ASAH1

引言 众所周知，葡萄糖驱动胰岛素分泌，以维持能量稳态。目前对人胰岛素的研究通常只使用葡萄糖刺激，然而，另外两大宏量营养素--蛋白质和脂肪有没有可能也调节胰岛素的释放？迄今为止，非碳水化合物营养素对胰岛素分泌的影响还没有充分研究，针对人体的碳水化合物、蛋白质和脂肪的胰岛素分泌反应水平变化和机制仍未被揭示。 对营养素刺激人类胰岛素分泌的研究有助于扩大我们的临床认识，然而我们对于这个问题的有限了解大多来自于对啮齿动物的研究。全面了解营养素诱导的人类胰岛素分泌在2型糖尿病（T2D）和相关新兴研究中具有重要意义。 近日，来自加拿大英属哥伦比亚大学和温哥华海岸健康研究所的James D.Johnson、Jelena Kolic研究小组与加拿大阿尔伯塔大学等单位合作在Cell Metabolism杂志上发表题为Proteomic predictors of individualized nutrient-specific insulin secretion in health and disease的论文，这篇文章对来自非糖尿病（ND）和2型糖尿病（T2D）人类胰岛在三大主要宏量营养素包括碳水化合物、蛋白质和脂质影响下的胰岛素分泌动力学进行了综合分析，同时辅以转录组学和蛋白质组学分析，发现了胰岛的分子特征，并鉴定了不同的蛋白质簇，这些蛋白质簇可以预测胰岛素对碳水化合物、蛋白质和脂质的分泌反应，首次并排比较了人类胰岛和干细胞来源的胰岛之间营养反应和蛋白质组。这是目前最大的人类胰岛数据集，揭示了营养反应的多样性及机制。这一资源有望为糖尿病患者提供更多的个性化治疗建议。 2016年至2022年间，作者系统地测量了来自140名供体的胰岛对碳水化合物（15或6 mM葡萄糖）、氨基酸（5 mM亮氨酸）和脂质（1.5 mM油酸/棕榈酸混合物）的胰岛素分泌反应。证实碳水化合物是最强的胰岛素促分泌剂，其次是氨基酸，然后是脂肪。T2D患者的胰岛在葡萄糖和脂肪酸刺激下胰岛素分泌减少，但氨基酸刺激下的胰岛素分泌未受影响。 图1: ND和T2D胰岛之间的差异丰度蛋白质(Credit: Cell Metabolism) 为了更好地了解ND和T2D患者的胰岛中分泌反应与基因表达变化之间的关系，作者对这些胰岛进行了全面的转录组和蛋白质组分析。通过RNA测序和质谱法分析了20,000多个mRNA和近8,000个蛋白质，数据确定了与葡萄糖刺激的胰岛素分泌相关的关键蛋白在T2D中减少，包括例如ARG2、NAT8L、HADH、PPM1K、AMACR、PFKFB2、G6PC2、ERO1B、OTULINL、ATP2A3和CASR等（图1）。不同个体的胰岛对营养素刺激的胰岛素分泌反应存在显著个体差异，特别是一些胰岛对脂肪酸的反应比葡萄糖更强烈，这一发现挑战了长期以来认为饮食脂肪对胰岛素释放的影响可以忽略不计的传统观点。 作者利用共表达网络分析揭示了与胰岛功能紧密相关的蛋白质网络（图2），确定了18个关键蛋白质模块，其中9个模块与胰岛功能直接相关。与营养素刺激的胰岛素分泌呈正相关的模块参与细胞骨架、线粒体代谢、胰岛素加工等关键过程。在蛋白质运输和定位中起作用的模块与KCl刺激的胰岛素分泌呈正相关，但与脂质刺激的胰岛素分泌呈负相关，表明人类胰岛中的这些蛋白质网络模块支持对刺激的差异化响应。 图2: 共表达网络分析确定的18个不同的蛋白质模块（方块内显示Pearson相关系数和p值，红色阴影表示正相关，蓝色阴影表示负相关，黑框表示显著相关）(Credit: Cell Metabolism) 接下来，作者研究分析了mRNA和蛋白质丰度与胰岛特征的关联，特别是与功能的关系。发现胰岛供体的年龄与涉及溶酶体降解、细胞衰老、炎症、线粒体功能障碍等蛋白质的丰度呈正相关，例如PPT、SCARB2、ASAH1、P2X436和NADPH、FDXR等。此外，蛋白质丰度与糖化血红蛋白水平相关，有趣的是这类蛋白还涉及癌症的进展，例如YIF1A、UBAP1、STX5等，这可能和血糖控制不佳与多种癌症风险增加有关。 胰岛功能分析揭示了脂质是胰岛素释放的重要调节因子，并鉴定了与总体胰岛分泌能力相关的关键蛋白质。作者建立了一个在线资源，可以显示检测到的蛋白质、供体特征、胰岛分离参数和胰岛功能之间的关系（www.humanislets.com）。在基础葡萄糖条件下，约8%的供体胰岛对脂质的胰岛素分泌反应异常强烈，对脂质高反应的胰岛WNT4降低，表明胰岛可能处于较不成熟状态；同时，HNF1B、SULT1A1和PDGFRA表达升高。约9%的供体胰岛对亮氨酸的胰岛素分泌反应强于高葡萄糖。 治疗糖尿病非常有潜力的一种疗法是利用胚胎干细胞分化为β细胞替代物，胚胎干细胞衍生的胰岛样团簇与人类胰岛相比，其基础胰岛素释放水平仅为1/10，其基础胰岛素释放及葡萄糖、氨基酸反应性低于人类胰岛，它可能更偏好非葡萄糖底物。蛋白质组学分析揭示了干细胞胰岛样团簇与人类胰岛的显著差异，包括未成熟β细胞标记物的增加及脂肪酸转运和代谢相关蛋白的上调，人胚胎干细胞衍生的胰岛样团簇具有脂质响应性。 高胰岛素血症与多种健康问题相关，这项研究在人胰岛中确定了碳水化合物、蛋白质和脂质三种营养素的胰岛素分泌反应，搭建了在线数据平台：www.humanlislands.com作为胰岛深层表型网络资源。作者的证据还表明营养特异性胰岛素分泌在普通人群中存在异质性，9%的供体胰岛对氨基酸有反应，8%的供体胰岛对脂肪酸的反应大于其葡萄糖刺激的胰岛素反应，这意味着这些胰岛与高葡萄糖引起的胰岛素分泌反应的胰岛截然不同，作者通过干细胞中的观察，推测这种脂质反应性与β细胞不成熟有关。这项研究为针对胰岛对碳水化合物、蛋白质和脂质响应的多样性提供个性化营养治疗奠定了基础。 图3：三大主要宏量营养素包括碳水化合物、蛋白质和脂质对人胰岛分泌反应的影响研究(Credit: Cell Metabolism) 参考文献 https://doi-org.libproxy1.nus.edu.sg/10.1016/j.cmet.2024.06.001 责编|探索君 排版|探索君 文章来源|“BioArt” End 往期精选 围观 一文读透细胞死亡(Cell Death) | 24年Cell重磅综述（长文收藏版） 热文 Cell | 是什么决定了细胞的大小？ 热文 Nature | 2024年值得关注的七项技术 热文 Nature | 自身免疫性疾病能被治愈吗？科学家们终于看到了希望 热文 CRISPR技术进化史 | 24年Cell综述

导读：呼吸道合胞病毒（RSV）是一种全球性流行的呼吸道感染病原体，易感人群主要是婴幼儿、老年人等免疫力低的人群。在早期疫苗开发中，接种RSV灭活疫苗带来的疾病加重现象，导致RSV疫苗的开发异常缓慢。直至近年来，RSV疫苗的开发逐渐取得积极数据，包括辉瑞的Abrysvo和GSK的Arexvy。这无疑激励了应用更先进的生物技术去开发RSV疫苗，如mRNA疫苗，其中进展最快的是Moderna的mRNA-1345。本期文章，菌菌将从专利角度，解析Moderna针对RSV mRNA疫苗、以及mRNA-1345建立的技术护城河。01 Moderna疫苗mRNA-1345疫苗简介呼吸道合胞病毒（RSV）病毒基因组编码11个蛋白质，其中黏附蛋白（Glycoproteins Protein, 简称G蛋白）和融合蛋白（Fusion Protein，简称F蛋白）是RSV膜表面主要糖蛋白，是刺激机体产生中和抗体的关键病毒抗原。RSV疫苗通常以F蛋白或G蛋白作为靶点。Moderna开发的mRNA-1345编码F蛋白的融合前构象（即pre F），使用的LNP递送系统与新冠疫苗mRNA-1273相同。02 RSV mRNA疫苗的相关专利概述Moderna在专利WO2017070622A1中描述了RSV mRNA疫苗的序列组成和递送系统，该发明评估了多个抗原靶点，包括RSV G蛋白、F蛋白（pre F）、以及其他抗原/抗原片段。RSV mRNA疫苗的递送系统为包括阳离子脂质、PEG修饰的脂质、固醇和非阳离子脂质的脂质纳米颗粒（LNP）。结果表明，RSV mF疫苗（F蛋白）和RSV mDS-CAVl mRNA疫苗（pre F）蛋白的中和抗体效价高于DS-CAV1蛋白/ADJU-PHOS疫苗（mRNA+佐剂）。WO2019148101A1和WO2021155243A1专利中，Moderna公开了RSV mRNA疫苗的非编码序列特征：专利公开了2种可选的5’ UTR，如下图所示。经比对，第二条与新冠疫苗mRNA-1345的5’ UTR序列相同。同样，2条3’ UTR序列中，第二条与mRNA-1345一致。并且在专利中，2和4是搭配使用的。图1：Moderna RSV mRNA疫苗的UTR序列此外，Moderna在专利WO2019202035A1中公开了多种可选的UTR序列，用于RSV mRNA疫苗的设计。其中5’ UTR的序列来源包括但不限于以下基因：HSD17B4, RPL32, ASAH1 , ATP5A1 , MP68, NDUFA4, NOSIP, RPL31 , SLC7A3, TUBB4B和UBQLN2；3’ UTR的序列来源包括以下基因：PSMB3, ALB7, alpha-globin, CASP1 , COX6B1 , GNAS, NDUFA1和RPS9等。专利同时列举了多种UTR序列、poly(A)序列和编码抗原的组合。Moderna在专利WO2022221336A1中详细描述了RSV mRNA疫苗的序列元件、核苷酸修饰和LNP递送系统的组分和比例。专利中介绍了这款疫苗是当前的热门品种mRNA-1345，相关信息菌菌将在下文详述。03 mRNA-1345疫苗专利WO2022221336A1Moderna在专利WO2022221336A1中详细描述了mRNA-1345疫苗的序列、LNP组成以及该疫苗的临床疗效。mRNA-1345疫苗是LNP包封的mRNA，专利公开了其mRNA序列，从5’端至3’端依次为：5' 帽结构（m7G）、5’ UTR（同图1列举的第1条序列）、抗原编码序列（融合前RSV F糖蛋白三聚体形式，其细胞质尾部结构域缺失）、3’ UTR（同图1列举的第3条序列）和poly(A)尾（长度为100 nt的多聚腺苷酸尾巴）。mRNA-1345经N1-甲基假尿嘧啶修饰。图2：mRNA-1345抗原编码序列示意图LNP组成方面，包括15-20 mol%中性脂质，35-45 mol%胆固醇，0.5-5 ml% PEG修饰脂质和45-55 mol%可电离脂质。据CDC最新公开信息，mRNA-1345与新冠疫苗mRNA-1273的递送系统是一致的。图3：mRNA-1345中阳离子脂质结构04 总结整体来看，Moderna建立了丰富的mRNA疫苗矩阵，涵盖mRNA抗原编码序列和UTR等非编码序列、mRNA核苷酸修饰和LNP递送系统等。从Moderna在RSV疫苗领域公开的详细进展来看，他们早期评估筛选了有效的RSV抗原，即pre F作为目标抗原，作为mRNA-1345编码序列的来源。非编码序列方面，Moderna在多个专利布局中描述了适用于RSV疫苗的UTR序列，包括后续新冠疫苗mRNA-1273使用的UTR序列、以及相似序列。识别微信二维码，添加生物制品圈小编，符合条件者即可加入生物制品微信群！请注明：姓名+研究方向！版权声明本公众号所有转载文章系出于传递更多信息之目的，且明确注明来源和作者，不希望被转载的媒体或个人可与我们联系(cbplib@163.com)，我们将立即进行删除处理。所有文章仅代表作者观点，不代表本站立场。

Virtual Webinar to Take Place October 11, 2023 at 2:00pm ETPRINCETON, N.J., Oct. 04, 2023 (GLOBE NEWSWIRE) -- Acasti Pharma Inc. (Nasdaq: ACST) (Acasti or the Company), a late-stage, biopharma company advancing GTX-104, its novel formulation of nimodipine that addresses the high unmet medical needs for a rare disease, aneurysmal subarachnoid hemorrhage (aSAH), today announced that it will host a virtual KOL event titled GTX-104: a potential new treatment standard for rare and life-threatening aSAH on Wednesday, October 11, 2023 at 2:00pm ET. To register, click here. The event will feature W. Taylor Kimberly, MD, PhD (Massachusetts General Hospital) who will discuss the high unmet medical need and current treatment landscape for patients suffering from aSAH, a medical emergency in which bleeding occurs over the surface of the brain in the subarachnoid space between the brain and skull. The event will highlight Acasti's lead clinical asset, GTX-104, a novel formulation of nimodipine currently in Phase 3 targeting aSAH as an intravenous infusion. Acasti leadership will provide insight into trial design, market dynamics, and future directions. A live question and answer session will follow the formal presentations. About W. Taylor Kimberly, MD, PhD Dr. Kimberly is Chief of the Division of Neurocritical Care, and a stroke and critical care neurologist in the Department of Neurology at Massachusetts General Hospital. Clinically, he primarily cares for patients in the Neuroscience ICU as part of an integrated and multi-disciplinary team, coordinating care with Neurosurgery and Neuroendovascular specialists. He also sees Neuro ICU patients in follow-up in outpatient clinic as part of the NeuroRecovery clinical team. He serves on several hospital-based and national committees that focus on clinical guideline development and care improvement. Dr. Kimberly’s research group is located in the Center for Genomic Medicine (Kimberly Lab), and studies metabolomic and neuroimaging biomarkers of subarachnoid hemorrhage, stroke and cerebral edema. The goal of his research is to identify novel pathways and candidate therapeutic targets for the treatment of acute brain injury. Dr. Kimberly has co-led multi-site, randomized, placebo-controlled trials in the prevention of brain edema, and currently co-leads an international phase 3 trial evaluating the safety and efficacy of intravenous glibenclamide for the prevention of brain edema after large hemispheric stroke. About Acasti Acasti is a late-stage biopharma company with drug candidates addressing rare and orphan diseases. Acasti's novel drug delivery technologies have the potential to improve the performance of currently marketed drugs by achieving faster onset of action, enhanced efficacy, reduced side effects, and more convenient drug delivery. Acasti's lead clinical assets have each been granted Orphan Drug Designation by the FDA, which provides seven years of marketing exclusivity post-launch in the United States, and additional intellectual property protection with over 40 granted and pending patents. Acasti's lead clinical asset, GTX-104, is an intravenous infusion targeting aneurysmal Subarachnoid Hemorrhage (aSAH), a rare and life-threatening medical emergency in which bleeding occurs over the surface of the brain in the subarachnoid space between the brain and skull. Forward-Looking Statements Statements in this press release that are not statements of historical or current fact constitute "forward-looking statements" within the meaning of the U.S. Private Securities Litigation Reform Act of 1995, as amended, Section 27A of the Securities Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934, as amended, and "forward-looking information" within the meaning of Canadian securities laws (collectively, "forward-looking statements"). Such forward looking statements involve known and unknown risks, uncertainties, and other factors that could cause the actual results of Acasti to be materially different from historical results or from any future results expressed or implied by such forward-looking statements. In addition to statements which explicitly describe such risks and uncertainties, readers are urged to consider statements containing the terms "believes," "belief," "expects," "intends," "anticipates," "estimates", "potential," "should," "may," "will," "plans," "continue", "targeted" or other similar expressions to be uncertain and forward-looking. Readers are cautioned not to place undue reliance on these forward-looking statements, which speak only as of the date of this press release. The forward-looking statements in this press release, including statements regarding GTX-104's potential to bring enhanced treatment options to patients suffering from aSAH are based upon Acasti's current expectations and involve assumptions that may never materialize or may prove to be incorrect. Actual results and the timing of events could differ materially from those anticipated in such forward-looking statements as a result of various risks and uncertainties, including, without limitation: (i) the success and timing of regulatory submissions of the planned Phase 3 safety study for GTX-104; (ii) regulatory requirements or developments and the outcome and timing of the proposed IND application for GTX-104; (iii) changes to clinical trial designs and regulatory pathways; (iv) legislative, regulatory, political and economic developments; and (v) actual costs associated with Acasti's clinical trials as compared to management's current expectations. The foregoing list of important factors that could cause actual events to differ from expectations should not be construed as exhaustive and should be read in conjunction with statements that are included herein and elsewhere, including the risk factors detailed in documents that have been and are filed by Acasti from time to time with the Securities and Exchange Commission and Canadian securities regulators. All forward-looking statements contained in this press release speak only as of the date on which they were made. Acasti undertakes no obligation to update such statements to reflect events that occur or circumstances that exist after the date on which they were made, except as required by applicable securities laws. Acasti Contact:Prashant KohliChief Executive OfficerTel: 450-686-4555Email: info@acastipharma.comwww.acasti.com Investor Relations:LifeSci AdvisorsMike MoyerManaging DirectorPhone: 617-308-4306Email: mmoyer@lifesciadvisors.com