Lion Corp.

导读：递送是mRNA疫苗最具挑战性的难题之一。mRNA分子尺寸较大且具有亲水性，这使得其不能轻易地穿过构成细胞膜的疏水性脂质双分子层，因此，mRNA分子进入细胞是一个非常低效的过程。人们为促进mRNA进入细胞，将mRNA与阳离子脂质或聚合物偶联，这些阳离子脂质或聚合物一方面可以中和mRNA的大量负电荷，另外一方面又可以为其提供穿过细胞膜所需的疏水性。 尽管这些脂质制剂已经使用了很长时间，但它们的初始组合物并不是最佳的并且对于体内mRNA递送效率相当低。然而，脂质纳米颗粒（LNP）的出现彻底改变了这一领域。目前，LNP 是最先进的递送技术之一，Onpattro 于 2018 年获得批准就证明了这一点，Onpattro 是一种由 LNP 递送的基于小干扰 RNA (siRNA) 的疗法，用于治疗转甲状腺素蛋白淀粉样变性。新冠mRNA疫苗使得LNP技术以前所未有的速度大规模制备，用于新冠mRNA疫苗的LNP 通常由四种成分组成，包括可电离脂质、磷脂、胆固醇和聚乙二醇化脂质。 新冠自复制RNA疫苗ARCT154由CSL和Arcturus Therapeutics联合开发，并且已经获得日本厚生劳动省的批准用于18岁及以上成年人的初始接种和加强接种。saRNA是一种可在细胞内自我复制的基于甲病毒基因组的特殊RNA分子。与常规mRNA分子相比，saRNA分子存在一些特有的属性。首先，从分子大小来说，saRNA分子要比mRNA长3-4倍；其次，saRNA还编码病毒复制酶。复制酶可以扩增转染细胞中的 saRNA，减少疫苗接种所需的RNA量，并且复制过程中产生的双链RNA可增强适应性免疫的干扰素I反应。为了开发出保护效果更好，安全性更高的自复制saRNA疫苗，我们非常有必要对现有的LNP递送系统进行优化或者探索效率更高的其他递送平台。 根据其主要成分和机制，用于saRNA的递送系统分为脂质递送系统和非脂质递送系统两大类。本期文章，菌菌汇总了脂质递送系统与自复制saRNA相关的研究，非脂质递送系统将在下期文章中分享，敬请关注~ 01  脂质纳米颗粒（LNP） 早在2012年，Andrew J. Geall等人首次采用LNP递送saRNA，相关研究成果发表在PNAS上：Nonviral delivery of self-amplifying RNA vaccines。在saRNA-LNP制剂配方中，DSPC:胆固醇:PEG-DMG 2000:DLinDMA（可电离阳离子脂质）摩尔百分比为 10:48:2:40，同时，N:P 摩尔比（DlinDMA 上的氮与 RNA 上的磷酸盐）为8:1。采用LNP包封编码呼吸道合胞病毒RSV F蛋白的saRNA，免疫小鼠，RSV F蛋白 saRNA-LNP疫苗可在小鼠体内高效触发F蛋白特异性的血清抗体和细胞免疫反应。 02  纳米结构脂质载体-NLC NLC(Nanostructured Lipid Carrier)系统代表一种水包油乳液（oil in water）和固相纳米颗粒的混合配方，曾经被开发出来用于亲脂性小分子药物的递送。纳米颗粒核心由液体油相，比如三十碳六烯（squalene）和固相脂质（三肉豆蔻酸甘油酯，Dynasan 114）构成。表面活性剂（Tween 80）和阳离子脂质（DOTAP）包围纳米颗粒核心。携带负电荷的RNA分子被阳离子脂质吸附在纳米颗粒表面。 2018年，Neal Van Hoeven在Molecular Therapy发表文章：A Nanostructured Lipid Carrier for Delivery of a Replicating Viral RNA Provides Single, Low-Dose Protection against Zika，他们制备出一种寨卡病毒自复制mRNA疫苗，仅仅接种一针剂量时10ng的疫苗，就可以保护小鼠免于寨卡病毒感染造成的死亡。该疫苗采用高度稳定的纳米结构脂质载体NLC递送系统，并且与samRNA分开贮存，在接种前，可将samRNA和NLC递送系统即时混合，再进行注射。在这项研究中，我们可以看到NLC在25°存放9个月，其粒径大小依然保持不变，包封在颗粒表面的RNA会免于核酸酶的降解。 2022年3月14号，AAHI合作者Corey Casper在Molecular Therapy发表文章：A flexible, thermostable nanostructured lipid carrier platform for RNA vaccine delivery Alana，他们发现液体NLC单独系统在冷藏温度（2-8°）可以存放1年，NLC/RNA冻干体系可以在室温下（25°）存放8个月，在冷藏温度存放21个月，耐温稳定的NLC系统可以极大拓宽mRNA疫苗的运输和存放条件。 2022年3月23号，AAHI合作者Corey Casper在BioRxiv上传文章：A self-amplifying RNA vaccine against COVID-19 with long-term room-temperature stability，他们开发出一款编码Spike 抗原的自复制新冠mRNA疫苗，利用可冻干且耐温的纳米结构脂质载体NLC包封samRNA-Spike 。在小鼠模型中，证实该款疫苗可以刺激机体产生强烈的体液免疫和细胞免疫。最大的特色是，用NLC包封samRNA可极大改善mRNA稳定性，使得samRNA疫苗在室温下存储长达6-8个月，依然具备良好的免疫原性，能够刺激产生高滴度的血清中和抗体。 相比于LNP递送系统，NLC系统不需要特殊设计的，具有专利保护的可电离脂质，而是用到商业化的非常容易获得的DOTAP，因此制备简单，产量放大迅速。此外，NLC系统是独立制备的，可以大量贮存，能够与不同长度的RNA即时混合接种，这样在面对不断涌现的变异株时，将会大大加快疫苗研发应对的速度。 03  脂质无机纳米颗粒-LION LION 是一种高度稳定的阳离子角鲨烯乳液，具有嵌入疏水油相中的 15 nm 超顺磁性氧化铁 (Fe O (4) ) 纳米粒子，并含有 DOTAP，可与 saRNA 分子静电缔合在纳米粒子的表面，避免了包封过程。LION可独立制备，因为它在 4°C 下稳定，并且随后可以通过简单的混合步骤便可与saRNA发生结合。 2020年，Erasmus等人在Science Translational Medicine发表文章：An Alphavirus-derived replicon RNA vaccine induces SARS-CoV-2 neutralizing antibody and T cell responses in mice and nonhuman primates，他们开发了一种编码新冠S蛋白的自复制RNA疫苗，采用脂质无机纳米颗粒 (LION) 配制而成，旨在增强疫苗的稳定性、递送和免疫原性。在小鼠中进行基于 VEEV 的 LION-saRNA 疫苗单次肌内免疫和在猕猴的五个不同部位进行单次肌内注射后， LION-saRNA 诱导了强烈的抗体反应。这些抗体中和 SARS-CoV-2 的滴度与从 COVID-19 恢复期的人类中报告的滴度相当。小鼠和猕猴的初免/加强疫苗接种也诱导了特异性的 T 细胞反应。这种LION-saRNA 疫苗还在老年小鼠中诱导了强烈的免疫反应，表明对老年人具有保护的潜力。 与目前批准的基于 LNP 的 mRNA 疫苗相比，LION 技术具有多个优势。配方稳定性的提高使得分销和储存变得更加容易，特别是在基础设施不准备维持冷链的国家。saRNA-LION 疫苗已在印度进行临床试验评估，最近在该国获得批准，名称为 HDT-301。这是一个重要的里程碑，因为它是第一个被批准用于人类使用的saRNA 疫苗(EUA)。LION平台的另一个优点是能够快速适应新出现的VOC，甚至适应未来由不同病原体引起的流行病，因为该制剂可以很容易地与其他saRNA分子复合。 图片来源：HDT Bio公司官网(一家总部位于西雅图的临床阶段生物制药开发公司,其AMPLIFY疫苗平台是第一个在世界各地获得监管授权的自我复制 RNA ) 增加交叉中和活性的一种策略是设计多价疫苗。这可能需要增加每剂 saRNA 的总量，这可能会损害疫苗的安全性，因为一些使用 LNP 传递的 saRNA 的临床研究已显示出剂量限制性反应原性。在一项生物分布研究中，LNP 递送的 saRNA 被证明可以分布到远处的器官，而 LION 配制的 saRNA 则定位于注射的肌肉和引流淋巴结 (dLN)。结果，接种 LION/saRNA的小鼠表现出仅限于注射部位的先天免疫反应的激活，没有全身炎症或显着的体重减轻。这些结果表明，在 saRNA 的背景下，不同于 LNP 的替代递送平台（例如 LION）可能更适合以较低的反应原性引发最佳免疫。 图片来源：A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation. 04  鱼精蛋白-LPR 另一种为 saRNA 递送而开发的技术是基于脂质体-鱼精蛋白-RNA (LPR) 纳米粒子。 LPR 纳米颗粒采用自组装方法制备，该方法基于 saRNA 与鱼精蛋白通过静电相互作用缩合，然后进行阳离子脂质体封装和表面聚乙二醇化。 LPR 纳米颗粒经过进一步优化，可有效靶向 dLN，并通过 TLR 信号传导刺激先天免疫。该平台用于在小鼠体内传递编码 RBD 的基于 VEEV 的复制子。给小鼠皮下注射 2 μg LPR-saRNA 会诱导 dLN 中抗原的高表达和延长，并上调多种先天免疫信号通路，例如 IFN-I 和 STING。与 LPR-mRNA 疫苗相比，抗原表达水平显着更高，并且持续至少两周，而非复制 RNA 的情况仅持续四天。 图片来源：Self-replicating RNA nanoparticle vaccine elicits protective immune responses against SARS-CoV-2. 05  总结 自复制RNA疫苗被视作为mRNA疫苗的下一代革新技术平台，能够以极低的剂量实现更长久的免疫保护效果。自复制RNA相比常规mRNA分子量更大，对于递送系统带来巨大的挑战。 本文归纳总结了当前用于saRNA的脂质递送系统的特点和优势，包括LNP、NLC、LION以及LPRP。其中脂质纳米颗粒（LNP）递送系统的有效性已在新冠mRNA疫苗、saRNA疫苗的大规模接种中得到了验证。 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。

Program Now Open to Applications for Startups in this Year's Focus Areas: Women's Health; Nutrition and Food; Senior Care and Inclusive Tech SAN FRANCISCO, April 7, 2023 /PRNewswire/ -- Scrum Studio Inc., together with Japanese corporations across several industries, has launched the second year of Well-BeingX, a global open innovation program that aims to help create and grow new business opportunities with startups developing products and solutions that enhance well-being. Today, the program will start accepting applications from startups to participate in the second year of the program. Well-BeingX will be recruiting startups in categories related to health and wellness with a particular focus on Women's Health, Nutrition and FoodTech, and Senior Care and Inclusive Tech. Building on the program's success last year when 37 wellness startups were selected from over 10 countries, Well-BeingX is welcoming two new local governments, Fukui Prefecture and Kashima City, to its second-year program. Well-BeingX 2023 Participants Include: Partner Companies: Sumitomo Life Insurance Company, Tokyo Tatemono, Japan Tobacco, Hakuhodo, Lion Corporation Municipalities: Kobe City, Shibuya Ward, Osaka Chamber of Commerce and Industry, Chuo Ward, Fukui Prefecture, Kashima City Resource Partners: CIC Japan LLC "According to McKinsey & Co, the market for wellness products and services is $1.5 trillion with expected annual growth of 5 to 10 percent, as today's global consumers are continuing to realize the importance of health and well-being" said Michael Proman, managing director and partner, Scrum Ventures. "This is why we are more excited than ever to kick off year two of Well-BeingX. Last year's program was a success as we hosted hundreds of introductions of well-being startup founders to mentors and major Japanese corporations, resulting in co-development initiatives that address the most critical challenges in well-being, including fertility, stress reduction, lifesaving services for office workers and childcare." The application period opens today and ends June 14, 2023. Startups will be chosen by June when the official program kicks off, including access to industry executives and business development opportunities. The program runs from June to December. Interested startups are invited to apply here: The selected startups will have opportunities to co-create businesses with participating corporations and government organizations. Participants in the program will also be considered for investment by Scrum Ventures, the San Francisco-based VC firm that is associated with Scrum Studio. About Scrum Studio, Inc. Scrum Studio, Inc. is an open innovation platform formed to bridge the gap between iconic Japanese corporations with startups from around the world. The platform focuses on three pillars to create new business opportunities: acceleration, incubation, and connection. Its vertical studio programs include SmartCityX, Food Tech Studio - Bites!, and Well-BeingX. About Scrum Ventures Scrum Ventures is an early-stage venture firm based in San Francisco with a growing Tokyo office. With entrepreneurial expertise and a strong network in Japan, the firm actively supports portfolio companies' global growth and success. Media Contact: Mary Magnani CodePR [email protected] SOURCE Scrum Ventures

Emotional analysis technology developed by software solutions provider Opsis Emotion AI will be piloted over the next two years in a programme targeting more than 4,300 seniors in Singapore. The programme will see a collaboration between Opsis and social services agency Lions Befrienders (LB). Through the use of Opsis' emotion analysis software, counsellers from LB are expected to be able to diagnose seniors more accurately for mental health conditions such as anxiety, stress and depression. WHAT IT DOES Leveraging artificial intelligence technology, responses and facial expressions captured on video are analysed by the software to determine a person's mental state. It will then present heat maps of that person's positive and negative emotions. According to Andrew Ow, CEO and co-founder of Opsis, the tool can aid in detecting mental health issues, particularly in reticent people who may not be open to discussing their mental health conditions. "The objective is to assist healthcare professionals deal more effectively with mental health issues in people who are suffering in silence. With this tool, we could help prevent behavioural and psychological deterioration, and improve quality of life," he said. He added that Opsis' software could also help provide early warnings of suicidal tendencies, helping care professionals detect mental health issues through tele-counselling in order to address them early. WHY IT MATTERS By 2030, one in four Singaporeans will be aged over 65 years, and by 2050, the number is expected to rise to one in two. In present day, LB serves close to 7,600 seniors in rental flats. The social service agency said that the Opsis software will supplement existing mental health screening protocol by allowing counsellors to provide more accurate virtual care to elderly patients. The programme’s pilot phase with Lion Befrienders is intended to see counsellors match data derived from the software with their own findings to determine overall efficacy. Should this community pilot initiative be successful, the tool may be used to serve other beneficiaries in the social services sector, such as people with special needs. THE LARGER TREND In an interview with MobiHealthNews, Ow mentioned that Opsis is currently working with two public hospitals specialising in neuroscience along with with six social service agencies to aid care professionals in identifying signs of depression, anxiety, cognitive decline, psychosis, hallucinations and pain. "[One of the goals] of these partnerships is to aid self-help management and behavioural therapy with pre/post metrics over a period of time that helps track a person's healing progress," Ow said. He added that annotated data helps identify signs of depression, anxiety, grief, and pain, and can be used as a tool to help people create awareness of their emotional states and regulate their emotions better. "Our annotated datasets from the clinical, health, counselling and psychological fields are used to train our machine-learning algorithm to recognise emotions at a minute scale that can be used to aid assessment and tracking of mental health wellness," Ow explained. Technology that analyses facial expressions has been around since at least 2003, and has evolved significantly since. Another tech company in this space is Massachusetts-based emotions analytics company Affectiva. In 2016, the company raised $14m for its emotion analytics platform, with the aim of expanding globally.