2a cells, in turn altering the microfilament cytoskeleton. F-actin was restored when the RhoA/ROCK pathway was inhibited, suggesting that the process of BPA-induced neuronal cytoskeletal degradation is linked to the RhoA/ROCK signaling cascade.

项与 Y-27632 (Mitsubishi) 相关的新闻（医药）

2024-12-18

·Business Wire

Aurion Biotech Announces Positive Topline Data for Phase 1/2 Clinical Trial of AURN001, an Allogeneic Cell Therapy Product Candidate for the Treatment of Corneal Edema Secondary to Corneal Endothelial Dysfunction

SEATTLE & CAMBRIDGE, Mass. & TOKYO--( BUSINESS WIRE )-- Aurion Biotech , Inc. (Aurion Biotech), whose mission is to restore vision to millions of patients with life-changing regenerative therapies, today announced topline data from its Phase 1/2 clinical trial (CLARA) of AURN001, an allogeneic cell therapy product candidate for the treatment of corneal edema secondary to corneal endothelial dysfunction. AURN001 is a combination cell therapy product candidate comprised of allogeneic human corneal endothelial cells ( neltependocel ) and a rho kinase inhibitor (Y-27632). AURN001 is intended to be administered to the anterior chamber of the eye as a one-time procedure. The CLARA Phase 1/2 clinical trial ( NCT06041256 ) is a prospective, multi-center, randomized, double-masked, parallel-arm dose-ranging clinical trial designed to assess the safety, tolerability, and efficacy of AURN001 for the treatment of corneal edema secondary to corneal endothelial dysfunction. Ninety-seven subjects were randomized at US and Canadian sites to each of the following five arms: AURN001: 1.0 × 10 6 neltependocel + 100 μM Y-27632 (high cell dose) AURN001: 5.0 × 10 5 neltependocel + 100 μM Y-27632 (medium cell dose) AURN001: 2.5 × 10 5 neltependocel + 100 μM Y-27632 (low cell dose) 100 μM Y-27632 1.0 × 10 6 neltependocel At baseline across all arms, the mean age was 71.4 years, with 55% being female subjects. Baseline mean BCVA was 53.5 letters, using the Early Treatment Diabetic Retinopathy Standard (ETDRS) visual acuity test (20/85 Snellen), and CCT was 676.6 microns. The primary endpoint was the proportion of responders with a > 15-letter improvement ( > 3-line gain) from baseline in BCVA at six months. Key secondary endpoints included (i) change from baseline in BCVA and in CCT at six months and (ii) safety and tolerability. For the primary endpoint, a dose-dependent response was observed in the three AURN001 arms, with a statistically significant improvement in the high-dose AURN001 arm (50% of responders, p=0.020), as compared to the Y-27632-only arm (14.3%). A key secondary endpoint, change in BCVA at six months, showed a statistically significant improvement for the high-dose AURN001 arm (p=0.002), as compared to the Y-27632-only arm (using the full analysis set population (LOCF, LS mean 2 )). Additionally, a dose response was observed in the three AURN001 arms. For the key secondary endpoint, change in CCT at six months, there was a statistically significant improvement for the high-dose AURN001 arm (p=0.012), as compared to the Y-27632-only arm (using the full analysis set population (LOCF, LS Mean)). Separately, an improvement in patient-reported quality of life, assessed using the Visual Function Questionnaire (VFQ-25), was noted with the greatest benefit being seen in the high-dose AURN001 arm. Doses from all five treatment arms were generally well tolerated with favorable safety profiles. There was no dose relationship observed in frequency of types of adverse events (AEs). There were no ocular serious adverse events (SAEs) reported and two non-ocular SAEs (hip fracture and femur fracture). The most frequently reported (>3%) ocular TEAEs were ocular hypertension (9.3%), conjunctival hemorrhage (5.2%), eye pain (4.1%), and cystoid macular edema (3.1%) and the most frequently reported non-ocular TEAE was COVID-19 (3.1%). “We are thrilled with the topline results of the CLARA trial,” said Michael Goldstein, M.D., M.B.A., president and chief medical officer of Aurion Biotech. “We were especially pleased that in the high-dose AURN001 arm at six months, there was a statistically significant improvement in the primary endpoint. Based on these findings, combined with the generally favorable safety profile in the CLARA trial, we look forward to bringing the high dose of AURN001 forward into our proposed Phase 3 pivotal trials.” “We believe that today’s news is another important step forward in the clinical development of our investigational allogeneic cell therapy, AURN001, to help restore vision,” said Greg Kunst, chief executive officer of Aurion Biotech. “We look forward to presenting full results from the CLARA trial at future medical conferences.” Topline data from the CLARA study follows Aurion Biotech’s announcement in June 2024 that the U.S. Food and Drug Administration (FDA) granted AURN001 both Breakthrough Therapy Designation (BTD) and Regenerative Medicine Advanced Therapy (RMAT) designation, and Aurion’s commercial launch of its cell therapy in Japan, under the trade name, Vyznova TM , in September 2024. More information about Aurion Biotech’s recent progress can be found on its website: www.aurionbiotech.com/news . About Corneal Endothelial Dysfunction Corneal edema secondary to corneal endothelial dysfunction is a sight-threatening condition affecting millions of people throughout the world. When corneal endothelial cells die or degrade, they do not regenerate in vivo . If left untreated, corneal endothelial cell loss can cause corneal edema (swelling) and loss of vision. Although corneal transplants are used to treat corneal endothelial dysfunction, these procedures are complex, invasive and time-consuming. In addition, it is estimated that there is only one healthy donor cornea available for every 70 diseased eyes. 3 As such, there remains a significant unmet need for patients with corneal endothelial disease for alternative treatment options that are effective, straightforward and minimally invasive, without being limited by donor cornea supply. About Aurion Biotech, Inc. Aurion Biotech’s mission is to restore vision to millions of patients with life-changing regenerative therapies. It received the prestigious Prix Galien award for best start-up in biotech in 2022. Aurion Biotech is advancing AURN001, an investigational single administration, allogeneic cell therapy to treat corneal edema secondary to corneal endothelial dysfunction. Aurion Biotech developed and has launched the first commercially available corneal endothelial cell therapy in Japan. To learn more, visit www.aurionbiotech.com . 1 Although the study was not powered to show statistical significance, descriptive statistical analyses are provided in this topline readout. 2 LOCF: Last Observed Carry Forward statistical analysis, LS Mean: Least Squares Mean statistical analysis 3 JAMA Ophthalmology . 2016;134(2):167-173. doi:10.1001/jamaophthalmol.2015.4776.

细胞疗法临床结果临床3期突破性疗法

2024-12-18

·EndPoints

Aurion’s eye cell therapy improves vision in small study, plots Phase 3 trial

Aurion Biotech said its cell therapy for treating the cornea improved vision at the highest dose, which it plans to further study in a Phase 3 trial beginning later next year. The Seattle-based biotech is developing an allogeneic cell therapy for what’s called corneal edema secondary to corneal endothelial dysfunction — a condition where key cells of the outer layer of the eye are lost, resulting in swelling and cloudy or blurry vision. Typically, the condition is treated with corneal transplantation from a donor, but Aurion hopes its therapy can be used to treat many more patients than traditional cell transplants. Aurion president and chief medical officer Michael Goldstein said that while typical transplants are from one person to another, Aurion believes its cell therapy made from one donor can treat up to 1,000 patients, though its clinical trial used cells from several donors. After it is made, the cell therapy lasts for up to several days at a time and is given alongside a small molecule drug called a rho kinase inhibitor. “We can essentially solve the worldwide demand for the need for endothelial corneal tissue,” Goldstein said. The company enrolled 97 patients in a Phase 1/2 study conducted in the US and Canada and split them into five arms: three received different doses of the cell therapy called neltependocel plus the rho kinase inhibitor dubbed Y-27632; one arm received only the rho kinase inhibitor; and one received only the cell therapy. At the highest dose of one million cells, 50% of patients were considered responders compared to 14% in the group that just received the small molecule drug, according to Aurion, which said the difference was statistically significant with a p-value of 0.02. Responders were patients at six months with a greater than 3-line or 15-letter improvement in best corrected visual acuity, or BCVA, which is measured on a common vision chart test. Aurion noted that the study was powered to show statistical significance. The company also said the response was dose-dependent, meaning that the proportion of patients who responded at the lower doses was also lower. Aurion said that the high dose also showed a benefit on key secondary endpoints and a measure of patient-reported outcomes. The most common adverse event after treatment in the eye was ocular hypertension, which occurred in 9.3% of patients, followed by conjunctival hemorrhage at 5.2% and eye pain at 4.1%. Aurion licensed the cell therapy from Shigeru Kinoshita and his research institution Kyoto Prefectural University of Medicine in Japan, where it received commercial approval last year and launched commercially under the brand name Vyznova. The approved treatment in Japan also uses one million cells, which is the experimental dose that Aurion is moving forward in its North American study. Goldstein said that Aurion is planning to begin a pivotal study in North America in the latter half of 2025. The biotech expects to enroll about 100 patients and measure the proportion of responders for the primary endpoint, similar to the Phase 1/2 study.

细胞疗法临床3期临床结果上市批准免疫疗法

2024-11-13

·转化医学网

【Cell子刊】攻克癌症新视角：中山大学符立梧团队革新循环肿瘤细胞类器官培养技术

2024年11月9日，中山大学肿瘤防治中心符立梧团队在期刊《Trends in Cell Biology》上发表了题为“The culture and application of circulating tumor cell-derived organoids”的研究论文。研究结果表明，循环肿瘤细胞衍生的类器官（CTCDOs）对于开发患者衍生模型、建立生物银行、探索CTCs的生物学特性，以及推进转移研究等方面具有不可估量的价值，使得个性化治疗策略成为可能。 https://www.cell.com/trends/cell-biology/fulltext/S0962-8924(24)00210-1#au0030 循环肿瘤细胞衍生类器官在癌症研究中的应用 01 循环肿瘤细胞（CTCs）是转移的前体，并且与肿瘤复发强烈相关；因此，它们在涉及转移机制、个性化癌症治疗以及微转移扩散和进展的研究中，扮演着至关重要的角色，同时也在预防转移方面发挥作用。在临床实践中，基于CTC的液体活检比侵入性活检更受青睐，因为它们使用方便、风险低、成本效益高，且不具有组织特异性。 CTCDOs是从患者血液中分离出的CTCs衍生而来，保留了个体原发癌和CTCs的独特遗传和表型特性。与患者衍生的类器官(PDOs)相比，CTCDOs的侵入性更小，更安全。研究表明，来自转移部位的类器官与其原发部位衍生的对应物相比，表现出更高的转移倾向，这表明CTCDOs在更好地模拟肿瘤行为和转移机制方面具有潜力。从转移瘤衍生的PDXs，主要用于研究晚期转移的进展。与耗时、劳动密集和成本高昂的异种移植模型相比，CTCDOs能更好地反映血液中CTCs的特性，为早期转移的潜在机制提供进一步的洞见。CTCDOs还可以用来确定如何杀死CTCs，预防CTC重新激活，或促使CTCs进入休眠状态，为临床上预防（微）转移，提供新的目标。由于血液中CTCs的浓度低，培养CTCDOs具有挑战性；此外，不同癌症类型的CTCs在大小、形状和其他特征上，也有所不同。在这篇综述中，团队提供了CTCDOs发展和应用的最新概述。通过这篇综述，团队旨在指导CTCDOs在临床上的建立和实施。图形摘要 CTCDOs的维持与扩展 02 维持CTCDOs具有挑战性，因为CTCs的活性较差且体外培养条件苛刻。一些CTCDO培养是在一致的条件下维持的，而在其他培养中，维持阶段的因素则会进行调整。一项研究成功培养了14个CTCDOs，其中大多数维持了6至8周，2个扩展并维持超过1个月，尽管仅生成了一个稳定的细胞系。建议在前列腺CTCDOs的第1次传代后，去除Y-27632。CTCDOs的传代过程，与其他肿瘤衍生类器官的维持过程相似。循环肿瘤细胞（CTC）衍生类器官（CTCDO）的潜在临床应用。 CTCDOs开发中的局限性和挑战 03 探索不同的起始材料，可能会改善结果。大多数CTC培养样本来源于患者的外周血，但异种移植瘤提供了扩展CTC的替代方法。已经有成功的从PDX小鼠血液中的微量中分离CTCs，同时，PDXs中的CTCs已被用来生成CTC衍生的异种移植瘤（CTCDXs）。一些CTCDX模型已经成功建立，导致转移。PDXs或CTCDXs，可以用来增加类器官培养的CTC数量。此外，骨髓也可能是一个可行的起始材料，因为它可能作为肿瘤细胞的存活庇护所。骨髓脂肪细胞（BMAs）可能作为肿瘤细胞的直接能量来源，增强肿瘤细胞的增殖。随着类器官的生长，类器官中心的代谢产物积累，以及营养缺乏、与其他细胞的直接接触，也会影响它们的功能和稳定性。因此，控制CTCDO传代的频率也很重要。尽管取得了显著进展，构建完全功能的CTCDOs，仍有很长的路要走。当前发展循环肿瘤细胞（CTC）衍生类器官（CTCDO）培养系统的限制和挑战。总结 04 1. CTCDOs在癌症研究和个性化医疗中具有巨大潜力，尽管存在分离、培养和稳定性方面的挑战。需要解决的关键问题，包括扩大样本来源、改进培养系统、确保长期稳定传代和建立共培养系统。 2. CTCDOs可以作为研究肿瘤转移机制和开发新治疗策略的强大工具。CTCDOs有助于为个体患者开发个性化治疗策略，并可用于生成异种移植瘤。 3. CTCDOs在建立癌症模型和识别驱动基因方面具有潜力，得益于先进技术的支持。CTCDOs保留了CTCs的遗传多样性，有助于研究表面标志物和基因突变。 4. CTCDOs是进行药物相关研究的更经济、更简单的工具，适合高通量药物筛选。CTCDOs可用于评估药物治疗的潜力，包括药物敏感性测试。癌症治疗需要根据肿瘤的演变和新突变不断调整，CTCDOs有助于加速新药开发。CTCDOs可以作为基于最新遗传分析开发和筛选实验药物的有价值工具，以解决治疗后的药物抗性、复发和转移问题。参考资料： 1.Ring, A. ... Biology, vulnerabilities and clinical applications of circulating tumour cells Nat. Rev. Cancer. 2023; 23:95-111 2.Nikanjam, M. ... Liquid biopsy: current technology and clinical applications J. Hematol. Oncol. 2022; 15:131

临床研究

100 项与 Y-27632 (Mitsubishi) 相关的药物交易

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研发状态

10 条进展最快的记录,

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适应症	最高研发状态	国家/地区	公司	日期
心绞痛	临床前	日本	Kyushu University	-
心绞痛	临床前	日本	Mitsubishi Tanabe Pharma Corp.	-
哮喘	临床前	美国	Vanderbilt University School of Medicine	-
勃起功能障碍	临床前	美国	Georgia Health Sciences University	-
高血压	临床前	日本	Kyushu University	-
高血压	临床前	日本	Mitsubishi Tanabe Pharma Corp.	-
脑血管疾病	药物发现	日本	Mitsubishi Pharma Corp.	1997-12-05
肿瘤	药物发现	日本	Mitsubishi Pharma Corp.	1997-12-05