Cav2.1

点击图片查看OTC2023类器官前沿应用与3D培养论坛会后报告，OTC2024论坛征集投稿做报告，详询：王晨 180 1628 8769文章来源： EngineeringForLife患者来源的癌症类器官（PDOs）在个性化治疗选择和改善患者预后方面具有相当大的前景。然而，生成足够数量的PDOs在标准培养平台上测试治疗方法是一项挑战。这一挑战对于胰腺导管腺癌（PDAC）尤其严重，因为大多数患者被诊断为晚期不可切除肿瘤，并且患者的组织以针头活检的形式存在。为此，美国梅奥医学中心Alexander Revzin及其团队研究了微流控装置的开发和特性，用于测试使用有限数量的组织或PDOs的治疗。结果表明，微流控PDOs在表型和基因型上与金标准基质胶类器官相似，且具以下优点，包括1）形状均匀；2）最小细胞数要求；3）不依赖基质胶。通过测试PDO对几种化疗的反应证明了微流控PDOs的实用性，包括糖原合酶激酶（GSKI）抑制剂。此外，微流控类器官培养物被用于测试由NK细胞结合一种新型生物制剂组成的免疫治疗的有效性。总之，本研究的微流控装置为癌症活检的个性化肿瘤学提供相当大的好处，并可能在未来发展成为化疗或免疫治疗的辅助诊断。相关研究内容以“Microfluidic Organoid Cultures Derived from Pancreatic Cancer Biopsies for Personalized Testing of Chemotherapy and Immunotherapy”为题于2023年11月29日发表在《Advanced Science》上。图1 本研究概念示意图本研究采用微流控培养概念，描述来自患者活检的PDAC类器官的微流控培养。研究设备包含微孔阵列（直径250μm），这些微孔被低结合，以促进细胞聚集成球状体（图1）。图2 以单细胞消化作为起始材料，在微流控装置中形成PDAC类器官图2A显示了该研究的工作流程——PDAC活检分离，在基质胶上扩展类器官，然后收集并播种到微流控装置中。在播种后2天形成球体，培养7天后球体的直径增加（图2B）。与基质胶培养物相比，微流控装置中的球状体分布更均匀（图2C）。为了将微流控类器官与金标准基质胶培养物作为基准，对两种培养物进行免疫荧光染色，结果显示出两种培养物之间的相似性（图2D、E）。用于与微流控培养物进行比较的定制微孔阵列如图2F所示。比较类器官大小随时间的变化发现，微流控装置和微孔阵列的增殖动力学相似（图2G）。与微流控类器官培养相比，微孔阵列培养中的PDAC类器官的增殖能力明显较低，且PDAC特异性标记物CK19和Pdx1的表达水平也较低（图2H）。图3 基质胶和微流控类器官培养物的转录组学比较在主成分分析和比较样本与样本之间的距离时，该装置和基质胶类器官比从原发肿瘤、PDX和转移瘤中产生的其他PDAC PDO样本更相似（图3A、B）。在基质胶和装置类器官培养物中富集的差异表达基因火山图如图3E所示。通过对生物过程进行基因本体论分析（图3F）和对所策划的基因集进行基因集富集分析（图3G），发现基质胶类器官培养物对与缺氧相关的途径进行富集，表明与微流控类器官培养物相比氧合不足。图4 在微流控装置中培养完整的类器官新型装置有一个由常闭阀门保护的侧注射端口（图4A）。比较来自三个不同患者的类器官，分别为PDO-001、PDO-002和PDO-003，在微流控装置和基质胶培养的图像乳图4B所示。微流控类器官培养表现出基质胶中观察到的特征，PDO-001和PDO-002表现出腔/导管结构（图4C、D），PDO-003形成紧凑的球状体（图4E）。3例患者的微流体类器官培养物均对上皮标记物E-钙粘蛋白、PDAC标记物CK19和增殖标记物Ki67的染色强阳性（图4F-H）。以上结果证明来自三种不同PDAC活检的完整类器官可以在新型微流控装置中培养，该装置具有增殖标记和上皮/癌症表型表达，与金标准基质胶类器官培养物相似。图5 在微流控培养中测试针对MEK和RAF信号通路的化疗将PDAC类器官对分子靶向治疗的反应与GEM进行比较，GEM是一种抑制DNA合成并导致癌细胞凋亡的一线化疗药物（化学结构见图5A）。在微流控装置中单细胞2天形成球状体，之后给药7天（流程见图5A）。药物治疗前（第0天）和药物治疗后（第7天）微流控类器官培养物的亮场和活/死染色图像如图5B所示。2.5 μM En与2.5 μM Bi对肿瘤消退的效果与单独使用5 μM Bi相似，但对较高浓度的效果有差异（图5B-D）。相比于En，Bi是一种更有效的治疗方法，这些类器官对GEM的敏感性更高（图5E）。图6 使用三种患者特异性微流控类器官培养物的GSKI和GEM检测联合治疗联合化疗的时间线如图6A所示。观察到联合治疗的患者特异性差异（图6B-G）。在暴露于药物的PDO- 001类器官中观察到有限的细胞毒性（图6B、C），而PDO-002和PDO-003在所有浓度范围内都显示出对联合治疗的反应都增强（图6D-G）。图7 使用微流控类器官培养物测试NK细胞的免疫治疗实验时间线如图7A所示，抗B7-H3 TriKE增强NK细胞对肿瘤细胞杀伤的机制如图7B所示。对于3种PDOs，癌症类器官的单培养在微流控装置内持续生长（图7C，72 h）。为定量评估类器官生长，每24小时追踪单个类器官直径，将24、48和72小时的直径归一化到0小时的初始大小，并绘制所有PDAC样本每种条件下的平均类器官直径（图7D）。所有癌症类器官24小时大小减小60%。研究结果表明，抗B7-H3 TriKE提高了NK细胞对癌症类器官的溶细胞活性，也强调了微流控癌症类器官培养的效用。综上所述，本研究中开发并表征了新型的PDAC类器官微流控培养物，证明了微流控装置在类器官增殖和癌症表型表达以及免疫细胞的激活和浸润方面优于微孔阵列格式。RNA测序分析显示，微流控类器官培养物与金标准基质胶培养物相似，在微流控形式下具有更好的氧合和启动趋化因子信号和免疫细胞浸润。总之，本研究描述了一种新的微流控类器官培养平台，用于检测患者的特异性反应。该平台有助于以类似于金标准基质胶培养的方式维持癌症类器官，能够通过针活检获得的少量类器官进行药物检测，非常适合测试免疫治疗。在未来，该微流控平台可能与生物传感能力集成，用于定量评估单个类器官，并可能发展成为个性化癌症治疗的辅助诊断。文章来源：https://doi.org/10.1002/advs.202303088点击图片查看OTC2023类器官前沿应用与3D培养论坛会后报告，OTC2024论坛征集投稿做报告，详询：王晨 180 1628 8769

12月21日的《热心肠日报》，我们解读了 10 篇文献，关注：脂代谢，线粒体，肥胖，消化疾病，病毒组，母婴，肠脑轴，菌群-免疫互作，褪黑素，微生物生态学。 Nature：肠细胞处理膳食脂质，线粒体有何重要作用？ Nature——[64.8] ① 肠上皮特异性敲除DARS2、SDHA或COX10以造成肠上皮线粒体功能缺陷，可导致小鼠近端小肠肠细胞中出现大脂滴积累，影响存活；② 无脂喂养可抑制DARS2缺陷肠细胞的脂滴积累，表明脂滴源自膳食脂肪；③ 代谢追踪分析表明，肠上皮缺乏DARS2损害小鼠从肠道向外周器官运输膳食脂质的能力；④ DARS2缺陷使肠细胞中成熟的乳糜微粒减少，并破坏高尔基体，表明线粒体功能障碍可阻碍乳糜微粒从内质网向高尔基体的运输，进而导致膳食脂质在大脂滴中积累。 【主编评语】 线粒体功能障碍会导致氧化磷酸化不足和代谢缺陷，几乎可影响所有细胞类型，引发疾病。然而，线粒体在肠上皮细胞中的作用尚不清楚。Nature发表的一项最新研究，通过小鼠研究揭示了线粒体在肠上皮细胞中对膳食脂肪的运输和处理的关键作用，这些发现有助于理解线粒体病患者的肠道功能异常。（@mildbreeze） 【原文信息】 Mitochondrial dysfunction abrogates dietary lipid processing in enterocytes 2023-12-20, doi: 10.1038/s41586-023-06857-0 Science子刊：肠道菌群对肥胖及其改善的影响（综述） Science Translational Medicine——[17.1] ① 肠道菌群包含数万亿个细菌、真菌、病毒和古细菌，肥胖患者中其丰度及多样性改变；② 啮齿动物研究明确将肠道菌群与能量处理联系起来，并表明肠道菌群在肥胖发展中的作用；③ 目前人体研究通过前瞻性、多种族纵向队列研究以表明肠道菌群与肥胖之间的因果关系；④ 研究设计的标准化、宏基因组学方法和数据库的应用、扩大研究人群以及延长随访时间是未来研究要素；⑤ 超生理剂量的代谢物、粪菌移植和个性化益生菌或可用于对抗肥胖及其不利影响。 【主编评语】 啮齿动物研究表明肠道菌群与肥胖之间存在联系，但在人体中因果关系的证据仍然很少。近日，发表在Science Translational Medicine上的这篇综述，总结了肠道菌群与肥胖发展之间联系的证据，并概述了如何从机制上了解肠道菌群在能量稳态中的作用以减少全球肥胖流行的影响。（@圆圈儿） 【原文信息】 Evidence for the contribution of the gut microbiome to obesity and its reversal 2023-11-22, doi: 10.1126/scitranslmed.adg2773 Nature Reviews：肠道菌群与脂肪组织的“对话”调控超重/肥胖（综述） Nature Reviews Gastroenterology and Hepatology——[65.1] ① 脂肪组织分为白色、棕色和米色，在体内的能量储存、产热和新陈代谢中有不同的作用；② 环境因素对能量代谢有重大影响，其中饮食、运动和睡眠是主要因素；③ 肠道细菌参与肠道和脂肪组织之间的双向通讯，影响能量代谢、营养吸收、食欲和脂肪组织功能；④ 脂肪组织拥有自己独特的微生物群，这些微生物群因代谢健康和其他因素而异，对此深入研究可以提供新的见解；⑤ 将肠道微生物群研究从动物模型转化为人类应用面临着方法学和生物学方面的挑战。 【主编评语】 全球约40%的人口受到超重或肥胖的影响，超重和肥胖的特征是当能量摄入超过能量消耗时产生过多的脂肪堆积。一种合理的控制能量消耗的方法是调节白色脂肪组织（WAT）和/或棕色脂肪组织（BAT）的产热途径。在能够影响宿主代谢和能量平衡的不同环境因素中，肠道微生物群现在被认为是一个关键的参与者。近期发表于Nature Reviews Gastroenterology and Hepatology的一项综述讨论了WAT和BAT的一般生理学，并介绍了肠道细菌和多种微生物衍生的代谢物、它们的受体和信号通路如何调节脂肪组织的发育及其代谢能力。最后，通过提出具体的关键例子来描述从实验到临床的关键挑战。（@注册营养师陈彬林） 【原文信息】 Gut microbiota in overweight and obesity: crosstalk with adipose tissue 2023-12-08, doi: 10.1038/s41575-023-00867-z Lancet子刊：哪些非饮食方式可以改善肠道健康？ Lancet Gastroenterology & Hepatology——[35.7] ① 胃肠道环境与中枢神经系统存在双向调节关系，压力可能通过调节生物系统，如肠道菌群、循环细胞因子、其他炎症和免疫标志物等，诱发肠道炎症和心理健康症状；② 心理疗法可减轻肠易激综合征患者的肠道症状，减压疗法可帮助乳糜泻患者更好地坚持无麸质饮食，正念疗法或可提高肠易激综合征患者的生活质量；③ 生活方式可能对维持肠道健康有效，如有规律切充足的睡眠或运动；④ 接触自然的户外活动也可对肠道健康具有有益作用。 【主编评语】 除了膳食饮食以外，还有什么其他有方法可以潜在改善和维持肠道健康呢？Lancet Gastroenterology & Hepatology《食物问题》专栏中进行了归纳，良好的身心健康状态、高质量生活方式以及户外活动均对肠道菌群具有潜在的积极作用。当饮食治疗难以实施时，以上或可作为辅助干预方法。（@RZN） 【原文信息】 How can I improve my gut health via non-dietary means? 2023-12-06, doi: 10.1016/S2468-1253(23)00412-0 Nature子刊：肠道病毒组直接与学龄前哮喘风险相关 Nature Medicine——[82.9] ① 纳入哥本哈根儿童哮喘前瞻性研究2010（COPSAC2010）母婴队列的647名1岁儿童，进行肠道病毒组分析；② 特定的温和肠道噬菌体分类群被发现与哮喘的后期发展有关，特别是19个caudoviral家族的联合丰度对这种关联有显著贡献；③ 哮喘相关的病毒组和细菌组特征结合分析，其对哮喘风险具有加性影响，这意味着独立的病毒组-哮喘相关性；④ 病毒组相关的哮喘风险由宿主TLR9 rs187084基因变体调节，这表明噬菌体和宿主免疫系统之间存在直接相互作用。 【主编评语】 栖息在肠道中的噬菌体群落对细菌种群的结构和功能有着重要影响，但它们在生命早期的作用以及与健康和疾病的关系尚不清楚。Nature medicine近期发表的文章，发现婴儿肠道病毒组直接与宿主免疫系统之间相互作用，不依赖于对细菌结构和功能的影响，与学龄前哮喘风险相关联。（@章台柳） 【原文信息】 The infant gut virome is associated with preschool asthma risk independently of bacteria 2023-12-15, doi: 10.1038/s41591-023-02685-x Cell子刊：母亲来源的噬菌体更能稳定定植于肠道 Cell Host and Microbe——[30.3] ① 从28名足月婴儿和24名早产儿及其母亲身上收集的819个粪便样本进行宏基因组分析，发现早期生命中噬菌体的丰富度随着时间的推移而增加，并在3岁时达到类似成人的复杂性；② 大约9%早期噬菌体的定植（主要通过母体传播并感染拟杆菌）可持续3年，该现象在足月儿比早产儿更普遍；③ 尽管罕见，但具有终止密码子重新分配的噬菌体比未重新编码的噬菌体更有可能持续存在，并且通常在3年内显示出编码框内重新分配的终止密码子的增加。 【主编评语】 噬菌体是肠道微生物组的关键组成部分，但噬菌体在婴儿肠道中的定植过程尚不清楚。Cell Host and Microbe近期发表的文章，对婴儿期到3岁的肠道噬菌体进行分析，发现母体来源、终止密码子重新分配、宿主CRISPR-Cas基因座流行率和不同的噬菌体群体有助于稳定的病毒定植。（@章台柳） 【原文信息】 Infant gut DNA bacteriophage strain persistence during the first 3 years of life 2023-12-13, doi: 10.1016/j.chom.2023.11.015 Cell子刊：婴儿肠道菌群如何影响认知能力？ Cell Host and Microbe——[30.3] ① 本研究发现婴儿复合认知和肠道微生物群组成的关联最早在6月龄时就建立起来了；② 复合认知高于中位数（Inf-aboveCC）相比低于中位数（Inf-belowCC）的婴儿的肠道微生物群落具有更高的多样性和均匀度；③ 此外，Inf-aboveCC小鼠肠道富含已知与认知有关的Phocaeicola、拟杆菌和双歧杆菌；④ 认知能力较好婴儿的肠道微生物群的移植促进了受体小鼠的记忆，并调节了小鼠鼻周皮质（perirhinal cortex，PRC）的组氨酸代谢组。 【主编评语】 肠道微生物群与神经发育之间存在关系，但其潜在机制尚未确定。近期发表于Cell Host and Microbe的一项研究利用代谢组学等方法揭示了婴儿认知和肠道微生物群之间的联系，进一步分析发现肠道菌群可能通过调节组氨酸代谢影响婴儿的认知能力。（@注册营养师陈彬林） 【原文信息】 Infant gut microbiota contributes to cognitive performance in mice 2023-12-04, doi: 10.1016/j.chom.2023.11.004 Science子刊：新生儿肠道菌群影响疫苗接种后的T细胞免疫 Science Advances——[13.6] ① 暴露于HIV但未感染的婴儿（iHEU）对卡介苗（BCG）反应各不相同，不同反应者具有不同的肠道菌群组成，高反应者的粪便中富含婴儿双歧杆菌（BI），而低反应者的粪便中则富含多形拟杆菌（BT）；② BI和BT对新生小鼠免疫系统的发育有不同的影响，与灌胃BT的幼鼠相比，口服BI的新生无菌或SPF幼鼠表现出明显更多的BCG特异性T细胞；③ BI和BT对小鼠粪便代谢组，结肠转录组以及人类结肠细胞转录组的影响均存在差异。 【主编评语】 卡介苗接种通常可在新生儿中引起良好的TH1应答。新生儿极易受到感染，而且对大多数疫苗的反应较差，研究人员假设早期肠道微生物群影响疫苗T细胞反应。近期发表于Science Advances的一项研究发现，在生命的最初几天的肠道微生物群影响免疫发育，包括抗原特异性T细胞对新生儿疫苗接种的反应性。对新生儿卡介苗接种反应良好的婴儿粪便的婴儿双歧杆菌相对丰度较高，而反应较差者的多形拟杆菌更高。（@注册营养师陈彬林） 【原文信息】 Bifidobacterium infantis associates with T cell immunity in human infants and is sufficient to enhance antigen-specific T cells in mice 2023-12-08, doi: 10.1126/sciadv.ade1370 国内团队iMeta：褪黑素如何改善创伤性结肠损伤 iMeta——[N/A] ① 本研究旨在揭示TCI疾病的进展机制，并挖掘有效干预策略，以延长伤员的创伤后生存时间；② SERPINA3N蛋白在创伤性结肠损伤小鼠的肠上皮中表达显著上调；③ 逆转SERPINA3N蛋白表达水平的异常上调可减轻创伤性结肠损伤小鼠肠道稳态失调，延长个体生存时间；④ 褪黑素是具有口服活性的SERPINA3N蛋白表达抑制剂，预防性给予褪黑素治疗是延长个体创伤后生存时间的有效方法；⑤ 预防性给予MLT治疗有望成为改善TCI伤员预后的一种新型干预策略。 【主编评语】 解放军总医院第一医学中心卫勃团队在iMeta发表研究，基于创伤性结肠损伤（Traumatic colon injury, TCI）疾病小鼠实验模型，联合微生物组和蛋白质组学分析，探讨了TCI介导伤员快速死亡的微观机制。随后借助一系列体内外实验，明确了肠上皮SERPINA3N蛋白在TCI快速进展中的关键作用。为推动该研究成果的现实转化，作者测试了褪黑素（MLT）作为SERPINA3N蛋白表达抑制剂在延长TCI创伤后生存时间中的有效性。这种预防性干预方案可为灾害救援和军事行动中TCI伤员的治疗提供一种新思路、新选择。（@刘永鑫-农科院-宏基因组） 【原文信息】 Melatonin supplementation protects against traumatic colon injury by regulating SERPINA3N protein expression 2023-10-24, doi: 10.1002/imt2.141 肠道菌群介导的层级生态演化动力学（观点） Trends in Ecology and Ecolution——[16.8] ① 生态演化动力学是肠道菌群组装的核心；② 肠道菌群对宿主表型的影响使其成为生态演化动力学中的重要一环；③ 肠道菌群演化驱动因素包括：宿主因素（免疫、肠道环境）、微生物互作、环境因素；④ 肠道菌群生态演化动力学的3种途径：优先效应演化（生态位垄断）、代谢途径演化（互养式互作）、抗性演化（演化性群落挽救）；⑤ 在层级化生态演化动力学中，肠道菌群和宿主群落间存在自下而上和自上而下的反馈回路，影响它们的生态和演化轨迹。 【主编评语】 Trends in Ecology and Ecolution发表的这篇观点文章，探讨了肠道菌群介导的层级生态演化（eco-evo）动力学，包括其主要概念、驱动因素和途径，以及研究方法。（@mildbreeze） 【原文信息】 Hierarchical eco-evo dynamics mediated by the gut microbiome 2023-10-18, doi: 10.1016/j.tree.2023.09.013 感谢本期日报的创作者：mildbreeze，圆圈儿，湖人总冠军，RZN，章台柳，注册营养师陈彬林，Jadon，刘永鑫-农科院-宏基因组，Leo 点击阅读过去10天的日报： 12-20 | Nature重磅：从微生物暗物质中挖掘40万个新基因家族 12-19 | 周文/贺艳娟/贾伟/陈翔Cell子刊：从肠菌入手，改善多发性骨髓瘤治疗 12-18 | 《自然·综述》年度回顾：晚期大肠癌治疗的新选择 12-17 | Lancet子刊双文齐发：维生素D对心血管病和骨折的预防效果存疑 12-16 | 肠道细胞知多少？多文聚焦小细胞中的大学问 12-15 | 今日Science：菌群如何“排挤”病原体？生态学原理探究竟 12-14 | 今日Nature和Cell齐发，肠道菌群研究再掀小高潮 12-13 | 国内团队Cell子刊突破：调肠菌，改善肝癌患者手术预后！ 12-12 | 蜱虫叮咬使人对红肉过敏？JAMA解析α-gal综合征 12-11 | 《自然·综述》年度回顾：于君等详解癌症菌群研究新突破

Orum’s new program in immuno-oncology achieves simultaneous blockade of PD-1/PD-L1 pathway and delivery of a Cbl-b inhibitor using anti-PD-1-Cbl-bi conjugates and demonstrates enhanced T cell activation in vitro compared to anti-PD-1 alone, even in the presence of immunosuppressive factors ORM-6151 shows superior tolerability and robust single-dose efficacy, both in vitro and in vivo compared to CC-90009 or Mylotarg™, suggesting the potential for an improved therapeutic option in AML that appropriately balances efficacy with safety and tolerability of a clinically validated GSPT1 degradation mechanism Orum has identified pharmacodynamic biomarkers as predictors of efficacy to support clinical development of ORM-5029 in HER2-expressing solid tumors BOSTON & DAEJEON, South Korea--(BUSINESS WIRE)-- Orum Therapeutics, a clinical-stage private biotechnology company pioneering cell-specific targeted protein degradation (TPD² ™) and targeted protein stabilization (TPS² ™), today announced new preclinical data for three of Orum’s therapeutic programs: ORM-5029, which is a proprietary GSPT1 degrader conjugated to HER2-targeting antibody pertuzumab; ORM-6151, which delivers GSPT1 degrader to CD33+ tumors; and its new immuno-oncology program for a Cbl-b inhibitor (Cbl-bi) conjugated to anti-PD-1 antibody pembrolizumab. The data were presented in three posters at the 2023 American Association for Cancer Research (AACR) Annual Meeting. “We are pleased to present new data supporting clinical development of two programs from our TPD² GSPT1 Degrader Platform, which was developed to improve the therapeutic window and realize the full potential of GSPT1 degraders through precision delivery to cancer cells via antibody drug conjugates,” said Peter Park, Ph.D., Chief Scientific Officer of Orum Therapeutics. “We are also excited to unveil at AACR our new TPS² platform, which has the potential to achieve the full promise of Cbl-b inhibitors through T cell-specific delivery. With this novel approach, we show that we can limit unwanted systemic exposure, including the risk of toxicity associated with c-Cbl inhibition, prolong the activation of exhausted PD-1+ T cells, and provide resistance to negative environmental cues such as TGF-β or regulatory T cells. We believe our approach of harnessing the power of protein degraders and stabilizers with the precision of antibody targeting will improve the treatment of cancer for more patients.” Details of the data presented are as follows: Title: A novel antibody-enabled Dual-precision Targeted Protein Stabilization (TPS²) that augments anti-tumor immune response by targeting Cbl-b inhibitor to exhausted T cells while blocking checkpoint molecule, PD-1 Presenter: Joanne Lim, Ph.D. Abstract Number: 4436 Summary: Cbl-b inhibition results in stabilization of intracellular proteins downstream of the T cell activation pathways. Using a murine model well-established to be refractory to anti-PD-1 antibody treatment, we demonstrated that anti-PD-1-Cbl-b inhibitor approach increased effector T cell markers in the tumor and delayed tumor growth. Presented Data: The delivery of Cbl-b inhibitor (Cbl-bi) conjugated to anti-PD-1 antibody enhances the saturating, maximum activation of anti-PD-1 antibody by 300% in PD-1/PD-L1 blockade assay. Anti-PD-1-Cbl-bi conjugates enhance T cell responses of mixed lymphocyte reaction in a dose-dependent manner and overcome suppressive effects of TGF-β and Treg cells compared to antibody alone in vitro. Anti-PD-1-Cbl-bi conjugates enhance ex vivo activation of tumor-associated T cells from human cancer patients compared to antibody alone or when treated with antibody and unconjugated Cbl-bi. Anti-PD-1-Cbl-bi conjugates delay the growth of melanoma tumors in hPD-1 transgenic mice, with corresponding increase in intratumoral transcript levels of CD3e, granzyme B, and perforin in the tumor. Title: ORM-6151: A first-in-class CD33-antibody enabled GSPT1 degrader for AML Presenter: James Palacino, Ph.D. Abstract Number: 2700 Summary: ORM-6151 shows robust single-dose efficacy, both in vitro and in vivo with superiority or comparability to standard of care (SOC) agents. In addition, in vitro testing demonstrated superior tolerability to healthy bone marrow progenitor cells, suggesting better tolerability. Responses in representative TP53 wild-type and mutant AML models demonstrated comparable activity, indicating the potential for responses in a large percentage of AML patients with poor treatment options. Presented Data: ORM-6151 shows robust in vitro activity in p53 wild-type and mutant cell models. In ex-vivo AML blasts, ORM-6151 demonstrates efficacy superior to CC-90009, a small-molecule GSPT1 degrader, or Mylotarg, an FDA-approved treatment for AML. Compared to CC-90009 or Mylotarg, ORM-6151 exhibits minimal cytotoxic activity to healthy hematopoietic progenitor cells in vitro. A single treatment of ORM-6151 at 3 mg/kg regresses MV4-11 tumors in vivo mouse model with 9/9 complete response (CR), and a single dose of 1 mg/kg produced equivalent antitumor effect as SOC doublet given at optimal repeated dose. Title: Development of RNAscope multiplex-based assay for exploratory pharmacodynamic biomarkers assessment in breast cancer patients from Phase I clinical trial of ORM-5029, a potent GSPT1 degrader Presenter: Shikha Saini, Ph.D. Abstract Number: 2118 Summary: To fulfill the need to develop pharmacodynamic biomarkers as predictors of efficacy to support clinical development of ORM-5029, Orum identified a novel GSPT1 antibody for detection of depletion following treatment with ORM-5029. GSPT1 depletion and subsequent activation of the integrated stress response are both seen following in vitro or in vivo treatment with ORM-5029. Degree of pharmacodynamic (PD) change correlates with depth and duration of in vivo response. Orum is currently developing a multiplexed (IF/ISH/ISH) assay to track PD responses in patients, with an aim to better predict responses and efficacious doses. Presented Data: Orum identified the depletion of GSPT1 as a target-proximal pharmacodynamic biomarker in ORM-5029 treated in vitro and in vivo samples. Loss of GSPT1 drives activation of integrated stress response and upregulation of ATF3 and DDIT3 mRNA and these responses are also seen following treatment with ORM-5029, in both in vitro BT-474 cells and in vivo HCC1568 mouse model. PD responses correlate with degree and duration of response in an in vivo model treated with ORM-5029. The posters are available to download through the links below: PD-1-Cbl-bi TPS² preclinical poster: ORM-6151 preclinical poster: ORM-5029 predictive biomarker poster: About Orum’s TPS² Approach Orum’s Dual-precision Targeted Protein Stabilization (TPS²) approach combines novel payloads with the precise cell delivery mechanism of antibodies to increase the levels of intracellular target proteins in a cell-type-specific manner for oncology and immuno-oncology. The first program applying the TPS² approach is a Cbl-b inhibitor (Cbl-bi) conjugated to anti-PD-1 antibody. By delivering Cbl-bi specifically to exhausted T cells, the conjugate is designed to restore effector T cell function, where PD-1 checkpoint blockade alone is insufficient, while also enabling T cells to overcome the immunosuppressive mechanisms in a tumor microenvironment. About Orum’s GSPT1 Platform Using the TPD² Approach Orum’s GSPT1 platform uses the company’s unique Dual-precision Targeted Protein Degradation (TPD²) approach to build novel targeted protein degraders combined with the precise tumor cell delivery mechanisms of antibodies to generate innovative, first-in-class, cell-specific TPDs for the treatment of cancer. Orum has developed new molecular glue degrader payloads to specifically degrade an intracellular target protein within cancer cells via the E3 ubiquitin ligase pathway. Conjugated to antibodies, the payloads are designed to be delivered specifically to cancer cells and degrade the intracellular target protein GSPT1 and cause tumor cell death. About Orum Therapeutics Orum Therapeutics is a private, clinical-stage biotech pioneering the development of tumor-directed targeted protein degraders (TPD²) and stabilizers (TPS²) with the precision of antibody targeting to improve cancer treatment for more patients. The first therapeutic candidates from the TPD² GSPT1 platform, ORM-5029 and ORM-6151, are in clinical development for the treatment of solid tumors and hematologic cancers, respectively. The first program applying the TPS² approach is a Cbl-b inhibitor conjugated to anti-PD-1 antibody. Orum is located in Boston, US, and Daejeon, South Korea. For more info, visit . Forward-Looking Statements This press release contains forward-looking statements that are based on the current expectations and beliefs of Orum Therapeutics, Inc. (“Orum”). Statements in this release regarding matters that are not historical facts, including, but not limited to, statements relating to the potential for clinical efficacy, potency, and tolerability of Orum’s products; the potential for conducting and successfully completing clinical trials, translation of preclinical and non-human data to the clinical and human contexts, obtaining favorable trial results, conducting later-phase trials, obtaining regulatory approval of product candidates for sale, successfully commercializing product candidates, and improving the cancer treatment for more patients; the types of cancer that might be treated by Orum products; the successful prediction of patient responses and efficacious doses; and the preferability of Orum products to others available are forward-looking statements. These forward-looking statements are based on management’s expectations and assumptions as of the date of this release and are subject to numerous risks and uncertainties, which could cause actual results to differ materially from those expressed or implied by such statements. These risks and uncertainties include, without limitation, the uncertain and continually changing impacts and expected duration of the COVID-19 pandemic; the uncertainty of success in research and development activities; risks related to clinical trials, including potential delays, safety issues, or negative results; competition from alternative therapies; the risk that Orum may not be able to maintain and enforce its intellectual property; the risk that product candidates may not be successfully commercialized or adopted; the availability of financing; and risks related to the recruitment and retention of key employees, fluctuating markets and economic conditions, health care reform, prices, and reimbursement rates. The forward-looking statements in this presentation speak only as of the date of this release, and Orum undertakes no obligation to update or revise any of the statements. 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