Potentiating Natural Killer Cell Anti-Myeloma Effects

描述（由申请人提供）：数据表明多发性骨髓瘤是一个主要的健康问题。美国大约有54,000名骨髓瘤患者，2007年将有20,000名新患者被诊断为骨髓瘤，每年有近11,000名患者死于这种疾病。治愈率和10年生存率仍然很低，这表明需要新的治疗方法。我们已经在体外和临床试验中证明，来自单倍体相同供体的杀伤细胞免疫球蛋白样受体（KIR）配体（- l）不匹配的NK细胞可以杀死化疗耐药骨髓瘤。然而，我们只能在30%的患者中找到kirl不匹配的供体。我们建议采用三管齐下的方法：a)在自体环境中应用NK细胞治疗，使所有患者都能接受治疗；b)提高NK细胞的临床疗效。我们假设我们可以通过激活和扩增NK细胞以及调节NK细胞效应物与骨髓瘤靶点之间的相互作用来克服hla - I类诱导的自体NK细胞对骨髓瘤细胞的抑制。在特异性目标1中，我们将确定NK细胞的剂量和效力是否可以通过扩增和激活骨髓瘤患者的NK细胞来可靠地增加。如果我们要克服骨髓瘤的负担，NK细胞的扩增是重要的。如果没有扩张，NK细胞就会太少，无法根除所有的骨髓瘤。NK细胞的“增压”将克服自体骨髓瘤传递的任何抑制信号。我们将用转染了膜结合il - 15和共刺激分子4-1BB-L的K562细胞刺激NK细胞。在特异性目标2中，我们将评估激活的自体NK细胞的作用是否可以通过人源化的CS1抗体标记骨髓瘤细胞而增强。CS1是一个CD2受体家族分子，在骨髓瘤中表达，而在正常组织中不表达，因此具有骨髓瘤特异性杀伤作用。这种抗体将有效地“标记”骨髓瘤细胞，使其免受adc介导的NK细胞杀伤。在特异性目的3中，我们将确定激活的自体NK细胞的作用是否可以通过蛋白酶体抑制下调骨髓瘤的抑制配体而增加。由于骨髓瘤细胞上的HLA I类与NK细胞上的抑制性受体之间的相互作用，NK细胞通常不会杀死自体骨髓瘤。我们已经证明，在用蛋白酶体抑制剂硼替佐米治疗后，我们可以在体外和体内下调骨髓瘤上的hla - I类，并且这转化为自体NK细胞杀死骨髓瘤。因此，我们将评估硼替佐米与活化NK效应剂联合使用时是否存在潜在的协同或加性效应。这种临床方法可以应用于标准风险骨髓瘤患者，一旦我们证明了增强NK细胞治疗对高危或复发骨髓瘤患者的疗效，就可以获得更好的生长控制和更持久的缓解。此外，这项研究可能会导致其他NK细胞敏感的恶性肿瘤的更有效的治疗。公共卫生相关性：多发性骨髓瘤是一种目前无法治愈的骨髓癌。在美国大约有54000名骨髓瘤患者。这项拨款提案描述了三种利用患者自身免疫细胞摧毁癌症的新方法。这种治疗方法也可能对治疗其他癌症有用。

DESCRIPTION (provided by applicant): Data indicate that multiple myeloma is a major health problem. There are approximately 54,000 myeloma patients in the USA, 20,000 new patients will be diagnosed with myeloma in 2007, and nearly 11,000 patients succumb to their disease yearly. The cure rate and 10-year survival remains low indicating the need for new therapies. We have shown that chemo-resistant myeloma can be killed by killer-cell immunoglobulin- like receptor (KIR) ligand (-L) mismatched NK cells from a haplo-identical donor in vitro and in a clinical trial. However, we could find a KIR-L mismatched donor for only 30% of patients. We propose to use a 3-pronged approach to: a) apply NK cell therapy in the autologous setting making therapy possible for all patients and b) enhance the clinical efficacy of NK cells. We hypothesize that we can overcome the inhibition of autologous NK cells induced by HLA-class I on myeloma cells by activating and expanding the NK cells and by modulating the interaction between NK cell effectors and myeloma targets. In Specific Aim 1 we will determine if NK cell dose and potency can be reliably increased by expanding and activating NK cells from myeloma patients. Expansion of NK cells is important if we are to overcome the myeloma burden. Without expansion, there will be too few NK cells to eradicate all myeloma. `Supercharging' of the NK cells will overcome any inhibitory signals delivered by autologous myeloma. We will stimulate the NK cells with K562 cells transfected with membrane-bound IL15 and the co-stimulatory molecule 4-1BB-L. In Specific Aim 2 we will evaluate whether the action of activated autologous NK cells can be enhanced by flagging myeloma cells with a humanized antibody to CS1. CS1 is a CD2 receptor family molecule expressed by myeloma but not by normal tissues, therefore conferring myeloma-specific killing. This antibody will effectively `flag' myeloma cells for ADCC-mediated killing by NK cells. In Specific Aim 3 we will ascertain if the action of activated autologous NK cells can be increased by down regulating inhibitory ligands on myeloma by proteasome inhibition. NK cells do not normally kill autologous myeloma due to the interaction between HLA class I on myeloma cells with inhibitory receptors on NK cells. We have demonstrated that we can down regulate HLA-class I on myeloma in vitro and in vivo after treatment with the proteasome inhibitor bortezomib and that this translates into killing of myeloma by autologous NK cells. Hence, we will evaluate the existence of potential synergistic or additive effects when combining bortezomib with activated NK effectors. This clinical approach can be applied to patients with standard-risk myeloma to obtain even better growth control and more durable remissions once we have demonstrated the efficacy of enhanced NK cell therapy in patients with high-risk or relapsing myeloma. In addition, this research could lead to more efficacious treatment for other NK cell sensitive malignancies. PUBLIC HEALTH RELEVANCE: Multiple myeloma is a form of bone marrow cancer that is currently incurable. There are approximately 54,000 myeloma patients in the USA. This grant proposal describes 3 new ways in which the patient's own immune cells can be used to destroy the cancer. Such treatment may also be useful to treat other cancers.