Development of Natural Killer (NK) Cell Line-Derived Extracellular Vesicles as a New Treatment for Cancer

Vanquish Bio是一家免疫治疗公司，利用独特的天然药物重新想象癌症的治疗 杀伤(NK)细胞来源的胞外小泡(EVS)。这个STTR的目标是开发一种使用天然药物的疗法 免疫系统产品用于靶向摧毁骨髓瘤肿瘤细胞。多发性骨髓瘤(MM) 约占血液系统癌症的10%，在中国有约32,110例新病例和~12,960例死亡 2019年的美国。尽管治疗包括干细胞移植和/或化疗，但患者 不可避免地会故态复萌。需要新的创新来延长生存时间，增加缓解的持久性，并 为不符合一线治疗条件的患者提供选择。我们开发了NK3.3，这是一种来源于 来自一位健康的捐赠者。这些细胞可以杀死一系列的肿瘤类型。使用最小残留病小鼠模型 对于人类多发性骨髓瘤，注射NK3.3 EVS可防止肿瘤复发长达100天，而不是21天 不接受治疗。当NK3.3在培养中生长时，它们释放EV作为小的膜结合结构 还能够杀死许多不同的肿瘤细胞，而不损害正常细胞。NK3.3电动汽车还能杀死癌症干细胞 细胞(CSC)；这些细胞对标准化疗耐药，并导致转移和复发。NK 电动汽车可以大量生产、冷冻、储存和解冻，而不会失去功能。其他 使用NK3.3电动汽车的优势包括：1)耐缺氧的肿瘤微环境，2)能够 通过血脑屏障，3)稳定，4)低毒。电动汽车不受血型或 组织相容，因此是一种免疫治疗方式，可以是一种普遍的治疗方法。我们的目标 是开发一种新的输液疗法产品，将提高MM的存活率，并减少副作用 复发/难治或病情稳定的患者。 第一阶段是生成NK3.3衍生的电动汽车，并展示对MM的有效性，并延长 缓解，毒副作用最小。 具体目标1：确定最大限度地提高NK3.3 EVS的肿瘤杀伤活性的激活剂。NK电动汽车将成为 分别从激活细胞因子IL-2、IL-12和IL-15培养的NK3.3细胞中分离到 组合。我们的目标是制备NK EV，在治疗后72小时内杀死90%的≥肿瘤细胞， 低于10%的正常细胞死亡。 特定目标2：确定NK EVS对MM的低毒概念验证疗效 免疫缺陷小鼠。我们将对MM免疫缺陷患者给予不同浓度的NK EV 病情稳定的小鼠。目标是将≥50%的小鼠的缓解时间延长到21天以上。毒性将是 测量可见的定性迹象和行为，以及体重、器官重量和 血液化学。我们将证明生产更有效的具有抗MM活性的NK EV制剂的可行性 以概念验证为导向的NK EV治疗对MM的疗效和毒性数据。

Vanquish Bio is an immunotherapy company reimagining treatment of cancer with the use of unique natural killer (NK) cell derived extracellular vesicles (EVs). The goal of this STTR is to develop a therapy using natural products of the immune system for targeted destruction of myeloma tumor cells. Multiple myeloma (MM) represents ~10% of hematological cancers, and there were ~32,110 new cases and ~12,960 deaths in the U.S. in 2019. Despite treatments that include stem cell transplantation and/or chemotherapy, patients inevitably relapse. New innovations are needed that extend survival, increase durability of remission and provide options to patients ineligible for front-line therapy. We developed NK3.3, a human NK cell line derived from a healthy donor. These cells kill an array of tumor types. Using a minimal residual disease mouse model of human MM, administration of NK3.3 EVs prevented tumor recurrence for up to 100 days, versus 21 days without treatment. As NK3.3 grow in culture, they release EVs as small membrane-bound structures that are also capable of killing many different tumor cells without harming normal cells. NK3.3 EVs also kill cancer stem cells (CSC); these are resistant to standard chemotherapy and responsible for metastasis and recurrence. NK EVs can be produced in large quantities, frozen, stored and then thawed, without loss of function. Other advantages of using NK3.3 EVs include: 1) resistance to the hypoxic tumor microenvironment, 2) ability to cross the blood-brain barrier, 3) stability and 4) low toxicity. EVs are not restricted by blood group or histocompatibility and are therefore an immunotherapeutic modality that can be a universal treatment. Our goal is to develop a new infusion therapy product that will improve survival rates with less side effects for MM patients with relapsed/refractory or stable disease. Phase I is to generate NK3.3-derived EVs and demonstrate efficacy against MM and extend durability of remission, with minimal, toxic side effects. Specific Aim 1: Identify activators that maximize tumor killing activity of NK3.3 EVs. NK EVs will be isolated from NK3.3 cells cultured with activating cytokines IL-2, IL-12 and IL-15 individually and in combinations. Our goal is to prepare NK EVs that kill ≥90% of tumor cells within 72 hours of treatment, with less than 10% normal cell death. Specific Aim 2: Determine proof-of-concept efficacy with low toxicity of NK EVs in MM-bearing immunodeficient mice. We will administer different concentrations of NK EVs in MM-bearing immunodeficient mice with stable disease. The goal is to extend remission beyond 21 days in ≥50% of mice. Toxicity will be measured for qualitative visible signs and behavior, and quantitative variation in body weight, organ weight and blood chemistry. We will prove feasibility of producing more potent NK EV preparations with anti-MM activity and proof-of-concept oriented efficacy and toxicity data of NK EV therapy against MM.