MT2004-25: ALLOGENEIC NATURAL KILLER CELLS WITH RELAPSED ACUTE MYELOGENOUS LEUKE

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。这个GCRC应用程序将重点回答NK细胞生物学中的几个重要问题。本研究要验证的主要假设是抑制性NK细胞受体在癌症和移植的治疗中发挥作用。现在已知NK细胞表达识别I类MHC分子的抑制性受体。这些受体与MHC配体结合提供抑制信号，保护目标不被裂解。因此，我们认为，与自体NK细胞相比，携带I类MHC配体错配受体的异体NK细胞可能提供更有效的抗肿瘤治疗。这将在临床试验中进行测试（子研究1）。子研究1将检验NK细胞的稳态扩张需要清除淋巴间隙的假设，而NK细胞的稳态扩张是有效所必需的。亚研究1将在AML和肾细胞癌两个患者群体中进行分层。在预后不良的AML患者中，将使用具有抗急性白血病活性的住院化疗的高强度预备方案。这种程度的强度在肾细胞癌中没有临床依据。因此，一种改良的、强度较低的门诊化疗方案将被测试。此外，在NK细胞发育过程中，同种异体移植后NK细胞重建的获得是未知的。子研究2将重点回答这个问题，假设移植后早期重建的NK细胞受体将表现出受体的缺乏，这将与生理相关，并与临床结果相关。了解这一过程在移植中可能允许操纵供体移植物或供体选择，以加强癌症的最佳治疗。亚研究1：“MT2003-01：急性髓性白血病和肾细胞癌复发患者的同种异体自然杀伤细胞”。这是一项针对晚期AML患者的临床试验，这些患者在标准诱导化疗后未能达到缓解或患有转移性肾细胞癌。该试验是在接近完成其研究目标的GCRC（第679号方案）中进行的I期临床试验的直接外推。从I期细胞剂量升级研究中，我们得出结论，单倍体同种异体NK细胞输注是安全的，低剂量环磷酰胺和高剂量甲基强的松龙的制备方案不能抑制受体混合淋巴细胞的反应性，供体细胞可以在受体外周血中发现约5天，但不会长期存在，也不会在体内扩增。本研究的主要假设之一是同种异体NK细胞要想有效，它们必须在给予IL-2的两周内在体内扩增。在I期研究中未能实现这一目标可能是由于1)制备方案的抑制不足和2)淋巴间隙的清除不足，目前已知淋巴间隙在体内过继转移淋巴细胞的稳态扩张中很重要。这项临床试验将解决这两个假设。选择AML作为本研究的目标人群是基于最近的骨髓移植研究，这些研究表明，单倍体移植保护AML复发的能力与错配的NK细胞受体及其假定的MHC I类配体有关。本研究的患者将接受一种预备方案，该方案使用两种活性白血病药物：环磷酰胺和氟达滨，其免疫抑制作用更强。选择包括肾细胞癌是基于显示同种异体移植后活性的新数据，以及这种肿瘤历史上具有免疫应答性的事实。在GMP条件下，选择单倍体相同的NK细胞并在GCRC细胞治疗核心中激活。作为了解同种异体单倍体NK细胞作用的一部分，将在几个时间点对供体NK细胞和患者血液进行实验室研究，以了解它们的持久性、扩张性和免疫反应。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This GCRC application will focus on answering several important questions in NK cell biology. The main hypothesis to be tested in this study is that inhibitory NK cell receptors play a role in the therapy of cancer and transplantation. It is now well known that NK cells express inhibitory receptors that recognize class I MHC molecules. Engagement of these receptors by their MHC ligands provides an inhibitory signal which protects targets from being lysed. Therefore, it follows that therapy will allogeneic NK cells bearing receptors mismatched for class I MHC ligands may provide more effective anti-tumor therapy than autologous NK cells. This will be tested in a clinical trial (Sub-Study 1). Sub-Study 1 will test the hypothesis that clearance of lymphoid space is required for homeostatic expansion of NK cells, which will be required for efficacy. Sub-Study 1 will be stratified among two patient populations, AML and renal cell carcinoma. In poor prognosis AML patients, an intense preparative regimen using in-patient chemotherapy that has activity against acute leukemia will be used. This degree of intensity is not clinically warranted in renal cell carcinoma. Therefore, a modified, less intense regimen using out-patient chemotherapy will be tested. In addition, during NK cell development, the acquisition of NK cell reconstituting after allogeneic transplant is unknown. Sub-study 2 will focus on answering this question with the hypothesis that NK cell receptors reconstituting early after transplant will exhibit a paucity of receptors which will be of physiologic relevance and correlate wil clinical outcomes. Understanding this process in transplantation may allow maipulation of donor grafts or donor selection to enhance optimal therapy of cancer. Sub-Study 1: "MT2003-01: Allogeneic natural killer cells in patients with reapse of acute myelogenous leukemia and renal cell carcinoma". This is a clinical trial in advanced patients with AML who fail to achieve remission after standard induction chemotherapy or have metastatic renal cell carcinoma. This trial is a direct extrapolation of a phase I clinical trial performed in the GCRC (Protocol #679) which is nearing completion of its study goals. From that phase I cell dose escalation study, we conclude that haploidentical allogeneic NK cell infusions are safe, that the preparative regimen of low-dose cytoxan and high-dose methylprednisolone is unable to suppress recipient mixed lymphocyte reactivity and that donor cells can be found in recipient peripheral blood for approximately five days but that do not persist long term and do not expand in vivo. One of the main hypotheses for this study is that for allogeneic NK cells to be efficacious they must expand in vivo during the two week period of IL-2 administration. Failure to achieve this goal in the phase I study may be due to 1) inadequate suppression of the preparative regimen and 2) inadequate clearance of lymphoid space, now known to be important in homeostatic expansion of adoptively transferred lymphocytes in vivo. This clinical trial will address both of those hypotheses. The choice of AML as a target population for this study is based on recent bone marrow transplant studies showing that the ability of haploidentical transplants to protect against AML reapse is associated with mismatched NK cell receptors and their presumed MHC class I ligands. Patients on this study will receive a preparative regimen which is more immunosuppressive using two active leukemia drugs: cyclophosphomide and fludarbine. The choice of including renal cell carcinoma is based on new data showing activity after allogeneic transplant and the fact that this tumor is historically immune responsive. Haploidentical NK cells will be selected and activated in the GCRC Cell Therapy Core under GMP conditions. As a part of understanding the role of allogeneic haploidentical NK cells, laboratory studies will be performed on donor NK cells and patient blood at several time points to understand their persistence, expansion and immunologic response.