Viral priming and targeting NK cells against solid tumor malignancies

描述（由申请人提供）：在过去的20年里，我在自然杀伤（NK）细胞生物学、免疫疗法的发展和血液恶性肿瘤和急性髓性白血病（AML）的造血细胞移植（HCT）领域获得了nci的持续资助。我的团队是NK细胞过继移植的先驱，拥有世界上最大的经验，已经注入了200个单倍相同的NK细胞产品来治疗血液和实体肿瘤恶性肿瘤患者。最近，我们通过鉴定巨细胞病毒（CMV）诱导的NK细胞中独特的功能表型，描述了人类NK细胞生物学的新范式。这些长寿命，高度分化的NKG2C+/CD27+细胞，我们称之为“适应性”NK细胞，被培养和富集表达自我抑制性杀伤细胞免疫球蛋白样受体（KIR）。它们代表了人类在cmv感染小鼠中描述的类似记忆的Ly49H+ NK细胞。此外，我们发现扩增的NK细胞亚群选择性地单独或联合缺乏近端信号分子FcϵR1γ、EAT-2和SYK。重要的是，它们具有增强细胞因子产生和存活的表观遗传学启动，并通过CD16介导有效的抗体依赖性细胞毒性（ADCC）。本次杰出研究者奖的首要目标是制定策略，以提高内源性NK细胞在实体肿瘤恶性肿瘤患者中的抗肿瘤活性。目标是开发“现成的”试剂来激活NK细胞，克服抑制性受体信号传导，并将其靶向特定的肿瘤抗原。我的团队已经开发了几种新的NK细胞靶向药物，包括双特异性杀伤接合物（bispecific killer接合物，BiKEs），通过将scFv抗cd16与针对肿瘤抗原的scFv融合，以及与行业合作伙伴开发的针对肿瘤抗原的IL-15/IL-15Rα-Fc复合物。在本提案中，我们将使用几种策略评估这些药物的治疗潜力。首先，实体瘤患者的流行病学遗传学研究将确定巨细胞病毒与人乳头瘤病毒（HPV）暴露、病毒诱导的“适应性”NK细胞的发展、癌症发展风险和对标准治疗的反应之间的关系。其次，我们将评估一种新的实体肿瘤抗原，RHAMM，在肿瘤和“癌化”微环境中表达。第三，我们将使用一种新的异种模型，我们将用我们独特的药物测试针对RHAMM的“适应性”NK细胞的抗肿瘤功效。第四，在临床前评估后，我们将对治疗恶性肿瘤的最佳药物进行I期临床试验，并将测试病毒疫苗在诱导或增强“适应性”NK细胞功能方面的作用。最后，我将以我长期的指导经验为基础，利用这些研究作为培训未来转化科学家的平台。这里提出的创新研究得到了初步数据的很好支持，代表了NK细胞生物学中一个相当令人兴奋的新兴领域。利用先天“适应性”NK细胞靶向实体瘤的治疗方法的成功开发将对癌症免疫治疗领域产生重大影响。

DESCRIPTION (provided by applicant): During the last 2 decades I have had continuous NCI-funding in the areas of natural killer (NK) cell biology, development of immunotherapies and hematopoietic cell transplantation (HCT) for hematologic malignancies and acute myeloid leukemia (AML). My team, who pioneered adoptive transfer of NK cells, has the largest world experience having infused >200 haploidentical NK cell products to treat patients with hematologic and solid tumor malignancies. Recently we described a new paradigm in human NK cell biology by identifying a unique functional phenotype in NK cells induced by cytomegalovirus (CMV). These long-lived, highly differentiated NKG2C+/CD27+ cells, which we call "adaptive" NK cells are educated and enriched for the expression of self-inhibitory killer-cell immunoglobulin-like receptors (KIR). They represent the human equivalent of the memory-like Ly49H+ NK cells described in CMV-infected mice. Further, we identified expanded NK cell subsets selectively lacking the proximal signaling molecules FcϵR1γ, EAT-2 and SYK individually or in combination. Importantly, they are epigenetically primed for enhanced cytokine production and survival, and mediate potent antibody-dependent cellular cytotoxicity (ADCC) through CD16. The overarching goal of this Outstanding Investigator Award is to develop strategies to enhance the anti-tumor activity of endogenous NK cells in patients with solid tumor malignancies. The objective is to develop "off the shelf" reagents to activate NK cells, overcome inhibitory receptor signaling, and target them to specific tumor antigens. My group has developed several novel NK cell targeting agents, including bi-specific killer engagers (BiKEs), created by fusing scFv anti-CD16 with scFv for tumor antigens and IL-15/IL-15Rα-Fc complexes targeted to tumor antigens developed with an industry partner. In this proposal, we will evaluate the therapeutic potential for these agents using several strategies. First, epidemiologic genetic studies in patients with solid tumors will define the relationship between CMV and human papillomavirus (HPV) exposure, the development of virally-induced "adaptive" NK cells and the risk of cancer development and response to standard therapies. Second, we will evaluate a novel solid tumor antigen, RHAMM, expressed both on tumors and in the "cancerized" microenvironment. Third, we will use a novel xenogeneic model we will test the anti-tumor efficacy of "adaptive" NK cells targeted to RHAMM with our unique agents. Fourth, after preclinical evaluations, we will conduct phase I clinical trials of the optimal agents to treat sold tumor malignancies and will test the role of viral vaccines in inducing or enhancing "adaptive" NK cell function. Lastly, I will build on my long track record of mentorship to use these investigations as a platform to train future translational scientists. The innovative studies proposed here are well supported by preliminary data and represent an emerging area of considerable excitement in NK cell biology. The successful development of therapies to harness innate "adaptive" NK cells to target solid tumors will have a substantial impact the field of cancer immunotherapy.