Phase I study humanized 3F8 MoAb (IND 112594) and NK cells (IND BB-13399) for neuroblastoma

项目总结/摘要 神经母细胞瘤（NB）是儿童最常见的颅外实体瘤。虽然密集 诱导化疗和积极的手术提高了年轻患者的缓解率， 在老年患者和40%的化疗耐药NB患者中不太令人印象深刻。 在诊断时年龄> 18个月的儿童中，转移性NB的长期无复发生存率仅为 ~20%。这些令人失望的结果是追求新的治疗方法的令人信服的理由。 一种替代方法是用鼠3F8免疫治疗，鼠3F8是针对人免疫缺陷病毒的IgG3单克隆抗体。 GD2糖脂抗原普遍存在于NB细胞上。然而，它的主要缺点是 中和抗体的发展，这可能会限制其有效性。3F8给药可提供 对缓解期NB患者生存有利，但对软组织耐药NB患者无效 或患有进行性疾病的患者。这可能部分是由于其依赖于抗体依赖性 细胞介导的细胞毒性（ADCC），后者主要由自然杀伤（NK）细胞介导， 在大量预先治疗的患者中耗尽。从健康同种异体来源连续转移NK细胞 可以恢复和增强3F8在这些低风险患者中的作用。 自然杀伤（NK）细胞是具有通过多种途径进行抗肿瘤活性的能力的淋巴细胞。 途径，包括通过CD16 Fc受体的接合的ADCC。NK细胞的能力， 然而，细胞毒性反应是由其细胞表面受体，特别是抑制性免疫球蛋白决定的 类受体（KIR），其对自身MHC I类分子具有特异性。为了最大限度地发挥收养的效果， 转移NK细胞，应该选择最有可能产生 NK同种异体反应性和实现最高的功能性NK反应。这可以通过选择 基于供体和患者的HLA和KIR基因型来确定供体。 该提案提出了一种新的免疫治疗方法，用于治疗高风险的 神经母细胞瘤3F8单克隆抗体与过继转移的NK细胞的组合， 适当选择的供体应增加ADCC、抗体效力和肿瘤根除。的 高效人源化3F8抗体的可用性可以增加肿瘤应答，同时避免肿瘤细胞的增殖。 中和抗体的发展是如此常见的鼠类类似物。因为这些 免疫学方法从未在实体瘤或儿科中进行过联合研究 人群，一项I期研究，以检查人源化3F8与递增剂量的 NK细胞是必要的，也是该提案的首要目标。NK和3F8的潜在毒性 治疗，毒性监测计划，剂量递增计划，预期结果和停止规则， 提出了第二个目的是寻求供体-受体KIR/HLA免疫遗传学和供体FcR之间的相关性。 多态性与（1）体外NK活化和针对NB靶细胞的细胞毒性功能，以及（2）与 患者对治疗的反应。安全性和可行性的证明 NK细胞与单克隆抗体不仅会导致更有效的治疗方法，为穷人， 预后NB患者，但将广泛适用于其他癌症人群，如淋巴瘤和 目前用单克隆抗体治疗的乳腺癌。

PROJECT SUMMARY/ABSTRACT Neuroblastoma (NB) is the most common extracranial solid tumor of childhood. While intensive induction chemotherapy and aggressive surgery have improved remission rates in young patients, results have been less impressive in older patients and in the 40% of patients with chemoresistant NB. Metastatic NB in children > 18 months of age at diagnosis carries a long-term relapse-free survival of only ~20%. These disappointing results are compelling reasons for pursuing novel therapeutic approaches. One alternative approach has been immunotherapy with murine 3F8, an IgG3 monoclonal antibody to the GD2 glycolipid antigen ubiquitously present on NB cells. Its major disadvantage, however, is the development of neutralizing antibodies, which can limit its effectiveness. 3F8 administration can confer a survival benefit to NB patients in remission, but it is not effective for patients with resistant soft tissue NB or patients with progressive disease. This may be due in part to its dependence on antibody-dependent cell-mediated cytotoxicity (ADCC), the latter mediated largely by natural killer (NK) cells, which are depleted in heavily pre-treated patients. Adoptive transfer of NK cells from a healthy allogeneic source can restore and enhance 3F8 effects in these poor-risk patients. Natural killer (NK) cells are lymphocytes that have the capacity for antitumor activity via multiple pathways, including ADCC through engagement of the CD16 Fc receptor. The capacity of an NK cell for cytotoxic response, however, is dictated by its cell surface receptors, specifically the inhibitory killer-Ig like receptors (KIR) that are specific for self-MHC class I molecules. To maximize the effects of adoptively transferred NK cells, one should select donors from whom there is the greatest likelihood of engendering NK alloreactivity and of achieving highest functional NK response. This can be accomplished by selecting donors based on the HLA and KIR genotypes of the donor and the patient. This proposal presents a novel immunotherapeutic approach for the treatment of high-risk neuroblastoma. The combination of 3F8 monoclonal antibody with adoptively transferred NK cells from appropriately selected donors should increase ADCC, antibody efficacy, and tumor eradication. The availability of a highly effective humanized 3F8 antibody may increase tumor response while avoiding the development of neutralizing antibodies that are so common to murine analogs. Because these immunologic approaches have never been studied in combination for a solid tumor or in the pediatric population, a phase I study to examine the safety of humanized 3F8 combined with escalating doses of NK cells is necessary and represents the first aim of the proposal. Potential toxicities of NK and 3F8 therapies, a toxicity monitoring plan, dose escalation plan, the expected outcome and stopping rules are presented. The second aim seeks to correlate donor-recipient KIR/HLA immunogenetics and donor FcR polymorphism with (1) NK activation and cytotoxic function against NB target cells in vitro, and (2) with patient response to treatment. Demonstration of safety and feasibility of combining adoptively transferred NK cells with monoclonal antibody will not only result in a more potent therapeutic approach for poor- prognosis NB patients, but will be broadly applicable to other cancer populations such as lymphoma and breast carcinoma currently treated with monoclonal antibodies.